HK1073791A - Benzimidazoles useful as protein kinase inhibitors - Google Patents

Description

Benzimidazoles as protein kinase inhibitors

Application information

This application claims the benefit of U.S. provisional application 60/344,636 filed on 9/11/2001.

Technical Field

The present invention relates to substituted benzimidazole compounds of general formula (I):

wherein R is₁、R₂、R₃、R₄And X_aIs defined hereinafter. The compounds of the invention are useful as inhibitors of the Tec kinase family, including Itk kinase, and are therefore useful in the treatment of diseases and pathological conditions including inflammation, immune disorders, and allergic disorders. The invention also relates to processes for the preparation of such compounds and to pharmaceutical compositions comprising such compounds.

Background

Protein kinases play an important role in the activity of signaling that allows cells to produce cellular responses, such as activation, growth, and differentiation, in response to extracellular signals. Protein kinases transmit their signals by phosphorylating specific residues in target proteins. Those protein kinases that specifically phosphorylate tyrosine residues are called protein tyrosine kinases. Protein tyrosine kinases can be divided into two broad classes: for example, the Epidermal Growth Factor (EGF) receptor (S.Iwashhita and M.Kobayashi, 1992, CellularSignaling, 4, 123-ion 132) and the non-receptor for cytosol (C.Chan et al, 1994, Ann.Rev.Immunol., 12, 555-ion 592).

Interleukin-2 induced T cell kinase (Itk), a member of the Tec family of protein tyrosine kinases, also known as T cell specific kinases (Tsk), is predominantly expressed in T-lymphocytes (EMT), and additionally includes Txk, Tec, Btk, and Bmx. Tec family members are described based on the presence of pleckstrin homology regions (PH), proline rich Tec homology regions (TH) and Src homology SH3, SH2 and SH1 kinase regions located at the C-terminus from the N-terminus, respectively (S.Gibson et al, 1993, Blood, 82, 1561-1572; J.D.Silicono et al, 1992, Proc.Nat.Acad.Sci.89, 11194-11198; N.Yamada et al, 1993, Biochem and biophysis Res.Comm., 192, 231-240).

Itk is expressed in T cells, mast cells and natural killer cells. By stimulating the T Cell Receptor (TCR) in T cells and by activating the high affinity IgE receptor in mast cells. Lck, a member of the src tyrosine kinase family, phosphorylates Y511 in the kinase domain activation loop of Itk upon stimulation of the receptor in T cells (S.D. Heyeck et al, 1997, J.biol. chem, 272, 25401-. For phosphorylation and activation of PLC-. gamma.activated Itk and Zap-70 are required (S.C.Bunnell et al 2000, J.biol.chem., 275, 2219-one 2230). PLC-gamma catalyzes the formation of inositol 1, 4, 5-triphosphates and diacylglycerols, leading to calcium mobilization and PKC activation, respectively. These activities activate numerous downstream pathways and ultimately lead to degranulation (mast cells) and cytokine expression (T cells) (T. kawakami et al, 1999, j. leukcyte biol., 65, 286-.

Knock out at ItkThe role of Itk in T cell activation has been demonstrated in mice. CD4 from Itk knockout mice⁺The proliferative response of T cells was reduced in either mixed lymphocyte responses or after stimulation by ConA or anti-CD 3 (x.c. liao and d.r. littman, 1995, Immunity, 3, 757-. In addition, a few IL-2 produced after TCR stimulation of T cells from Itk knockout mice also resulted in reduced proliferation of these cells. In a further study, Itk lacks CD4⁺T cells decrease cytokine levels including IL-4, IL-5 and IL-13 by TCR, even after priming under induction conditions (D.J. Fowell, 1999, Immunity, 11, 399-.

The role of Itk in PLC-. gamma.activation and in calcium mobilization was also demonstrated in T cells of these knockout mice, which apparently impairs IP₃While there is no extracellular calcium influx after TCR stimulation (K.Liu et al, 1998, J.exp.Med.187, 1721-. These studies described above indicate that Itk plays an important role in the activation of T cells and mast cells. Therefore, inhibitors of Itk would be therapeutically effective in diseases mediated by inappropriate activation of these cells.

T cells have been fully demonstrated to play an important role in regulating immune responses (Powrie and dCofman, 1993, Immunology Today, 14, 270-. Indeed, activation of T cells is often the first sign of immune disease. After activation of the TCR, calcium influx is required to activate the T cells. After activation, T cells produce cytokines, including IL-2, 4, 5, 9, 10, and 13, which allow T cells to proliferate, differentiate, and exert effector functions. Clinical studies with IL-2 inhibitors have shown that interference with T cell activation and proliferation effectively suppresses immune responses in vivo (Waldmann, 1993, Immunology Today, 14, 264-. Thus, agents that inhibit the activation of T lymphocytes and the consequent production of cytokines are therapeutically useful for selectively inhibiting the immune response in patients in need of such immunosuppression.

Mast cells play a key role in asthma and allergic diseases by releasing pro-inflammatory mediators and cytokines. Fc epsilon RI, a high affinity receptor for IgE, antigen-mediated aggregation, activates mast cells (d.b. corry et al, 1999, Nature, 402, B18-23). This activation results in a series of signaling events that release mediators including histamine, proteases, leukotrienes and cytokines (j.r. gordon et al, 1990, Immunology Today, 11, 458-. These mediators increase vascular permeability, mucus production, bronchoconstriction, tissue degradation and inflammation, and thus they play an important role in the etiology and symptoms of asthma and allergic diseases.

Recent published data for mice using The Itk knockout showed that The number of memory T cells increased in The absence of Itk (A.T. Miller et al, 2002, The Journal of Immunology, 168, 2163-. One strategy to improve the vaccination approach is to increase the number of memory T cells produced (s.m. kaech et al, Nature Reviews Immunology, 2, 251-.

All documents cited in this application are incorporated by reference in their entirety.

Summary of The Invention

It is therefore an object of the present invention to provide a compound of formula (I):

wherein R is₁、R₂、R₃、R₄And X_aAs defined below.

It is another object of the invention to provide a method of inhibiting the Tec kinase family, including Itk kinase, and methods of treating diseases or conditions associated with the activity of these kinases by administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I).

It is also an object of the present invention to provide pharmaceutical compositions and processes for preparing compounds of formula (I) as described hereinafter.

Description of The Preferred Embodiment

In its broadest general embodiment of the present invention, there is provided a compound of formula (I):

wherein:

R₁is hydrogen or alkyl;

R₂selected from aryl and heteroaryl, each R₂Optionally substituted by one or more R_aSubstitution;

R₃is optionally substituted by one or more R_bSubstituted branched or unbranched C_1-10An alkyl chain is arranged on the base,

or R₃Is the following group:

-(CH₂)_n-L-R₆wherein L is selected from the group consisting of a direct bond, -NH-C (O) -, -O-C (O) -, -C (O) -, and-S (O)_m-, where m is 0, 1 or 2 and said radicals are optionally substituted by one or more R_bSubstitution;

wherein R is₆Independently selected from hydroxy, alkyl, alkoxy, alkylthio, aryl C_0-5Alkyl, aryloxy C_0-5Alkyl, heteroaryl C_0-5Alkyl, cycloalkyl C_0-5Alkyl, heterocyclic radical C_0-5Alkyl and amino, said amino being optionally substituted by acyl, alkyl, alkoxycarbonyl, cycloalkyl C_0-5Alkyl, aryl C_0-5Alkyl, heteroaryl C_0-5Alkyl or heterocyclyl radicals C_0-5Alkyl is mono-or di-substituted;

n is 1 to 10;

R₄is the following group:

wherein R is₄Covalently linked to the 5-or 6-position indicated by formula (I);

R₅selected from aryl radicals C_0-5Alkyl, heteroaryl C_0-5Alkyl, cycloalkyl C_0-5Alkyl and heterocyclic radical C_0-5Alkyl radical, each R₅Optionally substituted by one or more R_cSubstitution;

