HK1113569B - Pyrazolo[1,5-a]pyrimidines useful as inhibitors of protein kinases - Google Patents

Description

Pyrazolo [1, 5-a ] pyrimidines useful as protein kinase inhibitors

Technical Field

The present invention relates to compounds useful as inhibitors of protein kinases, particularly Tec family kinases, Aurora kinases and c-Met. 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 of making the compounds of the invention.

Background

Tec family non-receptor tyrosine kinases play a central role in antigen-receptor signaling, such as TCR, BCR and fcepsilon receptors, and are essential for T-cell activation. Deletion of Itk in mice results in reduced proliferation and secretion of T-cell receptor (TCR) -induced cytokines IL-2, I L-4, IL-5, IL-10, and IFN- γ. Itk has been implicated in allergic asthma and atopic dermatitis.

Tec family kinases are also essential for B-cell development and activation. Patients with Btk mutations have significant B-cell developmental block, resulting in almost complete depletion of B lymphocytes and plasma cells, severe reduction in Ig levels, and significant suppression of humoral responses to recall antigens. Btk deletion in mice has a clear effect on B-cell proliferation induced by anti-IgM and suppresses the immune response of thymus-independent type II antigens.

Tec kinases also play a role in mast cell activation through the high affinity IgE receptor (fcsri). Itk and Btk are expressed in mast cells and activated by fceri cross-coupling. Btk deficient murine mast cells, after cross-coupling to fceri, have reduced degranulation and reduced production of proinflammatory cytokines. Btk deficiency also results in a decrease in macrophage effector function.

Aurora proteins are three highly related families of serine/threonine kinases (termed Aurora-a, -B and-C) that are required for the progression of cells through the mitotic phase of the cell cycle. In particular, Aurora-a plays a crucial role in centrosome maturation and segregation, mitotic spindle formation and reliable chromosome segregation. Aurora-B is a chromosomal messenger protein that plays a central role in regulating chromosome alignment on metaphase plates, spindle assembly checkpoints, and the correct completion of cytokinesis.

Overexpression of Aurora-A (Aurora-2), Aurora-B (Aurora-1), or Aurora-C has been observed in some human cancers, including colorectal, ovarian, gastric, and invasive ductal adenocarcinomas.

Several studies have now demonstrated that Aurora-a or-B in human cancer cell lines is reduced or inhibited by siRNA, dominant negative or neutralizing antibodies by the progression of disruption through mitosis, with accumulation of 4NDNA cells. In some cases, this is followed by internal doubling and cell death.

The c-Met receptor tyrosine kinase is overexpressed in a number of human cancers and amplified during the transition between primary tumors and metastases. Various cancers in which c-Met is implicated include, but are not limited to, gastric adenocarcinoma, renal carcinoma, small cell lung carcinoma, colorectal carcinoma, colon carcinoma, prostate carcinoma, brain carcinoma, liver carcinoma, pancreatic carcinoma, and breast carcinoma. c-Met has also been implicated in atherosclerosis, pulmonary fibrosis, allergic disorders, autoimmune disorders and conditions associated with organ transplantation.

Therefore, there is an urgent need to develop compounds useful as protein kinase inhibitors. Specifically, there will be a need to develop compounds that are useful as inhibitors of the Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) and Aurora family protein kinases and c-Met, and in particular, are currently inadequate for the treatment of most disorders involving their activation.

Summary of The Invention

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

These compounds and pharmaceutically acceptable compositions thereof are useful in the treatment or prevention of various diseases, disorders, or conditions. The compounds provided by the invention are also useful in kinase studies in biological and pathological phenomena; the study of intracellular signal transduction pathways mediated by such kinases; and comparative evaluation of novel kinase inhibitors.

Detailed description of the invention

The present invention relates to compounds of formula I:

or a pharmaceutically acceptable salt thereof, wherein

R is optionally substituted- (C ═ Q) R^2aOr optionally substituted Y;

y is a 5-10 membered monocyclic or bicyclic heterocyclyl, aryl or heteroaryl ring;

q is a heteroatom selected from O, N or S; and

R^2ais C_1-6Aliphatic radical, aryl radical, heteroaryl radical, OR⁵Or N (R)⁵)₂；

R¹Is H or C_1-6An aliphatic group;

z is a bond or C_1-6An aliphatic group wherein up to two methylene units of the aliphatic chain are optionally and independently replaced in a chemically stable arrangement with a heteroatom selected from O, N or S;

R³and R⁴Each independently of the others being H, halogen, C_1-6Aliphatic radical, C_1-6Alkoxy, N (R)⁵)₂、CN、NO₂Or U_m-V, wherein m is 0 or 1;

v is H, aryl, heteroaryl, cycloaliphatic, heterocyclic or C_1-12An aliphatic group wherein up to two methylene units of the alkylene chain are optionally and independently replaced in a chemically stable arrangement by a heteroatom selected from O, N or S; v is optionally substituted by R⁸Substitution;

u is C_1-12An alkylene chain wherein up to two methylene units of the chain are optionally and independently replaced by-NR⁵-、-O-、-S-、-CO₂-、-OC(O)-、-C(O)CO-、-C(O)-、-C(O)NR⁵-、-NR⁵CO-、-NR⁵C(O)O-、-SO₂NR⁵-、-NR⁵SO₂-、-C(O)NR⁵NR⁵-、-NR⁵C(O)NR⁵-、-OC(O)NR⁵-、-NR⁵NR⁵-、-NR⁵SO₂NR⁵-、-SO-、-SO₂-、-PO-、-PO₂-or-POR⁵-replaced by a chemically stable arrangement;

R⁵is H, C_1-4Haloalkyl, -C (O) COR⁶、-C(O)R⁶、-C(O)OR⁶、-C(O)N(R⁶)₂、-SO₂R⁶、C_0-6Alkyl-heterocyclyl radical, C_0-6Alkyl-heteroaryl, C_0-5Alkyl-aryl, C_0-6Alkyl-cycloaliphatic radicals or C_1-6An aliphatic group wherein up to two methylene units of the aliphatic chain are optionally and independently replaced in a chemically stable arrangement with a heteroatom selected from O, N or S;

R⁶is H, C_1-6Alkoxy radical, C_1-4Haloalkyl, C_0-6Alkyl-heterocyclyl radical, C_0-6Alkyl-heteroaryl, C_0-6Alkyl-aryl, C_0-6Alkyl-cycloaliphatic radicals or C_1-6Aliphatic radicals in which up to two members of the aliphatic chain are presentThe methyl units are optionally and independently replaced by a heteroatom selected from O, N or S in a chemically stable arrangement; or two R⁶The groups together with the atoms to which they are attached are optionally joined to form a 5-10 membered carbocyclic or heterocyclic ring;

R⁸is halogen, -OR⁶、-N(R⁶)₂、-SR⁶、NO₂、CN、-COOR⁶、-C(O)N(R⁶)₂、-SO₂R⁶、-SO₂N(R⁶)₂、-NR⁶C(O)R⁶、-C(O)R⁶、-OC(O)R⁶、-NR⁶C(O)O-R⁶、-NR⁶SO₂-R⁶、-C(O)NR⁶N(R⁶)₂、-NR⁶C(O)N(R⁶)₂、-OC(O)N(R⁶)₂、-NR⁶N(R⁶)₂、-NR⁶SO₂N(R⁶)₂Or C_1-12An aliphatic radical in which up to three methylene units of the aliphatic chain may optionally be replaced by-C (O) N (R)⁶)-、-NR⁶CO(R⁶)-、-O-、-NR⁶-or-S-interrupted;

the conditions are as follows:

when Z is a bond, R is not 2- (phenylamino) -pyrimidin-4-yl, wherein phenyl is optionally substituted;

when Z is a bond, R is not- (C ═ O) N (R)^a)₂、-(C＝O)R^bOR- (C ═ O) OR^bWherein R is^aIs H, C_1-6Aliphatic radical, C_3-10Cycloaliphatic, aryl, heteroaryl, heterocyclic radical, C_0-6Alkyl- (C ═ O) N (R)^a)₂、C_0-6alkyl-SOR^b、C_0-6alkyl-SO₂R^b、C_0-6alkyl-CO₂R^b、C_0-6alkyl-CO₂H、C_0-6alkyl-OR^b、C_0-6alkyl-OH, C_0-6alkyl-N (R)^a)₂、C_0-6Alkyl- (C ═ O) -C_0-6Alkyl-OR^bOr C_0-6Alkyl- (C ═ O) -C_0-6alkyl-OH, and

R^bis C_1-6Aliphatic radical, C_3-10Cycloaliphatic, aryl, heteroaryl or heterocyclic groups;

when Z is a bond, R is not substituted by C_0-6Alkyl- (C ═ O) N (R)^a)₂、C_0-6alkyl-SO_nR^b、C_0-6alkyl-SO_nH、C_0-6alkyl-CO₂R^b、C_0-6alkyl-CO₂H、C_1-6alkyl-OR^b、C_1-6alkyl-OH, C_1-6alkyl-N (R)^a)₂、C_0-6Alkyl- (C ═ O) -C_0-6alkyl-OR^b、C_0-6Alkyl- (C ═ O) -C_0-6alkyl-OH;

wherein n is 0, 1 or 2; r^aAnd R^bIs as defined above;

when Z is a bond, R³Is not an optionally substituted dihydropyrimidinone or dihydropyridone ring;

when Z is a bond, R is not optionally substituted phenyl.

In some embodiments, Z is a bond. In some embodiments, R¹Is H.

In some embodiments, Y is represented by the formula:

wherein

Q is a heteroatom selected from O, N or S;

R⁷is halogen, -OR⁶、-N(R⁶)₂、-SR⁶、NO₂、CN、-COOR⁶、-C(O)N(R⁶)₂、-SO₂R⁶、-SO₂N(R⁶)₂、-NR⁶C(O)R⁶、-C(O)R⁶、-OC(O)R⁶Or C_1-12An aliphatic radical in which up to three methylene units of the aliphatic chain may optionally be replaced by-C (O) N (R)⁶)-、-NR⁶CO(R⁶)-、-O-、-NR⁶-or-S-interrupted;

is a single or double bond;

R^2bis carbon or a heteroatom selected from O, N or S;

R^2band Q together with the carbon atom to which they are attached form a 3-7 membered saturated or unsaturated monocyclic ring having 0-4 heteroatoms selected from O, N or S, or an 8-10 membered saturated or unsaturated bicyclic ring having 0-6 heteroatoms selected from O, N or S. In a further embodiment, R^2bIs carbon.

