Pyridones useful as inhibitors of kinases

The present invention relates to compounds useful as inhibitors of protein kinases. The invention also provides pharmaceutically acceptable compositions comprising said compounds and methods of using the compositions in the treatment of various disease, conditions, or disorders. The invention also provides processes for preparing the compounds of the invention and intermediate compounds useful in these processes.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to compounds useful as inhibitors of protein kinases. The invention also provides pharmaceutically acceptable compositions comprising the compounds of the invention and methods of using the compositions in the treatment of various disorders. The invention also provides processes for preparing the compounds of the invention and intermediate compounds useful in these processes.

BACKGROUND OF THE INVENTION

The search for new therapeutic agents has been greatly aided in recent years by a better understanding of the structure of enzymes and other biomolecules associated with diseases. One important class of enzymes that has been the subject of extensive study is protein kinases.

Protein kinases constitute a large family of structurally related enzymes that are responsible for the control of a variety of signal transduction processes within the cell. (See, Hardie, G. and Hanks, S.The Protein Kinase Facts Book, I and II, Academic Press, San Diego, Calif.: 1995). Protein kinases are thought to have evolved from a common ancestral gene due to the conservation of their structure and catalytic function. Almost all kinases contain a similar 250-300 amino acid catalytic domain. The kinases may be categorized into families by the substrates they phosphorylate (e.g., protein-tyrosine, protein-serine/threonine, lipids, etc.). Sequence motifs have been identified that generally correspond to each of these kinase families (See, for example, Hanks, S. K., Hunter, T.,FASEB J.1995, 9, 576-596; Knighton et al.,Science1991, 253, 407-414; Hiles et al.,Cell1992, 70, 419-429; Kunz et al.,Cell1993, 73, 585-596; Garcia-Bustos et al.,EMBO J.1994, 13, 2352-2361).

In general, protein kinases mediate intracellular signaling by effecting a phosphoryl transfer from a nucleoside triphosphate to a protein acceptor that is involved in a signaling pathway. These phosphorylation events act as molecular on/off switches that can modulate or regulate the target protein biological function. These phosphorylation events are ultimately triggered in response to a variety of extracellular and other stimuli. Examples of such stimuli include environmental and chemical stress signals (e.g., osmotic shock, heat shock, ultraviolet radiation, bacterial endotoxin, and H2O2), cytokines (e.g., interleukin-1 (IL-1) and tumor necrosis factor α (TNF-α)), and growth factors (e.g., granulocyte macrophage-colony-stimulating factor (GM-CSF), and fibroblast growth factor (FGF)). An extracellular stimulus may affect one or more cellular responses related to cell growth, migration, differentiation, secretion of hormones, activation of transcription factors, muscle contraction, glucose metabolism, control of protein synthesis, and regulation of the cell cycle.

Many diseases are associated with abnormal cellular responses triggered by protein kinase-mediated events as described above. These diseases include, but are not limited to, autoimmune diseases, inflammatory diseases, bone diseases, metabolic diseases, neurological and neurodegenerative diseases, cancer, cardiovascular diseases, allergies and asthma, Alzheimer's disease, and hormone-related diseases. Accordingly, there has been a substantial effort in medicinal chemistry to find protein kinase inhibitors that are effective as therapeutic agents.

The Tec family of non-receptor tyrosine kinases plays a central role in signaling through antigen-receptors such as the TCR, BCR and Fcε receptors (reviewed in Miller A, et al., Current Opinion in Immunology 14; 331-340 (2002). Tec family kinases are essential for T cell activation. Three members of the Tec family, Itk, Rlk and Tec, are activated downstream of antigen receptor engagement in T cells and transmit signals to downstream effectors, including PLC-γ. Deletion of Itk in mice results in reduced T cell receptor (TCR)-induced proliferation and secretion of the cytokines IL-2, IL-4, IL-5, IL-10 and IFN-γ (Schaeffer et al, Science 284; 638-641 (1999)), Fowell et al, Immunity 11; 399-409 (1999), Schaeffer et al Nature Immunology 2, 12; 1183-1188 (2001))). The immunological symptoms of allergic asthma are attenuated in Itk−/− mice. Lung inflammation, eosinophil infiltration and mucous production are drastically reduced in Itk−/− mice in response to challenge with the allergen OVA (Mueller et al, Journal of Immunology 170: 5056-5063 (2003)). Itk has also been implicated in atopic dermatitis. This gene has been reported to be more highly expressed in peripheral blood T cells from patients with moderate and/or severe atopic dermatitis than in controls or patients with mild atopic dermatitis (Matsumoto et al, International archives of Allergy and Immunology 129; 327-340 (2002)).

Splenocytes from Rlk−/− mice secrete half the IL-2 produced by wild type animals in response to TCR engagement (Schaeffer et al, Science 284; 638-641 (1999)), while combined deletion of Itk and Rlk in mice leads to a profound inhibition of TCR-induced responses including proliferation and production of the cytokines IL-2, IL-4, IL-5 and IFN-γ (Schaeffer et al Nature Immunology 2, 12; 1183-1188 (2001)), Schaeffer et al, Science 284; 638-641 (1999)). Intracellular signaling following TCR engagement is effected in Itk/Rlk deficient T cells; inositol triphosphate production, calcium mobilization, MAP kinase activation, and activation of the transcription factors NFAT and AP-1 are all reduced (Schaeffer et al, Science 284; 638-641 (1999), Schaeffer et al Nature Immunology 2, 12; 1183-1188 (2001)).

Tec family kinases are also essential for B cell development and activation. Patients with mutations in Btk have a profound block in B cell development, resulting in the almost complete absence of B lymphocytes and plasma cells, severely reduced Ig levels and a profound inhibition of humoral response to recall antigens (reviewed in Vihinen et al Frontiers in Bioscience 5:d917-928). Mice deficient in Btk also have a reduced number of peripheral B cells and greatly decreased levels of IgM and IgG3. Btk deletion in mice has a profound effect on B cell proliferation induced by anti-IgM, and inhibits immune responses to thymus-independent type II antigens (Ellmeier et al, J Exp Med 192:1611-1623 (2000)).

Tec kinases also play a role in mast cell activation through the high-affinity IgE receptor (FcεRI). Itk and Btk are expressed in mast cells and are activated by FcεRI cross-linking (Kawakami et al, Journal of Immunology; 3556-3562 (1995)). Btk deficient murine mast cells have reduced degranulation and decreased production of proinflammatory cytokines following FcεRI cross-linking (Kawakami et al. Journal of leukocyte biology 65:286-290). Btk deficiency also results in a decrease of macrophage effector functions (Mukhopadhyay et al, Journal of Immunology; 168, 2914-2921 (2002)).

Accordingly, there is a great need to develop compounds useful as inhibitors of protein kinases. In particular, it would be desirable to develop compounds that are useful as inhibitors of Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) protein kinases, particularly given the inadequate treatments currently available for the majority of the disorders implicated in their activation.

SUMMARY OF THE INVENTION

It has now been found that compounds of this invention, and pharmaceutically acceptable compositions thereof, are effective as inhibitors of protein kinases. In certain embodiments, these compounds are effective as inhibitors of Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) protein kinases. These compounds have the formula I as defined herein or a pharmaceutically acceptable salt thereof.

These compounds and pharmaceutically acceptable compositions thereof are useful for treating or preventing a variety of diseases, disorders or conditions, including, but not limited to, an autoimmune, inflammatory, proliferative, or hyperproliferative disease or an immunologically-mediated disease. The compositions are also useful in methods for preventing thrombin-induced platelet aggregation. The compounds provided by this invention are also useful for the study of kinases in biological and pathological phenomena; the study of intracellular signal transduction pathways mediated by such kinases; and the comparative evaluation of new kinase inhibitors.

