ES2457754T3 - Azaindoles useful as protein kinase inhibitors - Google Patents

Abstract

A compound of formula (I): **Formula** or a pharmaceutically acceptable salt thereof; wherein: Ring A is an optionally substituted five membered ring selected from: **Formula** or **Formula** x is 0, 1, or 2; each occurrence of R1 is halogen, CN, NO2 or UmR; R2 is independently selected from Tn-R' R3 is independently halogen -CN, NO2 or VpR'; R4 is VpR', where p is 0 and R' is a six membered fully unsaturated monocyclic ring with 0-1 nitrogen atoms; R5 is independently halogen -CN, NO2 or VpR'; each occurrence of T, U or V is independently a C1-6 alkylidene chain, where the C1-6 alkylidene chain is optionally substituted with a C1-6 aliphatic group or -OH, and up to two units of methylene of the C1-6 alkylidene chain are optionally and independently substituted with -NR-, -S-, -O-, -CS-, -CO2-, -OCO-, -CO-, -COCO-, -CONR- , -NRCO-, -NRCO2-, -SO2NR-, -NRSO2-, -CONRNR-, -NRCONR-, -OCONR-, -NRNR-, NRSO2NR-, -SO-, or -SO2-; m, n and p are each independently 0 or 1; each occurrence of R is independently hydrogen or a C1-6 aliphatic group optionally substituted with a C1-3 aliphatic group; and each occurrence of R' is independently hydrogen, a C1-6 aliphatic 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 a saturated, partially unsaturated, or fully unsaturated 8-12 membered bicyclic ring system having 0-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein the 3- 8 membered and the 8-12 membered bicyclic ring are optionally substituted with halogen, -O(C1-6 aliphatic) or C(O)NH2.

Description

Azaindoles useful as protein kinase inhibitors

Technical Field of the Invention

The present invention relates to compounds useful as protein kinase inhibitors. 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 methods for preparing the compounds of the invention.

Background of the invention

The search for new therapeutic agents has helped considerably in recent years through a better understanding of the structure of enzymes and other biomolecules associated with diseases. An important class of enzymes that have been the subject of extensive study is that of protein kinases.

Protein kinases constitute a large family of structurally related enzymes that are responsible for controlling a wide 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, CA: 1995). It is thought that protein kinases have evolved from a common ancestral gene due to the conservation of their structure and catalytic function. Almost all kinases contain a conserved catalytic domain of 250-300 amino acids. Kinases can be classified in families by the phosphorylating substrates (eg, tyrosine protein, serine / threonine protein, lipids etc.). Sequence motifs that generally correspond to each of these kinase families have been identified (see, e.g., Hanks, SK, Hunter, T., FASEB J. 1995, 9, 576 596; Knighton et al., Science 1991, 253, 407-414; Hiles et al., Cell 1992, 70, 419-429; Kunz et al., Cell 1993, 73, 585-596; Garcia-Bustos et al., EMBO J. 1994, 13, 2352-2361).

In general, protein kinases participate in intracellular signaling by transferring phosphoryl 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 biological function of the target protein. These phosphorylation events are triggered, ultimately, in response to several extracellular and other stimuli. Examples of these stimuli include various environmental and chemical stress signals (e.g., oxidative stress, thermal 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., macrophage granulocyte colony stimulating factor (GM-CSF) and fibroblast growth factor (FGF)). An extracellular stimulus can affect one or more cellular responses related to growth, migration and cell differentiation, hormone secretion, activation of transcription factors, muscle contraction, glucose metabolism, control of protein synthesis and cell cycle regulation .

Many diseases are associated with abnormal cellular responses triggered by the events mediated by protein kinases, as described above. These diseases include, among others, 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, considerable efforts have been made in medical chemistry to identify 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 TCR, BCR and Fcε receptors (reviewed in Miller A, et al. Current Opinion in Immunology 14; 331-340 (2002 ) The Tec family of kinases is essential for the activation of T lymphocytes. Three members of the Tec family, Itk, Rlk and Tec, are activated downstream of antigen receptor binding in T lymphocytes and transmit signals to effectors. downstream, including PLC-γ Itk deletion in mice results in a reduction of TCR-induced proliferation of T-lymphocyte receptors and the secretion of 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. Pulmonary inflammation, eosinophil infiltration and mucus production is dramatically reduced in Itk - / - mice in response to exposure to the OVA allergen (Mueller et al, Journal of Immunology 170: 5056-5063 (2003)). LTK has also been implicated in atopic dermatitis. It has been reported that this gene is expressed more in peripheral blood T lymphocytes of patients with moderate and / or severe atopic dermatitis than in controls or patients with atopic dermatitis (Matsumoto et al, International archives of Allergy and Immunology 129; 327 - 340 (2002)).

Splenocytes from TRlk - / - mice secrete half of the IL-2 produced by wild animals in response to TCR binding (Schaeffer et al, Science 284; 638-641 (1999)), while the combined deletion of Itk and Rlk in Rlk mice leads to a profound inhibition of TCR-induced responses, including the proliferation and production of 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 after TCR binding is performed on T lymphocytes deficient in Itk / Rlk; inositol triphosphate production, calcium mobilization, activation of

5 MAP kinase and the activation of NFAT and AP-1 transcription factors are all reduced (Schaeffer et al, Science 284; 638-641 (1999), Schaeffer et al Nature Immunology 2.12; 1183-1188 (2001) ).

The Tec family of kinases is also essential for the development and activation of B lymphocytes. Patients with Btk mutations have a deep blockage of B lymphocyte development, resulting in an almost complete absence of B lymphocytes and plasma cells, intensely reduced levels of Ig and a profound inhibition of the humoral response to memory antigens (reviewed in Vihinen et al Frontiers in Bioscience 5: d917-928). Btk-deficient mice also have a reduced number of peripheral B lymphocytes and considerably decreased levels of IgM and IgG3. Btk deletion in mice has a profound effect on the proliferation of anti-IgM-induced B lymphocytes and inhibits immune responses to

15 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 activated by crosslinking of FcεRI (Kawakami et al, Journal of Immunology; 3556-3562 (1995)). Btk-deficient murine mast cells have reduced degranulation and decreased production of proinflammatory cytokines after FcεRI cross-linking (Kawakami et al. Journal of leukocyte biology 65: 286-290). Btk deficiency also results in a decrease in macrophage effector functions (Mukhopadhyay et al, Journal of Immunology; 168, 2914-2921 (2002))

Janus kinases (JAK) are a family of tyrosine kinases consisting of JAK1, JAK2, JAK3 and TYK2. The

25 JAK play a critical role in cytokine signaling. Current substrates under the JAK family of kinases include the signal transducer and the protein transcription activator (STAT), JAK / STAT signaling has been implicated in the mediation of many abnormal immune responses such as allergies, asthma, autoimmune diseases such as transplant rejection, rheumatoid arthritis, amyotrophic lateral sclerosis and multiple sclerosis, as well as in solid and hematological malignancies, such as leukemia and lymphomas. The pharmaceutical intervention in the JAK / STAT route has been reviewed [Frank Mol. Med. 5, 432 456 (1999) & Seidel, et al, Oncogene 19, 2645-2656 (2000)].

JAK1, JAK2 and TYK2 are expressed ubiquitously, while JAK3 is predominantly expressed in hematopoietic cells. JAK3 binds exclusively to the gamma chain (γc) of the common cytokine receptor and is

35 activated by IL-2, IL-4, IL-7, IL-9 and IL-15. In fact, it has been shown that proliferation and survival of murine mast cells induced by IL-4 and IL-9 depends on JAK3-and γc- signaling [Suzuki et al, Blood 96. 2172-2808 (2000)].

Cross-linking of high affinity immunoglobulin (Ig) E receptors of sensitized mast cells leads to a release and proinflammatory mediators, including a series of vasoactive cytokines that results in acute or immediate allergic hypersensitivity reactions (type 1) [Gordon et al, Nature 346, 274-276 (1990) & Galli, N. Engl. J. Med., 328, 257-265 (1993)]. A crucial role has been established for JAK3 in mast cell responses mediated by IgE receptor in vitro and in vivo [Malaviya, et al, Biochem. Biophys Res. Commun. 257, 807-813 (1999)]. In addition, the prevention of the reactions of

Type 1 hypersensitivity, including anaphylaxis, mediated by mast cell activation through inhibition of JAK2 [Malaviya et al, J. Biol. Chem. 274.27028-27038 (1999)]. Directing JAK3 inhibitors to mast cells modulated mast cell degranulation in vitro and prevented anaphylactic reactions mediated by IgE antigen / receptor in vivo.

A recent study described the success of targeting JAK3 for immune suppression and allograft acceptance. The study demonstrated a dose-dependent survival of a buffalo heart allograft in Wistar Furth receptors after administration of JAK3 inhibitors, indicating the possibility of regulating unwanted immune responses in graft disease against the host [Kirken , Transpl. Proc. 33, 3268-3270 (2001)].

55 The phosphorylation of STAT mediated by IL-4 has been implicated as a participating mechanism in the early and late stages of rheumatoid arthritis (RA). The regulation by increase of proinflammatory cytokines in the synovium and synovial fluid of RA is a characteristic of the disease. It has been shown that IL-4 mediated activation of the IL-4 / STAT pathway is mediated by Janus kinases (JAK 1 and 3) and that JAK kinases associated with IL-4 are expressed in the synovium of RA. [Muller-Ladner, et al, J. Immunol. 164, 3894-3901 (2000)].

Familial amyotrophic lateral sclerosis (ELAF) is a fatal degenerative disorder that affects approximately 10% of ALS patients. Survival rates of mice with ELAF were increased after treatment with a specific JAK3 inhibitor. This suggested that JAK3 plays a role in ELAF [Trieu, et al,

65 Biochem. Biophys Res. Commun. 267, 22-25 (2000)].

The signal transducer and transcription activator (STAT) proteins are activated by, among others, the JAK family kinases. The results of a recent study suggested the possibility of intervening in the JAK / STAT signaling pathway by targeting JAK family kinases specific for the treatment of leukemia [Sudbeck, et al, Clin. Cancer Res. 5, 1569-1582 (1999)]. It was shown that specific compounds

5 of JAK3 inhibit the clonogenic growth of cell lines expressing JAK-3 DAUDI, RAMOS, LC1; 19, NALM-6, MOLT-3 and HL-60.

In animal models, TEL / JAK2 fusion proteins have induced myeloproliferative disorders and, in hematopoietic cell lines, the introduction of TEL / JAK2 resulted in the activation of STAT1, STAT3, 10 STAT5 and independent cytokine growth [Schwaller , et al, EMBO J. 17, 5321-5333 (1998)].

Inhibition of JAK 3 and TYK 2 nullify tyrosine phosphorylation of STAT3 and inhibited fungoid mycosis cell growth, a form of cutaneous T lymphocyte lymphoma. These results involved the family of JAK kinases in the constitutively activated JAK / STAT pathway that is present in fungal fungal infections [Nielsen,

15 et al, Proc. Nat. Acad. Sci. U.S.A. 94, 6764-6769

(1997)]. Similarly, it was shown that STAT3, STAT5, JAK1 and JAK2 were constitutively activated in mouse T lymphocyte lymphoma initially characterized by overexpression of LCK, so that it is also implied that the JAK / STAT pathway in cell growth abnormal [Yu, et al, J. Immunol. 159, 5206-5210 (1997)].

20 In addition, activation of STAT3 mediated by IL-6 was blocked by a JAK inhibitor, which leads to sensitization of myeloma cells to apoptosis [Catlett-Falcone, et al, Immunity 10,105-115 (1999)] . WO 2004/016609 describes substituted pyrrolipyridines that are considered to have an activity as inhibitors of ltk and, thus, are useful as pharmaceuticals, in particular for the treatment of allergic rhinitis and asthma. WO 00/43393 A1 describes compounds that will inhibit tyrosine enzymes

25 kinases, which are considered useful in the treatment of tyrosine kinase dependent diseases, such as angiogenesis, cancer, atherosclerosis and diabetic retinopathy.

Accordingly, there is a great need to develop useful compounds as protein kinase inhibitors. In particular, it would be desirable to develop compounds that are useful as inhibitors of the Tec family (e.g. 30, Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rlk) and the JAK family kinase proteins, in particular given the inadequate treatments currently available for most of the disorders involved in its activation.

Summary of the invention

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

These compounds and pharmaceutically acceptable compositions thereof are useful for treating or preventing various diseases, disorders or conditions, including, among others, autoimmune, inflammatory, proliferative or hyperproliferative diseases or a disease mediated by the immune system. The compositions are also useful in methods to prevent platelet aggregation induced by

45 thrombin The compounds provided by the present invention are also useful for the study of kinases in biological and pathological phenomena; the study of intracellular signal transduction pathways mediated by said kinases and the comparative evaluation of new kinase inhibitors.