In other embodiments, R is

In further embodiments, Q is N or O.

In other embodiments, R is Y. In a further embodiment, Y is 5-10 membered heteroaryl or heterocyclyl. In a further embodiment, Y is an optionally substituted 5-6 membered heteroaryl or heterocyclyl. In further embodiments, Y is optionally substituted with R⁷A substituted pyridine ring. In still further embodiments, Y is optionally substituted with R⁷A substituted 2-pyridine ring. In some embodiments, R⁷Is halogen, OR⁶、-N(R⁶)₂、-SR⁶、NO₂、CN、-OC(O)R⁶、-NR⁶C(O)R⁶、-SO₂N(R⁶)₂or-NR⁶SO₂-。

In other embodiments, R³And R⁴Each independently is U_m-V. In further embodiments, V is optionally substituted with R⁸Substituted aryl, heteroaryl, cycloaliphatic or heterocyclic. In a further embodiment, R³Is H, R⁴Is U_m-V; u is-C (O) NR⁵(ii) a m is 1. At R³And/or R⁴In other embodiments, m is 0 and V is aryl or heteroaryl. In a further embodiment, V is unsubstituted phenyl. In other embodiments, V is substituted with R⁸A substituted phenyl group. In other embodiments, V is substituted with R⁸A substituted heteroaryl group. In a further embodiment, V is substituted with R⁸A substituted heteroaryl group. In a further embodiment, V is substituted with R⁸A substituted pyridyl group. In a further embodiment, R⁸Is halogen, OH, CN, NH₂、OR⁶Or C_1-12An aliphatic group. In a further embodiment, R⁸Is C_1-12An aliphatic radical, wherein up to three methylene units of the aliphatic chain may optionally be replaced by-C (O) NR⁶、-NR⁶CO-or a heteroatom selected from O, N and S. In a further embodiment, R⁸is-C (O) OR⁶or-C (O) N (R)⁶)₂. In a further embodiment, R⁶Is H, C_1-6Alkyl-heterocyclyl radical, C_1-6Alkyl-heteroaryl, C_1-6Alkyl-aryl, C_1-6Alkyl-cycloaliphatic radicals.

In some embodiments, Z is C_1-6Alkyl wherein zero methylene units are replaced by a heteroatom selected from O, N or S; v is aryl or heteroaryl. In a further embodiment, Z is-CH₂-。

In some embodiments, the present invention provides a compound selected from the group consisting of:

in some embodiments, the present invention provides a composition comprising a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent. In further embodiments, the composition further comprises an ingredient for treating an autoimmune, inflammatory, proliferative, hyperproliferative disease, or immunologically-mediated disease, including rejection of transplanted organs or tissues and acquired immunodeficiency syndrome (AIDS).

The invention also provides a method of inhibiting Tec family kinase activity in a patient or a biological sample comprising administering to said patient or contacting said biological sample with a compound of the invention or a composition comprising said compound. In a further embodiment, the method comprises inhibiting Itk activity. The invention also provides a method of treating or lessening the severity of a disease or condition selected from: an autoimmune, inflammatory, proliferative, or hyperproliferative disease, or an immunologically-mediated disease, comprising administering to a patient in need thereof a compound of the present invention or a composition comprising said compound. In a further embodiment, the method comprises administering to said patient an additional therapeutic agent selected from the group consisting of a component for treating an autoimmune, inflammatory, proliferative, hyperproliferative disease, or immunologically-mediated disease, including rejection of transplanted organs or tissues and Acquired Immune Deficiency Syndrome (AIDS), wherein said additional therapeutic agent is appropriate for the disease being treated; the additional therapeutic agent is administered with the composition as a single dosage form or separately from the composition as part of multiple dosage forms.

In a further embodiment, the disease or condition is asthma, acute rhinitis, allergy, atrophic rhinitis, chronic rhinitis, membranous rhinitis, seasonal rhinitis, sarcoidosis, farmer's lung, fibroid lung, idiopathic interstitial pneumonia, rheumatoid arthritis, seronegative spondyloarthropathies (including ankylosing spondylitis, psoriatic arthritis, and reiter's disease), behcet's disease, sjogren's syndrome, systemic sclerosis, psoriasis, systemic sclerosis, atopic dermatitis, contact dermatitis and other eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigus, epidermolysis bullosa, urticaria, vascular dermatitis, vasculitides, erythroderma, cutaneous eosinophilia, uveitis, alopecia, vernal conjunctivitis clusion, celiac disease, proctitis, chronic rhinitis, membranous rhinitis, seasonal rhinitis, sarcoidosis, sjogren's syndrome, systemic sclerosis, atopic dermatitis, and other eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigoitis, pemphigus vulgaris, eosinophilic gastroenteritis, mastocytosis, pancreatitis, crohn's disease, ulcerative colitis, food-related allergy, multiple sclerosis, atherosclerosis, acquired immunodeficiency syndrome (AIDS), lupus erythematosus, systemic lupus, erythema, hashimoto's thyroiditis, myasthenia gravis, type I diabetes, nephrotic syndrome, eosinophilic fasciitis, hyper IgE syndrome, leprosy, sexil syndrome and idiopathic thrombocytopenic purpura, post-angioplasty restenosis, tumors, atherosclerosis, systemic lupus erythematosus, allograft rejection, including, without limitation, acute and chronic allograft rejection, e.g., secondary to post-transplantation of the kidney, heart, liver, lung, bone marrow, skin and cornea; and chronic graft-host disease.

In other embodiments, the invention provides compounds of formula I

Or a pharmaceutically acceptable salt thereof, wherein

R is- (C ═ Q) R^2aCN or Y; wherein

Y is a 5-10 membered monocyclic or bicyclic heterocyclyl, aryl or heteroaryl ring; each Y is independently and optionally substituted with 0-5J^YSubstitution;

q is O, NH, NR' or S;

r' is C_1-6Alkyl, optionally halogenated with 0 to 4 occurrences, C_1-6Aliphatic radical, NO₂、NH₂、-N(C_1-6Alkyl), -N (C)_1-6Alkyl radical)₂、SH、-S(C_1-6Alkyl), OH, -O (C)_1-6Alkyl), -C (O) (C)_1-6Alkyl), -C (O) NH₂、-C(O)N(C_1-6Alkyl) or-C (O) N (C)_1-6Alkyl radical)₂Substitution;

R^2ais C_1-6Aliphatic radical, C_6-10Aryl, 5-10 membered heteroaryl, 5-10 membered heterocyclyl, OR⁵Or N (R)⁵)₂(ii) a Each R^2aIndependently and optionally by 0-5J^2aSubstitution;

R¹is H, -C (O) (C)_1-6Alkyl), -C (O) O (C)_1-6Alkyl), -C (O) NH₂、-C(O)N(C_1-6Alkyl), -C (O) N (C)_1-6Alkyl radical)₂Or C_1-6An aliphatic group; each R¹Optionally halogenated groups present 0 to 4 times, C_1-6Haloalkyl, C_1-6Aliphatic radical, NO₂、NH₂、-N(C_1-6Alkyl), -N (C)_1-6Alkyl radical)₂、SH、-S(C_1-6Alkyl), OH or-O (C)_1-6Alkyl) substitution;

z is a bond or C_1-6Aliphatic radical ofWherein at most two methylene units of the aliphatic chain are optionally and independently replaced by a heteroatom selected from O, N or S in a chemically stable arrangement;

R³and R⁴Each independently of the others being H, halogen, C_1-6Alkoxy, N (R)⁵)₂、CN、NO₂Or U_m-V, wherein m is 0 or 1;

v is H, C_6-10Aryl, 5-10 membered heteroaryl, C_3-10Cycloaliphatic, 5-to 10-membered heterocyclic radical or C_1-12An aliphatic group wherein up to two methylene units of the alkylene chain are optionally and independently replaced in a chemically stable arrangement by a heteroatom selected from O, N or S; v is optionally substituted by 0-4R⁸Substitution;

u is C_1-12An alkylene chain wherein up to two methylene units of the chain are optionally and independently replaced by-NH-, -NR⁵-、-O-、-S-、-CO₂-、-OC(O)-、-C(O)CO-、-C(O)-、-C(O)NH-、-C(O)NR⁵-、-C(＝N-CN)、-NHCO-、-NR⁵CO-、-NHC(O)O-、-NR⁵C(O)O-、-SO₂NH-、-SO₂NR⁵-、-NHSO₂-、-NR⁵S₂-、-NHC(O)NH-、-NR⁵C(O)NH-、-NHC(O)NR⁵-、-NR⁵C(O)NR⁵-、-OC(O)NH-、-OC(O)NR⁵-、-NHNH-、-NHNR⁵-、-NR⁵NR⁵-、-NR⁵NH-、-NHSO₂NH-、-NR⁵SO₂NH-、-NHSO₂NR⁵-、-NR⁵SO₂NR⁵-、-SO-、-SO₂-、-PO-、-PO₂-or-POR⁵-substituted; u is optionally substituted by 0-6J^USubstitution;

R⁵is C_1-4Haloalkyl, -C (O) COR⁶、-C(O)R⁶、-C(O)OR⁶、-C(O)N(R⁶)₂、-SO₂R⁶、C_0-6Alkyl-heterocyclyl radical, C_0-6Alkyl-heteroaryl, C_0-6Alkyl-aryl, C_0-6Alkyl-cycloaliphatic estersGroup or C_1-6An aliphatic radical wherein up to three methylene units of the aliphatic chain are optionally and independently replaced by-NR' -, -O-, -S-, -CO₂-、-OC(O)-、-C(O)CO-、-C(O)-、-C(O)NR″-、-NR″CO-、-NR″C(O)O-、-SO₂NR″-、-NR″SO₂-、-C(O)NR″NR″-、-NR″C(O)NR″-、-OC(O)NR″-、-NR″NR″-、-NR″SO₂NR″-、-SO-、-SO₂-、-PO-、-PO₂-or-POR "-is replaced by a chemically stable arrangement; each R⁵Independently and optionally by 0-5J^R5Substitution; or two R⁵The groups together with the atoms to which they are attached are optionally joined to form a 5-10 membered carbocyclic or heterocyclic ring; wherein said ring is optionally substituted with 0-4J';