Also provided by this invention are processes for preparing compounds of this invention and intermediate compounds useful in these processes.

DETAILED DESCRIPTION OF THE INVENTION

This invention describes compounds of Formula I:

or a pharmaceutically accepted salt thereof, whereineach R3and R4is independently H, halogen or C1-4aliphatic optionally substituted with halogen, C1-2aliphatic, OCH3, NO2, NH2, CN, NHCH3, SCH3, or N(CH)2.R2is a 3-8-membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-12 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring system having 0-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; R2is optionally substituted with JR;each X1and X2is independently —C(O)—, —NR—, or —SO2— wherein one of X1or X2is —NR— and the other of X1or X2is —C(O)— or —SO2—;R is H, unsubstituted C1-6aliphatic;R1is -T-Q;T is a bond or C1-6aliphatic, wherein up to three methylene units of the chain are optionally and independently replaced by G or G′ wherein G is —NR5—, —O—, —S—, —SO—, SO2—, —CS—, or —CO—; G′ is cyclopropyl, C≡C, or C═C; T is optionally substituted with JT;Q is independently hydrogen, a C1-6aliphatic group, a 3-8-membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-12 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring system having 0-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; Q is optionally substituted with JQ;R5is optionally substituted R, C6-10aryl, C3-10cycloaliphatic, 5-14 membered heteroaryl, or 5-14 membered heterocyclyl; or two R5groups, together with the atom(s) to which they are attached, form an optionally substituted 3-7 membered monocyclic or 8-14 membered bicyclic ring;
the optional substituents JR, JT, and JQare defined herein.

Certain embodiments of this invention provide that

Other embodiments of this invention provide that

R3is H, X1is —NR—, R is H, and X2is —C(O)—; thena) R1is not H, C1-6alkyl, O(C1-6alkyl), CH(CH3)OC(═O)CH3, or imidazole;b) when T is —CH2—, Q is not 3-OH-phenyl, 4-OH-phenyl, 4-pyridyl, 3-NO2-phenyl, OH, OC(═O)CH3, or —C(═O)CH3;c) when T is —CH2CH2—, Q is not 2-pyridyl or —COOH;
when R2is 2,4-pyrimidyl, R3and R4are H, X1is —NR—, R is H, and X2is —C(O)—, thena) R1is not methyl, NHCH3, or —NHC(═O)NH2;
when R2is 4-pyridyl, R3and R4are H, X1is —NR—, R is H, and X2is —SO2—, thena) when T is a bond, Q is not optionally substituted C6-10aryl or C5-10heteroaryl;
when R2is 4-thiazolyl, R3is H, R4is CH3, X1is —C(O)—, X2is —NR—, R is H, thena) when T is —CH2CH2—, Q is not N(CH3)2;
when R2is optionally substituted phenyl, R3is H, X1is —NR—, R is H, and X2is —C(O)—, then,a) when T is C1aliphatic wherein 1 methylene unit of the chain is replaced by G; G is —NR5—; and R5is H; then Q is not 2,6-di-isopropylphenyl;b) when T is —O—CH2—, Q is not unsubstituted phenyl;c) when T is a bond, Q is not CH3;
when R2is unsubstituted phenyl, R3is H, X1is —C(O)—, X2is —NR—, R is H, thena) when T is a bond, Q is not CH3or CH2CH3;b) when T is —CH2CH2—, Q is not unsubstituted phenyl or N(CH2CH3)2;c) when T is —CH2CH2CH2—, Q is not N(CH2CH3)2;d) R1is not NH2or

As described herein, compounds of the invention may optionally be substituted with one or more substituents, such as are illustrated generally above, or as exemplified by particular classes, subclasses, and species of the invention. It will be appreciated that the phrase “optionally substituted” is used interchangeably with the phrase “substituted or unsubstituted.” In general, the term “substituted”, whether preceded by the term “optionally” or not, refers to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically feasible compounds. The term “stable”, as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and preferably their recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, a stable compound or chemically feasible compound is one that is not substantially altered when kept at a temperature of 40° C. or less, in the absence of moisture or other chemically reactive conditions, for at least a week.

The term “optionally interrupted” refers to the replacement of one atom within an alkylidene chain with another atom. Unless otherwise specified, the second atom can replace the first atom at any position, including terminal atoms. For example, a C1-3alkyl chain optionally interrupted with —O— can form —OCH2CH3, —CH2—OCH3, or CH2CH2OH. Unless otherwise specified, the terminal groups are bonded to hydrogen on the terminal side.

The term “aliphatic” or “aliphatic group”, as used herein, means a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as “carbocycle” “cycloaliphatic” or “cycloalkyl”), that has a single point of attachment to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-20 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-10 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-8 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-6 aliphatic carbon atoms, and in yet other embodiments aliphatic groups contain 1-4 aliphatic carbon atoms. In some embodiments, “cycloaliphatic” (or “carbocycle” or “cycloalkyl”) refers to a monocyclic C3-C8hydrocarbon or bicyclic C8-C12hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule wherein any individual ring in said bicyclic ring system has 3-7 members. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

It should be understood that ring systems herein may be linearly fused, bridged, or spirocyclic.

The term “heteroaliphatic”, as used herein, means aliphatic groups wherein one or two carbon atoms are independently replaced by one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon. Heteroaliphatic groups may be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and include “heterocycle”, “heterocyclyl”, “heterocycloaliphatic”, or “heterocyclic” groups.

The term “heterocycle”, “heterocyclyl”, “heterocycloaliphatic”, or “heterocyclic” as used herein means non-aromatic, monocyclic, bicyclic, or tricyclic ring systems in which one or more ring members are an independently selected heteroatom. In some embodiments, the “heterocycle”, “heterocyclyl”, “heterocycloaliphatic”, or “heterocyclic” group has three to fourteen ring members in which one or more ring members is a heteroatom independently selected from oxygen, sulfur, nitrogen, or phosphorus, and each ring in the system contains 3 to 7 ring members.

The term “alkoxy”, or “thioalkyl”, as used herein, refers to an alkyl group, as previously defined, attached to the principal carbon chain through an oxygen (“alkoxy”) or sulfur (“thioalkyl”) atom.

The terms “haloalkyl”, “haloalkenyl” and “haloalkoxy” means alkyl, alkenyl or alkoxy, as the case may be, substituted with one or more halogen atoms. The term “halogen” means F, Cl, Br, or I.

The term “aryl” used alone or as part of a larger moiety as in “aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refers to monocyclic, bicyclic, and tricyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. The term “aryl” may be used interchangeably with the term “aryl ring”.

The term “heteroaryl”, used alone or as part of a larger moiety as in “heteroaralkyl” or “heteroarylalkoxy”, refers to monocyclic, bicyclic, and tricyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic, at least one ring in the system contains one or more heteroatoms, and wherein each ring in the system contains 3 to 7 ring members. The term “heteroaryl” may be used interchangeably with the term “heteroaryl ring” or the term “heteroaromatic”.