Detailed description of the invention

1. General description of the compounds of the invention:

The present invention relates to a compound of formula I:

or a pharmaceutically acceptable salt thereof; in which: Ring A is an optionally substituted five-member ring selected from:

or

x is 0, 1, or 2;

Each occurrence of R1 is halogen, CN, NO2 or UmR; R2 is independently selected from Tn-R '; R3 is independently halogen -CN, NO2 or VpR '; R4 is VpR ', where p is 0 and R' is a completely unsaturated six-membered monocyclic ring with 0 -1 nitrogen atoms;

R5 is independently halogen -CN, NO2 or VpR '; each occurrence of T, U or V is, independently, a C1-6 alkylidene chain, in which the C1-6 alkylidene chain is optionally substituted with a C1-6 or -OH aliphatic group, and up to two units Methylene of the C1-6 alkylidene chain are optionally and independently substituted by -NR-, -S-, -O-, -CS-, -CO2-, -OCO-, -CO-, -COCO-, -CONR -, -NRCO-, -NRCO2-, -SO2NR-, -NRSO2-, -CONRNR-, -NRCONR-,

OCONR-, -NRNR-, NRSO2NR-, -SO-, or -SO2-; m, n and p are each independently 0 or 1; each occurrence of R is independently hydrogen or a C1-6 aliphatic group optionally substituted with a C1-6 aliphatic group; and each occurrence of R is, independently, hydrogen, a C1-6 aliphatic group, a 3-8 membered, saturated, partially unsaturated or completely unsaturated monocyclic ring having

30 0 -3 heteroatoms independently selected from nitrogen, oxygen or sulfur, or an 8-12 membered bicyclic ring system, saturated, partially unsaturated or completely unsaturated having 0 -5 heteroatoms independently selected from nitrogen, oxygen or sulfur, wherein the 3-8 membered monocyclic ring and the 8-12 membered bicyclic ring are optionally substituted with halogen, -O (C1-6 aliphatic) or C (O) NH2;

In another embodiment, the compounds of the present invention do not include the compounds listed in claim 9 on pages 152-166 of W02004 / 078756 A2, which is incorporated herein by reference.

40 2. Compounds and Definitions:

The compounds of the present invention include those generally described above and are further illustrated by the classes, subclasses and species disclosed herein. As used herein, the following definitions will apply unless otherwise indicated. For the purposes of the present invention, chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version,

Handbook of Chemistry and Physics, 75th Ed. In addition, the general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito 1999, and "March's Advanced Organic Chemistry", 5th Ed., Ed .: Smith, MB and March, J., John Wiley & Sons, New York: 2001, all of whose contents are incorporated herein by reference.

As described herein, the compounds of the invention may be optionally substituted with one or more substituents, as generally illustrated above, or as exemplified by specific classes, subclasses and species of the invention. It will be appreciated that the term "optionally substituted" is used interchangeably with the phrase "substituted or unsubstituted." In general, the term "substituted", whether or not preceded by the term "optionally", refers to radical substitution. of hydrogen in a given structure with the radical of a specified substituent. Unless otherwise indicated, an optionally substituted group may have a substituent in 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 the same or different in each position. The substituent combinations provided by the present invention are preferably those that result in the formation of stable or chemically viable compounds. The term "stable", as used herein, refers to compounds that are not substantially altered when subjected to conditions that allow their production, detection and, preferably, their recovery, purification and use for one or more of the purposes disclosed in this document. In some embodiments, a stable compound or chemically viable compound is one that is not substantially altered when stored at a temperature of 40 ° C or less, in the absence of moisture or other chemically reactive conditions, for at least one week.

As described herein, a specified number of atoms includes any integer therein. For example, a group that has 1-4 atoms could have 1, 2, 3 or 4 atoms.

The term "aliphatic" or "aliphatic group", as used herein, means a linear (ie, unbranched) or branched chain, substituted or unsubstituted hydrocarbon chain that is fully saturated or that contains one or more units. of unsaturation or a monocyclic hydrocarbon or bicyclic hydrocarbon that is fully saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as "cycloaliphatic" or "cycloalkyl" carbocycle) having a single point binding 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, groups aliphatic contain 1-8 aliphatic carbon atoms In other embodiments, aliphatic groups contain 1-6 car atoms aliphatic bonds and in other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In some embodiments, "cycloaliphatic" (or "carbocycle" or "cycloalkyl") refers to a monocyclic C3-C8 hydrocarbon or bicyclic C8-C12 hydrocarbon that is completely saturated or that contains one or more unsaturation units but is not aromatic , which has a single point of attachment to the rest of the molecule, in which any single ring in said bicyclic ring system has 3-7 members. Suitable aliphatic groups include, among others, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and hybrids thereof such as (cycloalkyl) alkyl, (cycloalkenyl) alkyl

or (cycloalkyl) alkenyl.

The term "heteroaliphatic," as used herein, means aliphatic groups in which one or two carbon atoms are independently substituted for one or more of oxygen, sulfur, nitrogen, phosphorus or silicon. Heteroaliphatic groups can be substituted or unsubstituted, branched or unbranched, cyclic groups

or acyclics, and include "heterocycle", "heterocyclyl", "heterocycloaliphatic" or "heterocyclic".

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 a heteroatom. independently selected. 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. Suitable heterocycles include, among others, 3 -1 H -benzimidazol-2-one, 3- (1-alkyl) -benzimidazol-2-one, 2-tetrahydrofuranyl, 3 tetrahydrofuranyl, 2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl, 2-morpholino, 3 -morpholino, 4-morpholino, 2-thiomorpholino, 3-thiomorpholino, 4-thiomorpholino, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 1-tetrahydropyrazinyl, 2-tetrahydropyrazinyl, 3-tetrahydropyrazinyl, 1-piperidinyl, 2-piperidinyl, 2-piperidinyl , 1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazolidinyl, 2-imidazolidinyl, 2-imidazolidinyl, 2-imidazolidinyl , 4-imidazolidinyl, 5-imidazolidinyl, indolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolane, benzoditian and 1,3-dihydro-imidazol-2-one.

The term "heteroatom" means one or more of oxygen, sulfur, nitrogen, phosphorus or silicon (including any oxidized form of nitrogen, sulfur, phosphorus or silicon; the quaternized form of any basic nitrogen or a substitutable nitrogen of a heterocyclic ring, by Example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR + (as in N-substituted pyrrolidinyl)).

The term "unsaturated," as used herein, means that a remainder has one or more units of unsaturation.

The term "alkoxy," or "thioalkyl," as used herein, refers to an alkyl group as defined above, attached to the main carbon chain through an oxygen atom ("alkoxy" ) or sulfur ("thioalkyl").

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 , in which at least one ring in the system is aromatic and in which each ring in the system contains 3 to 7 ring members. The term "aryl" can be used interchangeably with the term "aryl ring".

The term "heteroaryl" used alone or as part of a larger moiety as in "heteroaralkyl", "heteroarylalkoxy" refers to monocyclic, bicyclic and tricyclic ring systems having a total of five to fourteen ring members, in which at least one ring in the system is aromatic, at least one ring in the system contains one or more heteroatoms and in which each ring in the system contains 3 to 7 ring members. The term "heteroaryl" can be used interchangeably with the term "heteroaryl ring" or the term "heteroaromatic." Suitable heteroaryl rings include, but are not limited to, 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4 -imidazolyl, 5imidazolyl, benzimidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl , 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (e.g., 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g., 5-tetrazolyl) , triazolyl (e.g., 2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, benzofuryl, benzothiophenyl, indolyl e.g., 2-indolyl), pyrazolyl (e.g., 2-pyrazolyl ), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,3,4- thiadiazolyl, 1,2,5thiadiazolyl, purinyl, pyrazinyl, 1,3,5-triazinyl, quinolinyl (e.g., 2-q uolinolinyl, 3-quinolinyl, 4-quinolinyl) and isoquinolinyl (e.g. eg, 1-isoquinolinyl, 3-isoquinolinyl or 4-isoquinolinyl).

An aryl group (including aralkyl, aralkoxy, aryloxyalkyl and the like) or heteroaryl (including heteroaralkyl and heteroarylalkoxy and the like) may contain one or more substituents. Suitable substituents on the unsaturated carbon atom of an aryl or heteroaryl group are selected from halogen, -R °; -OR °; -SR °; 1,2-methylenedioxy; 1,2-ethylenedioxy; phenyl (Ph) optionally substituted with R °; = O (Ph) optionally substituted with R °; - (CH2) 1 -2 (Ph), optionally substituted with R °; -CH = CH (Ph), optionally substituted with R °; -NO2; -CN; -N (R °) 2; -NR ° C (O) R °; -NR ° C (S) R °; -NR ° C (O) N (R °) 2; -NR ° C (S) N (R °) 2; -NR ° CO2R °; -NR ° NR ° C (O) R °; -NR ° NR ° C (O) N (R °) 2; -NR ° NR ° CO2R °; -C (O) C (O) R °; -C (O) CH2C (O) R °; -CO2R °; -C (O) R °; -C (S) R °; -C (O) N (R °) 2; -C (S) N (R °) 2; -OC (O) N (R °) 2; OC (O) R °; -C (O) N (OR °) R °; -C (NOR °) R °; -S (O) 2R °; -S (O) 3R °; -SO2N (R °) 2; -S (O) R °; -NR ° SO2N (R °) 2; -NR ° SO2R °; -N (OR °) R °; -C (= NH) -N (R °) 2; or - (CH2) 0 -2NHC (O) R ° in which each independent occurrence of R ° is selected from hydrogen, an aliphatic C1-6 group, an unsubstituted 5-6 membered heterocyclic or heteroaryl ring, phenyl, -O (Ph), or CH2 (Ph), or, despite the above definition, two independent occurrences of R °, in the same substituent or in different substituents, together with the atom (s) to which each R ° group is attached , they 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. Optional substituents in the aliphatic group of R ° are selected from NH2, NH (C1-4 aliphatic), N (C1-4 aliphatic), halogen, C1-4 aliphatic, OH, O (C1-4 aliphatic), NO2, CN, CO2H, CO2 (C1-4 aliphatic), O (C1-4 haloaliphatic), or C1-4 haloaliphatic, in which each of the above C1-4 aliphatic groups of R ° is not substituted.

An aliphatic or heteroaliphatic group or a non-aromatic heterocyclic ring may contain one or more substituents. Suitable substituents 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), or = NR *, in which each R * is independently selected of hydrogen or an optionally substituted C1-6 aliphatic. Optional substituents in the aliphatic group of R * are selected from NH2, NH (C1-4 aliphatic), N (C1-4 aliphatic), halogen, C1-4 aliphatic, OH, O (C1-4 aliphatic), NO2, CN, CO2H, CO2 (C1-4 aliphatic), O (C1-4 haloaliphatic), or halo (C1-4 aliphatic), in which each of the above C1 - 4 aliphatic groups of R * is not substituted.

Optional nitrogen substituents of a non-aromatic heterocyclic 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, or -NR + SO2R +; wherein R + is hydrogen, an optionally substituted C1-6 aliphatic, optionally substituted phenyl, -O (Ph) optionally substituted, -CH2 (Ph) optionally substituted, - (CH2) 1 -2 (Ph) optionally substituted; -CH = CH (Ph) optionally substituted; or an unsubstituted 5-6 membered heteroaryl or heterocyclic ring having one to four heteroatoms independently selected from oxygen, nitrogen or sulfur or, despite the above definition, two independent occurrences of R +, in the same substituent or in different substituents, together with the one or more atoms to which each R + group is attached, they form a 5-8 membered heterocyclyl, aryl or heteroaryl ring or a 3-8 membered cycloalkyl ring having 0 -3 heteroatoms selected in a way independent of nitrogen, oxygen or sulfur. Optional substituents in the aliphatic group or the R + phenyl ring are selected from NH2, NH (C1-4 aliphatic), N (C1-4 aliphatic), halogen, C1-4 aliphatic, OH, O (C1-4 aliphatic) , NO2, CN, CO2H, CO2 (C1-4 aliphatic), O (C1-4 haloaliphatic), or halo (C1-4 aliphatic), in which each of the aliphatic groups

5 C1 - 4 above R + is not substituted.

The term "alkylidene chain" refers to a linear or branched carbon chain that can be completely saturated or has one or more units of establishment and has two points of attachment to the rest of the molecule, in which one or more units of Methylene can be substituted, optionally or independently, with a group, including, among others, CO, CO2, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO2, NRCONR, SO, SO2, NRSO2, SO2NR, NRSO2NR, O, S ; or NR.

The term "protecting group", as used herein, refers to an agent used to temporarily block one or more desired reactive sites in a multifunctional compound. In certain embodiments, a protecting group has one or more, preferably all, of the following characteristics: a) reacts selectively with a good yield to give a protected substrate that is stable to the reactions that occur in one or more of the other reactive sites; and b) it can be selectively removed in good yield by reagents that do not attack the regenerated functional group. Examples of protective groups are detailed in Greene, TW, Wuts, P. G in "Protective Groups in Organic Synthesis", Third Edition, John Wiley & Sons, New York: 1999, all of whose contents are incorporated herein by reference. The term "nitrogen protecting group", as used herein, refers to an agent used to temporarily block one or more desired reactive nitrogen sites in a multifunctional compound. Preferred nitrogen protecting groups also possess the above example characteristics and certain example nitrogen protecting groups are also detailed in Chapter 7 in

25 Greene, T.W., Wuts, P. G in "Protective Groups in Organic Synthesis", Third Edition, John Wiley & Sons, New York: 1999, all of whose contents are incorporated herein by reference.