R₇is hydrogen, alkenyl or alkyl;

or R₅And R₇Together with the nitrogen atom to which they are attached form: a 4-7 membered monocyclic or 8-14 membered bicyclic ring, wherein each monocyclic or bicyclic ring optionally contains an additional 1-3 heteroatoms selected from N, O and S, and each ring is aromatic or nonaromatic, wherein each monocyclic or bicyclic ring is optionally substituted with one or more R_cSubstitution;

each R_a、R_bOr R_cIndependently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, aryloxy, alkoxy, alkylthio, acyl, alkoxycarbonyl, acyloxy, acylamino, sulfonylamino, aminosulfonyl, alkylsulfonyl, carboxyl, acylamino, hydroxyl, halogen, trifluoromethyl, nitro, nitrile and amino, optionally mono-or di-substituted with alkyl, acyl or alkoxycarbonyl, wherein any of the above R, if possible, is substituted with one or more halogen atoms_a、R_bOr R_cEach of which may be optionally halogenated; and

X_aand X_bIs oxygen or sulfur;

or a pharmaceutically acceptable derivative thereof.

In another embodiment, there is provided a compound of formula (I) as directly above and wherein:

R₁is hydrogen;

R₂a heteroaryl selected from phenyl, naphthyl, and from thienyl, furyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, pyrazolyl, pyrrolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyranyl, quinoxalyl, indolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, benzothienyl, quinolinyl, quinazolinyl, and indazolyl, each R₂Optionally substituted by one or more R_aSubstitution;

R₃is optionally substituted by one or more R_bSubstituted branched or unbranched C_1-10An alkyl chain is arranged on the base,

or R₃Is the following group:

-(CH₂)_n-L-R₆wherein L is selected from the group consisting of a direct bond, -O-C (O) -, -C (O) -, and-S (O)_m-, where m is 0, 1 or 2 and said radicals are optionally substituted by one or more R_bSubstitution;

wherein R is₆Independently selected from hydroxy, C_1-5Alkyl radical, C_1-5Alkoxy radical, C_1-5Alkylthio, phenyl, naphthyl, benzyl, phenethyl, heteroaryl C_0-5Alkyl radical, C_3-7Cycloalkyl radical C_0-5Alkyl, heterocyclic radical C_0-5Alkyl and amino, said amino being optionally substituted by C_1-5Acyl radical, C_1-5Alkyl radical, C_1-5Alkoxycarbonyl, aryl C_0-5Alkyl, heteroaryl C_0-5Alkyl or heterocyclyl radicals C_0-5Alkyl is mono-or di-substituted; wherein each said heteroaryl of this paragraph is selected from the group consisting of thienyl, furyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, pyrazolyl, pyrrolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, and pyranyl, and wherein each said heterocyclyl of this paragraph is selected from the group consisting of pyrrolidinyl, morpholinyl, thiomorpholinyl, dioxolanyl, piperidinyl, and pyranylAnd a piperazinyl group;

R₅selected from phenyl, naphthyl, benzyl, phenethyl, C_1-5Alkyl, heteroaryl C_0-5Alkyl wherein heteroaryl is selected from thienyl, furyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, pyrazolyl, pyrrolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl and pyranyl, C_3-7Cycloalkyl radical C_0-5Alkyl and heterocyclic radical C_0-5Alkyl wherein heterocyclyl is selected from aziridinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, tetrahydrofuranyl, dioxolanyl, piperidinyl and piperazinyl, each R₅Optionally substituted by one or more R_cSubstitution;

each R_a、R_bOr R_cIndependently selected from hydrogen, C_1-5Alkyl radical, C_2-5Alkenyl radical, C_2-5Alkynyl, C_3-8Cycloalkyl, phenyl, benzyl, phenoxy, C_1-5Alkoxy radical, C_1-5Alkylthio radical, C_1-5Acyl radical, C_1-5Alkoxycarbonyl group, C_1-5Acyloxy, C_1-5Amido, C_1-5Sulfonylamino, aminosulfonyl, C_1-5Alkylsulfonyl, carboxyl, amido, hydroxyl, halogen, trifluoromethyl, nitro, nitrile and amino, optionally substituted by C_1-5Alkyl radical, C_1-5Acyl or C_1-5Alkoxycarbonyl mono-or di-substituted, wherein, if possible, any of the above R_a、R_bOr R_cEach of which may be optionally halogenated;

R₇is C_3-10Alkenyl or C_1-5An alkyl group;

and

X_aand X_bIs oxygen.

In another embodiment, there is provided a compound of formula (I) as directly above and wherein:

R₂selected from phenyl, naphthyl, and from thienyl,Heteroaryl of furyl, isoxazolyl, oxazolyl, imidazolyl, thiadiazolyl, pyrazolyl, pyridyl, quinoxalinyl and benzothienyl, each R₂Optionally substituted by one or more R_aSubstitution;

R₆independently selected from hydroxy, C_1-5Alkyl radical, C_1-5Alkoxy, phenyl, benzyl, phenethyl, heteroaryl C_0-5Alkyl, heterocyclic radical C_0-5Alkyl radical, C_3-7Cycloalkyl and amino, said amino being optionally substituted by C_1-5Acyl radical, C_1-5Alkyl radical, C_1-5Alkoxycarbonyl, aryl C_0-5Alkyl or heteroaryl C_0-5Alkyl is mono-or di-substituted;

wherein each said heteroaryl of this paragraph is selected from thienyl, furyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, pyrazolyl, pyrrolyl and imidazolyl;

n is 1 to 6;

R₅selected from phenyl, naphthyl, benzyl, phenethyl, C_1-5Alkyl, heteroaryl C_0-5Alkyl wherein in this paragraph the heteroaryl is selected from the group consisting of thienyl, furyl, imidazolyl and pyridyl, C_3-7Cycloalkyl radical C_0-5Alkyl and heterocyclic radical C_0-5Alkyl wherein heterocyclyl is selected from aziridinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyridinyl, morpholinyl, thiomorpholinyl, piperidinyl, and piperazinyl, each R₅Optionally substituted by one or more R_cSubstitution;

and R₇Is propenyl or C_1-3An alkyl group.

In a further embodiment, there is provided a compound of formula (I) as directly above and wherein:

R₂a heteroaryl group selected from phenyl and from thienyl, furyl, isoxazolyl, thiadiazolyl, pyrazolyl and pyridyl, each R₂Optionally substituted by one or more R_aSubstitution;

R₃comprises the following steps:

-(CH₂)_n-C(O)-R₆or

-(CH₂)_n-R₆；

Wherein R is₆Independently selected from hydroxy, C_1-5Alkyl radical, C_1-5Alkoxy, phenyl, thienyl C_0-5Alkyl radical, C_3-7Cycloalkyl and amino, said amino being optionally substituted by C_1-5Alkyl or C_1-5Alkoxycarbonyl mono-or di-substituted;

R₅selected from phenyl, benzyl, phenethyl, and C_3-7Cycloalkyl radical C_0-5Alkyl radical, each R₅Optionally substituted by one or more R_cSubstitution;

each R_a、R_bOr R_cIndependently selected from C_1-5Alkyl radical, C_3-8Cycloalkyl, phenyl, C_1-5Alkoxy and amino, optionally substituted by C_1-5Alkyl radical, C_1-5Alkoxycarbonyl, amide, hydroxy, halogen, trifluoromethyl, nitro and nitrile groups, wherein any of the above R, if possible_a、R_bOr R_cEach of which may be optionally halogenated;

and R is₇Is C_1-3An alkyl group.

In a further embodiment, there is provided a compound of formula (I) as directly above and wherein:

R₂selected from phenyl, thienyl, furyl, isoxazolyl and pyridyl, each R₂Optionally substituted by one or more R_aSubstitution;

R₅selected from phenyl and cyclohexyl, each R₅Optionally substituted by one or more R_cSubstitution;

and

n is 2 to 5.