Optional substituents on the aliphatic group of Roor on the ring formed by 2 Rogroups are selected from NH2, NH(C1-4aliphatic), N(C1-4aliphatic)2, halogen, C1-4aliphatic, OH, O(C1-4aliphatic), NO2, CN, CO2H, CO2(C1-4aliphatic), O(haloC1-4aliphatic), and haloC1-4aliphatic, wherein each of the foregoing C1-4aliphatic groups of Rois unsubstituted;

An aliphatic or heteroaliphatic group, or a non-aromatic heterocyclic ring may contain one or more substituents. Suitable substituents (e.g JR, JT, and JQ) on the saturated carbon of an aliphatic or heteroaliphatic group, or of a non-aromatic heterocyclic ring are selected from those listed above for the unsaturated carbon of an aryl or heteroaryl group and additionally include the following: ═O, ═S, ═NNHR*, ═NN(R*)2, ═NNHC(O)R*, ═NNHCO2(alkyl), ═NNHSO2(alkyl), ═NOH, and ═NR*, where each R* is independently selected from hydrogen and an optionally substituted C1-6aliphatic. Optional substituents on the aliphatic group of R* are selected from NH2, NH(C1-4aliphatic), N(C1-4aliphatic)2, halogen, C1-4aliphatic, OH, O(C1-4aliphatic), NO2, CN, CO2H, CO2(C1-4aliphatic), O(halo C1-4aliphatic), and halo(C1-4aliphatic), wherein each of the foregoing C1-4aliphatic groups of R* is unsubstituted.

Optional substituents (e.g JR, JT, and JQ) on the nitrogen of a non-aromatic heterocyclic ring or on the nitrogen of the heteroaryl ring are selected from —R+, —N(R+)2, —C(O)R+, —CO2R+, —C(O)C(O)R+, —C(O)CH2C(O)R+, —SO2R+, —SO2N(R+)2, —C(═S)N(R+)2, —C(═NH)—N(R+)2, and —NR+SO2R+; wherein R+is hydrogen, an optionally substituted C1-6aliphatic, optionally substituted phenyl, optionally substituted —O(Ph), optionally substituted —CH2(Ph), optionally substituted —(CH2)2(Ph); optionally substituted —CH═CH(Ph); or an unsubstituted 5-6 membered heteroaryl or heterocyclic ring having one to four heteroatoms independently selected from oxygen, nitrogen, and sulfur, or, notwithstanding the definition above, two independent occurrences of R+, on the same substituent or different substituents, taken together with the atom(s) to which each R+group is bound, form a 5-8-membered heterocyclyl, aryl, or heteroaryl ring or a 3-8-membered cycloaliphatic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Optional substituents on the aliphatic group or the phenyl ring of R+are selected from NH2, NH(C1-4aliphatic), N(C1-4aliphatic)2, halogen, C1-4aliphatic, OH, O(C1-4aliphatic), NO2, CN, CO2H, CO2(C1-4aliphatic), O(halo C1-4aliphatic), and halo(C1-4aliphatic), wherein each of the foregoing C1-4aliphatic groups of R+is unsubstituted.

The term “alkylidene chain” refers to a straight or branched carbon chain that may be fully saturated or have one or more units of unsaturation and has two points of attachment to the rest of the molecule, wherein one or more methylene units may optionally and independently be replaced with a group including, but not limited to, CO, CO2, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO2, NRCONR, SO, SO2, NRSO2, SO2NR, NRSO2NR, O, S; or NR.

As detailed above, in some embodiments, two independent occurrences of Ro(or R+, or any other variable similarly defined herein), are taken together with the atom(s) to which each variable is bound to form a 5-8-membered heterocyclyl, aryl, or heteroaryl ring or a 3-8-membered cycloalkyl ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Exemplary rings that are formed when two independent occurrences of Ro(or R+, or any other variable similarly defined herein) are taken together with the atom(s) to which each variable is bound include, but are not limited to the following: a) two independent occurrences of Ro(or R+, or any other variable similarly defined herein) that are bound to the same atom and are taken together with that atom to form a ring, for example, N(Ro)2, where both occurrences of Roare taken together with the nitrogen atom to form a piperidin-1-yl, piperazin-1-yl, or morpholin-4-yl group; and b) two independent occurrences of Ro(or R+, or any other variable similarly defined herein) that are bound to different atoms and are taken together with both of those atoms to form a ring, for example where a phenyl group is substituted with two occurrences of ORo

these two occurrences of Roare taken together with the oxygen atoms to which they are bound to form a fused 6-membered oxygen containing ring:

It will be appreciated that a variety of other rings can be formed when two independent occurrences of Ro(or R+, or any other variable similarly defined herein) are taken together with the atom(s) to which each variable is bound and that the examples detailed above are not intended to be limiting.

Unless otherwise stated, structures depicted herein are also meant to include an N-oxide derivative or a pharmaceutically acceptable salt of each of the compounds of formula I.

According to one embodiment of this invention, T is C1-3aliphatic optionally interrupted with zero or one G groups wherein G is selected from O, NR5, and S.

In some embodiments, T is —C1-2aliphatic-G- wherein G is O or NR5, and G is bound to Q in a chemically stable arrangement. In other embodiments, G is bound to X2in a chemically stable arrangement. In yet other embodiments T is C1-3aliphatic optionally interrupted with zero G groups.

In some embodiments, T is C1-3aliphatic optionally interrupted with zero or one G′ groups. In other embodiments, T is C1-3aliphatic optionally interrupted with zero or one G or G′ groups.

In some embodiments, T is —CH2—; in other embodiments T is a bond.

According to one embodiment of the invention, each R3and R4is independently H. In some embodiments, both R3and R4are H.

According to some embodiments R2is a 5-8 membered monocyclyl optionally substituted with up to 5 JRgroups. In certain embodiments, R2is a 5-6 membered aryl or heteroaryl optionally substituted with up to 5 JRgroups. In other embodiments R2is a 5-6 membered heteroaryl optionally substituted with up to 5 JRgroups, preferably R2is a 6 membered heteroaryl having 1 or 2 nitrogen atoms wherein R2is optionally substituted with up to 5 JRgroups.

In some embodiments, R2is C3-8cycloaliphatic optionally substituted with up to five JRgroups. In other embodiments, R2is C3-8cycloalkyl optionally substituted with up to five JRgroups. In certain embodiments, R2is C3-8cycloalkenyl optionally substituted with up to five JRgroups. In other embodiments, R2is cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, or cycloheptenyl, optionally substituted with up to five JRgroups.

In some embodiments R2is a pyridine ring optionally substituted with up to 5 JRgroups. In some embodiments, R2is 2-pyridinyl, 3-pyridyl, or 4-pyridyl optionally substituted with up to five JRgroups. In certain embodiments, R2is a pyrimidine ring optionally substituted with up to five JRgroups. In some embodiments, R2is a 2,4 pyrimidinyl. In other embodiments, R2is a 5-membered heteroaryl ring optionally substituted with up to five JRgroups. In some embodiments, R2is thiophene or pyrazole optionally substituted with up to five JRgroups. In yet other embodiments R is phenyl optionally substituted with up to 5 JRgroups.

In some embodiments R2is optionally substituted with up to 5 JRgroups; in other embodiments, up to 3 JRgroups; in yet other embodiments, 0 or 1 JRgroups.

In certain embodiments, JRis selected from oxo or ═NOH.

In some embodiments, each JRis independently selected from optionally substituted 5-8 membered heterocyclyl, optionally substituted —NR(C1-4alkyl)N(Ro)2, optionally substituted —NR(C1-4alkyl)ORo, —N(Ro)2, or optionally substituted —NH(5-6 membered heterocyclyl). In certain embodiments JRis —NH(C1-4alkyl)N(Ro)2; in other embodiments —NH(C1-4alkyl)NHRoor —NH(C1-4alkyl)NH2; In some embodiments JRis —NR(CH2CH2)N(Ro)2; In other embodiments JRis —N(CH3)CH2CH2N(Ro)2;

In other embodiments, each JRis independently selected from optionally substituted —NH(5-6 membered heterocyclyl).

In some embodiments JRis optionally and independently substituted with Ro.

