WO2004099190A1

WO2004099190A1 - Novel substituted benzimidazole derivatives

Info

Publication number: WO2004099190A1
Application number: PCT/SE2004/000695
Authority: WO
Inventors: Britt-Marie Swahn
Original assignee: Astrazeneca Ab
Priority date: 2003-05-09
Filing date: 2004-05-06
Publication date: 2004-11-18
Also published as: SE0301371D0

Abstract

The invention provides a compound of Formula I (I) wherein n is 1 or 2; R1 is selected from hydrogen, halogen, CN, COOR6, CONR6R7 or CH2OR6; R2 is selected from hydrogen, halogen or CN; R3 is selected from hydrogen or halogen; R4 is selected from hydrogen, COOR6, CONR6 R7, NO2, NR6R7, or NR6COR7; R5 selected from hydrogen, NR6R7 or NR6COR7; as a free base or a pharmaceutically acceptable salt, solvate or solvate of salt thereof, a process for their preparation, pharmaceutical formulations containing said compounds and to the use of said compounds in therapy.

Description

NOVEL SUBSTITUTED BENZUvLTDAZOLE DERIVATIVES

TECHNICAL FIELD

The present invention relates to novel substituted benzimidazole derivatives, useful for treatment of various disorders. The invention relates to methods for producing these compounds. The invention also provides pharmaceutical compositions comprising the compounds of the invention and methods of utilizing these compositions in the treatment of various disorders.

BACKGROUND TO THE INVENTION;

Protein kinases are important components of intracellular signalling pathways and kinases are involved in the regulation of a variety of cellular functions. The MAP kinase signalling pathways are activated by engagement of a number of cell surface receptors. One of these pathways, the JNK pathway is activated specifically by stress or pro-inflammatory cytokines. Activators include LPS, the cytokines tumour necrosis factor (TNF- ) and Interleukin-1 (JL-1), osmotic shock, chemical stress and UV radiation (Cohen, P. Trends in Cell Biol. 7:353-361 1997). Targets of the JNK pathway include a number of transcription factors, such as but not exclusively c-jun and ATF-2 (Whitmarsh, A. and Davis, R. J. Mol. Med. 74:589-607 1998).

Three different genes: JNK1, JNK2 and JNK3; encode the JNK family of enzymes. Alternatively spliced forms of these genes can give rise to 10 distinct isoforms: four for JNK1, four for JNK2 and two for JNK3. (Gupta, S. et al EMBO J. 15:2760-2770 1996). JNK1 and JNK2 are ubiquitously expressed in human tissues whereas JNK3 is selectively expressed in the brain, heart and testis (Dong, C. et al. Science 270:1-4 1998).

JNKs 1, 2 and 3 have been selectively knocked out in mice both singulary and in combination by both gene deletion and/or transgenic expression of dominant negative forms of the kinases (Dong, C. et al Science 282:2092-2095 1998; Yang, D. et al Immunity 9:575-585 1998; Dong, C, et al Nature 405:91-942000; Yang, D. et al Nature 389:865- 870 1997). Mice with targeted disruption of the JNK3 gene develop normally and are protected from excitotoxin induced apoptosis of neurones. This finding suggests that specific inhibitors of JNK3 could be effective in the treatment of neurological disorders characterised by cell death such as Alzheimer's disease and stroke. Mice disrupted in either JNK1 or 2 also develop normally. Peripheral T cells from either type of mice can be activated to make BL2, but in both cases, there is a defect in Thl cell development. In the case of JNK1 -/- mice, this is due to an inability to make gamma interferon (a key cytokine essential for the differentiation of Thl cells). In contrast, JNK2 -/- mice produce interferon gamma but are unable to respond to the cytokine. Similar defects in T cell biology (normal IL2 production but a block in Thl cell differentiation) are seen in T cells disrupted in the MKK7 gene confirming this role for the JNK pathway in T cell differentiation (Dong, C, et al Nature 405:91-942000).

JNK also plays a major role in apoptosis of cells (Davis RJ. Cell. 103:239-2522000). JNK is essential for UN induced apoptosis through the cytochrome C mediated pathway (Tournier, C. et al Science 288:870-874 2000). Ischemia and ischemia coupled with re- perfusion as well as restricted blood flow itself have been shown to be accompanied by activation of JΝK. Cell death can be prevented with dominant negative forms of JΝK transfected into cells demonstrating a potential utility for JΝK in conditions characterised by stress-induced apoptosis.

Activation of the JΝK pathway has been observed in a number of human tumours and transformed cell lines (Davis RJ. Cell. 103:239-252 2000). Indeed, one of the major targets of JΝK, c-jun, was originally identified as an oncogene indicating the potential of this pathway to participate in unregulated cell growth. JΝK also regulates phosphorylation of p53 and thus modulates cell cycle progression (Chen T. et al Mol. Carcinogenesis 15:215- 226 1996). Inhibition of JΝK may therefore be beneficial in some human cancers.

Based on current knowledge of JΝK signalling, especially JΝK3, has been implicated in areas of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, ALS, Huntington's disease, traumatic brain injury, as well as ischemic and haemorrhaging stroke.

Thus, there is a high unmet medical need for JNK specific inhibitors useful in treating the various conditions associated with JNK activation.

DISCLOSURE OF THE INVENTION

It has been found that compounds of the Formula I, which are benzimidazole compounds, are particularly effective JNK inhibitors and thereby suitable in the treatment of the various conditions associated with JNK activation.