In a further embodiment, there is provided a compound of formula (I) as directly above and wherein:

R₂selected from phenyl, thiophen-2-yl, isoxazol-5-yl and pyridin-3-yl, each R₂Optionally substituted by one or more R_aSubstitution;

R₆independently selected from hydroxy, methyl, ethyl, C_1-3Alkoxy, phenyl, thienyl C_0-5Alkyl radical, C_3-7Cycloalkyl and amino, said amino being optionally substituted by C_1-5Alkyl or C_1-5Alkoxycarbonyl mono-or di-substituted;

and

each R_a、R_bOr R_cIndependently selected from C_1-3Alkoxy, amino, optionally substituted by C_1-3Alkyl, amido, hydroxyl, fluorine, chlorine, bromine, trifluoromethyl, nitro and nitrile mono-or di-substituted.

In any one of the preceding embodiments, there is provided a compound of formula (I), wherein:

R₄covalently attached at the 5-position indicated by formula (I), or in another embodiment, R₄Covalently linked to the 6-position indicated by general formula (I).

In another embodiment, representative compounds of the invention are provided as shown below, which may be prepared according to general schemes and synthetic examples:

andor

A pharmaceutically acceptable derivative thereof.

In another embodiment, representative compounds of the invention are provided as shown below, which are preferred and can be prepared according to the general scheme and the synthetic examples (working example):

andor

A pharmaceutically acceptable derivative thereof.

In all compounds disclosed herein above, if their nomenclature does not correspond to the structural formula, it is understood that these compounds are defined by their structural formula.

The present invention includes the use of all of the above compounds containing one or more asymmetric carbon atoms, which may occur as racemates or racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. All such isomeric forms of these compounds are expressly included in the present invention. Each stereoconfigured carbon may be in either the R or S configuration, or a mixture of these configurations.

Some compounds of formula (I) may exist in more than one tautomeric form. For example,

other tautomers will be apparent to those of ordinary skill in the art and the invention encompasses all such tautomeric forms and methods of making and using them.

Unless otherwise defined, all terms used herein are to be understood in their ordinary meaning as is known in the art.

Unless otherwise defined, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, acyl, alkoxycarbonyl, acyloxy, amido, alkylsulfonyl and other alkyl-containing groups may be understood as C1-10, branched or unbranched, optionally partially or fully halogenated if structurally possible. Other more specific definitions are as follows:

BOC or t-BOC is tert-butoxycarbonyl.

t-Bu is tert-butyl.

DMF is dimethylformamide.

EtOAc was ethyl acetate.

EtOH and MeOH are ethanol and methanol, respectively.

TFA is trifluoroacetic acid.

THF is tetrahydrofuran.

DMSO is dimethylsulfoxide.

TBTU is O- (1H-benzotriazol-1-yl) -N, N.N ', N' -tetramethyluronium tetrafluoroborate.

FMOC is 9-fluorenylmethoxycarbonyl.

The term "aroyl" as used in the present specification is understood to mean "benzoyl" or "naphthoyl".

The term "carbocycle" is understood to mean an aliphatic hydrocarbon group containing from 3 to 12 carbon atoms. Carbocycles include hydrocarbon rings containing 3 to 10 carbon atoms. These carbocycles may be aromatic or nonaromatic ring systems, and are optionally partially or completely halogenated. The non-aromatic ring system may be mono-or poly-unsaturated. Preferred carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, phenyl, indanyl, 2, 3-indanyl, indenyl (indenyl), benzocyclobutenyl, dihydronaphthyl, tetrahydronaphthyl, naphthyl, decahydronaphthyl, benzocyclopentyl, and benzocyclopentenyl. Certain terms used to denote cycloalkyl groups such as cyclobutyl (cyclobutylanyl) and cyclobutyl (cyclobutylyl) may be used interchangeably.

The term "heterocyclyl" refers to a stable nonaromatic 4-8 membered (preferably 5 or 6 membered) monocyclic or nonaromatic 8-11 membered bicyclic heterocyclyl group which may be saturated or unsaturated. Each heterocyclyl group consists of carbon atoms and one or more, preferably 1 to 4, heteroatoms selected from nitrogen, oxygen and sulfur. The heterocyclic group may be attached through any ring atom, which results in a stable structure. Unless otherwise specified, heterocyclyl includes, but is not limited to, pyrrolidinyl, morpholinyl, thiomorpholinyl, dioxolanyl, piperidinyl, piperazinyl, aziridinyl, and tetrahydrofuranyl.

The term "heteroaryl" is understood to mean an aromatic 5-8 membered monocyclic or 8-11 membered bicyclic ring containing 1-4 heteroatoms, such as N, O and S. Unless otherwise specified, such heteroaryl groups include, but are not limited to, thienyl, furyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, pyrazolyl, pyrrolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyranyl, quinoxalinyl, indolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, benzothienyl, quinolinyl, quinazolinyl, and indazolyl.

The term "heteroatom" as used herein is understood to mean a non-carbon atom such as O, N, S and P.

In all alkyl or cycloalkyl groups, one or more carbon atoms are optionally interrupted by heteroatoms: o, S and N, it being understood that if N is unsubstituted it is NH, it being understood that heteroatoms may replace either terminal or internal carbon atoms in the branched or unbranched carbon chain.

Substituted on carbon, e.g. methylene carbon, with groups such as oxo, or, if substituted on the ring, e.g. by carbonyl > C ═ O instead of methylene-CH₂-, give definitions such as alkoxycarbonyl, acyl and amide groups.

The term "aryl" as used herein is understood to mean an aromatic carbocyclic or heteroaryl group as defined above. Unless otherwise specified, each aryl or heteroaryl group includes derivatives thereof which are partially or fully hydrogenated. For example, the quinolyl group may include decahydroquinolyl and tetrahydroquinolyl, and the naphthyl group may include hydrogenated derivatives thereof such as tetrahydronaphthyl. Each aryl or heteroaryl group may be partially or fully halogenated. Other partially or fully hydrogenated derivatives of the aryl or heteroaryl compounds described herein will be apparent to those of ordinary skill in the art.

Similar terms to the above cyclic groups such as aryloxy or heteroarylamino may be understood to refer to aryl, heteroaryl, heterocyclyl groups as described above attached to the corresponding functional group.

As used herein, "nitrogen" and "sulfur" include any oxidized forms of nitrogen and sulfur as well as any basic nitrogen in the form of quaternary ammonium. For example, for e.g., -S-C_1-6Alkyl, if not otherwise limited, is understood to include-S (O) -C_1-6Alkyl and-S (O)₂-C_1-6An alkyl group.

The term "halogen" as used in this specification is to be understood as meaning bromine, chlorine, fluorine or iodine. Definitions "partially or fully halogenated" and "substituted with one or more halogen atoms" include, for example, mono-, di-or trihalo derivatives on one or more carbon atoms. Non-limiting examples may be haloalkyl groups, such as-CH₂CHF₂、-CF₃And the like.

The compounds of the present invention are meant only those compounds that are accepted by those skilled in the art and are intended to be "chemically stable". For example, those compounds having "dangling bonds" or "carbanions" are not the compounds that can be expected by the inventive methods disclosed herein.

The term "patient" refers to a warm-blooded animal, preferably a human.

The invention includes pharmaceutically acceptable derivatives of the compounds of formula (I). By "pharmaceutically acceptable derivative" is meant any pharmaceutically acceptable salt or ester, or any other compound that, upon administration to a patient, is capable of providing (directly or indirectly) a compound for use in the present invention or a pharmacologically active metabolite or pharmacologically active residue thereof. Pharmacologically active metabolites are understood to mean any compound of the invention which is capable of being metabolized enzymatically or chemically.

Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acids include hydrochloric, bromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, p-methylphenylsulfonic, tartaric, acetic, citric, sulfuric, acid, citric acid, and mixtures thereof,Methanesulfonic acid, formic acid, benzoic acid, malonic acid, naphthalen-2-ylsulfuric acid and benzenesulfonic acid. Other acids, such as oxalic, while not pharmaceutically acceptable acids per se, may be used as intermediates in the preparation of the salts to obtain compounds or addition salts of pharmaceutically acceptable acids thereof. Salts derived from suitable bases include alkali metals (e.g., sodium), alkaline earth metals (e.g., magnesium), ammonium, and N- (C)₁-C₄Alkyl radical)₄ ⁺A salt.

In addition, prodrugs using compounds of formula (I) also fall within the scope of the invention. Prodrugs include those compounds which are modified by simple chemical transformations to yield the compounds of the present invention. Simple chemical transformations include hydrolysis, oxidation, and reduction. In particular, when a prodrug is administered to a patient, the prodrug can be converted to the above disclosed compounds, thereby producing the desired pharmacological activity.

Methods of therapeutic use

The compounds of the invention are potent inhibitors of Tec kinase family activity, particularly Itk. Accordingly, in one embodiment of the present invention, there is provided a method of treating an immune disorder (immunological disorder) using a compound of the present invention. In another embodiment, methods of treating inflammatory diseases (inflammation disorders) using the compounds of the present invention are provided. In another embodiment, methods of treating allergic diseases (allergic disorders) using the compounds of the present invention are provided. In another embodiment, methods of using the compounds of the invention to promote memory cell generation for vaccines (vaccines) are provided. In yet another embodiment, methods of treating cell proliferative diseases (cell proliferative disorders) using the compounds of the invention are provided.

Without wishing to be bound by any theory, the compounds of the present invention modulate T cell and mast cell activation by effectively inhibiting Itk. Inhibition of T cell activation is therapeutically useful for selectively inhibiting immune function. Thus, inhibiting Itk is a very effective means of preventing and treating various types of immune diseases, including inflammatory diseases, autoimmune diseases, organ or bone marrow transplant rejection and other diseases associated with T cell mediated immune responses. In particular, the compounds of the invention are useful in the prevention or treatment of acute or chronic inflammation, allergy, contact dermatitis, psoriasis, rheumatoid arthritis, multiple sclerosis, type 1 diabetes, inflammatory bowel disease (inflamematous boweldase), Guillain-Barre syndrome (Guillain-Barre syndrome), crohn's disease, ulcerative colitis (ulcerous colitis), cancer, graft versus host disease (and other forms of organ or bone marrow transplant rejection) and lupus erythematosus (lupus erythematous).

In addition to being effective inhibitors of Itk, the compounds of the invention are effective inhibitors of Tec family kinases including Txk, Tec, Btk, and Bmx, and are therefore useful in the treatment of diseases modulated by the activity of one or more of these Tec family kinases.

Inhibitors of mast cell activation and degranulation prevent the release of allergic pro-inflammatory mediators and cytokines. Itk inhibitors may therefore be useful in the treatment of inflammatory and allergic diseases, including asthma, Chronic Obstructive Pulmonary Disease (COPD), Adult Respiratory Distress Syndrome (ARDS), bronchitis, conjunctivitis, dermatitis, and allergic rhinitis. Other T cell mediated diseases or mast cell mediated immune responses will be apparent to those of ordinary skill in the art and may also be treated using the compounds and compositions of the present invention.

Inhibitors of Itk and other Tec family kinases may be useful in combination with other therapeutic agents in the treatment of immune disorders, inflammation, proliferative disorders, and allergic disorders. Although not all are included, for example, administration is in combination with a steroid, a leukotriene antagonist, an antihistamine, cyclosporine, or rapamycin (rapamycin).

One strategy to improve the vaccination approach (vaccination) is to increase the number of memory T cells generated. As described in the background, if there is no Itk in the mouse, a greater number of memory cells are generated. Thus, it is within the scope of the invention to use the compounds of the invention in formulations for improved vaccination to generate higher numbers of memory T cells.

For therapeutic use, the compounds of the present invention may be administered in any conventional manner in any conventional dosage form. Routes of administration include, but are not limited to, intravenous, intramuscular, subcutaneous, intrasynovial (intrasynovally), infusion (infusion), sublingual (sublingually), transdermal, oral, topical, or inhalation. Preferred modes of administration are oral and intravenous.

The compounds of the present invention may be administered alone or in combination with adjuvants including other active ingredients which may enhance the stability of the inhibitor, and in certain embodiments may facilitate the administration of pharmaceutical compositions containing the same, may improve solubility and dispersibility, enhance inhibitory activity, provide adjunctive therapy, and the like. Advantageously, such combination therapy uses lower doses than conventional therapeutic amounts, thus avoiding the toxicity and side effects that may result from the use of these agents alone. The compounds of the present invention may be administered in conventional therapeutic regimens or in physical association with other adjuvants, to form separate pharmaceutical compositions. Advantageously, these compounds can then be administered together in a single dose. In some embodiments, pharmaceutical compositions comprising such combination compounds comprise at least about 5%, but more preferably at least about 20%, of a compound of formula (I) (w/w) or combination thereof. The optimum percentage (w/w) of the compounds of the invention may be adjusted, such adjustments being within the skill of the art. Alternatively, the compounds may be administered separately (either sequentially or simultaneously). Administration alone may allow for better flexibility in dosing.

As noted above, the dosage forms of the compounds of the present invention include pharmaceutically acceptable carriers and adjuvants known to those of ordinary skill in the art. Such carriers and adjuvants include, for example, ion exchangers, alumina, aluminum stearate, lecithin, serum albumin, buffers, water, salts or electrolytes and cellulose-based materials. Preferred dosage forms include tablets, capsules, caplets, liquids, solutions, suspensions, emulsions, lozenges, syrups, powders, extemporaneous powders, granules, suppositories, and transdermal patches. Methods for preparing these Dosage Forms are known (see, e.g., h.c. ansel and n.g. popofosh, Pharmaceutical Delivery Forms and Drug Delivery Systems, 5the d., Lea and Febiger (1990)). Dosage criteria and requirements are well known in the art and can be selected by one of ordinary skill in the art from those methods and processes that are commercially available to suit a particular patient. In some embodiments, the dosage standard is about 1-1000 mg/dose for a 70kg patient. Although one dose per day may be sufficient, up to 5 doses per day may be administered. For oral doses, 2000 mg/day is required. The requirement for low or high dosages is determined by specific factors, as will be appreciated by those skilled in the art. For example, the specific dosage and treatment regimen will depend on such factors as the general health of the patient, the severity of the patient's disease and the course of the disease or its tendency to develop, and the judgment of the treating physician.