In one embodiment of this invention, each X1and X2is independently —C(O)— or —NR— wherein one of X1or X2is —NR— and the other of X1or X2is —C(O)—.

In one embodiments of this invention, Q is a 3-8-membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-12 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring system having 0-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

In certain embodiments Q is C6-10aryl, C3-10cycloaliphatic, 5-14 membered heteroaryl, or 5-14 membered heterocyclyl. In other embodiments Q is C6-10aryl or 5-14 membered heteroaryl. In yet other embodiments Q is a 5-6 membered aryl or heteroaryl. In some embodiments, Q is 5-8 membered heterocyclyl; in certain embodiments, 5-6 membered heterocyclyl; In certain embodiments Q is phenyl.

In some embodiments R2is optionally substituted with up to 5 JQgroups; in other embodiments, up to 3 JQgroups; in yet other embodiments, 0 or 1 JQgroups.

In some embodiments, the variables are as depicted in the Table I compounds.

Accordingly, representative examples of compounds of formula I are depicted in Table I.

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

Scheme I above shows a general synthetic route that is used for preparing the compounds 3a of this invention when R1is as described herein. Compounds of formula 3a may be prepared by reaction of amrinone 1 with an acid chloride in pyridine according to step (a) of Scheme I. The reaction is amenable to a variety of acid chlorides.

Compounds I-1 to I-67 and I-82 to I-85 were prepared according to the general methods described in Scheme I.

Scheme II above shows a general synthetic route that is used for preparing the compounds 3b of this invention when R0is as described herein. Compounds of formula 3b may be prepared by reaction of 1-20 with an excess of amine in NMP according to step (a) of Scheme II. The reaction is amenable to a variety of amines.

Compounds I-68 to I-81 were prepared according to the general methods described in Scheme II.

Scheme III above shows a general synthetic route that is used for preparing the compounds 7 of this invention when R2is as described herein. Starting material 4, which may be prepared by methods described by Warner, et al,J. Med. Chem.1994, 37, 3090, is methylated according to step (a) of Scheme II. Compound of formula 6 is formed by reaction of the iodide 5 with bis(pinacolato)diboron in presence of palladium as a catalyst. The formation of derivatives 7 is achieved by treating the boronic ester derivatives 6 with a halide R2-Hal in the presence of palladium as a catalyst by using the Suzuki coupling methods that are well known in the art. The reaction is amenable to a variety of substituted halides R2-Hal.

Scheme IV above shows a general synthetic route that has been used for preparing compounds 10 of this invention when R1and R2are as described herein. Demethylation of 7 in acidic conditions leads to the formation of 8, which is deprotected according to step (b). Finally, compounds of formula 10 may be prepared by reaction of derivatives 9 with an acid chloride 2 in pyridine. The reaction is amenable to a variety of acid chlorides 2.

Scheme V above shows another general synthetic route that has been used for preparing compounds 10 of this invention when R1and R2are as described herein. Intermediates 11, obtained by deprotection of amines 7, react with an acid chloride 2 in pyridine. The reaction is amenable to a variety of acid chlorides 2. After demethylation of intermediates 12 in acidic medium, pyridones 10 are obtained.

Scheme VI above shows another general synthetic route that has been used for preparing compounds 10 of this invention when R1and R2are as described herein. Intermediate 13, obtained by deprotection of amine 6, reacts with an acid chloride 2 to form compounds of formula 14. The reaction is amenable to a variety of acid chlorides 2. The formation of derivatives 12 is achieved by treating the boronic ester derivatives 14 with a halide R2-Hal in the presence of palladium as a catalyst by using the Suzuki coupling methods that are well known in the art. The reaction is amenable to a variety of substituted halides R2-Hal. After demethylation of intermediates 12 in acidic medium, pyridones 10 are obtained.

Scheme VII above shows a general synthetic route that is used for preparing the compounds 16 of this invention when R2and R0are as described herein. Compounds of formula 16 may be prepared by reaction of 15 with an excess of amine in NMP according to step (a) of Scheme VII. The reaction is amenable to a variety of amines.

Compounds II-1 to II-182 were prepared according to the general methods described in Schemes III, IV, V, VI and VII.

Scheme VIII above shows a general synthetic route that has been used for preparing compounds 19 of this invention when R, R1and R2are as described herein. Starting material 17 may be prepared by methods substantially similar to those described in the literature by Church et alJ. Org. Chem.1995, 60, 3750. Compounds of formula 19 are prepared according to step (a) of scheme VIII.

Compounds III-1 to III-54 were prepared according to the general method described in Schemes VIII.

Scheme IX above shows a general synthetic route that is used for preparing the compounds 21 of this invention when R1and R2are as described herein. Compounds of formula 21 may be prepared by reaction of derivatives 9 with a sulfonyl chloride 20 in pyridine. The reaction is amenable to a variety of sulfonyl chlorides 20.

Compound IV-1 was prepared according to the general method described in Scheme IX.

This invention also provides compounds that can be used as intermediates to synthesize compounds of this invention. Additionally, this invention provides processes for using these intermediate compounds to prepare compounds of this invention.

Specifically, a compound 22 can be used as an intermediate compound in a process for preparing a compound 23. Compound 23 can then be carried on to a compound of formula I.

wherein:
R10is an amino protective group;
R11is H or C1-6alkyl or R10and R11together with the nitrogen atom to which they are bound form an amine protective group;
R12is a hydroxyl protecting group; and
R2is as defined herein.

In one embodiment, compound 22 is reacted with an appropriate compound comprising R2under appropriate reaction conditions to form compound 23. An example of an appropriate compound comprising R2is R2—X, wherein X is an appropriate leaving group, such as a halo group. Appropriate reaction conditions are coupling conditions that allow bond formation between a boronic ester (or boronic acid) and R2—X. Appropriate leaving groups and appropriate coupling conditions are known to skilled practitioners (see, e.g., March, supra).

Compound 23 can be prepared by treating the boronic ester derivative 22 with a halide R2-Hal in the presence of palladium as a catalyst by using coupling methods that are well known in the art, e.g., by using Suzuki coupling.

Scheme XI depicts an example of using Suzuki coupling conditions in a method of this invention. In Scheme XI, R10is a Cbz group, R11is hydrogen, and R12is a methyl group. Nevertheless, it should be understood that the reaction depicted in Scheme XI could be employed with compound 22 in the place of compound 6 and compound 23 in the place of 7.

Compound 23 can be also prepared by treating the boronic ester derivative 22 with a nitrogen containing saturated, partially unsaturated, or fully unsaturated monocyclic or bicyclic ring (as described in the R2definition) by reacting through a nitrogen atom in the ring, in the presence of copper as a catalyst, e.g., by using coupling methods that are well known in the art (see, Chemick et al.J. Org. Chem.2005, 1486) to provide 23 (wherein R2is a 3-8-membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having at least one nitrogen heteroatom; or an 8-12 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring system having having at least one nitrogen heteroatom and R2being optionally substituted with JR). Scheme XII depicts an example of cooper mediated coupling conditions in a method of this invention. In Scheme XII, R10is a Cbz group, R11is hydrogen, and R12is a methyl group. Nevertheless, it should be understood that the reaction depicted in Scheme XII could be employed with compound 22 in the place of compound 6 and compound 23 in the place of 7.

In a process of this invention, compound 23 (and related compounds, such as compound 7) is converted to a compound of formula I by methods known to skilled practitioners including, but not limited to, those disclosed herein. In certain embodiments, the hydroxyl protective group in compound 23 is removed and then the amino protective group is removed. The resulting amine is reacted with an appropriate R1containing intermediate to provide the compound of formula I. For specific examples of this embodiment, see Scheme IV and Scheme IX. In Scheme IV, R10is a Cbz group, R11is hydrogen, and R12is a methyl group. Nevertheless, it should be understood that the reaction depicted in Scheme IV could be employed with compound 23 in the place of compound 7.