As detailed above, in some embodiments two independent occurrences of R ° (or R +, or any other variable defined similarly herein) are taken together with the atom (s) to which each variable is attached 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. Examples of rings that are formed when two independent occurrences of R ° (or R +, or any other variable defined similarly herein) are taken together with the atom (s) to which each variable is attached include, among others, the following: a) two independent appearances of R ° (or R +, or any

35 another variable similarly defined herein) that are attached to the same atom and are taken together with said atom to form a ring, for example N (R °) 2, when both occurrences of R ° are taken together with the nitrogen atom to form a piperidin-1-yl, piperazin-1-yl or morflin-4-lo group; and b) two independent occurrences of R ° (or R +, or any other variable similarly defined herein) that are attached to different atoms and are taken together with said atoms to form a ring, for example in which a group phenyl is substituted with two occurrences of OR °

these two occurrences of R ° are taken together with the oxygen atoms to which they are attached to form a ring 45 containing fused 6-member oxygen.

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

Unless otherwise indicated, the structures represented herein are also intended to include all isomeric forms (e.g., enantiomeric, diastereomeric and geometric (or conformational) forms of the structure, for example the R and R configurations). S for each asymmetric center, the isomers

(Z) and (E) double bond and conformational isomers (Z) and (E). Thus, the unique stereochemical isomers, as well as the enantiomeric, diatestereomeric and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. Unless otherwise indicated, all these tautomeric forms of the compounds of the invention fall within the scope of the invention. Additionally, unless otherwise indicated, with the structures represented herein, it is also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, the compounds having the present structures, except for the replacement of hydrogen by deuterium or tritium, or the substitution of a carbon or carbon 13C or 14C are within the scope of the present invention. Such compounds are useful, for example, as analytical tools or probes in biological assays.

3. Description of example compounds

All descriptions of embodiments herein may be applied to compounds of formula I, II, III, IV, V, and VI.

In certain embodiments of the present invention, R5 is independently Vp-R '.

In other embodiments, one of R3 and R5 is Vp-R ', in which R' is a fully unsaturated (i.e. aromatic) monocyclic ring of 5 or 6 members having 0 -3 heteroatoms independently selected from

15 nitrogen, oxygen or sulfur, or a completely unsaturated 9 or 10-membered bicyclic ring system that has 0-5 heteroatoms independently selected from nitrogen, oxygen or sulfur.

In other embodiments, one of R3 and R5 is Vp-R ', in which R' is independently a C1-6 aliphatic group, a 3-6 membered monocyclic ring, saturated, partially unsaturated or completely unsaturated selected independently of nitrogen, oxygen or sulfur, or a saturated, partially unsaturated or completely unsaturated bicyclic ring system of 8 or 12 members having 0 -5 heteroatoms independently selected from nitrogen, oxygen or sulfur.

In other embodiments, R4 is VpR ', and R' is a completely unsaturated six-membered monocyclic ring with 0 25 -1 nitrogen heteroatoms;

In other embodiments, p is 0.

In yet other embodiments, p is 1.

In other embodiments, V is -NR-, -S-, or -O-.

In other embodiments, R3 is Vp-R ', where p is 0 and R' is hydrogen.

In other embodiments, R5 is halogen or Vp-R ', where p is 0 and R' is hydrogen or C1-6 aliphatic. In yet other embodiments, this C1-6 aliphatic is C1-3 alkyl.

Ring A is:

In certain embodiments, R1 is UmR. In other embodiments, R1 is UmR, where m is 0 and R is H or CH3.

In certain embodiments, R2 is TnR ', in which n is 1.1. In other embodiments, R2 is TnR ', in which n is 0.

In certain embodiments, T is -NR-, -O-, -CO-, -CONR-, or -NRCO- ..

In certain embodiments, T is -NR-. In certain embodiments, T is -O-. In certain of these embodiments, R 'is C1-6 aliphatic. In other of these embodiments, both R and R 'are H.

In certain embodiments, NR-and R 'is C1-6 aliphatic. In certain of these embodiments, R is C1-6 aliphatic. In some embodiments, both R and R 'are C1-6 alkyl.

In certain embodiments, T is a C1-6 alkylidene chain in which the alkylidene chain is attached to Ring A through a methylene unit. In some of these embodiments, T is - (C1-5 alkyl) NR-., In some embodiments, T is -CH2NR-. In some of these embodiments, R 'is C1-6 aliphatic.

In other embodiments, T is a C1-6 alkylidene chain in which 0 methylene units are substituted with the groups disclosed herein.

In yet other embodiments, R2 is an optionally substituted 5-7 membered N-linked heterocyclyl. In certain embodiments, said N-linked heterocyclyls are selected from morpholinyl, piperidinyl, pyrrolidinyl, and piperazinyl. In certain embodiments, said N-linked heterocyclyls are optionally and independently substituted with 0-4 occurrences of amino, alkylamino, dialkylamino or C1-6 alkyl.

As generally described above, another compound of the present invention has the formula II:

or a pharmaceutically acceptable salt thereof. In other embodiments, a compound of the present invention has the formula III:

or a pharmaceutically acceptable salt thereof. In yet other embodiments, a compound has the formula IV:

or a pharmaceutically acceptable salt thereof. In yet other embodiments, a compound has the formula V:

or a pharmaceutically acceptable salt thereof. In yet other embodiments, a compound has the formula VI:

or a pharmaceutically acceptable salt thereof.

It will be appreciated that for the compounds of formulas II-VI, the variables in the compounds of formulas II-VI are as defined in any of the embodiments herein.

As generally described above, preferred substituents and variables (e.g., R ’groups) are 15 as illustrated in the compounds represented in Table 1.

Accordingly, representative examples of the compounds of formula I are shown below in Tables 1 and 2.

20 Table 1. Examples of Compounds of Formula I:

Table 2. Examples of Compounds of Formula I:

5 4. General synthesis methodology:

The compounds of the present invention can be prepared, in general, by methods known from

those skilled in the art for analogous compounds, as illustrated in the following general scheme and the

following preparative examples. 10

The following abbreviations are used:

EtOH is ethanol

TA is room temperature 15 Ts is Tosilo

Ph is phenyl

DME is dimethyl ether

Bu is butyl

EDC is 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide 20 DMF is dimethylformamide

D / N is overnight

Et2O is ether

CDI is N, N'-Carbonyldiimidazole

CLEM is liquid chromatography with 25 P mass spectroscopy is a suitable protecting group

Scheme I

Reagents and conditions: (a) AlCl3, CH2Cl2, TA, 16 hours; (b) EtOH, microwave irradiation, 120 ° C, 10 min.

Scheme I above shows a general synthetic route that is used to prepare the compounds 5 of the present invention when from R1 to R5 are as described herein. Intermediates 3 are prepared

35 using Friedel-Craft acylation methods that are well known in the art. This reaction can be treated with various substituted chloroacetyl chlorides to form compounds of formula 3. Finally, the compound of formula 5 is obtained by cyclization of intermediate 3 according to step (b). The reaction can be treated with various of the substituted thioamides of formula 4.

Scheme II

Reagents and conditions: (a) Br2, CHCl3, 0 ° C at RT; (b) "BuLi, THF, TsCl; (c) PdCl2 (dppf) 2, dioxane, KOAc, 5 bis (pinacolato) diboro, 18 hours; (d) Pd (PPh3) 4, Na2CO3, DME, EtOH / H2O, microwave irradiation, 120 ° C, 2 hours; (e) 3N NaOH, MeOH.

Scheme II above shows a general synthetic route that is used to prepare the compounds 11 of the present invention when A, from R1 to R5 are as described herein. Intermediate 7 is prepared by sprouting the compound of structure 1 followed by the subsequent protection of intermediate 6 with a tosyl group. The boronic esters 8 are formed according to Scheme II, step (c). The formation of biaryl bond derivatives 10 is achieved by treating bromide 9 with boronic ester derivatives 8 in the presence of palladium as a catalyst using Suzuki coupling methods that are well known in the art. The reaction can be treated with various substituted heteroaryl or aryl bromides 9. Finally, the tosyl protecting group is

15 removes under basic conditions according to Scheme II, step (e), giving the compounds of structure 11.

Scheme III

Reagents and conditions: (a) Br2, CHCl3, 0 ° C at RT; (b) "BuLi, THF, TsCl; (c) Pd (PPh3) 4, Na2CO3, DME, EtOH / H2O, microwave irradiation, 120 ° C, 2 hours; (d) 3N NaOH, MeOH.

Scheme III above shows another general synthetic route that has been used to prepare the compounds 11 of the present invention when A, from R1 to R5 and x are as described herein. Intermediate 7 is prepared as above according to Scheme II. In this case, the formation of the biaryl bond derivatives 10 is achieved by treating the bromides 7 with a boronic acid derivative 12 in the presence of palladium.

5 as a catalyst using Suzuki coupling methods that are well known in the art. The reaction can be treated with various heteroaryl or aryl bromides of boronic acids 12. Again, the tosyl protecting group is removed under basic conditions according to Scheme III, step (d), giving the compounds of structure 11.

10 Scheme IV

Reagents and conditions: (a) Lawesson reagent, Toluene, 110 ° C, D / N; (b) EtOH, reflux, D / N; (c) EtOH, 1N NaOH, 12 hours; (d) EDC, HOBt, DMF, NHR'R, TA, D / N.

Scheme IV above shows a general synthetic route that has been used to prepare the compounds 18 of the present invention when R, R 'and R1 to R5 are as described herein. Starting materials 13 can be prepared by procedures substantially similar to those described in the literature by Schneller and Luo J. Org. Chem. 1980, 45, 4045. Derivatives 14 are formed by the reaction of the

20 compounds 13 with Lawesson reagent. Cyclization of compounds 14 in the presence of β-ketoesters 15 give intermediates 16. The reaction can be treated with several β-ketoesters 15. After deprotection of esters 16 under basic conditions derivatives 18 are formed by a reaction step of coupling well known to a person skilled in the art.

25 Scheme V

Reagents and conditions: (a) "BuLi, THF, PCl; (b) i) 'BuLi, Et2O, -78 ° C, 1h, ii) R'SSR'; (c) Deprotection conditions.

Scheme V above shows a general synthetic route that has been used to prepare the compounds 18 of the present invention when R 'is as described herein. The starting material 19 can be prepared by procedures described by Mazeas, et al, Heterocycles 1999, 50, 1065. Intermediate 20,

35 obtained by protecting 19 with a suitable protecting group (P), is treated with the R'SSR 'disulfide according to Scheme V, step (b). After deprotection of indazole 21, the compounds of formula 22 are formed.

Scheme VI

5 Reagents and conditions: (a) R1OH, NaOMe, CuBr, DMF, Heating, 2.5 hours.

Scheme VI above shows a general synthetic route that has been used to prepare the compounds 23 of the present invention when R 'is as described herein. The starting material 19 is treated with the appropriate alcohol R'OH according to Scheme VI, step (a).

Scheme VII

Reagents and conditions: (a) "BuLi, THF, PCl; (b) NHR'R, PdCl2 (dppf), NaO'Bu, THF, heating; or HNR'R, Cu, 15 K2CO3, nitrobenzene, heating; (c ) Check out conditions.

Scheme VII above shows a general synthetic route that has been used to prepare compounds 25 of the present invention when R and R 'is as described herein. Intermediate 20, obtained by protection of 19 with a suitable protecting group (P) is tarted with an RR'NH amine in the presence of

20 Palladium as a catalyst using the Buchwald-Hartwig cross coupling reaction well known in the art. This cross-coupling reaction could be achieved by treating intermediate 20 with an RR'NH amine in the presence of copper as a catalyst using the Ullman reaction well known in the art. Both reactions can be treated with various substituted amines. After deprotection of indazole 24, the compounds of formula 25 are formed.

Scheme VIII

Reagents and conditions: (a) R4B (OH) 2, Pd (PPh3) 4, EtOH, H2O, DME, 100 ° C, D / N; (b) Br2, CHCl3, 0 ° C at RT; (c) "BuLi, THF, TsCl; (d) Pd (PPh3) 4, Na2CO3, DME, EtOH / H2O, microwave irradiation, 120 ° C, 2 hours; (e) 3N NaOH, MeOH.

Scheme VIII above shows a general synthetic route that has been used to prepare compounds 30 of the

The present invention when A, from R1 to R4 and x are as described herein. The compound of structure 19 is treated with a boronic acid derivative R4B (OH) 2 in the presence of palladium as a catalyst using the Suzuki coupling method, which is well known in the art. The reaction can be treated with various boronic acids of aryl or substituted heteroaryl. Intermediate 27 is prepared by sprouting the compounds of structure 26 followed by the subsequent protection of intermediate 27 with a tosyl group. Another Suzuki cross coupling reaction is achieved according to Scheme VIII, step (d). Finally, the tosyl protecting group is removed under basic conditions according to step (e) of Scheme VIII, giving the compounds of structure 30.

Scheme IX

Reagents and conditions: (a) "BuLi, THF, PCl; (b) i) 'BuLi, Et2O, -78 ° C, 1h, ii) R'CHO; (c) Deprotection conditions.