Biological activity

Itk test

Itk was purified as GST-fusion protein. Kinase activity was measured using DELFIA (dissociation amplification lanthanide fluoroimmunoassay) which uses an anti-phosphotyrosine antibodies (anti-phosphotyrosine antibodies) labeled with europium chelate to detect phosphate transfer to atactic polymer, polyglulu₄∶Tyr₁(PGTYR). Screening the reagent, buffer and sample were dispensed for analysis (assay) using the Zymark Allegro UHTS system, while the plate was also washed. In kinase assay buffer (50 m)MHEPES，pH 7.0，25mM MgCl₂，5mM MnCl₂，50mM KCl，100μM Na₃VO₄0.2% BSA, 0.01% CHAPS, 200. mu.M TCEP). The test samples were initially dissolved in DMSO at a concentration of 1mg/mL and then pre-diluted with assay buffer in 384-well polypropylene microtiter plates for dose response (9 doses with a final concentration of 3g/mL, serially diluted 1: 3). mu.L volume/well of the matrix mix, containing 15. mu.M ATP and 9 ng/. mu.L of LPGTYR-biotin (CIS biointematic) in kinase buffer, was added to a 384-well white Plate (PIERCE) coated with neutravidin (neutravidin), followed by 20. mu.L/well of the test sample solution and 20. mu.L/well of the diluted enzyme (final concentration of 7 nM). Background wells were incubated with buffer instead of 20. mu.L of enzyme. The assay plates were incubated at room temperature for 30 minutes. After incubation, assay plates were washed three times with 100. mu.L of wash buffer (50mM Tris-HCl, pH 7.4, 150mM NaCl, 0.05% Tween 20, 0.2% BSA). To each well was added 50. mu.L aliquots of europium-labeled phosphotyrosine antibody (Eu) diluted in 50mM Tris-HCl, pH7.8, 150mM NaCl, 10. mu.M DTPA, 0.05% Tween 40, 0.2% BSA, 0.05% BGG (final concentration of 1nM)³⁺PT66, WallacCR04-100) and incubation at room temperature for 30 min. After incubation was complete, the assay plate was washed with 100. mu.L of wash buffer and 50. mu.L of DELFIA potentiating solvent (Wallac) was added to each well. After 15 minutes, time resolved fluorescence was measured by LJL's analyser (excitation at 360nm, emission at 620nm, EU 400 dichoric Mirror) after a lag time of 250. mu.s.

Preferred compounds of the invention are 1 micromolar or less active.

In order that the invention may be more fully understood, the following examples are set forth. These examples are intended to illustrate preferred embodiments of the invention and are not to be construed as limiting the scope of the invention in any way.

The examples that follow are illustrative examples, and as will be recognized by those skilled in the art, specific reagents or conditions may be modified as needed for individual compounds without undue experimentation. The starting materials used in the schemes below are either commercially available or can be readily prepared from commercially available starting materials by those skilled in the art.

Total synthesis method

The invention also provides a method for preparing the compound of the general formula I. In all schemes, unless otherwise defined, the R substituents in the following formulas have the same meaning as the R substituents in formula I of the present invention described above. Intermediates used in the preparation of the compounds of the invention are either commercially available or can be readily prepared by known methods by those skilled in the art.

The compounds of formula I can be prepared by the process outlined in scheme 1.

Scheme 1

According to said method, nitrobenzoic acid (II) bearing a leaving group X adjacent to the nitro group is reacted with a compound bearing R₅And R₇The amine of (a) is coupled. Suitable leaving groups include halogen, preferably fluorine. Conventional coupling conditions known in the art may be used, for example reacting II with an amine in a suitable solvent such as dichloromethane in the presence of a coupling agent such as 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDC). The resulting intermediate III is then reacted with a compound having R₃In the presence of a suitable base such as triethylamine in a suitable solvent such as DMSO to give IV. This step may be carried out after R is added, if desired₅R₇The coupling of NH to benzoic acid II is completed. The nitro group of IV is then reduced using methods known in the art, such as catalytic hydrogenation. Suitable catalysts include platinum or palladium on carbon. The reaction can be carried out both under a hydrogen atmosphere and in the presence of a hydrogen source such as ammonium formateThis is carried out in a suitable solvent, for example ethanol. The resulting aniline is then cyclized in a suitable solvent, such as ethanol, by treatment with cyanogen bromide to give 2-aminobenzimidazole VI. With the radical R₂Acylation of the acid halide of VI affords the desired product of formula I. Further modification of the R substituents by conventional means may give other desirable products of formula I. With R in the 6-position₄The compounds of the general formula I can be prepared analogously starting from p-nitrobenzoic acid derivatives corresponding to II.

The synthesis of 2-aminobenzimidazole using solid phase chemistry has been reported (J.Lee et al, Tetrahedron Letters, 2001, 42, 2635-. If desired, the compounds of formula I can be prepared using solid phase chemistry as described in scheme 2, below. As is known in the art, after each step described below, the reaction kettle should be evacuated while the resin is washed with a solvent such as DMF, followed by washing with methanol and methylene chloride. Completion of each step can be monitored by means known in the art, such as ninhydrin test or cleavage of a small sample of the resin and analysis by LC-MS.

Scheme 2

According to this process, it is possible to start from a suitable resin carrying amino groups, such as the Sieber amide resin (VII). Coupling of Fmoc protected amino acid (VIII) to the amine on the resin using a coupling agent such as 1, 3-Diisopropylcarbodiimide (DIC) or O- (1H-benzothiazol-1-yl) -N, N' -tetramethyluronium tetrafluoroborate (TBTU) in the presence of a suitable base such as diisopropylethylamine in a suitable solvent such as DMF affords IX. The Fmoc protecting group was removed by treating the resin with about 20% to 50% piperidine in DMF. Followed by the use of 4-fluoro-3-nitrobenzoic acid in the presence of a suitable base such as diisopropylethylamine in a suitable solvent such as DMSODeprotection affords X. R is then reacted in DMF using the same coupling conditions as for solid phase synthesis, e.g. in the presence of bromo-tri-pyrrolidinyl-phosphorus₅R₇NH reacts with X to bring R₅And R₇To the carboxylic acid of X to give XI.

Then, conditions suitable for solid phase synthesis are used, e.g. with 1M-2M SnCl₂Treatment of XI in DMF with an aqueous solution reduces the nitro group on XI to give XII. Treatment of XII with a solution of cyanogen bromide at about 1M in a suitable solvent such as EtOH: DMF from 1: 2 to 1: 3 affords resin-bound 2-aminobenzimidazole XIII. Using R in the presence of a suitable base such as diisopropylethylamine and dimethylaminopyridine in a suitable solvent such as dichloromethane₂C (O) Cl treatment of XIII to give XIV. Finally, XIV is treated under dissociation conditions, for example, a 5% strength solution of trifluoroacetic acid in dichloromethane, to provide the desired product of formula I (R)₃＝-R’C(O)NH₂) Or a precursor thereof, which may be further modified using methods conventional in the art.

Synthetic examples

EXAMPLE 1.4 Synthesis of ethyl- {5- (cyclohexyl-methyl-carbamoyl) -2- [ (thiophene-2-carbonyl) -amino ] -benzimidazol-1-yl } butanoate

To 4-fluoro-3-nitrobenzoic acid (6g, 0.32mmol) in CH₂Cl₂(30mL) to the solution was added 1- [3- (dimethylamine) propyl group]3-Ethylcarbodiimide hydrochloride (8g, 0.42mmol) followed by additional N-methylcyclohexylamine (4.2mL, 0.32 mmol). The mixture was stirred at room temperature for 6 hours. The resulting solution was washed with 1N HCl (10mL) and saturated sodium carbonate (10mL) in that order, and the organic layer was dried over magnesium sulfate. The solvent was removed by evaporation and the resulting mixture was purified by gas chromatography using 100% ethyl acetateThe oil was purified to give cyclohexyl-N-methyl-4-fluoro-3-nitro-benzamide (6.0g, 67%) m.p.65-67 ℃.

A stirred solution of the above amide (2g, 7.1mmol), ethyl 4-aminobutyrate hydrochloride (2.4g, 14.2mmol), and triethylamine (2.5mL, 18.0mmol) in DMSO (25mL) was heated to 80 ℃ for 8 hours. The reaction mixture was poured into a separatory funnel containing dichloromethane (150mL) and water (150 mL). The organic layer was washed with water (5X 50mL), dried over magnesium sulfate and the solvent evaporated to give 4- [4- (N-cyclohexyl-N-methyl-carbamoyl) -2-nitro-phenylamino ] -butyric acid ethyl ester (2.8g, 100%).

A reaction flask equipped with a nitrogen line and a stir bar was charged with 10% palladium on activated carbon (0.22g) and ethanol (5 mL). The above amide (2.2g, 5.6mmol) in ethanol (25mL) was added followed by ammonium formate (3.9g, 61.8mmol) and the mixture was stirred at room temperature for 32 h. The reaction mixture was filtered through celite, washed with ethanol, and the filtrate was concentrated to a volume of 25 mL. The resulting 4- [ 2-amino-4- (N-cyclohexyl-N-methyl-carbamoyl) -phenylamino ] -butyric acid ethyl ester solution is used directly in the next step.