In another embodiment, the amino protective group in compound 23 is removed and then the resulting amine is reacted with an appropriate R1containing intermediate to provide a compound X. A compound of formula I is provided by removing the hydroxyl protective group from compound X. For a specific example of this embodiment, see Scheme V. In Scheme V, R10is a Cbz group, R11is hydrogen, and R12is a methyl group. Nevertheless, it should be understood that the reaction depicted in Scheme V could be employed with compound X in the place of compound 11 and compound XX in the place of compound 12.

In embodiments wherein X1is —NR—, the amino protective group R11may be a group R1—X2—. As would be recognized, in such embodiments, it would not be necessary to remove the amino protective group and replace it with an R1containing group. Accordingly, to obtain a compound of formula I, compound 23 would be reacted under conditions suitable to remove the hydroxyl protective group (thus providing the compound of formula I). In embodiments where the R1C(═O)— group is incompatible with the boronic ester formation, the boronic ester could be formed with R10being Cbz and then the Cbz group could be replaced with an R1containing group after formation of the boronic ester. For a specific example of this embodiment, see Scheme VI. In Scheme VI, R10(in compound 14) is R1C(═O)—, R11is hydrogen, and R12is a methyl group. Nevertheless, it should be understood that the reaction depicted in Scheme VI could be employed with compound 22 in the place of compound 6.

Alternatively, R10in 23 may be converted to R1—X2—. That is, a functional group in R10could be converted to the desired R1containing group. Then, the hydroxyl protective group would be removed to provide the compound of formula I. For specific examples of this embodiment, see Scheme II and Scheme VII.

Compound 22 may be prepared by methods known to skilled practitioners including, but not limited to, methods disclosed herein. In one embodiment, an iodo compound 24 is reacted under conditions to form the boronic ester 22 (Scheme XIII). For a specific example of such conditions, see Scheme III. In Scheme III, R10is a Cbz group, R11is hydrogen, and R12is a methyl group. Nevertheless, it should be understood that the reaction depicted in Scheme XII could be employed with compound 22 in the place of compound 6 and compound 23 in the place of 7.

It should be understood that instead of using a boronic ester 22 in a process of this invention, the corresponding boronic acid, could be used (Scheme XIV). The boronic acid could be used as a starting material or generated in situ. Compound 25 may be prepared by known methods including, but not limited to, conversion of boronic ester 22 to boronic acid 25.

The protective groups protect the amino and hydroxyl functional groups from reacting under conditions for converting the boronic ester or acid to the R2group. Many amino protective groups and hydroxyl protective groups are known to skilled practitioners. Examples of such protective groups may be found in T. W. Greene and P. G. M. Wutz, “Protective Groups in Organic Synthesis”, 3rdEdition, John Wiley & Sons, Inc. (1999) and earlier editions of this book and J. W. F. McOmie, “Protective Groups in Organic Synthesis”, Plenum Press (1973).

Although certain exemplary embodiments are depicted and described above and herein, it will be appreciated that a compounds of the invention can be prepared according to the methods described generally above using appropriate starting materials by methods generally available to one of ordinary skill in the art.

As disclosed herein, the present invention provides compounds that are inhibitors of protein kinases, and thus the present compounds are useful for the treatment of diseases, disorders, and conditions including, but not limited to an autoimmune, inflammatory, proliferative, or hyperproliferative disease or an immunologically-mediated disease. Accordingly, in another aspect of the present invention, pharmaceutically acceptable compositions are provided, wherein these compositions comprise any of the compounds as described herein, and optionally comprise a pharmaceutically acceptable carrier, adjuvant or vehicle. In certain embodiments, these compositions optionally further comprise one or more additional therapeutic agents. Such additional therapeutic agents include, but are not limited to an agent for the treatment of an autoimmune, inflammatory, proliferative, hyperproliferative disease, or an immunologically-mediated disease including rejection of transplanted organs or tissues and Acquired Immunodeficiency Syndrome (AIDS).

It will also be appreciated that certain of the compounds of present invention can exist in free form for treatment, or where appropriate, as a pharmaceutically acceptable derivative thereof. According to the present invention, a pharmaceutically acceptable derivative includes, but is not limited to, pharmaceutically acceptable salts, esters, salts of such esters, or any other adduct or derivative which upon administration to a patient in need is capable of providing, directly or indirectly, a compound as otherwise described herein, or a metabolite or residue thereof.

As used herein, the term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. A “pharmaceutically acceptable salt” means any non-toxic salt or salt of an ester of a compound of this invention that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention or an inhibitorily active metabolite or residue thereof. As used herein, the term “inhibitorily active metabolite or residue thereof” means that a metabolite or residue thereof is also an inhibitor of a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) protein kinases kinase.

In certain embodiments, the composition comprises an effective amount of the compound of claim1, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. In a specific embodiment, the compound is present in an amount to detectably inhibit a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) protein kinase.

This invention also provides a pharmaceutical composition made by combining a compound of this invention and a pharmaceutically acceptable carrier, adjuvant, or vehicle and a process for making a pharmaceutical composition comprising combining a compound of this invention and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In yet another aspect, a method for the treatment or lessening the severity of a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk)-mediated diseases is provided comprising administering an effective amount of a compound, or a pharmaceutically acceptable composition comprising a compound to a subject in need thereof. The methods may employ a compound of Formula I or any of the other compounds of this invention:

or a pharmaceutically accepted salt thereof, whereineach R3and R4is independently H, halogen or C1-4aliphatic optionally substituted with halogen, C1-2aliphatic, OCH3, NO2, NH2, CN, NHCH3, SCH3, or N(CH)2.R2is a 3-8-membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-12 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring system having 0-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; R2is optionally substituted with JR;
each X1and X2is independently —C(O)—, —NR—, or —SO2— wherein one of X1or X2is —NR— and the other of X1or X2is —C(O)— or —SO2—;R is H, unsubstituted C1-6aliphatic;R1is -T-Q;T is a bond or C1-6aliphatic, wherein up to three methylene units of the chain are optionally and independently replaced by G or G′ wherein G is —NR5—, —O—, —S—, —SO—, SO2—, —CS—, or —CO—; G′ is cyclopropyl, C≡C, or C═C; T is optionally substituted with JT;Q is independently hydrogen, a C1-6aliphatic group, a 3-8-membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-12 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring system having 0-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; Q is optionally substituted with JQ; andR5is optionally substituted R, C6-10aryl, C3-10cycloaliphatic, 5-14 membered heteroaryl, or 5-14 membered heterocyclyl; or two R5groups, together with the atom(s) to which they are attached, form an optionally substituted 3-7 membered monocyclic or 8-14 membered bicyclic ring;
wherein the optional substituents JR, JT, and JQare defined herein.

In certain embodiments of the present invention an “effective amount” of the compound or pharmaceutically acceptable composition is that amount effective for a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk)-mediated disease. The compounds and compositions, according to the method of the present invention, may be administered using any amount and any route of administration effective for treating or lessening the severity of a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk)-mediated disease. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the particular agent, its mode of administration, and the like. The compounds of the invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The expression “dosage unit form” as used herein refers to a physically discrete unit of agent appropriate for the patient to be treated. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific effective dose level for any particular patient or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed, and like factors well known in the medical arts. The term “patient”, as used herein, means an animal, preferably a mammal, and most preferably a human.