10 Scheme IX above shows a general synthetic route that has been used to prepare the compounds 32 of the present invention when R 'is as described herein. Intermediate 20, obtained by protecting 19 with a suitable protecting group (P), is treated with the appropriate R'CHO aldehyde according to step (b) of Scheme IX. After deprotection of indazole 31, the compounds of

15 formula 32.

X scheme

20 Reagents and conditions: (a) "BuLi, THF, PCl; (b) i) 'BuLi, Et2O, -78 ° C, 1h, ii) R'CH2Br; (c) Deprotection conditions.

Scheme X above shows a general synthetic route that has been used to prepare compounds 32 of the

The present invention when R 'is as described herein. Intermediate 20, obtained by protecting 19 with a suitable protecting group (P), is treated with the appropriate aldehyde R'CH2Br according to step (b) of Scheme X. After deprotection of indazole 33, the compounds of formula 34.

30 Scheme XI

Reagents and conditions: (a) CDI, DMF; (b) P2S5, pyridine.

The above Scheme XI shows a general synthetic route that has been used to prepare the compounds 38 of the present invention when R2 A R5 is as described herein. Starting materials 35 can be prepared by procedures substantially similar to those described in the literature by Allegreti et al, Org. Proc. Res. Dev. 2003, 7, 209. Intermediates 35 react with amines 36 following step (a) of

40 Scheme XI. The reaction can be treated with various amines 36. Cyclization of compounds 37 in the presence of P2S5 gives the desired derivatives 38.

Scheme XII

Reagents and conditions: (a) AlCl3, CH2Cl2, TA, 16 hours; (b) NH2OH.HCl, EtOH, heating, 1 hour.

5 Scheme XII above shows a general synthetic route that has been used to prepare the compounds 41 of the present invention when R2 A R5 is as described herein. Intermediates 40 are prepared using Friedel-Craft acylation methods that are well known in the art. This reaction can be treated with various substituted derivatives 39 to form compounds of formula 40. The compounds of

Formula 41 is obtained by cyclization of intermediate 40 according to step (b).

Although certain exemplary embodiments have been represented and described above and herein, it will be appreciated that a compound of the invention can be prepared in accordance with the procedures generally described above using suitable starting materials by generally procedures

15 available to one skilled in the art.

In accordance with the foregoing, in another embodiment, the present invention provides methods for preparing a compound of the present invention substantially as described herein and, in particular, as described in the Schemes and Examples.

5. Uses, Formulation and Administration

Pharmaceutically Acceptable Compositions

As discussed above, the present invention provides compounds that are inhibitors of protein kinases and, therefore, the present compounds are useful for the treatment of diseases, disorders and conditions, including, among others, autoimmune, inflammatory, proliferative diseases. or hyperproliferative or a disease mediated by the immune system. Accordingly, in another aspect of the present invention pharmaceutically acceptable compositions are provided, in which 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.

It will also be appreciated that some of the compounds of the present invention may exist in free form for treatment or, where appropriate, as a pharmaceutically acceptable derivative thereof. In accordance with the present invention, a pharmaceutically acceptable derivative includes, among others, pharmaceutically acceptable salts, esters, salts of said esters or any other adduct or derivative which, upon administration to a patient in need thereof, can provide, directly or indirectly. , a compound as, on the other hand, has been described in

This document, or a metabolite or residue thereof.

As used herein, the term "pharmaceutically acceptable salt" refers to salts that are, within the scope of firm medical judgment, suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation. , allergic response and the like, and that are 45 proportional to a reasonable ratio of benefits / risks. A "pharmaceutically acceptable salt" means any non-toxic salt or salt of an ester of a compound of the present invention which, upon administration to a receptor, is capable of providing, directly or indirectly, a compound of the present invention or a metabolite active inhibitor or residue thereof. As used herein, the term "active metabolite inhibitor or residue thereof" means that a metabolite or residue thereof is also an inhibitor of the

50 protein kinases of the Tec family (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rl).

Pharmaceutically acceptable salts are well known in the art. For example, M. Berge, et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable organic and inorganic bases and acids. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of an amino group formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or with organic acids such as acetic acid, oxalic acid. , maleic acid, tartaric acid, citric acid, succinic acid

or malonic acid, or using other methods used in the art, such as ion exchange. Other salts

Pharmaceutically acceptable include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorrate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheatoate, heptane, heptane, heptane, heptane , iodhydrate, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, 3-phenylpropionate , pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, ptoluenesulfonate, undecanoate, valerate salts and the like. Salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium salts and N + salts (C 1-4 alkyl). The present invention also provides for the quaternization of any group containing basic nitrogen of the compounds described herein. By means of said quaternization, water or oil soluble or dispersible products can be obtained. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like. Other pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulphonate.

As described above, the pharmaceutically acceptable compositions of the present invention further comprise a pharmaceutically acceptable carrier, adjuvant or vehicle which, as used herein, includes any and all solvents, diluents or other liquid carriers, dispersions

or auxiliary suspensions, active surface agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as is suitable for the desired specific dosage form. Several vehicles used in the formulation of pharmaceutically acceptable compositions and known techniques for the preparation thereof are disclosed in Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980). Except with respect to any conventional vehicle medium it is incompatible with the compounds of the invention, such as producing any undesirable biological effect or, otherwise, interacting in a harmful manner with any other component (s) of the pharmaceutically acceptable composition, its use is contemplated within the scope of the present invention. Some examples of materials that can serve as pharmaceutically acceptable carriers include, among others, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid or potassium sorbate, partial mixtures of glycerides of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes , polyethylene-polyoxypropylene block polymers, wool fat, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; gum tragacanth powder; malt; jelly; talcum powder; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; Sesame oil; olive oil; corn oil and soybean oil; glycols, such as propylene glycol or polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen free water; isotonic saline solution; Ringer's solution; ethyl alcohol and phosphate buffer solutions, as well as other compatible non-toxic lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, there may also be preservatives and antioxidants in the composition, according to the formulator's judgment.

Uses of pharmaceutically acceptable compounds and compositions

In yet another aspect, there is provided a method for the treatment or reduction of the severity of a disease mediated by the Tec family (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rlk) comprising administering to a subject in need of an effective amount of a compound, or a pharmaceutically acceptable composition, comprising a compound. In certain embodiments of the present invention, an "effective amount" of the compound or a pharmaceutically acceptable composition is the effective amount for a disease mediated by the Tec family (eg, Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rl). The compounds and compositions according to the process of the present invention can be administered using an amount and any route of administration effective to treat or reduce the severity of a disease mediated by the Tec family (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rl). The exact amount required will vary from one subject to another depending on the species, the age and general condition of the subject, the severity of the infection, the specific agent, its mode of administration and the like. The compounds of the invention are preferably formulated in unit dosage form for ease of administration and uniformity of dosage. The term "unit dosage form", as used herein, refers to a physically small unit of the therapeutic agent suitable for the patient to be treated. However, it will be understood that the total daily use of the compounds and compositions of the present invention will be decided by the responsible physician within the scope of a sound medical judgment. The specific effective dose level therapeutically effective for any patient or organism will depend on various factors, including the disorder being treated and its severity, the activity of the specific compound used, the specific composition used; the age, body weight, general health status, sex and diet of the patient or subject; the time of administration, the route of administration and the rate of excretion of the specific compound used; duration of treatment, drugs used in combination or coinciding with the specific compound used and similar factors well known in medical techniques. The term "patient", as used herein, means an animal, preferably a mammal, and, most preferably, a human being.

The pharmaceutically acceptable compositions of the present invention can be administered to humans.

5 and other animals by oral, rectal, parenteral, intracisternal, intravaginal, intraperitoneal, topical (as in powders, ointments or drops), buccal, such as a nasal or oral nebulizer, or the like, depending on the location and severity of the infection to be treated In certain embodiments, the compounds of the invention may be administered orally or parenterally at dose levels of from about 0.01 mg / kg to about 50 mg / kg and, preferably, from about 1 mg / kg to about 25 mg / kg of the subject's body weight per day, one or more times per day to obtain the desired therapeutic effect.

Liquid dosage forms for oral administration include, among others, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, liquid dosage forms may contain inert diluent commonly used in the art, such as, for example,

Water or other solvents, solubilizing and emulsifying agents, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular oils of cotton, peanut, corn, germ, olive, castor and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and sorbitan fatty acid esters, and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.

Injectable preparations may be formulated, for example sterile injectable aqueous or oleaginous suspensions according to the known technique using suitable dispersing agents or wetting agents and

25 suspension The sterile injectable preparation may also be a sterile injectable solution or suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example in the form of a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution according to U.S.P. In addition, sterile fixed oils are conventionally employed in the form of a solvent or suspending medium. For this purpose any mild fixed oil including synthetic mono- or diglycerides can be used. In addition, fatty acids such as oleic acid are used in the preparation of injectable substances.

Injectable formulations can be sterilized by, for example, filtration through a bacteria retention filter or by incorporating sterilizing agents in the form of sterile solid compositions that can be

Dissolve or disperse in sterile water or other sterile injectable medium before use.

In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the subcutaneous or intramuscular injection compound. This can be achieved by using a liquid suspension of crystalline or amorphous material with poor water solubility. Therefore, the absorption rate of the compound depends on its dissolution rate which, in turn, may depend on the size of the crystal and the crystalline form. Alternatively, delayed absorption of a parenterally administered compound is achieved by dissolving or suspending the compound in an oily vehicle. Depot injectable forms are manufactured by forming microcapsular matrices of the compound in biodegradable polymers, such as polylactide-polyglycolide. Depending on the proportion between the compound the polymer, and the nature of the concrete polymer used,

45 can control the release rate of the compound. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations are also prepared by trapping the compound in liposomes or microemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferably suppositories that can be prepared by mixing the compounds of the present invention with suitable excipients or non-irritating vehicles, such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at room temperature. but liquids at body temperature and, therefore, melt in the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and

55 granules In said solid dosage forms, the active compound is mixed with at least one pharmaceutically acceptable inert carrier or excipient such as sodium citrate or dicalcium phosphate and / or) fillers or expanders such as starches, lactose, sucrose, glucose, mannitol and silicic acid ; b) binders, such as, for example, carboxymethyl cellulose, microcrystalline cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; c) humectants, such as glycerol; d) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; e) solution retarding agents, such as paraffin; f) absorption accelerators, such as quaternary ammonium compounds; g) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; h) absorbents, such as kaolin and bentonite clay; and i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate; and mixtures thereof. In the case of capsules, tablets and pills, the form of

Dosage may also comprise buffering agents.

Solid compositions of a similar type can also be used as fillers in hard and soft filled gelatin capsules, using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols, and the like. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and coatings, such as enteric coatings and other coatings well known in the art of pharmaceutical formulation. They may optionally contain opacifying agents, and they may also be of a composition such that they release the active ingredient (s) only or, preferably, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of including compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type can also be used as fillers in hard and soft filled gelatin capsules, using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols, and the like.

The active compounds may also be in microencapsulated form with one or more excipients as indicated above. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and coatings, such as enteric coatings, release control coatings and other coatings well known in the art of pharmaceutical formulation. In said solid dosage forms, the active compound may be mixed with at least one inert diluent, such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances apart from inert diluents, for example lubricants for tablet formation and other auxiliaries for tablet formation such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and may also be of a composition that release the active ingredients alone or, preferably, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of including compositions that can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, nebulizers, inhalers or patches. The active component is mixed under sterile conditions with a pharmaceutically acceptable carrier and with any necessary preservative or buffer, as required. Ophthalmic formulations, ear drops and eye drops are also contemplated within the scope of the present invention. Additionally, the present invention contemplates the use of transdermal patches that have the added advantage of providing controlled release of a compound in the body. Said dosage forms can be prepared by dissolving or dispersing the compound in the appropriate medium. Absorption powers can also be used to increase the flow of the compound in the skin. The velocity can be controlled by providing a velocity control membrane or by dispersing the compound in a polymeric matrix or gel.

As described in general above, the compounds of the invention are useful as protein kinase inhibitors. In one embodiment, the compounds and compositions of the invention are inhibitors of one or more kinases of the Tec family (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rlk) and, therefore, without wishing to be bound by theory, the compounds and compositions are particularly useful for treating or reducing the severity of a disease, condition or disorder in which the activation of one or more of a Tec family kinase (e.g., Tec , Btk, Itk / Emt / Tsk, Bmx, Txk / Rlk) is involved in the disease, condition or disorder. When the activation of the Tec family (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rlk) is involved in a particular disease, condition or disorder, the disease, condition and disorder can also be referred to as “Tec-mediated family disease

(e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rlk) or disease symptom. Accordingly, in another aspect, the present invention provides a method for treating or reducing the severity of a disease, condition or disorder in which the activation of one or more of the Tec family (e.g., Tec, Btk , Itk / Emt / Tsk, Bmx, Txk / Rl) is involved in the disease state.