R⁶is H, C_1-6Alkoxy radical, C_1-4Haloalkyl, C_0-6Alkyl-heterocyclyl radical, C_0-6Alkyl-heteroaryl, C_0-6Alkyl-aryl, C_0-6Alkyl-cycloaliphatic radicals or C_1-6An aliphatic group wherein up to two methylene units of the aliphatic chain are optionally and independently replaced in a chemically stable arrangement with a heteroatom selected from O, N or S; each R⁶Independently and optionally by 0-5J^R6Substitution; or two R⁶The groups together with the atoms to which they are attached are optionally joined to form a 5-10 membered carbocyclic or heterocyclic ring; wherein said ring is optionally substituted with 0-4J';

R⁸is halogen, C_1-4Haloalkyl, phenyl, 5-8 membered heterocyclyl, 5-6 membered heteroaryl, -OR⁶、-N(R⁶)₂、-SR⁶、NO₂、CN、-COOR⁶、-C(O)N(R⁶)₂、-SO₂R⁶、-SO₂N(R⁶)₂、-NR⁶C(O)R⁶、-C(O)R⁶、-OC(O)R⁶、-NR⁶C(O)O-R⁶、-NR⁶SO₂-R⁶、-C(O)NR⁶N(R⁶)₂、-NR⁶C(O)N(R⁶)₂、-OC(O)N(R⁶)₂、-NR⁶N(R⁶)₂、-NR⁶SO₂N(R⁶)₂Or C_1-12An aliphatic radical, wherein up to three methylene units of the aliphatic chain may optionally be replaced by-C (O) R⁶、-C(O)O-、-OC(O)-、-C(O)-、-C(O)N(R⁶)-、-NR⁶CO(R⁶)-、-O-、-NR⁶-or-S-interrupted; each R⁸Independently and optionally by 0-5J^R8Substitution;

each J^Y、J^2a、J^U、J^R5、J^R6、J^R8J 'and J' are independently selected from N (R)⁹)₂、SR⁹、OR⁹Halo, CN, NO₂、COOR⁹、C(O)R⁹、SO₂R⁹、SOR⁹、-X-CF₃、-X-SH、-X-OH、C_1-4Haloalkyl, C_6-10Aryl, -X- (C)_6-10Aryl), 5-10 membered heteroaryl, -X- (5-10 membered heteroaryl), C_3-10Cycloaliphatic radical, -X- (C)_3-10Cycloaliphatic), 5-10 membered heterocyclyl, -X- (5-10 membered heterocyclyl), or X;

x is C_1-12An aliphatic radical in which up to two methylene units of the alkylene chain are optionally and independently replaced by-NH-, -NR' -, -O-, -S-, -CO₂-、-OC(O)-、-C(O)CO-、-C(O)-、-C(O)NH-、-C(O)NR″-、-C(＝N-CN)、-NHCO-、-NR″CO-、-NHC(O)O-、-NR″C(O)O-、-SO₂NH-、-SO₂NR″-、-NHSO₂-、-NR″SO₂-、-NHC(O)NH-、-NR″C(O)NH-、-NHC(O)NR″-、-NR″C(O)NR″、-OC(O)NH-、-OC(O)NR″-、-NHNH-、-NHNR″-、-NR″NR″-、-NR″NH-、-NHSO₂NH-、-NR″SO₂NH-、-NHSO₂NR″-、-NR″SO₂NR″-、-SO-、-SO₂-、-PO-、-PO₂-or-POR "-is replaced by a chemically stable arrangement; wherein R' is H or C_1-6An aliphatic group;

each J^Y、J^2a、J^U、J^R5、J^R6J 'and J' are optionally and independentlyN (R) occurring 0-4 times in situ⁹)₂、SR⁹、OR⁹Halo, CN, NO₂、COOR⁹、C(O)R⁹、SO₂R⁹、SOR⁹、-X-CF₃、-X-SH、-X-OH、C_1-4Haloalkyl, C_6-10Aryl, -X- (C)_6-10Aryl), 5-10 membered heteroaryl, -X- (5-10 membered heteroaryl), C_3-10Cycloaliphatic radical, -X- (C)_3-10Cycloaliphatic), 5-10 membered heterocyclyl, -X- (5-10 membered heterocyclyl), or X;

R⁹is H, C_1-6Aliphatic radical, C_1-4Haloalkyl, C_6-10Aryl, -X- (C)_6-10Aryl), 5-10 membered heteroaryl, -X- (5-10 membered heteroaryl), C_3-10Cycloaliphatic radical, -X- (C)_3-10Cycloaliphatic), 5-10 membered heterocyclyl, -X- (5-10 membered heterocyclyl), or X, or wherein two R' s⁹Together with the atoms to which they are attached form a 5-10 membered heterocyclyl, wherein said heterocyclyl is optionally substituted with 0-4 occurrences of halo, CN, NO₂、-COOH、-COO(C_1-6Alkyl), -C (O) H, SO₂H、SO₂(C_1-6Alkyl group), C_1-6Halogenated aliphatic radical, NH₂、-NH(C_1-6Alkyl), -N (C)_1-6Alkyl radical)₂、SH、-S(C_1-6Alkyl), OH, -O (C)_1-6Alkyl), -C (O) (C)_1-6Alkyl), -C (O) NH₂、-C(O)NH(C_1-6Alkyl), -C (O) N (C)_1-6Alkyl radical)₂、-C(O)NH₂、-C(O)NH(C_1-6Alkyl), -C (O) N (C)_1-6Alkyl radical)₂、C_1-4Haloalkyl, C_6-10Aryl, -X- (C)_6-10Aryl), 5-10 membered heteroaryl, -X- (5-10 membered heteroaryl), C_3-10Cycloaliphatic radical, -X- (C)_3-10Cycloaliphatic), 5-10 membered heterocyclyl, -X- (5-10 membered heterocyclyl), or X.

In some embodiments of formula I', if R⁴Is H, then R³Is not H. In a further embodiment, R⁴Is H, R³Is not H. In other embodiments, Z is a bond. In thatIn other embodiments, R¹Is H.

In some embodiments of formula I', R is

In further embodiments, Q is N or O. In a further embodiment, R^2aIs OR⁵、N(R⁵)₂Or a 5-8 membered heterocyclic group. In a further embodiment, the 5-8 membered heterocyclyl contains 1-2 nitrogen atoms. In still further embodiments, the 5-8 membered heterocyclyl is optionally substituted with 0-4 occurrences of C_1-6Aliphatic radical, C_1-4Haloalkyl, CN, halo, OH, O- (C)_1-6Aliphatic group), NH₂、NH(C_1-6Aliphatic radical), N (C)_1-6Aliphatic radical)₂Benzyl, -CH₂- (pyridyl) or-CH₂-pyrrolidinyl substitution.

In some embodiments of formula I', R^2aIs N (R)⁵)₂. In a further embodiment, R⁵Is H or an optionally substituted group selected from 5-8 membered heterocyclyl, - (C)_1-6Alkyl) - (5-8 membered heterocyclyl), 5-6 membered heteroaryl, - (C)_1-6Alkyl) - (5-6 membered heteroaryl), phenyl, - (C)_1-6Alkyl) - (phenyl), C_3-10Cycloaliphatic radical, - (C)_1-6Alkyl group) - (C_3-10Cycloaliphatic radical) and C_1-6An aliphatic group wherein up to three methylene units of the aliphatic chain are optionally and independently replaced by-NR' -, -O-, or-S-in a chemically stable arrangement. In still further embodiments, R⁵Is H or an optionally substituted radical selected from pyrrolidinyl, piperidinyl, piperazinyl, -CH₂- (5-to 6-membered heteroaryl), phenyl, benzyl and C_1-6An aliphatic group wherein at most one methylene unit of the aliphatic chain is optionally and independently replaced by-NR' -, -O-, or-S-in a chemically stable arrangement.

In some embodiments of formula I', J^R5Is halo, CN, C_1-4Halo or optionally substituted group selected from phenyl, benzyl, 5-8 membered heterocyclyl, 5-6 membered heteroaryl, CH₂- (5-to 6-membered heteroaryl), CH₂- (5-to 8-membered heterocyclic group) and C_1-6An aliphatic radical wherein up to two methylene units of the alkylene chain are optionally and independently replaced by-NR' -, -O-, -S-, -C (O) -, -SO-, or-SO₂-replaced by a chemically stable arrangement. In a further embodiment, J^R5Is halogeno, CN, phenyl, benzyl, CH₂- (pyridyl), CH₂- (pyrrolidinyl) or C_1-6An aliphatic group wherein up to two methylene units of the alkylene chain are optionally and independently replaced by-NR' -, -O-, or-S-.

In some embodiments of formula I', R is Y. In a further embodiment, Y is an optionally substituted 5-10 membered heteroaryl or heterocyclyl. In a further embodiment, Y is an optionally substituted 5-6 membered heteroaryl or 5-8 membered heterocyclyl. In still further embodiments, Y is optionally substituted with 0-4J^YA substituted pyridine ring. In further embodiments, Y is optionally substituted with 0-4J^YA substituted 2-pyridine ring.

In some embodiments of formula I', J^YIs halo, CN, NO₂、C_1-6Halogenated aliphatic group, phenyl, benzyl, 5-6 membered heteroaryl, C_1-6Alkyl- (5-6 membered heteroaryl), C_3-10Cycloaliphatic radical, (C)_1-6Alkyl group) - (C_3-10Cycloaliphatic), 5-to 8-membered heterocyclic group, C_1-6Alkyl- (5-8 membered heterocyclyl) or C_1-12An aliphatic radical in which up to two methylene units of the alkylene chain are optionally and independently replaced by-NR-, -O-, -S-, -C (O) -, -SO-or-SO₂-replaced by a chemically stable arrangement. In a further embodiment, J^YIs halo, CN, NO₂、CF₃、C_1-6Aliphatic radical, phenyl, benzyl, -O-benzyl, piperidinyl, pyrrolidinyl, -NR (C)_1-6Alkyl), -O (C)_1-6Alkyl), -S (C)_1-6Alkyl), OH, SH or NH₂。

In some embodiments of formula I', R³And R⁴Each independently is U_m-V. In a further embodiment, V is H, C_6-10Aryl, 5-10 membered heteroaryl, C_3-10Cycloaliphatic or 5-10 membered heterocyclyl, optionally substituted with 0-4R⁸And (4) substitution. In a further embodiment, V is an optionally substituted group selected from phenyl, 5-6 membered heteroaryl or 5-8 membered heterocyclyl. In a further embodiment, V is an optionally substituted group selected from phenyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolidinyl, piperidinyl, or morpholinyl. In still further embodiments, V is an optionally substituted phenyl.