In one aspect, the invention thus relates to compounds of Formula I

(I) wherein: n is 1 or 2

R¹ is selected from hydrogen, halogen, CN, COOR⁶, CONR⁶R⁷ or CH₂OR⁶; R² is selected from hydrogen, halogen or CN; R³ is selected from hydrogen or halogen;

R⁴ is selected from hydrogen, COOR⁶, CONR⁶R⁷, NO₂, NR⁶R⁷ or NR⁶COR⁷; R⁵ selected from hydrogen, NR⁶R⁷, or NR⁶COR⁷;

R⁶ and R⁷ are each independently hydrogen, Ci-βalkyl, C₂-₆alkenyl, (C3-₈cycloalkyl)Co- ₆alkyl, Cι-₆fluoroalkyl, heteroarylCo-δalkyl or heterocycleCo.₆alkyl; and said Cι_₆alkyl, C₂-₆alkenyl, (C₃-₈cycloalkyl)C₀-₆alkyl, -efluoroalkyl, heteroarylC₀-₆alkyl or heterocycleC₀-₆alkyl, may be substituted with one or more B;

or R⁶ and R⁷ form together a 4-, 5- or 6- membered heterocyclic ring containing 1 or 2 heteroatoms independently selected from N, O and S, and said ring may be substituted with one or more B;

B is R⁸, -COOR⁸, -COR⁸, -NHCOR⁸, -NR⁸R⁹, -CONR⁸R⁹, -OR⁸, -SO₂NR⁸R⁹, CN, or halogen;

R⁸ and R⁹ each independently are hydrogen, Ci-βalkyl, Ci-βfluoroalkyl, or hydroxyCi-όalkyl or;

R⁸ and R⁹ form together a 4-, 5- or 6- membered heterocyclic ring containing 1 or 2 heteroatoms independently selected from N, O and S, and said ring may be substituted with one or more B;

as a free base or a salt thereof.

Prefarably, the compounds of formula I are present in the form as a pharmaceutically acceptable salt.

Listed below are definitions of various terms used in the specification and claims to describe the present invention.

For the avoidance of doubt it is to be understood that where in this specification a group is qualified by 'hereinbefore defined' or 'defined hereinbefore' the said group encompasses the first occurring and broadest definition as well as each and all of the preferred definitions for that group.

For the avoidance of doubt it is to be understood that in this specification 'C₀.₆' means a carbon group having 0, 1, 2, 3, 4, 5 or 6 carbon atoms. For the avoidance of doubt it is to be understood that in this specification ^ζC_\A means a carbon group having 1, 2, 3, 4, 5 or 6 carbon atoms.

In this specification, unless stated otherwise, the term "alkyl" includes both straight and branched chain alkyl groups. ^a-kyl may be methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, n-pentyl, i-pentyl, t-pentyl, neo-pentyl, and hexyl.

In this specification, unless stated otherwise, the term "C₃.₈ cycloalkyl" includes a non- aromatic, completely saturated cyclic aliphatic hydrocarbon group containing 3 to 8 atoms. Examples of said cycloalkyl include, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

In this specification, unless stated otherwise, the term "alkenyl" includes both straight and branched chain alkenyl groups but references to individual alkenyl groups such as

2-butenyl is specific for the straight chain version only. Unless otherwise stated, the term "alkenyl" advantageously refers to chains with 2 to 5 carbon atoms, preferably 3 to 4 carbon atoms. C₂.₆alkenyl may be, but is not limited to, ethenyl, propenyl, 2- methylpropenyl, butenyl and 2-butenyl.

In this specification, unless stated otherwise, the terms "heterocyclic ring" and "heterocycle" includes a 3- to 10-membered non-aromatic partially or completely saturated hydrocarbon group, which contains one or two rings and at least one heteroatom. Examples of said heterocycle include, but are not limited to pyrrolidinyl, pyrrolidonyl, piperidinyl, piperazinyl, morpholinyl, oxazolyl, 2-oxazolidonyl or tetrahydrofuranyl.

In this specification, unless stated otherwise, the expression "R⁶R⁷ form together a 4-, 5- or 6-membered heterocyclic ring containing 1 or 2 heteroatoms independently selected from N, O and S," include, but are not hmited to piperidinyl, piperazinyl and morpholinyl. In this specification, unless stated otherwise, the expression "R⁸R⁹form together a 4-, 5- or 6-membered heterocyclic ring containing 1 or 2 heteroatoms independently selected from N, O and S," include, but are not limited to piperidinyl, piperazinyl and morpholinyl.

In this specification, unless stated otherwise, the term "heteroaryl" may be a monocyclic heteroaromatic, or a bicyclic fused-ring heteroaromatic group. Examples of said heteroaryl include, but are not limited to, pyridyl, pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, pyrazolyl, benzofuryl, indolyl, isoindolyl, benzimidazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, tetrazolyl or triazolyl.

In this specification, unless stated otherwise, the term "halogeno" may be fluoro, chloro, bromo or iodo.

In this specification, unless stated otherwise, the term " -e fluoroalkyl" may be an alkyl substituted with one or more fluorine atoms. Examples of said fluoroalkyl include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl and trifluoroethyl.

In another aspect of the invention there are provided compounds according to formula I wherein n is 1.

In yet another aspect of the invention there are provided compounds according to formula I wherein R² is halogen. Preferably said halogen is chloro.

In yet another aspect of the invention there are provided compounds according to formula I wherein R³, R⁴, and R⁵ is hydrogen.

I-n yet another aspect of the invention there are provided compounds according to formula I wherein R is selected from hydrogen and COOR .

In one embodiment of this aspect there are provided compounds according to formula I wherein R¹ is COOR⁶. Said R⁶ may be Cj-ealkyl optionally substituted with B. In one class of this embodiment, B is -OR⁸ or NR⁸R⁹, wherein R⁸ may be Cι-₆alkyl, preferably methyl. If B is NR⁸R⁹, R⁸ and R⁹ may be Cι-₆alkyl, preferably methyl.

In yet another aspect of the invention there are provided compounds according to formula I, wherein n is 1 ; R¹ is hydrogen, or COOR⁶, R² is chloro; and R³, R⁴, and R⁵ are hydrogen.

In one embodiment of this aspect there are provided compounds according to formula I, wherein R¹ is COOR⁶, and said R⁶ is

substituted with B; said B being -OR⁸ or NR⁸R⁹.