Cyanogen bromide (0.9g, 8.4mmol) was added to the previously obtained solution, and the obtained solution was stirred at room temperature for 24 hours. The solvent was removed by evaporation and the residue was partitioned between EtOAc (20mL) and saturated sodium carbonate (10 mL). The organic layer was washed with water (10mL) and dried over magnesium sulfate. The solvent was removed by evaporation and the resulting violet oil was purified by gas chromatography using 5-50% methanol/dichloromethane to give 4- [ 2-amino-5- (cyclohexyl-methyl-carbamoyl) -benzimidazol-1-yl ] -butyric acid ethyl ester (0.7g, 32%).

To a stirred solution of the above-mentioned amidobenzimidazole (0.7g, 1.8mmol) in pyridine (10mL) was added 2-thiophenecarbonyl chloride (0.41mL, 3.8 mmol). The reaction was completed within 6 hours. Pyridine was removed by evaporation and the resulting orange solid was purified by gas chromatography using 1% methanol/dichloromethane to give the title compound (0.61g, 67%) m.p.82-84 ℃.

EXAMPLE 2.4 Synthesis of- {5- (cyclohexyl-methyl-carbamoyl) -2- [ (thiophene-2-carbonyl) -amino ] -benzimidazol-1-yl } butyric acid

To the stirred solution of 4- {5- (cyclohexyl-methyl-carbamoyl) -2- [ (thiophene-2-carbonyl) -amino ] -methanol (10mL) and water (10mL)]To a solution of ethyl (0.5g, 1.0mmol) benzimidazol-1-yl) butanoate (example 1) was added solid NaOH (0.12g, 3 mmol). The reaction was completed within 4 hours. The mixture was acidified with 1N HCl and then CH₂Cl₂(20mL) dilution. The organic layer was dried over magnesium sulfate, the solvent was removed by evaporation, and the residue was purified by gas chromatography using 10% methanol/dichloromethane to give the title compound (0.43g, 92%) m.p.256-258 ℃.

Example 3.1 Synthesis of- (3-carbamoyl-propyl) -2- [ (thiophene-2-carbonyl) -amino ] -1H-benzimidazole-5-carboxylic acid-N-cyclohexyl-N-methyl-amide

To a solution of 4- {5- (cyclohexyl-methyl-carbamoyl) -2- [ (thiophene-2-carbonyl) -amino ] -benzimidazol-1-yl } butyric acid (example 2) (0.05g, 11mmol) in DMF (5mL) was added 1-hydroxybenzotriazole hydrate (0.02g, 16mmol) followed by 1- [3- (dimethylamino) propyl ] -3-ethylcarbodiimide hydrochloride (0.03g, 16 mmol). The mixture was stirred for 1 hour, then aluminum hydroxide (5mL) was added. Stirring was continued for 48 hours. The solution was neutralized with 1M HCl and partitioned between EtOAc (10mL) and water (10 mL). The organic layer was washed with saturated sodium carbonate (10mL) and water (3X 10 mL). The solvent was removed by evaporation and the resulting oil was purified by gas chromatography with 5% methanol in dichloromethane to give the title compound (0.02g, 40%) m.p.112-115 ℃.

Example 4.1- (3-hydroxy-propyl) -2- (thiophene-2-carbonyl) amino-1H-benzimidazole-5-carboxylic acid cyclohexyl-methyl-amide synthesis

A stirred (25mL) DMSO solution of N-cyclohexyl-N-methyl-4-fluoro-3-nitro-benzamide (example 1) (1.5g, 5.3mmol) and 3-amino-1-propanol (0.82mL, 10.7mmol) was heated to 80 ℃ for 8 hours. The reaction mixture was poured into a separatory funnel containing dichloromethane (100mL) and water (100 mL). The organic layer was washed with water (5 × 50mL), dried over magnesium sulfate, and the solvent was evaporated to give N-cyclohexyl-4- (3-hydroxy-propylamino) -N-methyl-3-nitro-benzamide (1.9g, 79%).

A reaction flask equipped with a nitrogen line and a stir bar was charged with 10% palladium on activated carbon (0.19g) and ethanol (5 mL). The above amide (1.9g, 5.7mmol) in ethanol (15mL) was added followed by ammonium formate (3.9g, 62mmol) and the mixture was stirred at room temperature for 7 h. The reaction mixture was filtered through celite, washed with ethanol, and the filtrate was concentrated to a volume of 15 mL. The resulting 3-amino-N-cyclohexyl-4- (3-hydroxypropylamino) -N-methyl-benzamide solution was used directly in the next step.

Cyanogen bromide (0.9g, 8.6mmol) was added to the previously obtained solution, and the obtained solution was stirred at room temperature for 48 hours. The solvent was removed by evaporation and the residue was partitioned between EtOAc (20mL) and saturated sodium carbonate (10 mL). The organic layer was washed with water (10mL) and dried over magnesium sulfate. The solvent was removed by evaporation and the resulting violet oil was purified by gas chromatography using 5-50% methanol/dichloromethane to give 2-amino-1- (3-hydroxypropyl) -1H-benzimidazole-5-carboxylic acid N-cyclohexyl-N-methyl-amide (0.7g, 37%).

To a stirred solution of the above-mentioned amidobenzimidazole (0.7g, 2.1mmol) in pyridine (10mL) was added 2-thiophenecarbonyl chloride (0.68mL, 6.4 mmol). The reaction was completed within 6 hours. Pyridine was removed by evaporation and the resulting orange solid was purified by gas chromatography using 1% methanol/dichloromethane to give thiophene-2-carboxylic acid 3- {5- (cyclohexyl-methyl-carbamoyl) -2- [ (thiophene-2-carbonyl) -amino ] -benzimidazol-1-yl } -propyl ester (0.50g, 43%), m.p.92-94 ℃.

To a stirred solution of the above-mentioned propyl ester (0.4g, 0.72mmol) in methanol (5mL) and water (5mL) was added solid NaOH (0.12g, 2.9 mmol). The reaction was completed within 6 hours. The mixture was acidified with 1M HCl and then diluted with EtOAc (10 mL). The organic layer was dried over magnesium sulfate, the solvent was removed by evaporation, and the residue was purified by gas chromatography with 5% methanol/dichloromethane to give the title compound (0.09g, 28%) m.p.105-107 ℃.

EXAMPLE 5 Synthesis of 3- {5- (cyclohexyl-methyl-carbamoyl) -2- [ (thiophene-2-carbonyl) -amino ] -benzimidazol-1-yl } -propyl acetate

To a solution of 1- (3-hydroxy-propyl) -2- [ (thiophene-2-carbonyl) -amino ] -1H-benzimidazole-5-carboxylic acid cyclohexyl-methyl-amide (example 4) (0.07g, 0.16mmol) in THF (10mL) was added anhydrous acetic acid (0.02mL, 0.19mmol) followed by triethylamine (0.02mL, 0.17mmol), and the mixture was stirred at room temperature for 48H. The resulting solution was washed with 1M HCl (10mL), and the organic layer was dried over magnesium sulfate. The solvent was removed by evaporation and the resulting oil was purified by gas chromatography using 5% methanol/dichloromethane to give the title compound (0.05g, 65%) m.p.74-76 ℃.

The following example describes the synthesis of compounds of formula I using solid phase chemistry.