As described generally above, the compounds of the invention are useful as inhibitors of protein kinases. In one embodiment, the compounds and compositions of the invention are inhibitors of one or more of Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase, and thus, without wishing to be bound by any particular theory, the compounds and compositions are particularly useful for treating or lessening the severity of a disease, condition, or disorder where activation of one or more of a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase is implicated in the disease, condition, or disorder. When activation of Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) is implicated in a particular disease, condition, or disorder, the disease, condition, or disorder may also be referred to as a “Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk)-mediated disease” or disease symptom. Accordingly, in another aspect, the present invention provides a method for treating or lessening the severity of a disease, condition, or disorder where activation or one or more of Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) is implicated in the disease state.

Also without wishing to be bound by any particular theory, the compounds and compositions of this invention are particularly useful for treating or lessening the severity of a disease, condition, or disorder where activation of Itk kinase is implicated in the disease, condition, or disorder and are particularly useful for inhibiting Itk selectively over Btk and Rlk (see, Examples 14-16 and 18).

The activity of a compound utilized in this invention as an inhibitor of a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase may be assayed in vitro, in vivo or in a cell line. In vitro assays include assays that determine inhibition of either the phosphorylation activity or ATPase activity of activated Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase. Alternate in vitro assays quantitate the ability of the inhibitor to bind to a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase. Inhibitor binding may be measured by radiolabelling the inhibitor prior to binding, isolating the inhibitor/Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk), complex and determining the amount of radiolabel bound. Alternatively, inhibitor binding may be determined by running a competition experiment where new inhibitors are incubated with a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase bound to known radioligands.

The term “measurably inhibit”, as used herein means a measurable change in a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase activity between a sample comprising said composition and a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase and an equivalent sample comprising a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase in the absence of said composition.

The term “Tec family tyrosine kinases-mediated condition”, as used herein means any disease or other deleterious condition in which Tec family kinases are known to play a role. Such conditions include, without limitation, autoimmune, inflammatory, proliferative, and hyperproliferative diseases and immunologically mediated diseases including rejection of transplanted organs or tissues and Acquired Immunodeficiency Syndrome (AIDS).

For example, Tec family tyrosine kinases-mediated conditions include diseases of the respiratory tract including, without limitation, reversible obstructive airways diseases including asthma, such as bronchial, allergic, intrinsic, extrinsic and dust asthma, particularly chronic or inveterate asthma (e.g., late asthma airways hyper-responsiveness) and bronchitis. Additionally, Tec family tyrosine kinases diseases include, without limitation, those conditions by inflammation of the nasal mucus membrane, including acute rhinitis, allergic, atrophic thinitis and chronic rhinitis including rhinitis caseosa, hypertrophic rhinitis, rhinitis purulenta, rhinitis sicca and rhinitis medicamentosa; membranous rhinitis including croupous, fibrinous and pseudomembranous rhinitis and scrofulous rhinitis, seasonal rhinitis including rhinitis nervosa (hay fever) and vasomotor rhinitis, sarcoidosis, farmer's lung and related diseases, fibroid lung and idiopathic interstitial pneumonia.

Tec family tyrosine kinases-mediated conditions also include diseases and disorders of the gastrointestinal tract, including, without limitation, Coeliac disease, proctitis, eosinophilic gastro-enteritis, mastocytosis, pancreatitis, Crohn's disease, ulcerative colitis, food-related allergies which have effects remote from the gut, e.g., migraine, rhinitis and eczema.

Tec family tyrosine kinases-mediated conditions also include allograft rejection including, without limitation, acute and chronic allograft rejection following for example transplantation of kidney, heart, liver, lung, bone marrow, skin and cornea; and chronic graft versus host disease.

It will also be appreciated that the compounds and pharmaceutically acceptable compositions of the present invention can be employed in combination therapies, that is, the compounds and pharmaceutically acceptable compositions can be administered concurrently with, prior to, or subsequent to, one or more other desired therapeutics or medical procedures. The particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved. It will also be appreciated that the therapies employed may achieve a desired effect for the same disorder (for example, an inventive compound may be administered concurrently with another agent used to treat the same disorder), or they may achieve different effects (e.g., control of any adverse effects). As used herein, additional therapeutic agents that are normally administered to treat or prevent a particular disease, or condition, are known as “appropriate for the disease, or condition, being treated”.

Additional therapeutic agents that may be used in the methods of this invention include, but are not limited to, agents for the treatment of an autoimmune, inflammatory, proliferative, hyperproliferative disease, or an immunologically-mediated disease including rejection of transplanted organs or tissues and Acquired Immunodeficiency Syndrome (AIDS), wherein the additional therapeutic agent is appropriate for the disease being treated; and the additional therapeutic agent is administered together with said composition as a single dosage form or separately from said composition as part of a multiple dosage form.

For example, chemotherapeutic agents or other anti-proliferative agents may be combined with the compounds of this invention to treat proliferative diseases and cancer. Examples of known chemotherapeutic agents include, but are not limited to, For example, other therapies or anticancer agents that may be used in combination with the inventive anticancer agents of the present invention include surgery, radiotherapy (in but a few examples, gamma.-radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes, to name a few), endocrine therapy, biologic response modifiers (interferons, interleukins, and tumor necrosis factor (TNF) to name a few), hyperthermia and cryotherapy, agents to attenuate any adverse effects (e.g., antiemetics), and other approved chemotherapeutic drugs, including, but not limited to, alkylating drugs (mechlorethamine, chlorambucil, Cyclophosphamide, Melphalan, Ifosfamide), antimetabolites (Methotrexate), purine antagonists and pyrimidine antagonists (6-Mercaptopurine, 5-Fluorouracil, Cytarabile, Gemcitabine), spindle poisons (Vinblastine, Vincristine, Vinorelbine, Paclitaxel), podophyllotoxins (Etoposide, Irinotecan, Topotecan), antibiotics (Doxorubicin, Bleomycin, Mitomycin), nitrosoureas (Carmustine, Lomustine), inorganic ions (Cisplatin, Carboplatin), enzymes (Asparaginase), and hormones (Tamoxifen, Leuprolide, Flutamide, and Megestrol), Gleevec™, adriamycin, dexamethasone, and cyclophosphamide. For a more comprehensive discussion of updated cancer therapies see, http://www.nci.nih.gov/, a list of the FDA approved oncology drugs at http://www.fda.gov/cder/cancer/druglistframe.htm, and The Merck Manual, Seventeenth Ed. 1999, the entire contents of which are hereby incorporated by reference.

Other examples of agents the inhibitors of this invention may also be combined with include, without limitation: treatments for Alzheimer's Disease such as Aricept® and Excelon®; treatments for Parkinson's Disease such as L-DOPA/carbidopa, entacapone, ropinrole, pramipexole, bromocriptine, pergolide, trihexephendyl, and amantadine; agents for treating Multiple Sclerosis (MS) such as beta interferon (e.g., Avonex® and Rebif®), Copaxone®, and mitoxantrone; treatments for asthma such as albuterol and Singulair®; agents for treating schizophrenia such as zyprexa, risperdal, seroquel, and haloperidol; anti-inflammatory agents such as corticosteroids, TNF blockers, IL-1 RA, azathioprine, cyclophosphamide, and sulfasalazine; immunomodulatory and immunosuppressive agents such as cyclosporine, tacrolimus, rapamycin, mycophenolate mofetil, interferons, corticosteroids, cyclophosphamide, azathioprine, and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonian agents; agents for treating cardiovascular disease such as beta-blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers, and statins; agents for treating liver disease such as corticosteroids, cholestyramine, interferons, and anti-viral agents; agents for treating blood disorders such as corticosteroids, anti-leukemic agents, and growth factors; and agents for treating immunodeficiency disorders such as gamma globulin.