In another aspect of the invention there are provided compounds said compounds being:

5-[(5-chloro-3H-benzimidazol-2-yl)thio] - 1 ,3-dihydro-2H-indol-2-one; 2-Methoxyethyl 5-chloro-2-[(2-oxo-2,3-dihydro-lH-indol-5-yl)thio]-3H-benzimidazole-4- carboxylate; 2-(Oimethylamino)ethyl 5-chloro-2-[(2-oxo-2,3-dihydro-lH-indol-5-yl)thio]-3H- benzimidazole-4-carboxylate; as a free base or a salt thereof.

The present invention relates to the use of compounds of formula I as hereinbefore defined as well as to the salts thereof. Salts for use in pharmaceutical formulations will be pharmaceutically acceptable salts, but other salts may be useful in the production of the compounds of formula I. Such salts are possible, includes both pharmaceutically acceptable acid and base addition salts. A suitable pharmaceutically-acceptable salt of a compound of Formula I is, for example, an acid-addition salt of a compound of Formula I which is sufficiently basic, for example an acid-addition salt with an inorganic or organic acid such as hydrochloric.

Some compounds of formula I may have chiral centres and/or geometric isomeric centres (E- and Z- isomers), and it is to be understood that the invention encompasses all such optical, diastereoisomers and geometric isomers.

Certain compounds of the present invention may exist as tautomers. It is to be understood that the present invention encompasses all such tautomers. The present invention relates to novel benzimidazole derivatives, which are inhibitors of c- Jun N-terminal kinases (JNKs). JNKs have been implicated in mediating a number of disorders. The invention relates to methods for producing these inhibitors. The invention also provides pharmaceutical compositions comprising the inhibitors of the invention and methods of utilizing these compositions in the treatment of various disorders.

PHARMACEUTICAL COMPOSITIONS According to one aspect of the present invention.there is provided a pharmaceutical composition comprising a compound of formula I, as a free base or a pharmaceutically acceptable salt, solvate or solvate of salt thereof, for use in the prevention and/or treatment of conditions associated with c-Jun N-terminal kinases (JNKs).

The composition may be in a form suitable for oral administration, for example as a tablet, for parenteral injection as a sterile solution or suspension. In general the above compositions may be prepared in a conventional manner using pharmaceutically carriers or diluents. Suitable daily doses of the compounds of formula I in the treatment of a mammal, including man, are approximately 0.01 to 250 mg/kg bodyweight at peroral administration and about 0.001 to 250 mg/kg bodyweight at parenteral administration. The typical daily dose of the active ingredients varies within a wide range and will depend on various factors such as the relevant indication, the route of administration, the age, weight and sex of the patient and may be determined by a physician.

A compound of formula I, or a pharmaceutically acceptable salt, solvate or solvate of salt thereof, can be used on its own but will usually be administered in the form of a pharmaceutical composition in which the formula I compound/salt (active ingredient) is in association with a pharmaceutically acceptable diluent or carrier. Dependent on the mode of administration, the pharmaceutical composition may comprise from 0.05 to 99 %w (per cent by weight), for example from 0.10 to 50 %w, of active ingredient, all percentages by weight being based on total composition. A diluent or carrier includes water, aqueous polyethylene glycol, magnesium carbonate, magnesium stearate, talc, a sugar (such as lactose), pectin, dextrin, starch, tragacanth, microcrystalline cellulose, methyl cellulose, sodium carboxymethyl cellulose or cocoa butter.

A composition of the invention can be in tablet or injectable form. The tablet may additionally comprise a disintegrant and/or may be coated (for example with an enteric coating or coated with a coating agent such as hydroxypropyl methylcellulose).

The invention further provides a process for the preparation of a pharmaceutical composition of the invention which comprises mixing a compound of formula I, or a pharmaceutically acceptable salt thereof, as hereinbefore defined, with a pharmaceutically acceptable diluent or carrier.

An example of a pharmaceutical composition of the invention is an injectable solution containing a compound of the invention, or a a pharmaceutically acceptable salt, solvate or solvate of salt thereof, as hereinbefore defined, and sterile water, and, if necessary, either sodium hydroxide or hydrochloric acid to bring the pH of the final composition to about pH 5, and optionally a surfactant to aid dissolution.

Liquid solution comprising a compound of formula I, or a salt thereof, dissolved in water.

MEDICAL USE

The novel substituted substituted benzimidazole derivatives of the present invention have been found to have activity as medicaments. It has been found that the compounds of the present invention are well suited for inhibiting JNK. Accordingly, the compounds of the present invention are expected to be useful in the treatment of JNK-mediated condition, i.e. the compounds may be used to produce an inhibitory effect of JNK in mammals, including man, in need of such treatment.

The term "JNK-mediated condition", as used herein means any disease or other deleterious condition in which JNK is known to play a role. Such conditions include, without limitation, inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, cancer, infectious diseases, neurodegenerative diseases, allergies, reperfusion/ischemia in stroke, heart attacks, angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, thrombin-induced platelet aggregation, and conditions associated with prostaglandin endoperoxidase synthase-2.

Inflammatory diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, acute pancreatitis, chronic pancreatitis, asthma, allergies and adult respiratory distress syndrome.

Autoimmune diseases which may be treated or prevented by the compounds of this invention include, but are not Hmited to, glomerulonephritis, rheumatoid arthritis, systemic lupus erythematosus, scleroderma, chronic thyroiditis, Graves' disease, autoimmune gastritis, diabetes, autoimmune hemolytic anemia, autoimmune neutropenia, thrombocytopenia, atopic dermatitis, chronic active hepatitis, myasthenia gravis, multiple sclerosis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, psoriasis and graft vs. host disease.

Destructive bone disorders which may be treated or prevented by the compounds of this invention include, but are not limited to, osteoporosis, osteoarthritis and multiple myeloma-related bone disorder.

Proliferative diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, acute myelogenous leukemia, chronic myelogenous leukemia, metastatic melanoma, Kaposi's sarcoma, multiple myeloma and HTLV-1 mediated tumorigenesis. Angiogenic disorders which may be treated or prevented by the compounds of this invention include, but are not limited to, solid tumors, ocular neovasculization and infantile haemangiomas.

Infectious diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, sepsis, septic shock and Shigellosis.