Example 6: synthesis of 2-benzoylamino-1- (carbamoyl-ethyl) -1H-benzimidazole-5-carboxylic acid cyclohexylmethyl-amide

Sieber amide resin (100mg, 0.52mmol/g, 0.052mmol) was added to a solid phase shaker. DMF (20mL) was then added and the resin was allowed to swell for 10 minutes before the reagents were added. TBTU (83mg, 0.26mmol) and N, N-diisopropylethylamine (90mL, 0.52mmol) were added separately, followed by Fmoc-beta-alanine (81mg, 0.26 mmol). The container was then shaken at room temperature for 24 hours. After the vessel was drained, the resin was washed three times with DMF, MeOH, and dichloromethane (20mL portions, 10 min). Ninhydrin test negative at this point indicates that the reaction is complete.

The Fmoc group was removed under standard deprotection conditions: to the resin was added 20mL of a 1: 1 DMF: pyridine solution. The mixture was shaken at room temperature for 3 hours, drained and washed three times with DMF, MeOH, and dichloromethane (20mL portions, 10 min). Ninhydrin test positive at this point indicated removal of the Fmoc protecting group.

The resin was then swollen in DMSO (20mL) for 10 minutes. To the resin was added 4-fluoro-3-nitrobenzoic acid (48mg, 0.26mmol) and N, N-diisopropylethylamine (90mL, 0.52 mmol). The container was then shaken at room temperature for 24 hours. After draining the vessel, the resin was washed twice with DMF, DMF: H₂One O (1: 1) rinse and three rinses with methylene chloride (20mL portions). Ninhydrin test negative at this point indicates that the reaction is complete.

The resin was then swollen in DMF (20mL) for 10 min. To the swollen resin was added bromo-tri-pyrrolidinyl-phosphine hexafluorophosphate (PyBroP) (121mg, 0.26mmol), diisopropylethylamine (90mL, 0.52mmol), and N-methylcyclohexylamine (34mL, 0.26 mmol). The container was then shaken at room temperature for 24 hours. After the vessel was drained, the resin was washed three times with DMF, MeOH, and dichloromethane (20mL portions, 10 min). A small aliquot of the resin was then cleaved and analyzed by LC-MS to ensure completion of the reaction.

To the resin was added 2.0M SnCl₂Hydrate in DMF (20 mL). The container was then shaken at room temperature for 24 hours. After the vessel was drained, the resin was washed three times with DMF, MeOH, and dichloromethane (20mL portions, 10 min). A small aliquot of the resin was then cleaved and analyzed by LC-MS to ensure completion of the reaction.

To the resin was added 1.0M BrCN solution in 1: 3 EtOH: DMF (20 mL). The container was then shaken at room temperature for 24 hours. After the vessel was drained, the resin was washed three times with DMF, MeOH, and dichloromethane (20mL portions, 10 min). A small aliquot of the resin was then cleaved and analyzed by LC-MS to ensure completion of the reaction.

The resin was then swollen in dichloromethane (20mL) for 10 minutes. To the swollen resin were added 4-dimethylaminopyridine (31mg, 0.26mmol), diisopropylethylamine (90mL, 0.52mmol), and benzoyl chloride (30mL, 0.26 mmol). The container was then shaken at room temperature for 6 hours. After the vessel was drained, the resin was washed three times with DMF, MeOH, and dichloromethane (20mL portions, 10 min). A cleavage solution of 5% TFA in dichloromethane (20mL) was then added, shaken at room temperature for 3 hours, and the collected solution was concentrated in vacuo to afford the product as a yellow oil. The crude product is generally greater than 90% pure. Purification was accomplished by TLC prepared from 3% EtOH in dichloromethane to give the title compound as a white solid.

Claims (18)

1. A compound of the general formula (I):

wherein:

R₁is hydrogen or alkyl;

R₂selected from aryl and heteroaryl, each R₂Optionally substituted by one or more R_aSubstitution;

R₃is optionally substituted by one or more R_bSubstituted branched or unbranchedChained C_1-10An alkyl chain is arranged on the base,

or R₃Is the following group:

-(CH₂)_n-L-R₆wherein L is selected from the group consisting of a direct bond, -NH-C (O) -, -O-C (O) -, -C (O) -, and-S (O)_m-, where m is 0, 1 or 2 and said radicals are optionally substituted by one or more R_bSubstitution;

wherein R is₆Independently selected from hydroxy, alkyl, alkoxy, alkylthio, aryl C_0-5Alkyl, aryloxy C_0-5Alkyl, heteroaryl C_0-5Alkyl, cycloalkyl C_0-5Alkyl, heterocyclic radical C_0-5Alkyl and amino, said amino being optionally substituted by acyl, alkyl, alkoxycarbonyl, cycloalkyl C_0-5Alkyl, aryl C_0-5Alkyl, heteroaryl C_0-5Alkyl or heterocyclyl radicals C_0-5Alkyl is mono-or di-substituted;

n is 1 to 10;

R₄is the following group:

wherein R is₄Covalently linked to the 5-or 6-position indicated by formula (I);

R₅selected from aryl radicals C_0-5Alkyl, heteroaryl C_0-5Alkyl, cycloalkyl C_0-5Alkyl and heterocyclic radical C_0-5Alkyl radical, each R₅Optionally substituted by one or more R_cSubstitution;

R₇is hydrogen, alkenyl or alkyl;

or R₅And R₇Together with the nitrogen atom to which they are attached form: a 4-7 membered monocyclic or 8-14 membered bicyclic ring, wherein each monocyclic or bicyclic ring optionally contains an additional 1-3 heteroatoms selected from N, O and S, and each ring is aromatic or nonaromatic, wherein each monocyclic or bicyclic ring is optionally substituted with one or more R_cSubstitution;

each R_a、R_bOr R_cIndependently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,Aryl, aralkyl, aryloxy, alkoxy, alkylthio, acyl, alkoxycarbonyl, acyloxy, acylamino, sulfonylamino, aminosulfonyl, alkylsulfonyl, carboxyl, acylamino, hydroxyl, halogen, trifluoromethyl, nitro, nitrile and amino, optionally mono-or disubstituted by alkyl, acyl or alkoxycarbonyl, wherein any of the above R, if possible_a、R_bOr R_cEach of which may be optionally halogenated; and

X_aand X_bIs oxygen or sulfur;

or a pharmaceutically acceptable salt, ester, isomer, or tautomer thereof.

2. A compound according to claim 1, wherein

R₁Is hydrogen;

R₂a heteroaryl selected from phenyl, naphthyl, and from thienyl, furyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, pyrazolyl, pyrrolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyranyl, quinoxalyl, indolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, benzothienyl, quinolinyl, quinazolinyl, and indazolyl, each R₂Optionally substituted by one or more R_aSubstitution;

R₃is optionally substituted by one or more R_bSubstituted branched or unbranched C_1-10An alkyl chain is arranged on the base,

or R₃Is the following group:

-(CH₂)_n-L-R₆wherein L is selected from the group consisting of a direct bond, -O-C (O) -, -C (O) -, and-S (O)_m-, where m is 0, 1 or 2 and said radicals are optionally substituted by one or more R_bSubstitution;

wherein R is₆Independently selected from hydroxy, C_1-5Alkyl radical, C_1-5Alkoxy radical, C_1-5Alkylthio, phenyl, naphthyl, benzyl, phenethyl, heteroaryl C_0-5Alkyl radical, C_3-7Cycloalkyl radical C_0-5Alkyl, heterocyclic radical C_0-5Alkyl and aminoSaid amino group being optionally substituted by C_1-5Acyl radical, C_1-5Alkyl radical, C_1-5Alkoxycarbonyl, aryl C_0-5Alkyl, heteroaryl C_0-5Alkyl or heterocyclyl radicals C_0-5Alkyl is mono-or di-substituted; wherein each said heteroaryl of this paragraph is selected from thienyl, furyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, pyrazolyl, pyrrolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, and pyranyl, and wherein each said heterocyclyl of this paragraph is selected from pyrrolidinyl, morpholinyl, thiomorpholinyl, dioxolanyl, piperidinyl, and piperazinyl;