The activity of a compound used in the present invention as a kinase inhibitor of the Tec family (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rl) can be analyzed in vitro, in vivo or In a cell line. In vitro assays include assays that determine the inhibition of phosphorylation activity or ATPase activity of the Tec family kinase (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rl) activated . As an alternative, in vitro assays quantify the inhibitor's ability to bind to a Tec family kinase (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rl) activated. The inhibitor binding can be measured by radiolabeling the inhibitor before binding, isolating the inhibitor from the Tec family complex (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rl) and determining the amount of the radiolabel attached. Alternatively, inhibitor binding can be determined by performing a competition experiment in which new inhibitors are incubated with a Tec family kinase (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rl ) linked to known radioligands.

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

The term "tyrosine kinase mediated condition of the Tec family", as used herein, means any disease or other harmful condition in which it is known that the kinase of the Tec family 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).

5 For example, tyrosine kinase-mediated conditions of the Tec family include respiratory tract diseases, including, without limitation, reversible obstructive airway diseases, including asthma, such as bronchial, allergic, intrinsic, extrinsic and dust asthma, in particular chronic or habitual asthma (eg, hyperresponsiveness of the respiratory tract due to late asthma) and bronchitis. Additionally, tyrosine kinase diseases of the Tec family include, without limitation, conditions characterized by inflammation of the nasal mucous membrane, including acute, allergic rhinitis and chronic rhinitis, including gas rhinitis, hypertrophic rhinitis, purulent rhinitis, rhinitis dry and medicated rhinitis; membranous rhinitis, including croup, fibrinous and pseudomembranous rhinitis and scrofulous rhinitis, seasonal rhinitis, including nerve rhinitis (hay fever) and vasomotor rhinitis, sarcoidosis, farmer's lung and related diseases, fibroid lung

15 and idiopathic interstitial pneumonia.

Tyrosine kinase mediated conditions of the Tec family also include bone and joint diseases including, among others, rheumatoid arthritis (cloth formation), seronegative spondiolarthropathies (including ankylosing spondylitis, psoriatic arthritis and Reiter's disease), Behcet's disease, Sjogren's syndrome and systemic sclerosis.

Kinase-mediated conditions of the Tec family also include skin diseases and disorders, including, without limitation, psoriasis, systemic sclerosis, atopic dermatitis, contact dermatitis and other eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermolysis bullosa,

25 urticaria, angioedema, vasculitidis, erythema, cutaneous eosinophilias, uveitis, alopecia areata and vernal conjunctivitis.

Tyrosine kinase-mediated conditions of the Tec family also include diseases and disorders of the gastrointestinal tract including, among others, celiac disease, proctitis, eosinophilic gastroenteritis, mastocytosis, pancreatitis, Crohn's disease, ulcerative colitis, food-related allergies that have remote effects from the intestine, for example migraines, rhinitis and eczema.

Tyrosine kinase-mediated conditions of the Tec family also include diseases and disorders of other tissues and systemic diseases including, without limitation, multiple sclerosis, atherosclerosis, acquired immunodeficiency syndrome (AIDS), lupus erythematosus, systemic lupus erythematosus, Hashimoto's thyroiditis ,

35 myasthenia gravis, type I diabetes, nephrotic syndrome, eosinophilic fasciitis, hyper IgE syndrome, lepromatous leprosy, sezary syndrome and idiopathic thrombocytopenic purpura, restenosis after angioplasty, tumors (eg, leukemia, lymphomas), atherosclerosis and systemic lupus erythematosus.

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

It will also be appreciated that the pharmaceutically acceptable compounds and compositions of the present invention can be used in combination therapies, that is, the compounds and pharmaceutically compositions.

Acceptable can be administered together with, before or after one or more desired therapeutic or medical procedures. The specific combination of therapies (therapeutic and procedures) for use in a combination regimen will take into account the compatibility of the desired therapeutic and / or procedures and the therapeutic effect that is desired to be achieved. It will also be appreciated that the therapies employed can achieve a desired effect for the same disorder (for example, a compound of the invention can be administered together with another agent used to treat the same disorder) or can achieve different effects (e.g., control of any adverse effects). As used herein, additional therapeutic agents that are normally administered to try to prevent a particular disease or condition are known as "suitable for the disease or condition being treated."

For example, chemotherapeutic agents or other antiproliferative agents can be combined with the compounds of the present invention to treat proliferative diseases and cancer. Examples of known chemotherapeutic agents include, among others, for example, other therapies or anticancer agents that can be used in combination with the anticancer agents of the invention of the present invention include surgery, radiotherapy (in just a few examples, gamma radiation, radiotherapy with a neutron beam, electron beam radiotherapy, proton therapy, brachytherapy and systemic radioactive isotopes, to name a few), endocrine therapy, biological response modifiers (interferons, interleukins, and tumor necrosis factor (TBF) to name a few few), hyperthermia and cryotherapy, agents that mitigate any adverse effects (e.g., antiemetics) and other approved chemotherapeutic drugs, including, but not limited to, alkylating agents (mechloretamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide), antimetabolites (methotrexate) , purine antagonists and

65 pyrimidine antagonists (6-Mercaptopurine, 5-Fluorouracil, Citarabila, Gemcitabine), spindle poisons (Vinblastine, Vincristine, Vinorelbine, Paclitaxel), podophyllotoxins (Etoposide, Irinotecan, Topotecan), antibiotics (Doxorubicin, Mitomycin, Mitomycinomycin 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 FDA-approved cancer drugs at

5 http://www.fda.gov/cder/cancer/druglistframe.htm and the and The Merck Manual, Seventeenth Ed. 1999, all of whose content is incorporated herein by reference.

Other examples of agents with which the inhibitors of the present invention can also be combined include, without limitation: treatments for Alzheimer's disease such as Aricept® and Excelon®; treatments for Parkinson's disease such as L-DOPA / carbidopa, entacapone, ropinrol, pramipexole, bromocriptine, pergolide, trihexefendil and amantadine; agents for treating multiple sclerosis (MS) such as beta interferon (eg, Avonex® and Rebif®), Copaxone®, and mitoxantrone; asthma treatments 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, anticonvulsants, ion channel blockers, riluzole, and anti-Parkinsonian agents; agents for the treatment of cardiovascular diseases such as beta-blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers and statins; agents for treating liver diseases such as corticosteroids, cholestyramine, interferons and antiviral agents; agents for treating blood disorders such as corticosteroids, antileukemic agents and growth factors; and agents to treat immunodeficiency disorders such as gamma globulin.

The amount of additional therapeutic agent present in the compositions of the present invention will be no more than the amount that would normally be administered in a composition comprising said therapeutic agent as the sole active agent. Preferably, the amount of additional therapeutic agent in the compositions of the present disclosure will range from about 50% to 100% of the amount normally present in a composition comprising said agent as the only therapeutically active agent.

The compounds of the present invention or pharmaceutically acceptable compositions thereof can 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

The present invention as generally described above, and in classes and subclasses herein and a suitable vehicle for coating said implantable device. In yet another aspect, the present invention includes an implantable device coated with a composition comprising a compound of the present invention as generally described above, and in classes and subclasses herein and a suitable vehicle for coating said implantable device.

For example, vascular stents have been used to overcome restenosis (recessing of the vessel wall after injury). However, patients using stents or other implantable devices are at risk of clots or platelet activation. These unwanted effects can be prevented or mitigated by precoating the device with a pharmaceutically acceptable composition that

45 comprises a kinase inhibitor. Suitable coatings and general preparation of implantable coated devices are described in US Pat. 6,099,562; 5,886,026; and 5,304,121. The coatings are normally biocompatible polymeric materials such as hydrogel polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate and mixtures thereof. The coatings may optionally be covered by a suitable cover of fluorosilicone, polysaccharides, polyethylene glycol, phospholipids or combinations thereof to confer controlled release characteristics in the composition.

Another aspect of the invention relates to the inhibition of the activity of the Tec family (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rlk) in a biological sample or a patient, in which the procedure includes administering to

55, 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; biopsy material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears or other body fluids or extracts thereof.

Inhibition of the kinase activity of the Tec family (e.g., Tec, Btk, Itk / Emt / Tsk, Bmx, Txk / Rlk) in a biological sample is useful for various purposes known to one skilled in the art. Examples of such purposes include, among others, blood transfusion, organ transplantation, storage of biological samples and biological tests.

Synthesis Examples

65 As used herein, 1 H NMR is nuclear magnetic resonance. HPLC is high performance liquid chromatography. The term "Tr (min)" refers to the retention time in HPLC, in minutes, associated with the compound. Unless otherwise indicated, the HPLC method used to obtain the reported retention time is as follows:

5 Column: Ace 5 C8, 15 cm x 4.6 mm id Gradient: 0 -100% acetonitrile + methanol (50:50) (20 mM Tris phosphate at pH 7.0) Flow rate: 1.5 ml / min Detection: 225 nm

10 Example 1

5-Phenyl-1H-pyrrolo [2,3-b] pyridine

15 5-Bromo-1H-pyrrolo [2,3-b] pyridine (2 g, 10.15 mmol), feniboronic acid (1.24 g, 10.15 mmol) and palladium tetrakis (triphenylphosphine) (117 mg 0.10 mmol) were suspended in ethanol (5 ml), water (6 ml) and DME (22 ml) and heated at 100 ° C overnight. The solvent was removed in vacuo and the reaction was purified by column chromatography eluting with 30% ethyl acetate in gasoline, giving the title compound as a

20 off-white solid (1.51 g, 77%). MS (ES +) 195, (ES-) 193. δH (CDCl3) 6.60 (1H, s), 7.36-7.43 (2H, m), 7.68 (2H, d), 8.18 (1H, s), 8.62 (1H, s), 10.39 (1H, br s).

Example 2

2-Chloro-1- (5-Phenyl-1H-pyrrolo [2,3-b] pyridin-3-yl) -ethanone

5-Phenyl-1H-pyrrolo [2,3-b] pyridine (200 mg, 1.03 mmol) and aluminum chloride (412 mg, 3.09 mmol) were suspended in

30 DCM dry and stirred at room temperature for 1 hour. Chloroacetyl chloride (98 μl, 1.24 mmol) was added dropwise and the resulting amber solution was stirred at room temperature overnight. The reaction was quenched with methanol (5 ml) and stirred at room temperature for 2 hours. Then, the solvent was evaporated to give an orange oil. This was divided between DCM and water. The organic layer was concentrated in vacuo and the product was triturated with diethyl ether, giving the title compound as a beige solid (188 mg, 67%). MS (ES +) 271, (ES-) 269. δH

35 (CDCl3) 4.57 (2H, s), 7.44 (1H, t), 7.50 (2H, t), 7.70 (2H, d), 8.23 (1H, s), 8 , 70 (1H, s), 8.91 (1H, s), 11.59 (1 H, br s).

Example 3

Diethyl-4 [- (5-Phenyl-1 H -pyrrolo [2,3-b] pyridin-3-yl) -thiazol-2-yl] amine

2-Chloro-1- (5-phenyl-1H-pyrrolo [2,3-b] pyridin-3-yl) -ethanone (50 mg, 0.18 mmol) and 1,1-diethylthiourea (24 mg, 0, 18 mmol) were suspended / dissolved in ethanol (2 ml) and heated in the microwave at 120 ° C for 10 minutes. Mix

The crude reaction was purified by HPLC eluting with acetonitrile / water, to give the title compound as a cream-colored solid (9.5 mg, 15%). MS (ES +) 349, (ES-) 347. δH (CDCl3) 1.31 (6H, t), 3.59 (4H, q), 6.61 (1H, s), 7.38 (1H, t ), 7.50 (2H, t), 7.69 (2H, d), 7.78 (1H, s), 8.56-8.62 (2H, m), 8.94 (1H, br s ).

Several other compounds of formula I have been prepared by methods substantially similar to those

50 described herein, Example 3. The characterization data for these compounds are summarized in Table 3 below and include HPLC, CLE / MS (observed) and 1 H NMR data.

Table 3. Characterization data for certain compounds of formula I

Compound No. I

M + 1 (obs) Tr (min) 1H NMR

one

292 9.60 (CDCl3) 2.82 (3H, s), 7.26 (1H, s), 7.40 (1H, t), 7.52 (2H, t), 7.70 (2H, d), 7, 89 (1H, s), 8.53 (1H, s), 8.62 (1H, s), 9.07 (1H, br s)

2

349 10.72 (CDCl3) 1.31 (6H, t), 3.59 (4H, q), 6.61 (1H, s), 7.38 (1H, t), 7.50 (2H, t), 7, 69 (2H, t), 7.78 (1H, s), 8.56-8.62 (2H, m), 8.94 (1H, br s)

3

306 9.96 (CDCl3) 2.56 (3H, s), 2.75 (3H, s), 7.39 (1H, t), 7.46-7.53 (3H, m), 7.68 (2H, d ), 8.58 (1H, s), 8.61 (1H, s), 9.44 (1H, br s)

4

363 10.98 (CDCl3) 1.33 (6H, t), 2.47 (3H, s), 3.54 (4H, q), 7.36 (1H, t), 7.45-7.51 (3H, m ), 7.69 (2H, d), 8.60 (1H, s), 8.76 (1H, s), 9.14 (1H, br s)

5

307 8.74 (CDCl3) 2.40 (3H, s), 5.02 (2H, s), 7.34-7.39 (2H, m), 7.46 (2H, t), 7.65 (2H, d ), 8.51 (1H, s), 8.59 (1H, s), 9.96 (1H, br s)

6

364 10.17 (CDCl3) 1.48 (3H, t), 2.70 (3H, s), 4.50 (2H, q), 7.39 (1H, t), 7.48 (2H, t), 7, 61 (1H, s), 7.70 (2H, d), 8.65 (2H, s), 9.07 (1H, br s)

7

350 9.89 (CDCl3) 1.50 (3H, t), 4.54 (2H, q), 7.41 (1H, t), 7.52 (2H, t), 7.66-7.72 (3H, m ), 8.03 (1H, s), 8.55 (1H, s), 8.65 (1H, s), 9.26 (1H, br s)

9

363 9.78 (CDCl3) 3.55 -3.63 (4H, m), 3.86 -3.92 (4H, m), 6.76 (1H, s), 7.36-7.44 (1H, m) , 7.46-7.54 (1H, m), 7.83 (1H, s), 8.53 (1H, s), 8.59 (1H, s), 9.64 (1H, s)

12

390 8.63 (DMSO) 3.25-3.45 (4H, m), 3.90 (2H, br s), 4.76 (2H, br s), 7.42 (1H, t), 7.54 (2H , t), 7.79 (2H, d), 8.20 (1H, s), 8.36 (1H, s), 8.63 (2H, d), 8.82 (1H, br s), 12.20 (1H, s).