Niral diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, acute hepatitis infection (including hepatitis A, hepatitis B and hepatitis C), HIN infection and CMV retinitis.

Neurodegenerative diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, Alzheimer's disease, cognitive disorders, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, Frontotemporal dementia Parkinson's Type, Parkinson dementia complex of Gaum, HIV dementia, corticobasal degeneration, dementia pugilistica, Down's syndrome, postencephelatic parkinsonism, progressive supranuclear palsy, Pick's Disease, Niemann-Pick's Disease, epilepsy, a peripheral neuropathy, apoptosis-driven neurodegenerative disease that may be caused by traumatic injury, traumatic brain injury, cerebral ischemias, spinal cord injury, head trauma, ALS, acute hypoxia, ischemia, glutamate neurotoxicity, ischemic and haemorrhaging stroke and cancer.

"JNK-mediated conditions" also include ischemia/reperfusion in stroke, heart attacks, myocardial ischemia, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, hepatic ischemia, liver disease, congestive heart failure, pathologic immune responses such as that caused by T cell activation and thrombin-induced platelet aggregation.

In addition, compounds of the instant invention may be capable of inhibiting the expression of inducible pro-inflammatory proteins. Therefore, other "JNK-mediated conditions" which may be treated or prevented by the compounds of this invention include edema, analgesia, fever and pain, such as neuromuscular pain, headache, cancer pain, dental pain and arthritis pain. One embodiment of the invention relates to the use of the compounds of formula (I) in the treatment JNK mediated conditions selected from the group consisting of Alzheimer's disease, cognitive disorders, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, Frontotemporal dementia Parkinson's Type, Parkinson dementia complex of Gaum, HTV dementia, corticobasal degeneration, dementia pugilistica, Down's syndrome, postencephelatic parkinsonism, progressive supranuclear palsy, Pick's Disease, Niemann- Pick's Disease, epilepsy, a peripheral neuropathy, spinal cord injury, head trauma, ALS, traumatic brain injury, haemorrhaging stroke and cancer.

In a preferred embodiment the invention the condition is Alzheimer's Disease.

The present invention relates also to the use of the compound of formula (I) as defined hereinbefore, in the manufacture of a medicament for the treatment of JNK mediated condition and any other condition mentioned hereinbefore.

The present invention relates further to the use of the compound of formula (I) as defined hereinbefore, in the manufacture of a medicament for the treatment of Alzheimer's disease, cognitive disorders, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, Frontotemporal dementia Parkinson's Type, Parkinson dementia complex of Gaum, HTV dementia, corticobasal degeneration, dementia pugilistica, Down's syndrome, postencephelatic parkinsonism, progressive supranuclear palsy, Pick's Disease, Niemann- Pick's Disease, epilepsy, a peripheral neuropathy, spinal cord injury, head trauma, ALS, traumatic brain injury, haemorrhaging stroke and cancer.

The present invention relates to the use of the compound of formula (I) as defined hereinbefore, in the manufacture of a medicament for the treatment of Alzheimer's disease.

According to another embodiment, the invention provides a method of treatment of JNK mediated conditions and any other condition mentioned hereinbefore, comprising administering to a patient in need of such treatment, a therapeutically effective amount of the compound of formula (I). In the context of the present specification, the term "therapy" and "treatment" also includes "prevention" unless there are specific indications to the contrary. The terms "treat", "therapeutic" and "therapeutically" should be construed accordingly.

The term "patient", as used herein, means an animal, preferably a human.

In this specification, unless stated otherwise, the term 'inhibitor' means a compound that by any means, partly or completely, blocks the transduction pathway leading to the production of a response by the ligand.

The term "condition", unless stated otherwise, means any disorder and disease associated with JNK activity.

NON-MEDICAL USE

In addition to their use in therapeutic medicine, the compounds of formula I or salt thereof, are also useful as pharmacological tools in the development and standardisation of in vitro and in vivo test systems for the evaluation of the effects of JNK inhibitor related activity in laboratory animals such as cats, dogs, rabbits, monkeys, rats and mice, as part of the search for new therapeutics agents.

METHODS OF PREPARATION

The compounds of this invention may be prepared by methods known to those skilled in the art for analogous compounds, as illustrated by the general schemes and procedures below and by the preparative examples that follow.

All starting materials are commercially available or prepared by methods known to those skilled in the art for analogous compounds, earlier described in the literature.

An aspect of the invention relates to a process for the preparation of compounds of formula (I) comprising of reacting a compound of formula (II):

wherein R⁴ and R⁵ are as defined in formula (I) or are protected derivatives thereof; with a compound of formula (TTT):

(in)

wherein R¹, R² and R³ are as defined in formula (I) or are protected derivatives thereof and L' is a leaving group such as SO₂Me or halogen such as fluoro, chloro or bromo. This reaction may be performed in refluxing alcohol, such as isopropanol or sec-butyl alcohol, or in refluxing ethyl acetate for 2-48 h.

Intermediate compounds of formula (TT) can be prepared using known chemistry, as described in the preparative examples, for example according to the scheme below: The 5-mercapto-oxindole intermediate (II) can be prepared, for example according to the scheme below, by heating oxindole with chlorosulphonic acid ( Justus Liebigs Ann. Chem.;1896 (291); 328). The 5-chlorosulfonyl group can be reduced with a reducing agent such as triphenylphosphine to yield the 5-mercapto-oxindole intermediate (II). CiSO_gH

Intermediate compounds of formula (TTT) can be prepared using known chemistry, as described in the preparative examples, for example according to the scheme below: The nitro group can be reduced with hydrogen in the presence of a catalyst such as Pd or Pt on carbon. The diamino compound is reacted with carbon disulfide in an inert solvent such as dimethyl formarnide (Org. Synth. 30, 1950, 56) to yield the 2-mercapto compound. Methylation of the 2-mercapto compound can be performed with iodomethane in a solvent such as acetone or methylene chloride in the presence of a base such as potassium carbonate at room temperature (J. Chem. Soc. 1949, 3311-3312, Tetrahedron, 1995, 11515-11530). Oxidation of the 2-methylthio group to the 2-methanesulfonyl group can be performed with oxidizing agents such as m-chloroperoxy benzoic acid or oxone at room temperature (J. Chem. Soc. 1949, 3311-3312, J. Heterocycl. Chem.1995, 707-718) to yield (]H).