R₅selected from phenyl, naphthyl, benzyl, phenethyl, C_1-5Alkyl, heteroaryl C_0-5Alkyl wherein heteroaryl is selected from thienyl, furyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, pyrazolyl, pyrrolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl and pyranyl, C_3-7Cycloalkyl radical C_0-5Alkyl and heterocyclic radical C_0-5Alkyl wherein heterocyclyl is selected from aziridinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, tetrahydrofuranyl, dioxolanyl, piperidinyl, and piperazinyl, each R₅Optionally substituted by one or more R_cSubstitution;

each R_a、R_bOr R_cIndependently selected from hydrogen, C_1-5Alkyl radical, C_2-5Alkenyl radical, C_2-5Alkynyl, C_3-8Cycloalkyl, phenyl, benzyl, phenoxy, C_1-5Alkoxy radical, C_1-5Alkylthio radical, C_1-5Acyl radical, C_1-5Alkoxycarbonyl group, C_1-5Acyloxy, C_1-5Amido, C_1-5Sulfonylamino, aminosulfonyl, C_1-5Alkylsulfonyl, carboxyl, amido, hydroxyl, halogen, trifluoromethyl, nitro, nitrile and amino, optionally substituted by C_1-5Alkyl radical, C_1-5Acyl or C_1-5Alkoxycarbonyl mono-or di-substituted, wherein, if possible, any of the above R_a、R_bOr R_cEach of which may be optionally halogenated;

R₇is C_3-10Alkenyl or C_1-5An alkyl group;

and

X_aand X_bIs oxygen.

3. A compound according to claim 2, wherein

R₂Selected from phenyl, naphthyl, and heteroaryl selected from thienyl, furyl, isoxazolyl, oxazolyl, imidazolyl, thiadiazolyl, pyrazolyl, pyridyl, quinoxalinyl, and benzothienyl, each R₂Optionally substituted by one or more R_aSubstitution;

R₆independently selected from hydroxy, C_1-5Alkyl radical, C_1-5Alkoxy, phenyl, benzyl, phenethyl, heteroaryl C_0-5Alkyl, heterocyclic radical C_0-5Alkyl radical, C_3-7Cycloalkyl and amino, said amino being optionally substituted by C_1-5Acyl radical, C_1-5Alkyl radical, C_1-5Alkoxycarbonyl, aryl C_0-5Alkyl or heteroaryl C_0-5Alkyl is mono-or di-substituted; wherein each said heteroaryl of this paragraph is selected from thienyl, furyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, pyrazolyl, pyrrolyl and imidazolyl;

n is 1 to 6;

R₅selected from phenyl, naphthyl, benzyl, phenethyl, C_1-5Alkyl, heteroaryl C_0-5Alkyl wherein the heteroaryl in this paragraph is selected from thienyl, furyl, imidazolyl and pyridyl, C_3-7Cycloalkyl radical C_0-5Alkyl and heterocyclic radical C_0-5Alkyl wherein heterocyclyl is selected from aziridinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyridinyl, morpholinyl, thiomorpholinyl, piperidinyl, and piperazinyl, each R₅Optionally substituted by one or more R_cSubstitution;

and

R₇is propenyl or C_1-3An alkyl group.

4. A compound according to claim 3, wherein

R₂A heteroaryl group selected from phenyl and from thienyl, furyl, isoxazolyl, thiadiazolyl, pyrazolyl and pyridyl, each R₂Optionally substituted by one or more R_aSubstitution;

R₃comprises the following steps:

-(CH₂)_n-C(O)-R₆or

-(CH₂)_n-R₆；

Wherein R is₆Independently selected from hydroxy, C_1-5Alkyl radical, C_1-5Alkoxy, phenyl, thienyl C_0-5Alkyl radical, C_3-7Cycloalkyl and amino, said amino being optionally substituted by C_1-5Alkyl or C_1-5Alkoxycarbonyl mono-or di-substituted;

R₅selected from phenyl, benzyl, phenethyl and C_3-7Cycloalkyl radical C_0-5Alkyl, each optionally substituted by one or more R_cSubstitution;

each R_a、R_bOr R_cIndependently selected from C_1-5Alkyl radical, C_3-8Cycloalkyl, phenyl, C_1-5Alkoxy, amino, which is optionally substituted by C_1-5Alkyl radical, C_1-5Alkoxycarbonyl, amide, hydroxy, halogen, trifluoromethyl, nitro and nitrile groups, wherein any of the above R, if possible_a、R_bOr R_cEach of which may be optionally halogenated;

and R is₇Is C_1-3An alkyl group.

5. A compound according to claim 4, wherein

R₂Selected from phenyl, thienyl, furyl, isoxazolyl and pyridyl, each optionally substituted with one or more R_aSubstitution;

R₅selected from phenyl and cyclohexyl, each optionally substituted by one or more R_cSubstitution;

and

n is 2 to 5.

6. A compound according to claim 5, wherein

R₂Selected from phenyl, thiophen-2-yl, isoxazol-5-yl and pyridin-3-yl, each optionally substituted with one or more R_aSubstitution;

R₆independently selected from hydroxy, methyl, ethyl, C_1-3Alkoxy, phenyl, thienyl C_0-5Alkyl radical, C_3-7Cycloalkyl and amino, said amino being optionally substituted by C_1-5Alkyl or C_1-5Alkoxycarbonyl mono-or di-substituted;

and

each R_a、R_bOr R_cIndependently selected from C_1-3Alkoxy, amino, optionally substituted by C_1-3Alkyl, amido, hydroxyl, fluorine, chlorine, bromine, trifluoromethyl, nitro and nitrile mono-or di-substituted.

7. A compound according to any one of claims 1 to 6, wherein R₄Covalently bonded at the 5-position indicated by the general formula (I).

8. A compound according to any one of claims 1 to 6, wherein R₄Covalently linked to the 6-position indicated by general formula (I).

9. A compound selected from the group consisting of:

and

or a pharmaceutically acceptable salt, ester, isomer, or tautomer thereof.

10. A compound selected from the group consisting of:

and

or a pharmaceutically acceptable salt, ester, isomer, or tautomer thereof.

11. A pharmaceutical composition comprising a pharmaceutically effective amount of a compound according to claim 1 together with one or more pharmaceutically acceptable carriers and/or adjuvants.

12. A method of treating an immune disorder comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of claim 1.

13. A method of treating inflammatory diseases which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of claim 1.

14. A method of treating allergic diseases which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of claim 1.

15. A method of treating a disorder selected from the group consisting of chronic inflammation, contact dermatitis, psoriasis, rheumatoid arthritis, multiple sclerosis, type 1 diabetes, inflammatory bowel disease, guillain-barre syndrome, crohn's disease, ulcerative colitis, graft versus host disease, lupus erythematosus, asthma, Chronic Obstructive Pulmonary Disease (COPD), Adult Respiratory Distress Syndrome (ARDS), bronchitis, conjunctivitis, dermatitis, and allergic rhinitis, comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of claim 1.

16. A method of treating cancer comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of claim 1.

17. A method of administering a vaccine to an individual in need thereof, the method comprising administering the vaccine in combination with a pharmaceutically effective amount of a compound of claim 1.

18. A process for the preparation of a compound of the general formula (I) wherein R₂、R₃、R₅And R₇As defined in claim 1, the method comprising:

reacting nitrobenzoic acid (II) in which X is a leaving group with a compound carrying R₅And R₇In the presence of a coupling agent, in a suitable solvent to obtain a compound III;

will be transformed intoCompound III and with R₃In the presence of a suitable base in a suitable solvent to give IV;

reducing the nitro group of IV by catalytic hydrogenation in a suitable solvent using a suitable catalyst under an atmosphere of hydrogen or in the presence of a hydrogen source;

cyclization of compound V by treatment with cyanogen bromide in a suitable solvent to give 2-aminobenzimidazole VI, followed by treatment with a compound having R₂Acylating the acid halide of formula VI to give the desired product of formula I, and isolating the product.