In some embodiments of formula I', R³Is U_m-V; m is 1, U is C_1-6An aliphatic radical wherein up to two methylene units of the aliphatic chain are optionally and independently replaced by-C (O) -, -C (O) NR⁵or-C (O) O-. In a further embodiment, U is-C (O) NR⁵-. In still further embodiments, V is an optionally substituted phenyl or an optionally substituted pyridyl.

In some embodiments of formula I', R³Is U_m-V; m is 0 and V is aryl or heteroaryl.

In some embodiments of formula I', R⁸Is halogen, C_1-4Haloalkyl, phenyl, 5-8 membered heterocyclyl, 5-6 membered heteroaryl, -OR⁶、-N(R⁶)₂、-SR⁶、NO₂、CN、-COOR⁶、-C(O)N(R⁶)₂、-SO₂R⁶、-SO₂N(R⁶)₂、-NR⁶C(O)R⁶、-C(O)R⁶、-NR⁶SO₂-R⁶、-C(O)NR⁶N(R⁶)₂Or C_1-12An aliphatic radical, wherein up to three methylene units of the aliphatic chain may optionally be replaced by-C (O) R⁶、-C(O)O-、-OC(O)-、-C(O)-、-C(O)N(R⁶)-、-NR⁶CO(R⁶)-、-O-、-NR⁶-or-S-interrupted. In a further embodiment, R⁸is-OR⁶、-N(R⁶)₂、C(O)R⁶、-C(O)N(R⁶)₂Or a 5-7 membered heterocyclic group. In still further embodiments, R⁸is-C (O) N (R)⁶)₂Or C (O) R⁶。

In some embodiments of formula I', R⁶Is H or an optionally substituted group selected from 5-8 membered heterocyclyl, - (C)_1-6Alkyl) - (5-to 8-membered heterocyclyl), benzyl, - (C)_1-6Alkyl) - (5-8 membered heteroaryl) and C_1-6An aliphatic group wherein up to two methylene units of the aliphatic chain are optionally and independently replaced in a chemically stable arrangement by a heteroatom selected from O, N or S. In a further embodiment, R⁶Is H or an optionally substituted group selected from 5-8 membered heterocyclyl and C_1-6An aliphatic group wherein up to two methylene units of the aliphatic chain are optionally and independently replaced in a chemically stable arrangement by a heteroatom selected from O, N or S.

In some embodiments of formula I', J^R6Is selected from C_1-6Alkyl, halo, CN, OH, -O (C)_1-6Alkyl), NH₂、-N(C_1-6Alkyl) and-N (C)_1-6Alkyl radical)₂. In some embodiments of formula I ', J' is selected from C_1-6Aliphatic radical, halo radical, CN, OH, -O (C)_1-6Alkyl), NH₂、-N(C_1-6Alkyl), -N (C)_1-6Alkyl radical)₂、-C(O)OH、-C(O)O(C_1-6Alkyl), 5-6 membered heteroaryl, -CH₂- (5-6 membered heteroaryl), 5-6 membered heterocyclyl, -CH₂- (5-to 6-membered heterocyclic group) and C_1-6An aliphatic group wherein up to two methylene units of the aliphatic chain are optionally and independently replaced in a chemically stable arrangement by a heteroatom selected from O, N or S.

In some embodiments of formula I', Z is a bond and R is selected from

In certain embodiments, R is selected from

In a further embodiment, R is

In some embodiments of formula I', J^YIs selected from-X- (C)_6-10Aryl), -X- (5-10 membered heteroaryl), -X- (C)_3-10Cycloaliphatic), -X- (5-10 membered heterocyclyl) or X. In some embodiments, X is C_1-12An aliphatic radical wherein up to two methylene units of the alkylene chain are optionally and independently replaced by-NR' -, -O-, -S-, -C (O) -, -SO-, or-SO₂-substituted. In a further embodiment, X is C_1-12Aliphatic radicals in which at most one methylene unit is optionally and independently replaced by-NR' -. In still further embodiments, at least one-NR "-is directly linked to R.

In some embodiments of formula I', J^YIs an optionally substituted group selected from C_6-10Aryl, 5-10 membered heteroaryl, C_3-10Cycloaliphatic and 5-10 membered heterocyclic groups. In other embodiments, J^YIs halo, CN, NO₂、CF₃OR ', SR ' OR N (R ')₂。

In some embodiments of formula I', J wherein R is present 2 times^YSubstituted, one of which is J^YIs selected from-X- (C)_6-10Aryl), -X- (5-10)Membered heteroaryl), -X- (C)_3-10Cycloaliphatic), -X- (5-10 membered heterocyclyl) or X, another J^YSelected from H, halo, CN, NO₂、CF₃、OR″、SR″、N(R″)₂、C_6-10Aryl, 5-10 membered heteroaryl, C_3-10Cycloaliphatic or 5-10 membered heterocyclic group. In a further embodiment, J wherein R is present 2 times^YSubstituted, one of which is J^YSelected from X, another J^YSelected from H, halo, CN, NO₂、CF₃OR ', SR ' OR N (R ')₂。

In some embodiments, the present invention provides a compound of formula II:

or a pharmaceutically acceptable salt thereof, wherein ring a is Y.

In a further embodiment of formula II, if R is⁴Is H, then R³Is not H. In a further embodiment, R⁴Is H, R³Is not H.

In some embodiments, the present invention provides a compound of formula III:

or a pharmaceutically acceptable salt thereof, wherein

R³Is halogen, C_1-6Aliphatic radical, C_1-6Alkoxy, N (R)⁵)₂、CN、NO₂Or U_m-V；

Ring a is a 5-8 membered monocyclic heteroaryl ring.

In some embodiments of formula III, ring A has the formula

In a further embodiment, ring a has a structural formula selected from the group consisting of:

in some embodiments, the present invention provides a compound of formula IV:

wherein each Z¹And Z²Is CH or N, R³As defined in formula III.

In some embodiments of formula IV, Z²Is CH. In some embodiments, Z¹Is N or CH. In a further embodiment, Z¹Is N, Z²Is CH. In a further embodiment, Z¹And Z²Are all CH.

In some embodiments of formula IV, R³Is C_1-6An aliphatic group. In a further embodiment, R³Is C_1-3An alkyl group.

In some embodiments of formula IV, J^YIs optionally substituted-N (R)⁹)₂. In a further embodiment, J^YIs optionally substituted-NHR⁹. In a further embodiment, J^YIs optionally substituted-N (R)⁹)₂Wherein said two R are⁹Constituting an optionally substituted 5-8 membered heterocyclic group.

In some embodiments, the present invention provides a compound selected from table 5.

In some embodiments, the present invention provides a composition comprising a compound of formula I', II, III, or IV, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent. In a further embodiment, the composition comprises an additional therapeutic agent selected from the group consisting of an agent for treating autoimmune, inflammatory, proliferative, hyperproliferative diseases, or immunologically mediated diseases, including rejection of transplanted organs or tissues and Acquired Immune Deficiency Syndrome (AIDS).

In some embodiments, the present invention provides a method of inhibiting Tec family kinase activity in a patient or a biological sample, the method comprising administering to said patient or contacting said biological sample with a compound of formula I', II, III or IV or a pharmaceutically acceptable salt thereof or a composition comprising said compound. In a further embodiment, the method comprises inhibiting Itk activity.

In some embodiments, the present invention provides a method of treating or lessening the severity of a disease or condition selected from an autoimmune, inflammatory, proliferative, or hyperproliferative disease, or an immunologically-mediated disease in a patient in need thereof, comprising administering to said patient a compound of formula I', II, III, or IV, or a pharmaceutically acceptable salt thereof, or a composition comprising said compound. In a further embodiment, the method comprises administering to said patient an additional therapeutic agent selected from the group consisting of a component for treating an autoimmune, inflammatory, proliferative, hyperproliferative disease, or immunologically-mediated disease, including rejection of transplanted organs or tissues and Acquired Immune Deficiency Syndrome (AIDS), wherein said additional therapeutic agent is appropriate for the disease being treated; the additional therapeutic agent is administered with the composition as a single dosage form or separately from the composition as part of multiple dosage forms.

In some embodiments, the disease or condition to be treated is asthma, acute rhinitis, allergic, atrophic rhinitis, chronic rhinitis, membranous rhinitis, seasonal rhinitis, sarcoidosis, farmer's lung, fibroid lung, idiopathic interstitial pneumonia, rheumatoid arthritis, seronegative spondyloarthropathies (including ankylosing spondylitis, psoriatic arthritis, and Lett's disease), Behcet's disease, Sjogren's syndrome, systemic sclerosis, psoriasis, systemic sclerosis, atopic dermatitis, contact dermatitis and other eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigus, epidermolysis bullosa, urticaria, vascular dermatitis, vasculitides, erythroderma, cutaneous eosinophilia, uveitis, alopecia, vernal conjunctivitis clusteritis, celiac disease, chronic rhinitis, seasonal rhinitis, sarcoidosis, sjogren's syndrome, systemic sclerosis, and other eczematous dermatitis, Proctitis, eosinophilic gastroenteritis, mastocytosis, pancreatitis, crohn's disease, ulcerative colitis, food-related allergies, multiple sclerosis, atherosclerosis, acquired immunodeficiency syndrome (AIDS), lupus erythematosus, systemic lupus, erythema, hashimoto's thyroiditis, myasthenia gravis, type I diabetes, nephrotic syndrome, eosinophilic fasciitis, hyper IgE syndrome, leprosy nodular leprosy, sexil syndrome, and idiopathic thrombocytopenic purpura, post-angioplasty restenosis, tumors, atherosclerosis, systemic lupus erythematosus, allograft rejection, including, without limitation, acute and chronic allograft rejection, e.g., secondary to post-transplantation of the kidney, heart, liver, lung, bone marrow, skin, and cornea; and chronic graft-host disease.