It will be appreciated by those skilled in the art that in the processes of the present invention certain functional groups such as carboxylic acids or amino groups in the starting reagents or intermediate compounds may need to be protected by protecting groups. Thus, the preparation of the compounds of formula (I) may involve, at an appropriate stage, the addition and removal of one or more protecting groups.

The protection and deprotection of functional groups is fully described in 'Protective Groups in Organic Chemistry', edited by J. W. F. McOmie, Plenum Press (1973), and 'Protective Groups in Organic Synthesis', 2nd edition, T. W. Greene & P. G. M. Wuts, iley-Interscience (1991).

The compounds of formula (I) may be converted to a further compound of formula (I) using standard chemistry, for example, acylation of amine groups. These acylations can be performed by reacting the amine group with the corresponding acid in an inert solvent such as DMF in the presence of coupling reagents such as 1,3-dicyclohexylcarbodiimide, l-(3- climethylaminopropyl)-3-ethylcarbodiimide, HATU or TBTU.

The compounds of formula (I) may be converted to a further compound of formula (I) using standard chemistry, for example, alkylation of amine groups. These alkylations may be performed by reacting the amine group with an aldehyde in the presence of a reducing agent such as sodium triacetoxyborohydride or sodium cyanoborohydride.

The compounds of formula (I) may be converted to a further compound of formula (I) using standard chemistry, for example, amidation of carboxylic acids. These amidations can be performed by reacting the carboxylic acid group with the corresponding amine in an inert solvent such as DMF in the presence of coupling reagents such as 1,3- dicyclohexylcarbodiimide, l-(3-dimethylarninopropyl)-3-ethylcarbodiimide, HATU or TBTU.

EXAMPLES

The invention will now be described in more detail with the following examples that are not to be construed as limiting the invention.

All chemicals and reagents were used as received from suppliers. 1H and ¹³C nuclear magnetic resonance (NMR) spectra were recorded on a BRUKER DPX 400 (400 MHz) spectrometer using the following solvents and references.

CDC1₃ : 1H NMR TMS (0.0 ppm) and ¹³C the central peak of CDC1₃ (77.0). CD₃OD : 1H NMR 3.31 ppm (central peak) and ¹³C 49.0 ppm (central peak). DMSO-_fo 1H NMR 2.50 ppm (central peak) and ¹³C 39.51 ppm (central peak). LC-MS were recorded on a Waters Alliance 2790 + ZMD spectrometer equipped with software Mass Lynx 3.5. MS (ECI)

Flash column chromatography was carried out on silica gel 60 (230-400 mesh). Petroleum ether with boiling range 40-60°C was used.

Ambient temperature is defined as a temperature between 16 and 25 °C.

List of abbreviations DMF N,N-Dimemylformamide

EtOAc Ethyl acetate

MeOH Methanol

THF Tetrahydrofuran

DIPEA Diisopropylethylamine HATU O-(7-Azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate

TBTU O lH-Benzo1riazol-l-yl)-N,N,N',N'-tetramemyluronium tetrafluoroborate

ΗOBt 1-Ηydroxybenzotriazole hydrate

Example 1 5-[(5-chloro-3H-benzimidazol-2-yl)thio]-l,3-dihydro-2-!-r-indol-2-one

(i) 2-Oxoindoline-5-sulfonyl chloride

Oxindole (1.33g, 10 mmol) was slowly added to cooled clorosulfonic acide (2.7 ml). The mixture was slowly allowed to warm to room temperature and stirred lh at this temperature before heated to 68°C for 2h. After cooling to 0°C the reaction mixture was poured on ice to quench and the product was filtered of and washed with cold water. After drying 1.24 g (59%) of the title compound was obtained. 1H ΝMR (DMSO-d6): δ 3.61, (2H, s), 6.75 (1H, s), 7.50 (2H, d) 10.61 (1H, s, ΝH). ¹³C ΝMR (DMSO-d6): δ 36.1, 108.2, 122.3, 125.3, 125.4, 141.7, 144.3, 176.9. MS (ECI) m/z 232 (M+l).

(ii) 5-Mercapto-l,3-dihydro-2H-indol-2-one To a solution of 2-oxoindoline-5-sulfonyl chloride (940 mg, 4.0 mmol) in 12 ml THF triphenylphosphine (3.6 g, 14 mmol) was added followed by 0.5 ml of water. The solution was heated to 50°C for 90 min. After cooling to room temperature 15 ml water and 15 ml 2M NaOH were added and the aqueous phase was washed 2 times with 10 ml ethylacetat. Addition of ca. 15 ml 2M HC1 to the aq. phase (« pH 2) and filtration of the resulting product lead after rinsing with water and drying in vacuum to 607mg (92%) of the title compound. 1H NMR (DMSO-d6): δ 2.58 (2H, s), 5.17 (1H, s, NH), 6.82 (1H, s), 7.25 (2H, d), 10.31 (1H, s, SH). ¹³C NMR (DMSO-d6): δ 34.1, 108.2, 120.1, 124.3, 125.4, 126.9, 139.8, 174.3. MS (ECI) mJz 166 (M+l).

(iii) 5-[(5-chloro-3H-benzimidazol-2-yl)thio]-l,3-dihydro-2H-indol-2-one

5-Chloro-2-(methylsulfonyl)-3H-benzimidazole (23 mg, 0.1 mmol) and 5-mercapto-l,3- dihydro-2H-indol-2-one (16 mg, 0.1 mmol) were dissolved in 3 ml isopropanol heated to 68°C for 72h. After cooling to room temperature the solvent was removed and the crude product purified by preparative reversed phase ΗPLC to result 28 mg (91%) of the title compound.