14

391 9.20 (CDCl3) 3.81-3.90 (4H, m), 4.66 (2H, br s), 7.41 (1H, t), 7.54 (2H, t), 7.62-7, 69 (3H, m), 7.84 (1H, s), 8.55 (1H, s), 8.65 (1H, s), 9.11 (1H, br s).

16

404 8.89 (CDCl3) 1.18 -1.34 (2H, m), 1.73 -1.82 (2H, m), 1.98 -2.37 (3H, m), 2.58 -2.70 ( 2H, m), 3.20 (3H, s), 3.42 -3.53 (2H, m), 6.62 (1 H, s), 7.33 7.42 (1H, m), 7 , 45 -7.54 (2H, m), 7.65 -7.71 (2H, m), 7.80 (1H, s), 8.54 -8.59 (2H, m), 9.42 (1H, br s)

18

362 (DMSO) 2.81 -2.87 (4H, m), 3.35-3.44 (4H, m), 7.27 (1H, s), 7.37 (1H, t), 7.45 -7.52 (2H, m), 7.73 -7.77 (2H, m), 7.84 (1H, s), 8.54 (1H, s), 8.59 (1H, s), 11.88 (1H, s)

46

335 8.92 (DMSO) 2.18 (3H, s), 7.43 (1H, t), 7.49-7.56 (3H, m), 4.79-4.82 (2H, m), 7.94 (1H, s), 8.58 (1H, s), 8.71 (1H, s), 12.00 (1H, s), 12.19 (1H, s)

48

418 8.48 (DMSO): 1.40 (2H, t), 1.80 -1.95 (3H, m), 2.88 (2H, t), 3.22 -3.31 (4H, m), 7, 39 (1H, t), 7.51 (2H, t), 7.85 (2H, d), 8.15 (1H, s), 8.29 (2H, br s), 8.60 (1H, s), 8.78 (1H, s), 8.99 (1H, t), 12.17 (1H, s).

49

293 (DMSO) 6.95 (1H, s), 7.00 (1H, s), 7.37 (1H, t), 7.46-7.51 (2H, m), 7.75-7.79 (3H, m), 8.54 (1H, s), 8.59 (1H, s), 11.84 (1 H, s)

fifty

321 10.00 (DMSO) 3.13 (6H, s), 7.10 (1H, s), 7.39 (1H, t), 7.46-7.54 (2H, m), 7.75-7.78 (2H, m), 7.85 (1H, s), 8.54 (1H, s), 8.62 (1H, s), 11.88 (1H, s)

fifty

538 10.9 (CDCl3) 1.60 -1.80 (4H, m), 2.03 -2.16 (1H, m), 2.73-2.83 (2H, m), 3.21 (3H, s) , 3.2 -3.50 (2H, m), 4.10 -4.32 (2H, m), 5.15 (2H, s), 6.65 (1H, s), 7.26 -7 , 40 (6H, m), 7.44 -7.55 (2H, m), 7.65 -7.70 (2H, m), 7.80 (1H, s), 8.59 (2H, s ), 9.20 (1H, s)

52

365 10.0 (CDCl3) 3.25 (3H, s), 3.44 (3H, s), 3.68 -3.74 (2H, m), 3.75-3.80 (2H, m), 6.65 (1H, s), 7.36-7.44 (1H, m), 7.46-7.54 (2H, m), 7.65-7.72 (2H, m), 7.84 (1H , s), 8.58 (1H, s), 8.61 (1H, s), 9.51 (1H, br s)

53

389 11.22 (CDCl3) 0.28 -0.40 (2H, m), 0.53 -0.63 (2H, m), 0.93 -1.02 (3H, m), 1.15 -1.26 ( 1H, m), 1.72-1.85 (2H, m), 3.38-3.45 (2H, m), 3.45-3.55 (2H, m), 6.61 (1H, s), 7.32-7.42 (1 H, m), 7.42-7.53 (2H, m), 7.62-7.72 (2H, m), 7.78 (1H, s ), 8.60 (1H, s), 8.66 (1H, s), 9.21 (1H, br s)

54

379 10.6 (DMSO) 1.2 -1.3 (6H, m), 3.4 -3.6 (4H, m), 3.8 -3.9 (3H, s), 6.9 (1H, m) , 7.0 (1H, s), 7.3 (2H, m), 7.4 (1H, m), 7.8 -7.9 (1H, s), 8.5 -8.6 (1H , s), 8.7 (1H, s), 11.8 -11.9 (0.7H, s)

55

323 8.5 (DMSO) 3.7 -3.8 (3H, s), 6.8 -7.0 (3H, m), 7.2 -7.3 (2H, m), 7.3 -7.4 ( 1H, m), 7.7 -7.8 (1H, s), 8.5 -8.6 (2H, m), 11.8 (0.6H, s)

56

347 10.38 (CDCl3) 2.05 -2.14 (4H, m), 3.54 -3.61 (4H, m), 6.65 (1H, s), 7.36-7.43 (1H, m) , 7.46 -7.54 (2H, m), 7.76 -7.83 (2H, m), 7.86 (1H, s), 8.54 -8.59 (2H, m), 9 , 56 (1H, br s)

57

379 10.39 (CDCl3) 1.27 -1.33 (3H, m), 3.43 (3H, s), 3.55 -3.62 (2H, m), 3.67 -3.77 (4H, m) , 6.64 (1H, s), 7.35-7.42 (1H, m), 7.46-7.54 (2H, m), 7.66-7.71 (2H, m), 7 , 80 (1H, s), 8.60 (2H, s), 9.42 (1H, br s)

58

335 10.35 (CDCl3) 1.24 -1.34 (3H, m), 3.27 (3H, s), 3.55 -3.64 (2H, m), 6.64 (1H, s), 7.35 -7.41 (1H, m), 7.45 -7.53 (2H, m), 7.65 -7.72 (2H, m), 7.85 (1H, s), 8.58 -8 , 63 (2H, m), 9.57 (1H, br s).

59

376 9.82 (CDCl3) 2.41 (3H, s), 2.53 -2.63 (4H, m), 3.58-3.66 (4H, m), 6.72 (1H, s), 7.37 -7.43 (1H, m), 7.46 -7.53 (2H, m), 7.65 -7.71 (2H, m), 8.52 (1H, s), 8.60 (1H , s), 9.39 (1H, br s)

60

365 9.61 (CDCl3) 1.29 -1.35 (3H, m), 3.46 -3.53 (2H, m), 3.74 -3.81 (2H, m), 3.95-4.02 ( 2H, m), 6.66 (1H, s), 7.34-7.40 (1H, m), 7.47-7.54 (2H, m), 7.65-7.70 (2H, m), 7.78 (1H, s), 8.44 (1H, s), 8.61 (1H, s), 9.80 (1 H, br s)

61

363 9.61 (DMSO) 1.14 (6H, d, J = 6.8Hz), 2.72 -2.84 91H, m), 7.35-7.56 (4H, m), 7.75-7.82 (2H, m), 7.90 (1H, brs), 8.58 (1H, brs), 8.70 (1H, brs), 11.98 (1H, brs), 12.13 (1H, brs) .

62

363 10.97 (CDCl3) 0.80 -0.92 (6H, m), 1.25 -1.35 (3H, m), 3.41 -3.52 (2H, m), 4.25 -4.35 ( 1H. M), 6.60 (1H, s), 7.33-7.40 (1H, m), 7.45-7.52 (2H, m), 7.64-7.71 (2H, m), 7.81 (1H, s), 7.56 (1H, s), 7.60 (1H, s), 9.90 (1H, br s)

63

392 10.27 (CDCl3) 1.34 (3H, t), 2.46 (6H, s), 2.82-2.99 (2H, m), 3.52-3.59 (2H, q), 3.65 -3.80 (2H, m), 6.66 (1H, s), 7.35 -7.41 (1H, m), 7.43 -7.52 (2H, m), 7.63 -7 , 70 (2H, m), 7.80 (1H, s), 8.56 (1H, s), 8.60 (1H, s), 9.29 (1H, br s)

64

381 9.64 (DMSO) 1.14 (6H, d, J = 6.9Hz), 2.72-2.86 (1H, m), 7.30-7.40 (2H, m), 7.52 (1H, s), 7.79-7.91 (3H, m), 8.56 (1H, brs), 8.66 (1H, brs), 11.99 (1H, brs), 12.12 (1H, brs ).

65

353 8.98 (DMSO) 1.15 (6H, d, J = 6.8Hz), 2.72-2.86 (1H, m), 7.18 (1H, s), 7.53 (1H, s), 7 , 78 -7.89 (2H, m), 8.30 (1 H, s), 8.56 -8.62 (1H, m), 11.90 (1H, brs), 12.14 (1H, brs).

66

298 9.58 (DMSO) 1.21 (6H, t, J = 7.0Hz), 3.51 (4H, q, J = 7.0Hz), 7.08 (1H, s), 8.04 (1H, brs) , 8.65 (1H, brs), 8.91 (1H, brs), 12.48 (1H, brs).

67

405 10.68 (DMSO) 1.00 (6H, d), 1.40 (3H, t), 2.11 -2.23 (1H, m), 2.63 (2H, d), 4.34 (2H, q ), 7.34-7.41 (1H, m), 7.46-7.54 (3H, m), 7.73-7.75 (2H, m), 7.99 (1H, s), 8.56 (1H, s), 8.76 (1H, s), 11.95 (1H, s)

68

363 9.80 (DMSO) 1.42 (3H, t), 2.46 (3H, s), 4.35 (2H, q), 7.34-7.40 (1H, m), 7.51-7.56 (3H, m), 7.76-7.71 (2H, m), 8.01 (1H, s), 8.60 (1H, s), 8.74 (1H, s), 12.00 ( 1H, s)

69

349 10.59 (DMSO) 1.2 (6H, q), 3.5 (4H, t), 6.9 (1H, s), 7.4 (1H, m), 7.5 (2H, m), 7, 7 (1H, m), 7.9 (1H, s), 8.1 (2H, m), 8.5 (1H, d), 11.8 (NH, s)

70

392 9.9 (DMSO) 1.2 -1.3 (6H, m), 3.5 -3.6 (4H, m), 6.9 (1H, s), 7.5 -7.6 (3H, m) , 7.7 (1H, s), 7.9 -8.1 (3H, m), 8.4 -8.5 (1H, d), 10.6 (1H, s), 11.5 -11 , 6 (1H, s)

71

364 10.1 (DMSO) 1.2 -1.3 (6H, m), 3.5 (4H, m), 6.6 -6.7 (1H, d), 6.8 (1H, s), 6.8 6.9 (1 H, m), 7.2-7.3 (2H, m), 7.4 (1H, s), 7.8 (2H, m), 8.2 (1H, d), 9.0 (1H, s), 11.3 -11.4 (1H, s)

72

311 8.82 (CDCl3) 1.51 (3H, t), 2.48 (3H, s), 3.18 (1H, s), 4.42 (2H, q), 7.10 (1H, s), 7, 84 (1H, s), 8.51 (1H, s), 8.66 (1H, s), 9.91 (1H, br s)

73

406 7.73 (DMSO) 1.14 (6H, d, J = 6.8H2), 2.74-2.86 (1H, m), 7.42 (1H, brs), 7.58 (1H, s), 7 , 84 -7.94 (3H, m), 7.98 -8.12 (3H, m), 8.65 (1H, brs), 8.75 (1H, brs), 12.01 (1H, brs ), 12.15 (1H, brs).

74

330 8.32 evaron 1.23 (6H, t, J = 7.0Hz), 2.85 (3H, d, J = 4.4Hz), 3.52 (4H, q, J = 7.0Hz), 6.98 ( 1H, s), 7.90 (1H, brs), 8.50-8.60 (1H, m), 8.74 (1H, brs), 8.86 (1H, brs), 12.01 (1H , brs).