In some embodiments, the present invention provides a method of inhibiting c-Met kinase activity in a patient or a biological sample, comprising administering to said patient or contacting said biological sample with I, I', II, III, or IV compound or a pharmaceutically acceptable salt thereof, or a composition comprising said compound.

In some embodiments, the present invention provides a method of treating or lessening the severity of cancer in a patient in need thereof, comprising the step of administering to said patient a compound of formula I, I', II, III or IV, or a pharmaceutically acceptable salt thereof, or a composition comprising said compound. In further embodiments, the method further comprises administering a chemotherapeutic agent to said patient, wherein the chemotherapeutic agent is administered with said composition as a single dosage form or separately from said composition as part of multiple dosage forms.

In a further embodiment, the cancer is renal cancer. In other embodiments, the cancer is selected from glioblastoma, gastric cancer, or a cancer selected from colon, breast, prostate, brain, liver, pancreatic, or lung cancer. In a further embodiment, the cancer is gastric cancer. In other embodiments, the cancer is glioblastoma or a cancer selected from breast, colon, or liver cancer.

In other embodiments, the present invention provides a method of inhibiting tumor metastasis or lessening the severity thereof in a patient in need thereof comprising the step of administering to said patient a compound of formula I, I', II, III, or IV, or a pharmaceutically acceptable salt thereof, or a composition comprising said compound.

In some embodiments, the present invention provides a method of inhibiting Aurora kinase activity in a patient or a biological sample, comprising administering to said patient or contacting said biological sample with I, I', II, III, or IV compound or a pharmaceutically acceptable salt thereof, or a composition comprising said compound.

In some embodiments, the present invention provides a method of treating or lessening the severity of melanoma, myeloma, leukemia, lymphoma, neuroblastoma, or a cancer selected from colon cancer, breast cancer, gastric cancer, ovarian cancer, cervical cancer, lung cancer, Central Nervous System (CNS) cancer, renal cancer, prostate cancer, bladder cancer, or pancreatic cancer in a patient in need thereof, comprising the step of administering to said patient a compound of formula I, I', II, III, or IV, or a pharmaceutically acceptable salt thereof, or a composition comprising said compound.

The compounds of the present invention include those generally described above, further illustrated as major classes, minor classes, and species disclosed herein. As used herein, the following definitions will apply, unless otherwise indicated. For the purposes of the present invention, the chemical elements will be according to the CAS version of Handbook of Chemistry and Physics, 75^thEd is determined. In addition, the general principles of Organic Chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausaltito: 1999 and "March's advanced organic Chemistry", 5^th Ed.，Ed.：Smith，M.B.and March，J.，John Wiley &Sons, New York: 2001, the entire contents of which are incorporated herein by reference.

As described herein, the compounds of the invention may be optionally substituted with one or more substituents, such as those set forth in the summary above, or as exemplified by particular classes, subclasses, and species of the invention. It is to be understood that the phrase "optionally substituted" is used interchangeably with the phrase "substituted or unsubstituted". In general, the term "substituted", with or without the term "optionally" preceding, means that a hydrogen radical in a given structure is replaced by a radical designated as a substituent. Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group, and if more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at each position. Substituent combinations contemplated by the present invention are preferably those that form stable or chemically feasible compounds. The term "stable" as used herein means compounds that are substantially unchanged when subjected to the conditions used for their preparation, detection, preferably recovery, purification, and for one or more of the purposes disclosed herein. In some embodiments, a stable compound or chemically feasible compound is one that remains substantially unchanged in the absence of moisture or other chemically reactive conditions at a temperature of 40 ℃ or less for at least one week.

The term "optionally interrupted" denotes the replacement of one atom by another atom within the alkylene chain. Unless otherwise specified, a second atom may replace a first atom at any position, including the terminal atom. For example, C optionally interrupted by-O-)_1-3The alkyl chain can form-OCH₂CH₃、-CH₂-OCH₃Or CH₂CH₂And (5) OH. Unless otherwise specified, the terminal group is bonded to hydrogen on the terminal side.

As used herein, the term "aliphatic" or "aliphatic group" means a straight (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or that contains one or more units of unsaturation, or a monocyclic 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 a "carbocycle", "cycloaliphatic", or "cycloalkyl"), which has a single point of attachment to the remainder of the molecule. Unless otherwise specified, aliphatic groups contain 1-20 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-10 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-8 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-6 aliphatic carbon atoms, and in other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In some embodiments, "cycloaliphatic" (or "carbocycle" or "cycloalkyl") refers to monocyclic C₃-C₈Hydrocarbons or bicyclic radicals C₈-C₁₂A hydrocarbon, which is fully saturated or contains one or more units of unsaturation, but is not aromatic, which has a single point of attachment to the rest of the molecule, wherein any single ring in said bicyclic ring system is a 3-7 membered ring. Suitable aliphatic groups include, but are not limited to, linear or branched substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and hybrids thereof, such as (cycloalkyl) alkyl, (cycloalkenyl) alkyl, or (cycloalkyl) alkenyl.

The term "heteroaliphatic", as used herein, refers to an aliphatic group in which one or two carbon atoms are independently replaced by one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon. Heteroaliphatic groups may be substituted or unsubstituted, straight or branched chain, cyclic or acyclic, and include "heterocyclic", "heterocyclyl", "heterocycloaliphatic", or "heterocyclic" groups.

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

The term "heteroatom" means one or more oxygen, sulfur, nitrogen, phosphorus or silicon (including any oxidized form of nitrogen, sulfur, phosphorus or silicon; quaternized form of any basic nitrogen or heterocycle may be substituted for nitrogen, e.g., 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 the moiety has one or more units of unsaturation.

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

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

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

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

Aryl (including aralkyl, aralkoxy, aryloxyalkyl, and the like) or heteroaryl (including heteroaralkyl and heteroaralkoxy, and the like) may contain one or more substituents. Suitable substituents on the unsaturated carbon atoms of aryl or heteroaryl groups are selected from halogen, -R^o、-OR^o、-SR^o1, 2-methylenedioxy, 1, 2-ethylenedioxy, optionally substituted by R^oSubstituted phenyl (Ph), optionally substituted with R^osubstituted-O (Ph), optionally substituted with R^oSubstituted- (CH)₂)_1-2(Ph), optionally substituted by R^osubstituted-CH ═ CH (Ph), -NO₂、-CN、-N(R^o)₂、-NR^oC(O)R^o、-NR^oC(S)R^o、-NR^oC(O)N(R^o)₂、-NR^oC(S)N(R^o)₂、-NR^oCO₂R^o、-NR^oNR^oC(O)R^o、-NR^oNR^oC(O)N(R^o)₂、-NR^oNR^oCO₂R^o、-C(O)C(O)R^o、-C(O)CH₂C(O)R^o、-CO₂R^o、-C(O)R^o、-C(S)R^o、-C(O)N(R^o)₂、-C(S)N(R^o)₂、-OC(O)N(R^o)₂、-OC(O)R^o、-C(O)N(OR^o)R^o、-C(NOR^o)R^o、-S(O)₂R^o、-S(O)₃R^o、-SO₂N(R^o)₂、-S(O)R^o、-NR^oSO₂N(R^o)₂、-NR^oSO₂R^o、-N(OR^o)R^o、-C(＝NH)-N(R^o)₂Or- (CH)₂)_0-2NHC(O)R^oWherein each independently occurring R^oSelected from hydrogen, optionally substituted C₁-C₆Aliphatic radical, unsubstituted 5-6 membered heteroaryl or heterocycle, phenyl, -O (Ph) or-CH₂(Ph), or R independently occurs twice on the same substituent or different substituents, although as defined above^oAnd each R^oThe atoms to which the groups are bonded together 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. R^oIs selected from NH₂、NH(C_1-4Aliphatic radical), N (C)_1-4Aliphatic radical)₂Halogen, C_1-4Aliphatic radical, OH, O (C)_1-4Aliphatic group), NO₂、CN、CO₂H、CO₂(C_1-4Aliphatic radical), O (halogeno C_1-4Aliphatic group) or halogeno C_1-4Aliphatic radical, wherein R^oEach of the above C_1-4Aliphatic groups are unsubstituted.

The aliphatic or heteroaliphatic group or the non-aromatic heterocycle may contain one or more substituents. Suitable substituents on the saturated carbon atoms of the aliphatic or heteroaliphatic groups or of the non-aromatic heterocyclic rings are selected from those listed above for the aryl or heteroaryl unsaturated carbons, and additionally include the following groups: o, S, NNHR^*、＝NN(R^*)₂、＝NNHC(O)R^*、＝NNHCO₂(alkyl) ═ NNHSO₂(alkyl) or ═ NR^*Wherein each R is^*Independently selected from hydrogen or optionally substituted C_1-6An aliphatic group. R^*Is selected from NH₂、NH(C_1-4Aliphatic radical), N (C)_1-4Aliphatic radical)₂Halogen, C_1-4Aliphatic radical, OH, O (C)_1-4Aliphatic group), NO₂、CN、CO₂H、CO₂(C_1-4Aliphatic radical), O (halogeno C_1-4Aliphatic group) or halogeno C_1-4Aliphatic radical, wherein R^*Each of the above C^1-4Aliphatic groups are unsubstituted.