1H NMR (DMSO d6): δ 2.47 (2Η, s), 6.82 (1H, d), 7.25 (1H, d), 7.82 (3H, m), 7.5 (1H, m),

7.89 (1H, NH), 10.27 (1H, NH). MS (ECI) m/z 316 (M+l).

Example 2

2-Methoxyethyl 5-chloro-2-[(2-oxo-2,3-dihydro-lH-indol-5-yl)thio]-3-ff- benzimidazole-4-carboxylate (i) 2-Amino-6-chloro-3-nitrobenzonitήle

2,6-Dichloro-3-nitrobenzonitrile (lOOg) was suspended in methanol (1.2L) and the mixture saturated with gaseous ammonia. After 4 days the mixture was filtered to give 2-amino-6- chloro-3-nitrobenzonitrile as a bright yellow solid (60g). 1H NMR (d₆-DMSO) : δ 8.32 (1H, dd), 7.82 (2H, broad s), 6.96 (1H, dd).

(ii) 2, 3-Diamino-6-chlorohenzonitrile 2-Amino-6-chloro-3-nitrobenzonitrile (8g) in acetic acid (100ml) was treated with 5% palladium on carbon (lg) and the mixture hydrogenated at 20 atmospheres for 16hrs. The catalyst was filtered off and the acetic acid removed in vacuo to give 2,3-diamino-6- chlorobenzonitrile.

1H NMR (d₆-DMSO) : δ 6.63(lH,dd), 6.55 (1H, dd), 5.73 (2H, broad s), 5.13 (2H, broad s).

(iii) 5-Chloro-2-mercapto-3H-benzimidazole-4-carbonitrile

2,3-Diamino-6-chlorobenzonitrile (7g) was treated with a 50% solution of carbon disulfide in dimethyl formamide (100ml) and the solution heated in a sealed bomb at 70 °C for 16hrs. After cooling the bomb was vented and the mixture poured into water (500ml). The product was collected by filtration (8g) 1H NMR (d₆-DMSO) : δ 13.71 (1H, broad s), 13.11 (1H, broad s), 7.37 (2H, S)

(iv) 5-Chloro-2-(methylthio)-3H-benzimidazole-4-carbonitrile 5-Chloro-2-mercapto-3H-benzimidazole-4-carbonitrile (8g) in acetone (150ml) was treated with solid potassium carbonate (8g). The suspension was carefully titrated at room temperature with methyl iodide (0.5g aliquots) using lc-ms to ascertain disappearance of starting material and appearance of S-methylated material. When all the starting material had been consumed, the acetone was removed in vacuo and the residue partitioned between water and ethyl acetate to give a brown solid (8g).

1H NMR (de-DMSO) at 90°C: δ 12.81 (1H, broad s), 7.86 (1H, broad s), 7.4 (2H, broad ), 2.72 (3H, broad s)

(v) 5-Chloro-2-(methylthio)-3H-benzimidazole-4-carboxylic acid 5-Chloro-2-(methylthio)-3H-benzimidazole-4-carbonitrile (2.4g) in 10M sodium hydroxide (40ml) was refluxed for 16hrs. The reaction was cooled and quenched with 2M hydrochloric acid until it reached pH=7 and a precipitate formed. The carboxylic acid was isolated by filtration and air dried to give a brown solid (2g). MS: APCI(-ve) 242 (M-l)

(vi) 2-Methoxyethyl 5-chloro-2-(methylthio)-3H-benzimidazole-4-carboxylate 5-Chloro-2-(methylthio)-3H-benzimidazole-4-carboxylic acid (121 mg, 0.5 mmol), HATU (380 mg, lmmol) and DIPEA (348 μl, 2mmol) were dissolved in 3 ml dry DMF and stirred at room temperature for 10 min before 2-methoxy-ethanol (54 μl, lmmol) was added to the reactions mixture. After 48h the solvent was removed in vacuum and the crude product purified on silica (EtOAc/MeOH 9/1) to result 102 mg (68%) of the title compound. 1H NMR (CDCl₃-d3): δ 2.74, (3H, s), 3.55 (3H, s), 3.75 (2H, t) 4.48 (2H, t), 7.21 (IH, d), 7.64 (IH, d).

MS (ECI) m/z 301 (M+l).

(vii) 2-Methoxyethyl 5-chloro-2-[(2-oxo-2,3-dihydro-lH-indol-5-yl)thio]-3H- benzimidazole-4-carboxylate 2-Methoxyethyl 5-chloro-2-(methylthio)-3H-benzimidazole-4-carboxylate (12 mg, 0.04 mmol) was dissolved in 1 ml MeOΗ and Oxone^® (61 mg, lmmol) was added. The slurry was stirred at room temperature for 6h, filtered and washed with little MeOΗ. The solvent was removed and the crude product taken up in water. The resulting solution is neutralized with 2M NaOΗ and extracted with EtOAc (3 x 5 ml). The Organic phase is dried over Na₂SO₄ and concentrated to ca. 1/5 of its volume. To this solution 5-mercapto-l,3-dihydro- 2H-indol-2-one (7 mg, 0.04 mmol) and 1 ml isopropanol were added and the mixture heated to 68°C for 72h. After cooling to room temperature the solvent was removed and the crude product purified by preparative reversed phase ΗPLC to result 7 mg (42%) of the title compound. 1H NMR (MeOΗ d4): δ 2.16 (3Η, s), 2.28 (2H, s), 2.47 (2H, t), 3.26 (2H, t), 5.69 (IH, d), 6.01 (IH, d), 6.26 (3H, m). MS (ECI) m z 418 (M+l).

Example 3 2-(T imethylamino)ethyl 5 hloro-2 (2-oxo-2,3-dihydro-lH-indol-5-yl)thio]-3H- benzimidazole-4-carboxylate

(i) 2-(Dimethylamino)ethyl 5-chloro-2-(methylthio)-3H-benzimidazole-4-carboxylate 5-Chloro-2-(methylthio)-3H-benzimidazole-4-carboxylic acid (121 mg, 0.5 mmol), HATU (380 mg, lmmol) and DIPEA (348 μl, 2mmol) were dissolved in 3 ml dry DMF and stirred at room temperature for 20 min before 2-dimethylamino-ethanol (100 μl, lmmol) was added to the reactions mixture. After 48h the solvent was removed in vacuum and the crude product purified by silica gel chromatography (EtOAc/MeOH 9/1) to result 71 mg (45%) of the title compound.