75

297 9.86 (DMSO) 1.31 (6H, t, J = 7.0Hz), 3.16 (1H, s), 3.59 (4H, q, J = 7.0Hz), 6.60 (1H, s) , 7.80 (1H, brs), 8.49 (1H, brs), 8.55 (1H, brs), 9.75 (1H, brs).

Example 4

3-iodo-5-phenyl-1H-pyrrolo [2,3-b] pyridine

A solution of 5-phenyl-1H-pyrrolo [2,3-b] pyridine (2.96 g, 15.24 mmol, 1 eq.) In anhydrous DMF (60 ml), under stirring at room temperature, was treated with iodine (7.74 g, 30.50 mmol, 2 eq.) and then potassium hydroxide (3.20 g, 57.14 mmol, 3.75 eq.) The reaction mixture was stirred at room temperature for 15 hours before diluting with a mixture of aqueous sodium thiosulfate and ethyl acetate. The organic layer was separated, washed with a saturated aqueous solution of sodium chloride, dried over sodium sulfate and then concentrated in vacuo. The resulting oil was then dissolved in a mixture of DCM / MeOH and adsorbed on silica gel. Then, the material was loaded dry on a column and subjected to silica gel chromatography using a mixture of ethyl acetate (1 ). Petroleum ether (2) as eluent, giving 3-iodo-5-phenyl-1 H -pyrrolo [2,3-b] pyridine (1) (3.16 g, 65%) as a white solid. 1H NMR (400Mhz, DMSO) 7.34-7.41 (1H, m), 7.46-7.56 (2H, m), 7.70-7.80 (3H, m), 7.81 -7.89

10 (1H, m), 8.53-8.60 (1H, m), 12.21 (1H, brs).

Example 5

3-iodo-5-phenyl-1-tosyl-1H-pyrrolo [2,3-b] pyridine

A suspension of 60% of sodium hydride in mineral oil (79 mg, 1.98 mmol, 1.2 eq) in anhydrous DMF (30 ml), under stirring at room temperature, was treated with a solution of 3-iodine. 5-phenyl-1H-pyrrolo [2,3-b] pyridine (1) (530-20 mg, 1.66 mmol, 1.0 eq.) In DMF (5 ml). Then, the reaction mixture was stirred at room temperature for 1 hour before cooling to 0 ° C. A solution of p-toluenesulfonyl chloride (316 mg, 1.66 mmol, 1.0 eq.) In anhydrous DMF (5 ml) was then added and the reaction mixture was allowed to warm to room temperature for 15 hours. Then, the reaction mixture was diluted with a mixture of water and ethyl acetate, washed with a saturated aqueous solution of sodium chloride, dried over sodium sulfate, filtered and then concentrated to

25 empty. The resulting oil was then subjected to silica gel chromatography using a mixture of ethyl acetate (1): 40-60 petroleum ether (2) as eluent, giving 3-iodo-5-phenyl-1-tosyl-1H- pyrrolo [2,3-b] pyridine (2) (743 mg, 95%) as a white solid. 1H NMR (400 MHz, DMSO) 2.37 (3H, s), 7.39-7.56 (5H, m), 7.75 (2H, d), 7.91 (1H, s), 8 , 05 (2H, d), 8.20 (1H, s), 8.71 (1H, s).

30 Example 6

5-phenyl-3- (1H-pyrrol-2-yl) -1H-pyrrolo [2,3-b] pyridine

A mixture of 3-iodo-5-phenyl-1-tosyl-1H-pyrrolo [2,3-b] pyridine (2) (150 mg, 0.32 mmol, 1 eq.), Tetrakis (triphenylphosphine) palladium ( 0) (4 mg, 0.0035 mmol, 0.01 eq.) And 1- (tert-butoxycarbonyl) -1H-pyrrol-2-yl-2boronic acid (67 mg, 0.32 mmol, 1 eq.) Is Introduced in a microwave tube. Then, the mixture was treated with DME (4 ml), EtOH (0.86 ml), water (1.14 ml) and a solution of aqueous 2N sodium carbonate (0.63 ml). The tube was introduced in

40 the microwave and heated at 160 ° C for 40 minutes. The tube was allowed to cool to room temperature and diluted with water / ethyl acetate. The organic layer was separated, dried over sodium sulfate and concentrated in vacuo to give a gum. The gum was dissolved in DMSO and subjected to reverse phase chromatography using ACN / water as gradient eluent, giving 5-phenyl-3- (1 H -pyrrol-2-yl) -1-H-pyrrolo [2,3- b] pyridine (3) as a solid. 1 H NMR (400 MHz, DMSO) 6.10 -6.16 (1H, m), 6.45 -6.50 (1H, m), 6.78 -6.84 (1H, m), 7, 32 -7.54 (3H, m), 8.37 -8.42 (1H,

45 m), 8.52 -8.58 (1H, m), 11.05 (1H, brs), 11.75 (1H, brs).

Several of other compounds of formula I have been prepared by methods substantially similar to those described herein, Example 6. The characterization data for these compounds are summarized in Table 4 below and include HPLC, CLE / EM data (observed ) and 1 H NMR.

60 Table 4. Characterization data for certain compounds of formula I

No. of II

 Compound M + 1 (obs) Tr (min) 1H NMR

2

319 9.70 (DMSO) 2.55 (3H, s), 7.36-7.45 (1H, m), 7.47-7.59 (2H, m), 7.61-7.69 (1H, m) , 7.75 -7.85 (2H, m), 7.92 -7.99 (1H, m), 8.19 -8.25 (1H, m), 8.41 -8.48 91H, m ), 8.59-8.66 91H, m), 12.34 (1H, brs).

Example 7: ITK inhibition assay

The compounds were subjected to selective detection according to their ability to inhibit ltk using a radioactive phosphate incorporation assay. The tests were carried out in buffer consisting of 100 mM HEPES (pH 7.4), 10 mM MgCl2, 25 mM NaCl, 0.01% DE BSA and 1 mM DTT at 25 ° C. in the presence of Itk 30 nM. Final substrate concentrations were 15 μM [γ-33P] ATP (400 μCi 33P ATP / μmol ATP, Amersham Pharmacia Biotech / Sigma Chemicals) and 2 μM peptide (SAM68 Δ332-443). A stock buffer solution of the assay was prepared containing all the reagents listed above with the exception of ATP and the test compound of interest. 50 μl of the stock solution was introduced into a 96-well plate, followed by the addition of 1.5 μl of mother DMSO containing serial dilutions of the test compound (usually from a final concentration of 15 μM with dilutions in series by 2) in duplicate (final concentration of DMSO 1.5%). The plate was pre-incubated for 10 minutes at 25 ° C and the reaction was initiated by the addition of 50 µl of [γ-33P] ATP (final concentration 15 µM).

The reaction was stopped after 10 minutes by adding 50 µl of a mixture of TCA / ATP (20% TCA, 0.4 mM ATP). A 96-well GF / C Unifilter plate (Perkin Elmer Life Sciences, cat. No. 6005174) was pretreated with 50 μl of Milli Q water before the addition of the total reaction mixture (150 μl). The plate was washed with 200 μl of Milli Q water, followed by 200 μl of a mixture of TCA / ATP (5% TCA, 1 mM ATP). This wash cycle was repeated 2 more times. After drying, 30 µl of an Optiphase 'SuperMix' scintillation liquid cocktail (Perkin Elmer) was added to the well before counting the scintillation (1450 Microbeta Liquid Scintillation Counter, Wallac).

The IC50 data was calculated from nonlinear regression analysis of the initial velocity data using the Prism software package (GraphPad Prism version 3.0a for Macintosh, GraphPad Software, San Diego California, USA).

The tests were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 mM MgCl 2, 0.1% BSA and 1 mM DTT. The final substrate concentrations in the assay were [γ-33P] 7.5 μM ATP (400 μCi 33P ATP / μmol ATP, Amersham Pharmacia Biotech / Sigma Chemicals) and 3 μM peptide (SAM68 protein D332-443). The tests were carried out at 25 ° C in the presence of 50 nM ltk. A stock buffer solution of the assay was prepared containing all the reagents listed above with the exception of ATP and the test compound of interest. 50 μl of the stock solution was introduced into a 96-well plate, followed by the addition of 2 μl of mother DMSO containing serial dilutions of the test compound (usually from a final concentration of 50 μM with serial dilutions per 2) in duplicate (final concentration of DMSO 2%). The plate was pre-incubated for 10 minutes at 25 ° C and the reaction was initiated by the addition of 50 µl of [γ-33P] ATP (final concentration 7.5 µM).

The reaction was stopped after 10 minutes by the addition of 100 ml of 0.2M phosphoric acid + 0.01% TWEEN

20. A 96-well plate of multi-screen phosphocellulose filter (Millipore, Cat. No. MAPHN0B50) was pretreated with 100 µl of 0.2M phosphoric acid + 0.01% TWEEN 20 before adding 170 ml of the mixture If the test was stopped, the plate was washed with 4 x 200 μl of 0.2M phosphoric acid + 0.01% TWEEN 20. After drying, 30 μl of an Optiphase 'SuperMix' scintillation liquid cocktail (Perkin Elmer) was added to the well before counting the scintillation (Liquid scintillation counter 1450 Microbeta, Wallac).

Ki data (app) was calculated from nonlinear regression analysis of initial velocity data using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego California, USA). .).

Example 8: ITK inhibition assay (AlphaScreen ™):

The compounds were subjected to selective detection according to their ability to inhibit ltk using an AlphaScreen ™ phosphotyrosine assay at Vertex Pharmaceuticals. The tests were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 mM MgCl 2, 0.1% BSA and 1 mM DTT. The final substrate concentrations in the assay were 100 μM ATP (Sigma Chemicals) and 2 μM peptide (SAM68 Δ332-443 biotinylated). The tests were carried out at 25 ° C and in the presence of ltk (30 nM). A stock buffer solution of the assay was prepared containing all the reagents listed above with the exception of ATP and the test compound of interest. 25 μl of the stock solution was introduced into each well of a 96-well plate, followed by the addition of 1 μl of DMSO with serial dilutions of the test compound (usually from a final concentration of 15 μM) in duplicate ( final concentration of DMSO 2%). The plate was pre-incubated for 10 minutes at 25 ° C and the reaction was initiated by the addition of 25 µl of ATP (final concentration 100 µM). Background counts were determined by adding 5 µl of 500 mM EDTA to the control wells containing the test buffer and DMSO before starting with ATP.

The reaction was stopped after 30 minutes by diluting the reaction 225 times in MOPS buffer (20 mM MOPS (pH 7.0), 1 mM DTT, 10 mM MgCl 2, 0.1% BSA) containing 50 mM EDTA, to carry the final concentration of SAM68biotin at 9 nM.

AlphaScreen ™ reagents were prepared according to the instructions of the test kit manufacturers (AlphaScreen ™ phosphotyrosine) (P-Tyr-100), PerkinElmer catalog number 6760620C). In dim light, 20 μl of AlphaScreen ™ reagents were introduced into each well of a white semi-area of a 96-well plate (Coming Inc. -COSTAR 3693) with 30 μl of the stopped and diluted kinase reactions. The plates were incubated in the dark for 60 minutes before reading in a Fusion Alpha plate reader (PerkinElmer).

After removing the mean background values for all data points, Ki (app) data was calculated

15 from nonlinear regression analysis using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego California, USA).

In general, the compounds of the invention, including the compounds in Table 1 and Table 2, are effective for ITK inhibition.

Example 9: ITK inhibition assay (UV):

The compounds were subjected to selective detection according to their ability to inhibit ltk using a standard coupled enzyme assay (Fox et al., Protein Sci., (1998) 7, 2249). The tests were carried out in a

25 mixture of 20 mM MOPS (pH 7.0), 10 mM MgCl2, 0.1% BSA and 1 mM DTT, 2.5 mM phosphoenolpyruvate, 300 μM NADH, 30 μg / ml pyruvate kinase and 10 μg / ml of lactate dehydrogenase. The final substrate concentrations in the assay were 100 μM ATP (Sigma Chemicals) and 3 μM peptide (SAM68 Δ332-443 biotinylated). The tests were carried out at 25 ° C and in the presence of 100 nM ltk.

A stock buffer solution of the assay was prepared containing all the reagents listed above with the exception of ATP and the test compound of interest. 60 μl of the stock solution was introduced into each well of a 96-well plate, followed by the addition of 2 μl of mother DMSO containing serial dilutions of the test compound (usually from a final concentration of 15 μM). The plate was pre-incubated for 10 minutes at 25 ° C and the reaction was initiated by the addition of 5 µl of ATP. The initial reaction rates are

35 determined with a Molecular Devices SpectraMax Plus plate reader for a period of 10 minutes. The IC50 and Ki data were calculated from nonlinear regression analysis using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego California, USA).

In general, the compounds of the invention, including the compounds in Table 1 and Table 2, are effective for ITK inhibition.