Optional substituents on the non-aromatic heterocyclic nitrogen being selected from the group consisting of-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⁺)₂or-NR⁺SO₂R⁺(ii) a Wherein R is⁺Is hydrogen, optionally substituted C_1-6An aliphatic group, an optionally substituted phenyl group, an optionally substituted-O (Ph), an optionally substituted-CH₂(Ph), optionally substituted- (CH)₂)_1-2(Ph), optionally substituted-CH ═ CH (Ph), or unsubstituted 5-6 membered heteroaryl or heterocyclic ring, having one to four heteroatoms independently selected from oxygen, nitrogen or sulfur, or R, although as defined above, independently occurs twice on the same substituent or on different substituents⁺And each R⁺The atoms to which the groups are bonded together 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. R⁺Is selected from NH or an optional substituent on the phenyl ring₂、NH(C_1-4Aliphatic radical), N (C)_1-4Aliphatic radical)₂Halogen, C_1-4Aliphatic radical, OH, O (C)_1-4Aliphatic group), NO₂、CN、CO₂H、CO₂(C_1-4Aliphatic radical), O (halogeno C_1-4Aliphatic group) or halogeno C_1-4Aliphatic radical, wherein R⁺Each of the above C_1-4Aliphatic groups are unsubstituted.

The term "AAlkyl chain "means a straight or branched carbon chain which may be fully saturated or have one or more units of unsaturation, and has two points of attachment to the rest of the molecule, wherein one or more methylene units may optionally and independently be replaced by groups including, but not limited to, CO₂、COCO、CONR、OCONR、NRNR、NRNRCO、NRCO、NRCO₂、NRCONR、SO、SO₂、NRSO₂、SO₂NR、NRSO₂NR, O, S or NR.

As noted above, in some embodiments, two independent occurrences of R^o(or R)⁺Or any other variable similarly defined herein) together with the atom to which each variable is bonded 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. Two independently occurring R^o(or R)⁺Or any other variable similarly defined herein) along with the atoms to which each variable is bonded include, but are not limited to, the following: a) two independently occurring R^o(or R)⁺Or any other variable similarly defined herein) to the same atom and together with that atom form a ring, e.g. N (R)^o)₂In which two R are present^oTogether with the nitrogen atom, form piperidin-1-yl, piperazin-1-yl, or morpholin-4-yl; and b) two independent occurrences of R^o(or R)⁺Or any other variable similarly defined herein) to different atoms and together with these atoms form a ring, e.g.

OR in which the phenyl radical is present twice^oSubstitution, R of both occurrences^oTogether with the oxygen atoms to which they are bonded form a fused 6-membered oxygen containing ring:

it will be appreciated that two independent occurrences of R^o(or R)⁺Or any other variable similarly defined herein) may form a variety of other rings along with the atom to which each variable is bonded, and the above detailed examples are not intended to be limiting.

As described herein, a bond drawn from a substituent to the center of one ring within a polycyclic ring system (as shown below) represents the substitution of the substituent at any substitutable position in any ring within the polycyclic ring system. For example, panel a represents a possible substitution in any of the positions shown in panel b.

This also applies to polycyclic systems fused with alternative ring systems (which will be represented by dashed lines). For example, in scheme c, X is an optional substituent for ring a and ring B.

However, if two rings in a polycyclic ring system each have a different substituent drawn from the center of each ring, then each substituent represents only a substitution on the ring to which it is attached, unless otherwise specified. For example, in figure d, Y is only an optional substituent for ring a and X is only an optional substituent for ring B.

Unless otherwise specified, the disclosure is intended to includeThe depicted structure is also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational) forms of the structure; for example, the R and S configurations of each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Thus, single stereochemical isomers as well as enantiomeric, diastereomeric and geometric (or conformational) mixtures of these compounds are within the scope of the invention. Unless otherwise specified, all tautomeric forms of the compounds of the invention are within the scope of the invention. In addition, unless otherwise specified, the structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, except that hydrogen is replaced by deuterium or tritium or carbon is replaced by¹³C-or¹⁴C-enriched carbon instead of compounds having the structure of the present invention are within the scope of the present invention. Such compounds are useful, for example, as analytical tools or probes in biological assays.

The compounds of the invention can generally be prepared by methods known to those skilled in the art for analogous compounds, as illustrated by the following general schemes and the following preparation examples.

Scheme I

Reagents and conditions: (a) i) LDA, toluene, ii) o-ClC₄H₆CH₂SCN, toluene, 2 hours; (b) n is a radical of₂H₄.H₂O, EtOH, reflux, 16 h.

Scheme I above shows a general synthetic route for the preparation of compound 3 of the present invention, wherein R and Z are as described herein. Intermediate 2 can be prepared by methods substantially similar to those described in the literature: davis and Cava j.org.chem.1983, 48, 2774. Cyclization of malononitrile 2 with hydrazine hydrate gives the desired diaminopyrazole 3.

Scheme II

Reagents and conditions: (a) EtOH (in the case of 4),ⁱPrOH (in the case of 5), AcOH_cat.Microwave irradiation, 180 ℃ for 15 minutes.

Scheme II above shows a general synthetic route for the preparation of Compound 6 of the present invention, wherein Z, R and R³As described herein. Pyrazolo [1, 5-a ] is prepared by microwave-assisted cyclization of diaminopyrazole 3 with commercially available malondialdehyde 4 or equivalent 5]Pyrimidine 6. Derivative 5 can be prepared by the method described by copperol et al, j.het.chem.1974, 44, 51.

Table 1 below gives exemplary compounds prepared according to the general methods described in schemes I and II.

TABLE 1

Scheme III

Reagents and conditions：(a)EtOH，AcOH_cat.Reflux, 3Hours; (b) ArB (OH)₂，Pd(dppf)₂Cl₂，2M Na₂CO₃Microwave irradiation, 120 ℃ for 20 minutes.

Scheme III above shows a general synthetic route for the preparation of compound 9 of the present invention, wherein Z, R and Ar are as described herein. Pyrazolo [1, 5-a ] pyrimidine 8 is prepared by cyclization of diaminopyrazole 3 with commercially available 2-bromomalondialdehyde 7. Finally, the generation of biaryl derivative 9 is achieved by treating bromide 8 with the desired boronic acid derivative in the presence of a palladium (0) catalyst using microwave assisted Suzuki coupling methods well known in the art. This reaction is applicable to a variety of substituted aryl or heteroaryl boronic acids.

Exemplary compounds prepared according to the general procedure described in scheme III are given in table 2 below.

TABLE 2

Procedure IV

Reagents and conditions: (a) DMF, 160 ℃, 10 hours; (b) DDQ, 1, 4-dioxane, reflux, 2-3 hours.

Scheme IV above shows a general synthetic route for the preparation of Compound 12 of the present invention, wherein Z, R and R⁴As described herein. The cyclisation of the diaminopyrazole 3 is effected in the presence of beta-dimethylaminones 10 by a process substantially similar to that described in the literature: elnagdi and erian ball. chem. soc. jpn 1990, 63, 1854. This reaction is applicable to a variety of diaminopyrazoles 3 and beta-dimethylaminones 10. According toScheme IV step (b) oxidizes intermediate 11 with DDQ.

Table 3 below gives exemplary compounds prepared according to the general procedure described in scheme IV.

TABLE 3

Procedure V

Reagents and conditions: (a) AcOH, refluxing; (b) i) protective conditions, ii) POCl₃，90℃；(c)i)H₂Pd/C, NaOAc, EtOH, ii) deprotection conditions.

Scheme V above shows a general synthetic route for the preparation of compound 16 of the present invention, wherein Z, R, R³And R⁴As described herein. Intermediate 14 can be prepared by methods described in the literature: sofon et al, Pharmazie 1994, 49, 482 and Ram et al, Indian J. chem. Sect. B1995, 34, 514. After protection of the amine of formula 14, derivative 15 is obtained by methods well known to those skilled in the art. Finally, chloro derivative 15 is reduced according to scheme V, step (c) i), to deprotect the amine to provide a compound of structure 16.

Scheme VI

Reagents and conditions：(a)1N NaOH，MeOH；(b)EDC，HOBt，DCM/DMF，HNR⁵V。

Scheme VI above shows a general procedure for the preparation of compounds of formula 19 of the present invention, wherein Z, R, R⁵And V is as described herein. Each of the above steps is well known to those skilled in the art.

Table 4 below gives exemplary compounds prepared according to the general procedure described in scheme VI.

TABLE 4

Other compounds prepared according to the schemes and examples described herein are provided in table 5:

TABLE 5

As discussed above, the present invention provides compounds that are inhibitors of protein kinases and thus are useful in the treatment of diseases, disorders and conditions including, but not limited to, autoimmune, inflammatory, proliferative or hyperproliferative diseases or immunologically-mediated diseases. Thus, in another aspect of the invention, pharmaceutically acceptable compositions are provided, wherein these compositions comprise any of the compounds as described herein, and optionally 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 certain compounds of the invention can be present in free form for use in therapy, or as appropriate pharmaceutically acceptable derivatives thereof. According to the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, salts of such esters, or any other adduct or derivative capable of providing, directly or indirectly, a compound as described herein or a metabolite or residue thereof upon administration to a patient in need thereof.

The term "pharmaceutically acceptable salt" as used herein, means those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and lower animals without undue toxicity, irritation, allergic response and the like, commensurate with a reasonable benefit/risk ratio. "pharmaceutically acceptable salt" means any non-toxic salt or ester salt of a compound of the present invention that, upon administration to a recipient, is capable of providing, directly or indirectly, a compound of the present invention or an inhibitorily active metabolite or residue thereof. The term "its inhibitory active metabolite or residue" as used herein means that its metabolite or residue is also an inhibitor of a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/R1k) protein kinase, Aurora family kinase or c-Met.

Pharmaceutically acceptable salts are well known in the art. Pharmaceutically acceptable salts are described in detail, for example, in j.pharmaceutical Sciences, 1977, 66, 1-19, by s.m.berge et al, which is incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups 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 by other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipates, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, formates, fumarates, glucoheptanoates, glycerophosphates, gluconates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxyethanesulfonates, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, embonate, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, Propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts derived from suitable bases include alkali metals, alkaline earth metals, ammonium and N⁺(C_1-4Alkyl radical)₄And (3) salt. The invention also encompasses quaternization of any basic nitrogen-containing group of the compounds as disclosed herein. By means of such quaternization, products which are soluble or dispersible in water or oil 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, noneToxic ammonium, quaternary ammonium and amine cations, generated using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryl sulfonates.