1H NMR (MeOH-d4): δ 1.09, (6H, s), 1.39 (3H, s), 1.42 (2H, t) 3.17 (2H, t), 5.90 (IH, d), 6.24 (IH, d). MS (ECI) m/z 312 (M-l).

(ii) 2-(Dimethylamino)ethyl 5-chloro-2-[(2-oxo-2,3-dihydro-lH-indol-5-yl)thio]-3H- benzimidazole-4-carboxylate

2-(Dimethylamino)ethyl 5-chloro-2-(methylthio)-3H-benzimidazole-4-carboxylate (16 mg, 0.05 mmol) was dissolved in 2 ml EtOAc and Oxone^® (67 mg, 1. lmmol) was added. The slurry was stirred at room temperature for 6h, filtered and washed with little EtOAc. 2ml 2M NaOH and 2ml water were added to the EtOAc solution. The phases were separated and the aqueous phase extracted with EtOAc (3 x 5 ml). The organic phase is dried over Na₂SO₄ and concentrated to ca. 1/5 of its volume. To this solution 5-mercapto-l,3-dihydro- 2H-indol-2-one (8 mg, 0.05 mmol) was added and the mixture heated to 68°C for 72h.

After cooling to room temperature the solvent was removed and the crude product purified by preparative reversed phase ΗPLC to result 6 mg (28%) of the title compound. 1HNMR (MeOΗ-d4): δ 1.13 (6H, s),1.47 (2H, t), 1.91 (2H, s), 3.23 (2H, t), 5.62 (IH, d), 5.59 (IH, d), 6.13 (2H, m), 6.24 (IH, d). MS (ECI) m/z 431 (M+l).

Biological evaluation

The compounds of this invention may be assayed for their activity according to the following procedure:

A scintillation proximity assay (SPA) based on the inhibition of JNK3 catalyzed transfer of the γ-phosphate group of [γ- 33 P] ATP to biotinylated ATF2, has been set up to identify inhibitory compounds. The resulting 33 P-labeled biotinylated ATF2 is trapped on SPA beads surface coated with streptavidin.

The assay is performed in 96-well plates. Test compounds made up at 10 mM in DMSO and 1:3 serial dilutions are made in 100% DMSO. These serial dilutions are then diluted 1:10 in assay buffer (50 mM MOPS pH 7.2, 150 mM, NaCl, 0.1 mM EGTA, 1 mM DTT, 6.25 mM β-glycerolphosphate) and 10 μl are transferred to assay plates (results in 2% DMSO final concentration in assay). To each well with test compound a 2.4 μl JNK3/ATP enzyme solution (1.18 U/ml JNK3, 20 μM ATP, 2 mM Mg(Ac)₂, 0.01 % Brij-35 in assay buffer) was added. The mixture was pre-incubated for 10 minutes at ambient temperature. After this, 3.6 μl of a [γ-³³P] ATP-solution (0.20 μCi/μl [γ-³³P]ATP, 66.6 mM Mg(Ac)₂, 1 mM DTT, 50 mM MOPS pH 7.2, 150 mM NaCl, 0.1 mM EGTA) was added to each well followed by 10 μl of a ATF2 solution (60 μg/ml biotinylated ATF2 in assay buffer) to start the reaction. The reaction was allowed to proceed for 10 minutes at ambient temperature. After this, the reaction was terminated by the addition of 200 μl per well of stop buffer/bead mix (0.4 mg/ml streptavidin coated SPA-beads in 50 mM EDTA, pH 7.6). Plates were sealed with a plastic cover and centrifuged (2000 rpm, 5 minutes) to settle the beads followed by counting in a Wallac 1450 microbeta™. ;-

The IC₅₀ values were calculated as the concentration of test compound at which the ATF2 phosphorylation is reduced to 50% of the control value.

Results

Typical Ki values for the compounds of the present invention are in the range of about 0.001 to about 10,000 nM. Other values for Ki are in the range of about 0.001 to about

1000 nM. Further values for j are in the range of about 0.001 nM to about 300 nM.

List of abbreviations

SPA scintillation proximity assay

ATP adenosine triphosphate ATF Activating transcription factor

MOPS 3-[N-Morpholino]-propanesulfonic acid

EGTA Ethylene glycol-bis(β-arninoethylether)-N,N,N',N'-tetrααcetιc acid

DTT dithiothreitol

JΝK Jun Ν-terminal kinases MAP mitogen-activated protein

Claims

1. A compound of Formula I:

(I) wherein: n is 1 or 2

R⁴ is selected from hydrogen, COOR⁶, CONR⁶R⁷, NO₂, NR⁶R⁷, or NR⁶COR⁷; R⁵ selected from hydrogen, NR⁶R⁷ or NR⁶COR⁷;

R⁶ and R⁷ are each independently hydrogen, Ci-βalkyl, C₂-₆alkenyl, (C₃-₈cycloalkyl)Co.₆alkyl, Cι-₆fluoroalkyl, heteroarylCo-ealkyl or heterocycleCo.₆alkyl; and said -_δalkyl, C₂-₆alkenyl, (C₃-₈cycloalkyl)Co-₆alkyl, -efluoroalkyl, heteroarylCo-₆alkyl or heterocycleCo-₆alkyl, may be substituted with one or more B;

B is R⁸, -COOR⁸, -COR⁸, -NHCOR⁸, -NR⁸R⁹, -CONR⁸R⁹, -OR⁸, -SO₂NR⁸R⁹, CN, or halogen; R⁸ and R⁹ each independently are hydrogen, Ci-₆alkyl, Ci-efluoroalkyl, or hydroxyCι.₆alkyl, or;

R⁸ and R⁹ form together a 4-, 5- or 6- membered heterocyclic ring containing 1 or 2 heteroatoms independently selected from N, O and S, and said ring may be substituted with one or more B; as a free base or a salt thereof.