The compounds of this invention or pharmaceutically acceptable compositions thereof may also be incorporated into compositions for coating implantable medical devices, such as prostheses, artificial valves, vascular grafts, stents and catheters. Accordingly, the present invention, in another aspect, includes a composition for coating an implantable device comprising a compound of the present invention as described generally above, and in classes and subclasses herein, and a carrier suitable for coating said implantable device. In still another aspect, the present invention includes an implantable device coated with a composition comprising a compound of the present invention as described generally above, and in classes and subclasses herein, and a carrier suitable for coating said implantable device.

Vascular stents, for example, have been used to overcome restenosis (re-narrowing of the vessel wall after injury). However, patients using stents or other implantable devices risk clot formation or platelet activation. These unwanted effects may be prevented or mitigated by pre-coating the device with a pharmaceutically acceptable composition comprising a kinase inhibitor. Suitable coatings and the general preparation of coated implantable devices are described in U.S. Pat. Nos. 6,099,562; 5,886,026; and 5,304,121. The coatings are typically biocompatible polymeric materials such as a hydrogel polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate, and mixtures thereof. The coatings may optionally be further covered by a suitable topcoat of fluorosilicone, polysaccarides, polyethylene glycol, phospholipids or combinations thereof to impart controlled release characteristics in the composition.

Another aspect of the invention relates to inhibiting Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) activity in a biological sample or a patient, which method comprises administering to the patient, or contacting said biological sample with a compound of formula I or a composition comprising said compound. The term “biological sample”, as used herein, includes, without limitation, cell cultures or extracts thereof; biopsied material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.

Inhibition of Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase activity in a biological sample is useful for a variety of purposes that are known to one of skill in the art. Examples of such purposes include, but are not limited to, blood transfusion, organ-transplantation, biological specimen storage, and biological assays.

EXAMPLES

As used herein, the term “Rt(min)” refers to the HPLC retention time, in minutes, associated with the compound. Unless otherwise indicated, the HPLC method utilized to obtain the reported retention time is as follows:

A variety of other compounds of Formula I have been prepared by methods substantially similar to those described herein. The characterization data for these compounds is summarized in Table I-A below and includes HPLC, LC/MS (observed) and1H NMR data.

1H NMR data is summarized in Table I-A below wherein1H NMR data was obtained at 400 Mhz in deuterated DMSO, unless otherwise indicated, and was found to be consistent with structure. Compound numbers correspond to the compound numbers listed in Table 1.

(5-Iodo-2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid benzyl ester (3.68 g, 9.94 mmol) was dissolved in chloroform (50 mL) at room temperature under nitrogen in the dark (foil wrapped). Silver carbonate (3.70 g, 13.2 mmol) was added followed by iodomethane (6.2 mL, 99.4 mmol). The reaction mixture was allowed to stir at room temperature for 48 hours. The silver salts were removed by filtration through a pad of celite, washing with more chloroform and the filtrate concentrated in vacuo. The residue was purified by column chromatography to give the title compound as a white solid (3.14 g, 82% yield). MS (ES+) m/e=385.1H NMR (CDCl3) δH 3.97 (3H, s), 5.23 (2H, s), 7.15 (1H, br s), 7.35-7.47 (5H, m), 8.00 (1H, s), 8.64 (1H, br s).

[2-Methoxy-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-3-yl]-carbamic acid benzyl ester (506 mg, 1.32 mmol) was dissolved in methanol (10 mL). Pd(OH)2on carbon (51 mg, 10 mol %) was added and the reaction was degassed with nitrogen. The nitrogen atmosphere was replaced by hydrogen and the reaction mixture was stirred at room temperature for 5 hours. The palladium residue was removed by filtration through a path of Celite, rinsing with more methanol. The filtrate was concentrated in vacuo to give the title compound as an off-white solid (319 mg, 97% yield). MS (ES+) m/e=251.1H NMR (DMSO-d6) δH 1.27 (12H, s), 3.88 (3H, s), 4.89 (2H, br s), 7.13 (1H, s), 7.64 (1H, s).

A variety of other compounds of Formula II have been prepared by methods substantially similar to those described herein. The characterization data for these compounds is summarized in Table II-A below and includes HPLC, LC/MS (observed) and1H NMR data.

1H NMR data is summarized in Table II-A below wherein1H NMR data was obtained at 400 Mhz in deuterated DMSO, unless otherwise indicated, and was found to be consistent with structure. Compound numbers correspond to the compound numbers listed in Table 1.

A variety of other compounds of Formula III have been prepared by methods substantially similar to those described herein Example 12. The characterization data for these compounds is summarized in Table III-A below and includes HPLC, LC/MS (observed) and1H NMR data.

1H NMR data is summarized in Table III-A below wherein1H NMR data was obtained at 400 Mhz in deuterated DMSO, unless otherwise indicated, and was found to be consistent with structure. Compound numbers correspond to the compound numbers listed in Table 1.

ITK Inhibition Assay

Compounds were screened for their ability to inhibit Itk using a radioactive-phosphate incorporation assay. Assays were carried out in a mixture of 100 mM HEPES (pH 7.5), 10 mM MgCl2, 25 mM NaCl, 0.01% BSA and 1 mM DTT. Final substrate concentrations were 15 μM [γ-33P]ATP (400 mCi 33P ATP/mmol ATP, Amersham Pharmacia Biotech/Sigma Chemicals) and 2 μM peptide (SAM68 protein D332-443). Assays were carried out at 25° C. in the presence of 30 nM Itk. An assay stock buffer solution was prepared containing all of the reagents listed above, with the exception of ATP and the test compound of interest. 50 μL of the stock solution was placed in a 96 well plate followed by addition of 1.5 μL of DMSO stock containing serial dilutions of the test compound (typically starting from a final concentration of 15 μM with 2-fold serial dilutions) in duplicate (final DMSO concentration 1.5%). The plate was pre-incubated for 10 minutes at 25° C. and the reaction initiated by addition of 50 μL [γ-33P]ATP (final concentration 15 μM).

The reaction was stopped after 10 minutes by the addition of 50 μL of a TCA/ATP mixture (20% TCA, 0.4 mM ATP). A Unifilter GF/C 96 well plate (Perkin Elmer Life Sciences, Cat no. 6005174) was pretreated with 50 μL Milli Q water prior to the addition of the entire reaction mixture (150 μL). The plate was washed with 200 μL Milli Q water followed by 200 mL of a TCA/ATP mixture (5% TCA, 1 mM ATP). This wash cycle was repeated a further 2 times. After drying, 30 μL Optiphase ‘SuperMix’ liquid scintillation cocktail (Perkin Elmer) was added to the well prior to scintillation counting (1450 Microbeta Liquid Scintillation Counter, Wallac).

IC50 data were calculated from non-linear regression analysis of the initial rate data using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego Calif., USA).

Assays were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 nM MgCl2, 0.1% BSA and 1 mM DTT. Final substrate concentrations in the assay were 7.5 μM [γ-33P]ATP (400 mCi 33P ATP/mmol ATP, Amersham Pharmacia Biotech/Sigma Chemicals) and 3 μM peptide (SAM68 protein D332-443). Assays were carried out at 25° C. in the presence of 50 nM Itk. An assay stock buffer solution was prepared containing all of the reagents listed above, with the exception of ATP and the test compound of interest. 50 μL of the stock solution was placed in a 96 well plate followed by addition of 2 μL of DMSO stock containing serial dilutions of the test compound (typically starting from a final concentration of 50 μM with 2-fold serial dilutions) in duplicate (final DMSO concentration 2%). The plate was pre-incubated for 10 minutes at 25° C. and the reaction initiated by addition of 50 μL [γ-33P]ATP (final concentration 7.5 μM).