As noted above, the pharmaceutically acceptable compositions of the present invention additionally comprise a pharmaceutically acceptable carrier, adjuvant or vehicle, as described herein, including any and all solvents, diluents or other liquid excipients, dispersing or suspending aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like as appropriate for the particular dosage form desired. Remington's Pharmaceutical Sciences, SixteenthEdition, e.w. martin (Mack Publishing co., Easton, Pa., 1980) disclose various carriers for formulating pharmaceutically acceptable compositions and known techniques for their preparation. Except insofar as any conventional carrier medium is incompatible with the compounds of the invention, e.g., any other component that produces any undesirable biological effect or interacts in a deleterious manner with a pharmaceutically acceptable composition, its use is contemplated as falling within the scope of the present invention. Some examples of materials capable of serving as pharmaceutically acceptable carriers include, but are not limited to, 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 glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts; colloidal silicon dioxide; magnesium trisilicate; polyvinylpyrrolidone; a polyacrylate; waxes; polyethylene-polypropylene oxide-block polymers; lanolin; 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; crushed tragacanth; malt; gelatin; talc; 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; ringer's solution; ethanol; a phosphate buffer solution; and other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate; coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preserving and anti-oxidizing agents may also be present in the composition, according to the judgment of the person skilled in the art.

In another aspect, methods of treating or lessening the severity of a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) -mediated disease, Aurora family-mediated disease, or c-Met-mediated disease are provided comprising administering to a subject in need thereof an effective amount of a compound or a pharmaceutically acceptable composition comprising a compound. In certain embodiments of the invention, an "effective amount" of a compound or pharmaceutically acceptable composition is an amount effective to treat or reduce the severity of a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) -mediated disease, an Aurora family-mediated disease, or a c-Met-mediated disease. The compounds and compositions according to the methods of the invention can be administered in any amount and by any route of administration effective to treat or reduce the severity of a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) -mediated disease, Aurora family-mediated disease, or c-Met-mediated disease. The exact amount required will vary from subject to subject, depending on the species, age and general condition of the subject, the severity of the infection, the particular drug, the manner in which it is administered, and the like. The compounds of the present invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The expression "dosage unit form" as used herein denotes physically discrete pharmaceutical units, as appropriate for the patient to be treated. It will be understood, however, that the total daily amount of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific effective dosage level for any particular patient or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the particular compound employed; the specific composition employed; the age, weight, general health, sex, and diet of the patient; the time of administration, the route of administration, and the rate of excretion of the particular compound employed; the duration of the treatment; drugs used in combination or concomitantly with the specific compound employed; and other factors well known in the medical arts. The term "patient" as used herein means an animal, preferably a mammal, most preferably a human.

The pharmaceutically acceptable compositions of the present invention may be administered orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as powders, ointments, or drops), buccally, as an oral or nasal spray, and the like to humans and other animals, depending on the severity of the infection being treated. In certain embodiments, the compounds of the present invention may be administered orally or parenterally at a dosage level of from about 0.01mg/kg to about 50mg/kg, preferably from about 1mg/kg to about 25mg/kg, of the subject's body weight per day, one or more times a day, to achieve the desired therapeutic effect.

Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Injectable preparations, for example sterile injectable aqueous or oleaginous suspensions, may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. Among the acceptable carriers and solvents that may be employed are water, ringer's solution, U.S. p. and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-or di-glycerides. In addition, fatty acids, such as oleic acid, may be used in the preparation of injectables.

The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

In order to prolong the effect of the compounds of the invention, it is often desirable to delay absorption of the compounds following subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material which is poorly water soluble. The rate of absorption of a compound depends on its rate of dissolution, which in turn may depend on crystal size and crystal form. Alternatively, delayed absorption of the parenterally administered compound form is achieved by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are prepared by forming a microencapsulated matrix of the compound in a biodegradable polymer, such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the nature of the particular polymer employed, the release rate of the compound can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations can also be prepared by entrapping the compound in liposomes or microemulsions which are compatible with body tissues.

Rectal or vaginal compositions are preferably suppositories which can be prepared by mixing the compounds of the invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity to release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier, for example sodium citrate or dicalcium phosphate, and/or a) fillers or extenders, for example starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders, for example carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia, c) wetting agents, for example glycerol, d) disintegrants, for example agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) dissolution retarders, for example paraffin, f) absorption accelerators, for example quaternary ammonium compounds, g) wetting agents, for example cetyl alcohol and glycerol monostearate, h) absorbents, for example kaolin and bentonite, and i) lubricants, for example talc, calcium stearate, sodium silicate, and the like, Magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

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

The active compound may also be in microencapsulated form, containing one or more of the above-mentioned excipients. Solid dosage forms such as tablets, dragees, capsules, pills and granules can be provided with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms, the active compound may be mixed with at least one inert diluent, for example sucrose, lactose or starch. Such dosage forms may also contain, under normal circumstances, other substances in addition to inert diluents, such as tableting lubricants and other tableting aids, for example 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 they release the active ingredient(s) only, or preferentially, in a certain portion of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that may be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers, as appropriate. Ophthalmic formulations, ear drops and eye drops are also encompassed within the scope of the present invention. In addition, the present invention encompasses the use of transdermal patches, which have the added advantage of controlling the delivery of compounds to the body. Such dosage forms may be prepared by dissolving or dispersing the compound in the appropriate medium. Absorption enhancers may also be used to increase the flux of the compound across the skin. The rate can be controlled by providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

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

The activity of compounds useful as inhibitors of the Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase, Aurora family kinases, or c-Met in the present invention may be determined in vitro, in vivo, or in cell lines. In vitro assays include determining the inhibition of phosphorylation activity or ATPase activity of an activated Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase, an Aurora family kinase, or c-Met. A selective in vitro assay can quantify the ability of an inhibitor to bind to a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase, an Aurora family kinase, or c-Met. Binding of the inhibitor can be measured by radiolabelling the inhibitor prior to binding, isolating the inhibitor/kinase complex and determining the amount of radiolabel bound. Alternatively, binding of the inhibitor may be determined by incubating the novel inhibitor with a Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase, Aurora family kinase, or c-Met, which are known to bind to radioligands, in a competition assay.

The term "measurably inhibits" as used herein means that there is a measurable change in kinase activity between a sample containing a compound of the invention and the kinase concerned and an equivalent sample containing the kinase without the compound present.

The term "Tec family tyrosine kinase-mediated disorder" as used herein refers to any disease or other deleterious disorder in which a Tec family kinase is known to play a role. Such conditions include, without limitation, autoimmune, inflammatory, proliferative and hyperproliferative diseases and immunologically-mediated diseases, including rejection of transplanted organs or tissues and Acquired Immune Deficiency Syndrome (AIDS).

For example, Tec family tyrosine kinase-mediated disorders include respiratory diseases including, without limitation, reversible obstructive airway diseases including asthma, such as bronchial, allergic, intrinsic, extrinsic and dust asthma, particularly chronic or refractory asthma (e.g., late asthma airway hyperreactivity) and bronchitis. In addition, Tec family tyrosine kinase diseases include, without limitation, those conditions characterized by mucositis, including acute, allergic and atopic and chronic rhinitis, including rhinitis caseosa, hypertrophic, purulent, rhinitis sicca and rhinitis medicamentosa; membranous rhinitis, including croup, fibrinous and pseudomembranous rhinitis and scrofula rhinitis; seasonal rhinitis, including rhinitis nervosa (hay fever) and vasomotor rhinitis; sarcoidosis; farmer's lung and related diseases; a fibroid lung; and idiopathic interstitial pneumonia.

Tec family tyrosine kinase-mediated disorders also include bone and joint diseases including, without limitation, rheumatoid arthritis (pannus formation in), seronegative spondyloarthropathies (including ankylosing spondylitis, psoriatic arthritis, and reiter's disease), becker's disease, sjogren's syndrome, and systemic sclerosis.

Tec family kinase-mediated disorders also include diseases and conditions of the skin including, without limitation, psoriasis, systemic sclerosis, atopic dermatitis, contact dermatitis and other eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigus, epidermolysis bullosa, urticaria, xeroderma, vasculitis nodosa, erythroderma, cutaneous eosinophilia, uveitis, alopecia, cluster and vernal conjunctivitis.

Tec family tyrosine kinase-mediated conditions also include diseases and disorders of the gastrointestinal tract including, without limitation, celiac disease, proctitis, eosinophilic gastroenteritis, mastocytosis, pancreatitis, crohn's disease, ulcerative colitis, gastrointestinal remote food-related allergies such as migraine, rhinitis and eczema.

Tec family tyrosine kinase-mediated disorders also include diseases and conditions of other tissues and systemic diseases including, without limitation, multiple sclerosis, atherosclerosis, acquired immunodeficiency syndrome (AIDS), lupus erythematosus, systemic lupus, erythema, hashimoto's thyroiditis, myasthenia gravis, type I diabetes, nephrotic syndrome, eosinophilic fasciitis, hyper IgE syndrome, leprosy, sexil syndrome and idiopathic thrombocytopenic purpura, post-angioplasty restenosis, tumors (e.g., leukemia, lymphoma), atherosclerosis, and systemic lupus erythematosus.

Tec family tyrosine kinase-mediated disorders also include allograft rejection, including, without limitation, acute and chronic allograft rejection, e.g., secondary to post-transplantation of the kidney, heart, liver, lung, bone marrow, skin and cornea; and chronic graft-host disease.

According to another embodiment, the present invention provides a method of treating or preventing an Aurora-mediated disorder comprising the step of administering to a patient one of the above pharmaceutical compositions.

Preferably, the method is for treating or preventing a condition selected from the group consisting of: cancers, such as breast, colon, prostate, skin, pancreas, brain, genitourinary tract, lymphatic system, stomach, larynx and lung cancers, including lung adenocarcinoma and small cell lung cancer; stroke, diabetes, myeloma, hepatomegaly, cardiac hypertrophy, alzheimer's disease, cystic fibrosis and viral disease or any of the specific diseases or conditions mentioned above.

According to another embodiment, the present invention relates to a method of inhibiting c-Met kinase activity in a biological sample comprising the step of contacting said biological sample with a compound of the present invention or a composition comprising said compound.

According to another embodiment, the present invention relates to a method of inhibiting c-Met kinase activity in a patient comprising the step of administering to said patient a compound of the present invention or a composition comprising said compound.