2. A compound according to claim 1 as a pharmaceutically acceptable salt thereof.

3. A compound according to claim 1 or 2, wherein n is 1.

4. A compound according to any one of claims 1 to 3, wherein R² is halogen,

5. A compound according to claim 4, wherein said halogen is chloro.

6. A compound according to any one of claims 1 to 5, wherein R³, R⁴ and R⁵ are hydrogen.

7. A compound according to any one of claims 1 to 6, wherein R¹ is selected from hydrogen and COOR⁶.

8. A compound according to claim 7, wherein R¹ is COOR⁶.

9. A compound according to claim 8, wherein R⁶ is C βalkyl optionally substituted with B.

10. A compound according to claim 9, wherein B is -OR⁸ or NR⁸R⁹.

11. A compound according to claim 10, wherein R⁸ is -βalkyl, preferably methyl.

12. A compound according to claim 10, wherein R⁸ and R⁹ is d-βalkyl, preferably methyl.

13. A compound according to claim 1, wherein n is 1; R 1 i •s hydrogen, or COOR , R is chloro; and R³, R⁴, and R⁵ is hydrogen.

14. A compound according to claim 14, wherein R¹ is COOR⁶, and said R⁶ is -ealkyl 5 substituted with B ; said B being -OR⁸ or NR⁸R⁹.

15. A compound which is: 5-[(5-chloro-3H-benzimidazol-2-yl)thio]-l,3-dihydro-2H-indol-2-one; l o 2-Methoxyethyl 5 -chloro-2- [(2-oxo-2,3 -dihydro- lH-indol-5 -yl)thio]-3H-benzimidazole-4- carboxylate;

2-(Dimethylamino)ethyl 5-chloro-2-[(2-oxo-2,3-dihydro-lH-indol-5-yl)thio]-3H- benzimidazole-4-carboxylate;

15 as a free base or a salt thereof.

16. Processes for the preparation of a compound as defined in claims 1 or 15 comprising reacting a compound of formula (II):

20 wherein R and R⁵ are as defined in formula (I); with a compound of formula (HI):

(m) wherein R¹, R² and R³ are as defined in formula (I) and L' is a leaving group.

25

17. A pharmaceutical formulation comprising as active ingredient a therapeutically effective amount of the compound of any one of claims 1 to 15 in association with pharmaceutically acceptable carriers or diluents.

18. A pharmaceutical formulation comprising as active ingredient a therapeutically effective amount of the compound of any one of claims 1 to 15 for use in the prevention and/or treatment of conditions associated with JNK activation.

19. A compound according to any one of claims 1 to 15 for use in therapy.

20. Use of a compound according to any one of claims 1 to 15 in the manufacture of a medicament for the prevention and/or treatment of conditions associated with JNK activation.

21. Use of a compound according to any one of claims 1 to 15 in the manufacture of a medicament for the prevention and/or treatment of inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, cancer, infectious diseases, neurodegenerative diseases, allergies, reperfusion/ischemia in stroke, heart attacks, angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, thrombin- induced platelet aggregation, and conditions associated with prostaglandin endoperoxidase synthase-2.

22. Use of a compound according to any one of claims 1 to 15 in the manufacture of a medicament for the prevention and/or treatment of conditions selected from: central or peripheral neurological degenerative disorders including Alzheimer's disease, cognitive disorders, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, Frontotemporal dementia Parkinson's Type, Parkinson dementia complex of Gaum, RTV dementia, corticobasal degeneration, dementia pugilistica, Down's syndrome, postencephelatic parkinsonism, progressive supranuclear palsy, Pick's Disease, Νiemann- Pick's Disease, epilepsy, a peripheral neuropathy, spinal cord injury, head trauma, ALS, traumatic brain injury, haemorrhaging stroke and cancer.

23. The use according to claim 22, wherein said condition is Alzheimer's disease.

24. Use of a compound according to any one of claims 1 to 15 in the manufacture of a medicament for the prevention and/or treatment of conditions associated with inhibiting the expression of inducible pro-inflammatory proteins.

25. The use according to claim 24, wherein said condition is selected from edema, analgesia, fever and pain, such as neuromuscular pain, headache, cancer pain, dental pain and arthritis pain.

26. A method of treating or preventing conditions associated with JNK activation comprising the administration of a therapeutically effective amount of a compound of Formula I according to any one of claims 1 to 15 to a mammal in need thereof.

27. A method of treating or preventing conditions selected from of inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, cancer, infectious diseases, neurodegenerative diseases, allergies, reperfusion/ischemia in stroke, heart attacks, angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, thrombin-induced platelet aggregation, and conditions associated with prostaglandin endoperoxidase synthase-2, comprising the administration of a therapeutically effective amount of a compound of Formula I according to any one of claims 1 to 15 to a mammal in need thereof.

28. A method of treating or preventing conditions selected from central or peripheral neurological degenerative disorders including Alzheimer's disease, cognitive disorders, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, Frontotemporal dementia Parkinson's Type, Parkinson dementia complex of Gaum, HTV dementia, corticobasal degeneration, dementia pugilistica, Down's syndrome, postencephelatic parkinsonism, progressive supranuclear palsy, Pick's Disease, Niemann-Pick's Disease, epilepsy, a peripheral neuropathy, spinal cord injury, head trauma, ALS, traumatic brain injury, haemorrhaging stroke and cancer, comprising the administration of a therapeutically effective amount of a compound of Formula I according to any one of claims 1 to 15 to a mammal in need thereof.

29. The method according to claim 28, wherein said condition is Alzheimer's disease.

30. A method of treating or preventing associated with inhibiting the expression of inducible pro-inflammatory proteins comprising the administration of a therapeutically effective amount of a compound of Formula I according to any one of claims 1 to 15 to a mammal in need thereof.

31. The method according to claim 30, wherein the condition is selected from edema, analgesia, fever and pain, such as neuromuscular pain, headache, cancer pain, dental pain and arthritis pain.