The reaction was stopped after 10 minutes by the addition of 100 mL 0.2M phosphoric acid+0.01% TWEEN 20. A multiscreen phosphocellulose filter 96-well plate (Millipore, Cat no. MAPHN0B50) was pretreated with 10 μL 0.2M phosphoric acid+0.01% TWEEN 20 prior to the addition of 170 mL of the stopped assay mixture. The plate was washed with 4×200 μL 0.2M phosphoric acid+0.01% TWEEN 20. After drying, 30 μL Optiphase ‘SuperMix’ liquid scintillation cocktail (Perkin Elmer) was added to the well prior to scintillation counting (1450 Microbeta Liquid Scintillation Counter, Wallac).

Ki(app) data were calculated from non-linear regression analysis of the initial rate data using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego Calif., USA).

Compounds were screened for their ability to inhibit Itk using a standard coupled enzyme assay (Fox et al., Protein Sci., (1998) 7, 2249).

An assay stock buffer solution was prepared containing all of the reagents listed above, with the exception of ATP and the test compound of interest. 60 μl of the stock solution was placed in a 96 well plate followed by addition of 2 μl of DMSO stock containing serial dilutions of the test compound (typically starting from a final concentration of 15 μM). The plate was preincubated for 10 minutes at 25° C. and the reaction initiated by addition of 5 μl of ATP. Initial reaction rates were determined with a Molecular Devices SpectraMax Plus plate reader over a 10 minute time course. IC50 and Ki data were calculated from non-linear regression analysis using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego Calif., USA).

In general, compounds of the invention are effective for the inhibition of ITK. Preferred compounds showed Ki below 0.1 μM in the coupled enzyme assay (I-70, I-76, I-78, I-79, I-80). Preferred compounds showed Ki between 0.1 μM and 1 μM in the coupled enzyme assay (I-5, I-10, I-11, I-69, I-82, I-83, I-84, II-4, II-7, II-41).

BTK Inhibition Assay

Compounds were screened for their ability to inhibit Btk using a radioactive-phosphate incorporation assay at Vertex Pharmaceuticals. Assays were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 mM MgCl2, 0.1% BSA and 1 mM DTT. Final substrate concentrations in the assay were 50 μM [γ-33P]ATP (200 mCi 33P ATP/mmol ATP, Amersham Pharmacia Biotech, Amersham, UK/Sigma Chemicals) and 2 μM peptide (SAM68 D332-443). Assays were carried out at 25° C. and in the presence of 25 nM Btk. An assay stock buffer solution was prepared containing all of the reagents listed above, with the exception of the peptide and the test compound of interest. 75 μL of the stock solution was placed in a 96 well plate followed by addition of 2 μL of DMSO stock containing serial dilutions of the test compound (typically starting from a final concentration of 15 μM) in duplicate (final DMSO concentration 2%). The plate was preincubated for 15 minutes at 25° C. and the reaction initiated by addition of 25 μL peptide (final concentration 2 μM). Background counts were determined by the addition of 100 mL 0.2M phosphoric acid+0.01% TWEEN to control wells containing assay stock buffer and DMSO prior to initiation with peptide.

The reaction was stopped after 10 minutes by the addition of 100 mL 0.2M phosphoric acid+0.01% TWEEN. A multiscreen phosphocellulose filter 96-well plate (Millipore, Cat no. MAPHN0B50) was pretreated with 100 μL 0.2M phosphoric acid+0.01% TWEEN 20 prior to the addition of 170 mL of the stopped assay mixture. The plate was washed with 4×200 μL 0.2M phosphoric acid+0.01% TWEEN 20. After drying, 30 μL Optiphase ‘SuperMix’ liquid scintillation cocktail (Perkin Elmer) was added to the well prior to scintillation counting (1450 Microbeta Liquid Scintillation Counter, Wallac).

After removing mean background values for all of the data points, Ki(app) data were calculated from non-linear regression analysis using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego Calif., USA).

In general, compounds of the invention, including compounds in Table 1, are effective for the inhibition of Btk. Preferred compounds showed Ki above 0.5 μM in the radioactive incorporation assay (II-43, II-61, II-114, II-149). Preferred compounds showed Ki below 0.5 μM in the radioactive incorporation assay (II-51, II-58, II-61, II-63, II-77, II-78, II-80, II-112).

Compounds were screened for their ability to inhibit Btk using an AlphaScreen™ phosphotyrosine assay at Vertex Pharmaceuticals. Assays were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 mM MgCl2, 0.1% BSA and 1 mM DTT. Final substrate concentrations in the assay were 50 μM ATP (Sigma Chemicals) and 2 μM peptide (Biotinylated SAM68 D332-443). Assays were carried out at 25° C. and in the presence of 25 nM Btk. An assay stock buffer solution was prepared containing all of the reagents listed above, with the exception of peptide and the test compound of interest. 37.5 μL of the stock solution was placed in each well of a 96 well plate followed by 1 μL of DMSO containing serial dilutions of the test compound (typically starting from a final concentration of 15 μM) in duplicate (final DMSO concentration 2%). The plate was preincubated for 15 minutes at 25° C. and the reaction initiated by addition of 12.5 μL peptide (final concentration 2 μM). Background counts were determined by the addition of 5 μL 500 mM EDTA to control wells containing assay stock buffer and DMSO prior to initiation with Biotin-SAM68.

The reaction was stopped after 30 minutes by diluting the reaction 225-fold into MOPS buffer (20 mM MOPS (pH 7.0), 1 mM DTT, 10 mM MgCl2, 0.1% BSA) containing 50 mM EDTA to bring the final concentration of peptide to 9 nM.

AlphaScreen™ reagents were prepared according to the manufacturers instructions (AlphaScreen™ phosphotyrosine (P-Tyr-100) assay kit, PerkinElmer catalogue number 6760620C). Under subdued lighting, 20 μL of AlphaScreen™ reagents were placed in each well of a white half area 96 well plate (Corning Inc.—COSTAR 3693) with 30 μL of the stopped, diluted kinase reactions. Plates were incubated in the dark for 60 minutes prior to reading on a Fusion Alpha plate reader (PerkinElmer).

After removing mean background values for all of the data points, Ki(app) data were calculated from non-linear regression analysis using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego Calif., USA).

RLK Inhibition Assay

Compounds were screened for their ability to inhibit Rlk using a standard coupled enzyme assay (Fox et al.,Protein Sci., (1998) 7, 2249). Assays were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 mM MgCl2, 0.1% BSA and 1 mM DTT. Final substrate concentrations in the assay were 100 μM ATP (Sigma Chemicals) and 10 μM peptide (Poly Glu:Tyr 4:1). Assays were carried out at 30° C. and in the presence of 40 nM Rlk. Final concentrations of the components of the coupled enzyme system were 2.5 mM phosphoenolpyruvate, 300 μM NADH, 30 μg/ml pyruvate kinase and 10 μg/ml lactate dehydrogenase.

An assay stock buffer solution was prepared containing all of the reagents listed above, with the exception of ATP and the test compound of interest. 60 μl of the stock solution was placed in a 96 well plate followed by addition of 2 μl of DMSO stock containing serial dilutions of the test compound (typically starting from a final concentration of 7.5 μM). The plate was preincubated for 10 minutes at 30° C. and the reaction initiated by addition of 5 μl of ATP. Initial reaction rates were determined with a Molecular Devices SpectraMax Plus plate reader over a 10 minute time course. IC50 and Ki data were calculated from non-linear regression analysis using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego Calif., USA).