Example 10: BTK inhibition assay:

The compounds were subjected to selective detection according to their ability to inhibit BTJ using an assay of

45 incorporation of radioactive phosphate in Vertex Pharmaceuticals. The tests 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. The final substrate concentrations in the assay were 100 μM ATP (Sigma Chemicals) and 5 μM peptide (SAM68 Δ332-443). The tests were carried out at 25 ° C and in the presence of Btk (25 nM) and [γ-33P] ATP (100μCi 33P ATP / µmol ATP, Amersham Pharmacia Biotech, Amersham, United Kingdom). A stock buffer solution of the assay was prepared containing all the reagents listed above with the exception of SAM68 and the test compound of interest. 75 μl of the stock solution was introduced into a 96-well plate, followed by the addition of 1.5 μl of mother DMSO containing serial dilutions of the test compound (usually from a final concentration of 15 μM in duplicate ( final concentration of DMSO 1.5%.) The plate was pre-incubated for 15 minutes at 25 ° C and the reaction was initiated by the addition of 25 µl of SAM68 (final concentration 5 µM). Background counts were

55 determined by adding 50 µl of 20% TCA + 0.4 mM ATP to the control wells containing the test buffer and DMSO before starting with SAM68.

The reaction was stopped after 60 minutes by the addition of 50 µl 20% TCA / + 0.4 mM ATP. A 96-well GF / C Unifilter plate (Perkin Elmer Life Sciences, cat. No. 6005174) was pretreated with 50 μl of Milli Q water before the addition of the total reaction mixture (150 μl). The plate was washed with 200 μl of Milli Q water, followed by 200 μl of 5% TCA / 1 mM ATP. The water wash / TCA cycle was repeated 2 more times. After drying, 30 µl of an Optiphase 'SuperMix' scintillation liquid cocktail (Perkin Elmer) was added to the well before counting the scintillation (1450 Microbeta Liquid Scintillation Counter, Wallac).

65 After removing the average background values for all data points, Ki (app) data was calculated from nonlinear regression analysis using the Prism software package (GraphPad Prism version 3.0a for Macintosh, GraphPad Software, San Diego California, USA).

The compounds were subjected to selective detection according to their ability to inhibit Btk using an AlphaScreen ™ phosphotyrosine assay in Vertex Pharmaceuticals. The tests were carried out in a mixture of MOPS

20 mM (pH 7.0), 10 mM MgCl2, 0.1% BSA and 1 mM DTT. The final substrate concentrations in the assay were 50 μM ATP (Sigma Chemicals) and 2 μM peptide (SAM68 Δ332-443 biotinylated). The tests were carried out at 25 ° C and in the presence of Btk (25 nM). A stock buffer solution of the assay was prepared containing all the reagents listed above with the exception of SAM68-Biotin and the test compound of interest. 37.5 μl of the stock solution was introduced into each well of a 96-well plate, followed by the addition of 1 μl of DMSO with serial dilutions of the test compound (usually from a final concentration of 15 μM) per duplicated (final concentration of DMSO 2%). The plate was pre-incubated for 15 minutes at 25 ° C and the reaction was initiated by the addition of 12.5 μl of SAM68-Biotin (final concentration 2 μM). Background counts were determined by adding 5 µl of 500 mM EDTA to the control wells containing the test buffer and DMSO before starting with Biotin-SAM68.

The reaction was stopped after 30 minutes by diluting the reaction 225 times in MOPS buffer (20 mM MOPS (pH 7.0), 1 mM DTT, 10 mM MgCl 2, 0.1% BSA) containing 50 mM EDTA, to carry the final concentration of SAM68biotin at 9 nM.

AlphaScreen ™ reagents were prepared according to the instructions of the test kit manufacturers (AlphaScreen ™ phosphotyrosine) (P-Tyr-100), PerkinElmer catalog number 6760620C). In dim light, 20 μl of AlphaScreen ™ reagents were introduced into each well of a white semi-area of a 96-well plate (Coming Inc. -COSTAR 3693) with 30 μl of the stopped and diluted kinase reactions. The plates were incubated in the dark for 60 minutes before reading in a Fusion Alpha plate reader (PerkinElmer).

25 After eliminating the average background values for all data points, Ki (app) data was calculated from nonlinear regression analysis using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego California, USA).

In general, the compounds of the invention, including the compounds in Table 1 and Table 2, are effective for Btk inhibition.

Example 11: RLK inhibition assay:

The compounds were subjected to selective detection according to their ability to inhibit Rlk using a standard coupled enzyme assay (Fox et al., Protein Sci., (1998) 7, 2249). The tests were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 mM MgCl 2, 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). The tests were carried out at 30 ° C and in the presence of 40 nM Rlk. The 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.

A stock buffer solution of the assay was prepared containing all the reagents listed above with the exception of ATP and the test compound of interest. 60 μl of the stock solution was introduced into each well

45 of a 96-well plate, followed by the addition of 2 μl of mother DMSO containing serial dilutions of the test compound (usually from a final concentration of 7.5 μM). The plate was pre-incubated for 10 minutes at 30 ° C and the reaction was initiated by the addition of 5 µl of ATP. The initial reaction rates were determined with a Molecular Devices SpectraMax Plus plate reader for a period of 10 minutes. The IC50 and Ki data were calculated from nonlinear regression analysis using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego California, USA).

In general, the compounds of the invention, including the compounds in Table 1 and Table 2, are effective for RLK inhibition.

55 Example 12: JAK3 inhibition test:

The compounds were subjected to selective detection according to their ability to inhibit JAK using the assay shown below. The reactions were carried out in a kinase buffer containing 100 mM HEPES (pH 7.4), 1 mM DTT, 10 mM MgCl 2, 25 mM NaCl and 0.01% BSA.

Substrate concentrations in the assay were 5 μM ATP (200 uCi / μmol ATP) and 1 μM poly Glu: Tyr. The reactions were carried out at 25 ° C and 1 Nm JAK3.

To each well of a 96-well polycarbonate plate, 1.5 µl of a candidate JAK3 inhibitor was added

65 together with 50 μl of kinase buffer containing 2 μM poly (Glu) 4Tyr and 10 μM ATP. This was then mixed and 50 µl of kinase buffer containing the 2 nM JAK3 enzyme was added to start the reaction. After 20 minutes at room temperature (25 ° C), the reaction was stopped with 50 µl of 20% trichloroacetic acid (TCA) which also contained 0.4 mM ATP. The entire contents of each well were then transferred to a plate of 96-well fiberglass filter using a TomTek cell harvester. After washing, 60 µl of scintillation liquid was added and the incorporation of 33P was detected in a Perkin Elmer TopCount.

In general, the compounds of the invention, including the compounds in Table 1 and Table 2, are effective for the inhibition of JAK (eg, JAK-3).

Claims (21)

1. A compound of formula (I): or a pharmaceutically acceptable salt thereof; in which: Ring A is an optionally substituted five-member ring selected from: x is 0, 1, or 2; Each occurrence of R1 is halogen, CN, NO2 or UmR; R2 is independently selected from Tn-R 'R3 is independently halogen -CN, NO2 or VpR'; R4 is VpR ', where p is 0 and R' is a completely unsaturated six-membered monocyclic ring with 0 -1 nitrogen atoms; R5 is independently halogen -CN, NO2 or VpR '; each occurrence of T, U or V is, independently, a C1-6 alkylidene chain, wherein the C1-6 alkylidene chain is optionally substituted with a C1-6 or -OH aliphatic group, and up to two units of C1-6 alkylidene chain methylene are optionally and independently substituted with -NR-, -S-, -O-, -CS-, -CO2-, -OCO-, -CO-, -COCO-, -CONR- , -NRCO-, -NRCO2-, -SO2NR-, -NRSO2-, -CONRNR-, -NRCONR-, OCONR-, -NRNR-, NRSO2NR-, -SO-, or -SO2-; m, n and p are each independently 0 or 1; each occurrence of R is independently hydrogen or a C1-6 aliphatic group optionally substituted with a C1-3 aliphatic group; and each occurrence of R ’is, independently, hydrogen, a C1-6 aliphatic group, a 3-8 membered, saturated, partially unsaturated or completely unsaturated monocyclic ring having 30 0 -3 heteroatoms independently selected from nitrogen, oxygen or sulfur, or an 8-12 membered bicyclic ring system, saturated, partially unsaturated or completely unsaturated having 0 -5 heteroatoms independently selected from nitrogen, oxygen or sulfur, wherein the 3-8 membered monocyclic ring and the 8-12 membered bicyclic ring are optionally substituted with halogen, -O (C1-6 aliphatic) or C (O) NH2.

2.

 The compound of claim 1, wherein p is 0.

3.

 The compound of any one of claims 1-2, wherein p is 1.

The compound of claim 3, wherein V is -NR-, -S-or -O-.

5.

The compound of any one of claims 1-4, wherein R3 is Vp-R ', wherein p is 0 and R' is hydrogen.

6.

 The compound of any one of claims 1-5, wherein R5 is halogen or Vp-R ', wherein p is 0 and R' is hydrogen or C1-6 aliphatic.

The compound of claim 6, wherein R5 is halogen or Vp-R ', wherein p is 0 and R' is hydrogen or C1-3 alkyl.

8. The compound of claim 1, wherein R1 is UmR.

The compound of claim 1, wherein R2 is TnR ', wherein n is 1.

10. The compound of claim 9, wherein T is -NR-, -O-, -CO-, -CONR- or -NRCO-. 11. The compound of claim 1, wherein R2 is TnR ', wherein n is 0. 15 12. The compound of claim 1, which has a formula selected from or a pharmaceutically acceptable salt thereof. 13. The compound of claim 12, which has a formula selected from or a pharmaceutically acceptable salt thereof. 14. The compound of any one of claims 12-13, wherein R1 is UmR, wherein m is 0 and R is H or CH3.

fifteen.

 The compound of any one of claims 12-14, wherein R2 is TnR ', wherein n is 1. 5

16. The compound of claim 15, wherein T is -NR-, -O-, -CO-, -CONR- or -NRCO-.

17.

 The compound of claim 16, wherein T is -NR-. 18. The compound of claim 17, wherein R and R 'are both C1-6 aliphatic.

19. The compound of any one of claims 12-14, wherein R2 is TnR ', wherein n is 0. 20. The compound of claim 19, wherein R 'is an N-linked heterocyclyl selected from morpholinyl, piperidinyl, pyrrolidinyl and piperazinyl, wherein the N-linked heterocyclyl is optionally substituted with aliphatic C1-6.

twenty-one.

 The compound of claim 1 selected from:

22

 A composition comprising a compound of any one of claims 1-21, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

23. The composition of claim 22, further comprising an additional therapeutic agent selected from an agent for the treatment of an autoimmune, inflammatory, proliferative or hyperproliferative disease, or an immunologically mediated disease, including rejection of transplanted organs or tissues and Acquired immunodeficiency syndrome (AIDS). A method of inhibiting the kinase activity of the Tec family in a biological sample, wherein the method comprises contacting said biological sample with a compound according to any one of claims 1-21. 25. A composition comprising a compound according to any one of claims 1-21 20 for use in the treatment or reduction of the severity of a disease or condition selected from an autoimmune, inflammatory, proliferative or hyperproliferative disease or a disease mediated by the immune system. 26. The compound or composition for use according to claim 25, further comprising a An additional therapeutic agent selected from an agent for the treatment of an autoimmune, inflammatory, proliferative or hyperproliferative disease, or an immunologically mediated disease, including rejection of transplanted organs or tissues and acquired immunodeficiency syndrome (AIDS), wherein: said additional therapeutic agent is suitable for the disease being treated; and said additional therapeutic agent is administered together with said composition as a single dosage form 30 or, separately, from said composition as part of a multiple dosage form. 27. The compound or composition for the use of claim 25 or claim 26, wherein the disease or disorder is asthma, water rhinitis, allergy, atrophic rhinitis, chronic rhinitis, membranous rhinitis, seasonal rhinitis, sarcoidosis, Farmer's lung, fibroid lung, idiopathic interstitial pneumonia, rheumatoid arthritis, 35 seronegative spondyloarthropathy (including ankylosing spondylitis, psoriatic arthritis and Reiter's disease), Behcet's disease, Sjogren's syndrome, systemic sclerosis, psoriasis, systemic sclerosis, dermatitis, dermatitis by contact and other eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigus, bullous epidermolysis, urticaria, angioedemas, vasculitidis, erythemas, cutaneous eosinophilias, uveitis, alopecia, vernal conjunctivitis, erythrocyte encephalitis, erythrocyte encephalitis, erythrocyte encephalitis, erythrocyte encephalitis, erythrocyte encephalitis, erythrocytes, erythrocytes, erythrocytes, erythrocytes, erythrocytes, edema 40 pancreatitis, Crohn's disease, ulcerative colitis, food-related allergies, multiple sclerosis, atherosclerosis, acquired immunodeficiency syndrome (AIDS), lupus erythematosus, systemic lupus erythematosus, Hashimoto's thyroiditis, myasthenia gravis, type I diabetes, nephrotic syndrome , eosinophilic fasciitis, hyper IgE syndrome, lepromatous leprosy, Sezary syndrome and idiopathic thrombocytopenic purpura, restenosis after angioplasty, tumors, atherosclerosis, systemic lupus erythematosus, allograft rejection including, without limitation, acute and chronic chronic allograft rejection , kidney, heart, liver, lung, bone marrow, skin and cornea transplant, and chronic graft versus host disease.