The term "c-Met-mediated disease" or "c-Met-mediated disorder" as used herein refers to any disease state or other deleterious condition in which c-Met is known to play a role. The term "c-Met-mediated disease" or "c-Met-mediated disorder" also refers to those diseases or disorders that are alleviated by treatment with a c-Met inhibitor. Such conditions include, without limitation, cancers of the kidney, stomach, colon, brain, breast, prostate and lung, glioblastoma, atherosclerosis, pulmonary fibrosis, conditions associated with organ transplantation, allergic conditions and autoimmune conditions.

The term "c-Met" is synonymous with "cMet", "MET", "Met", or other names known to those skilled in the art.

According to one embodiment, the present invention relates to a method of treating or lessening the severity of cancer of the kidney, stomach, colon, brain, breast, prostate and lung, glioblastoma, atherosclerosis, pulmonary fibrosis, disorders associated with organ transplantation, allergic or autoimmune disorders in a patient in need thereof comprising administering to said patient a compound of the present invention or a composition thereof.

In an alternative embodiment, the present invention relates to a method of treating or lessening the severity of gastric or brain cancer in a patient in need thereof comprising administering to said patient a compound of the present invention or a composition thereof.

According to another embodiment, the present invention relates to a method of treating or lessening the severity of a renal cancer in a patient in need thereof comprising administering to said patient a compound of the present invention or a composition thereof.

According to another embodiment, the present invention relates to a method of treating or lessening the severity of gastric cancer in a patient in need thereof comprising administering to said patient a compound of the present invention or a composition thereof.

According to another embodiment, the present invention relates to a method of inhibiting tumor metastasis in a patient in need thereof comprising administering to said patient a compound of the present invention or a composition thereof.

The term "biological sample" as used herein refers to a sample that is external to a living organism, including without limitation cell cultures and extracts thereof; biopsy material obtained from a mammal or an extract thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.

Inhibition of protein kinase, e.g., c-Met, activity in a biological sample can be used for a variety of purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, blood transfusion, organ transplantation, biological sample storage, and biological assays.

Another embodiment of the present invention relates to a method of inhibiting protein kinase activity in a patient comprising administering to said patient a compound of the present invention or a composition comprising said compound. The protein kinases include, but are not limited to, the protein kinases listed above.

It will also be appreciated that the compounds and pharmaceutically acceptable compositions of the present invention may be used in combination therapy, that is, the compounds and pharmaceutically acceptable compositions may be administered simultaneously, prior to, or subsequent to one or more other desired therapeutic agents or pharmaceutical procedures. The particular combination of therapies (therapeutic agents or procedures) used in the combination regimen will take into account the compatibility of the desired therapeutic agent and/or procedure with the desired therapeutic effect to be achieved. It will also be appreciated that the therapies used may achieve the desired effect on the same condition (e.g., the compounds of the invention may be administered simultaneously with another drug used to treat the same condition), or they may achieve different effects (e.g., control of any side effects). As used herein, an additional therapeutic agent that is normally administered to treat or prevent a particular disease or condition is said to be "appropriate for the disease or condition being treated.

For example, chemotherapeutic agents or other antiproliferative agents may be combined with the compounds of the invention to treat proliferative diseases and cancer. Other therapies or anti-cancer agents that may be used in combination with the anti-cancer agents of the present invention include surgery, radiation therapy (some examples are gamma irradiation, neutron beam radiation therapy, electron beam radiation therapy, proton therapy, brachytherapy and systemic radioisotopes, to name a few), endocrine therapy, biological response modifiers (interferons, interleukins and Tumor Necrosis Factor (TNF), to name a few), hyperthermia and cryotherapy, drugs that attenuate any side effects (e.g., antiemetics) and other approved chemotherapeutic drugs including, but not limited to, alkylating drugs (nitrogen mustards, chlorambucil, cyclophosphamide, melphalan, ifosfamide), antimetabolites (methotrexate), purine and pyrimidine antagonists (6-mercaptopurine, 5-fluorouracil, cytarabine, gemcitabine), fusioninhibitors (vinblastine, gemcitabine), and other chemotherapeutic agents, Vincristine, vinorelbine, paclitaxel), podophyllotoxin (etoposide, irinotecan, topotecan), antibiotics (doxorubicin, bleomycin, mitomycin), nitrosoureas (nitrosourea mustard, cyclohexylnitrosourea), inorganic ions (cisplatin, carboplatin), enzymes (asparaginase), hormones (tamoxifen, leuprorelin acetate, flutamide, and megestrol), Gleevec^TMDoxorubicin, dexamethasone, and cyclophosphamide. For a more complete discussion of The most recent cancer therapies, see http:// www.nci.nih.gov/, FDA approved tumor drug list http:// www.fda.gov/cd/cancer/drug frame. htm and The Merck Manual, Seven Ed.1999, The entire contents of which are incorporated herein by reference.

Other examples of drugs that may also be combined with the inhibitors of the invention include, but are not limited to: therapeutic agents for Alzheimer's disease, e.g. AriceptAnd Excelon(ii) a Therapeutic agents for parkinson's disease, such as levodopa/carbidopa, entacapone, ropinirole, pramipexole, bromocriptine, pergolide, trihexyphenidyl, and amantadine; drugs for treating Multiple Sclerosis (MS), e.g. interferon-beta (e.g. Avonex)And Rebif)、CopaxoneAnd mitoxantrone; therapeutic agents for asthma, e.g. albuterol and Singulair(ii) a Drugs for the treatment of schizophrenia, such as reptile, visfate, serekon and haloperidol; anti-inflammatory agents, such as corticosteroids, TNF blockers, IL-1RA, azathioprine, cyclophosphamide, and sulfasalazine; immune modulating and immunosuppressive agents such as cyclosporine, tacrolimus, rapamycin, mycophenolate mofetil, interferon, corticosteroids, cyclophosphamide, azathioprine, and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anticonvulsants, ion channel blockers, riluzole, and antiparkinson agents; drugs for the treatment of cardiovascular diseases, such as beta-blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers and statins; drugs for treating liver diseases, such as corticosteroids, cholestyramine, interferons, and antiviral agents; drugs for treating hematological disorders, such as corticosteroids, antileukemic agents and growth factors; and drugs for the treatment of immunodeficiency disorders, such as gamma globulin.

The amount of additional therapeutic agent in the compositions of the present invention will not exceed the amount normally administered in compositions containing the therapeutic agent as the only active ingredient. Preferably, the amount of the additional therapeutic agent in the presently disclosed compositions will be from about 50% to 100% of the content in typical compositions containing the drug as the sole therapeutically active ingredient.

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

Vascular stents, for example, have been used to overcome restenosis (restenosis of the vessel wall after injury). However, patients using stents or other implantable devices are at risk for clot formation or platelet activation. These undesirable effects can be prevented or mitigated by pre-coating the device with a pharmaceutically acceptable composition comprising a kinase inhibitor. Suitable coatings and general methods of making coated implantable devices are described in U.S. Pat. nos. 6,099,562, 5,886,026, and 5,304,121. The coating is typically a biocompatible polymeric material such as hydrogel polymers, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate copolymers and mixtures thereof. The coating may optionally be further covered by a surface layer of a suitable fluorosilicone, polysaccharide, polyethylene glycol, phospholipid, or combinations thereof, to impart controlled release characteristics to the composition.

Examples

The term "Rt (min)" as used herein means the HPLC retention time in minutes associated with a compound. Unless otherwise indicated, the HPLC method used to obtain the reported retention times was as follows:

column: ace 5C 8, 15cm × 4.6mm id

Gradient: 0-100% acetonitrile + methanol (50: 50) (20mM Tris phosphate, pH 7.0)

Flow rate: 1.5ml/min

And (3) detection: 225nm

Example 1

2-pyridin-2-yl-malononitrile

To a solution of diisopropylamine (15.3mL, 109mmol) in toluene (500mL) was added dropwise 1.6M over a period of-1/2 hours at 0-5 deg.C under nitrogenⁿBuLi/THF (68.5mL, 109 mmol). The reaction mixture was stirred for an additional 15 minutes, then 2-pyridylacetonitrile (5.55mL, 49.6mmol) was added dropwise over 1 hour. A solution of 2-chlorobenzyl thiocyanate (Schlesinger J.Am.chem.Soc.1954, 76, 585) (20.0g, 109mmol) in toluene (100mL) was then added over a period of 1 hour. The reaction mixture was stirred for an additional 2 hours. Water was added and the layers were separated. The organic phase was extracted twice with 200mL of 2N NaOH. The aqueous phases were combined, cooled to 0 ℃ and acidified to-pH 1. The precipitated solid was filtered and dried to give the title compound as a pale tan solid (4.50g, 63% yield). Delta H (DMSO-d)₆)6.79(1H，t)，7.09(1H，d)，7.73(1H，t)，7.79(1H，dd)，12.95(1H，br s)。

Example 2

4-pyridin-2-yl-1H-pyrazole-3, 5-diamine

2-pyridin-2-yl-malononitrile (0.65g, 4.59mmol) and hydrazine hydrate (II)225 μ L, 4.59mmol) in EtOH (5mL) was heated to reflux and stirred for 16 h. The reaction mixture was cooled to room temperature and filtered. Et for solid obtained₂O wash to afford the title compound as a light tan solid (0.42g, 52%). Evaporating the filtrate to dryness, purifying by silica gel chromatography and eluting with NH₄OH∶MeOH∶CH₂Cl₂(0.5: 5: 95) to give another 63.3mg (8%) of the desired product. MS (ES)⁺)176，(ES^-)174.δH(DMSO-d₆)5.45(4H，br s)，6.98(1H，t)，7.57(1H，d)，7.69(1H，t)，8.46(1H，d)，10.55(1H，br s)。

Example 3

3-pyridin-2-yl-6-pyridin-4-yl-pyrazolo [1, 5-a ] pyrimidin-2-ylamine I-1

A mixture of 4-pyridin-2-yl-1H-pyrazole-3, 5-diamine (290mg, 1.66mmol) and 2-pyridin-4-yl-malondialdehyde (247mg, 1.66mmol) was placed in ethanol (5mL) containing a catalytic amount of acetic acid (6 drops). The reaction mixture was subjected to microwave irradiation at 140 ℃ for 15 minutes. The crude mixture was cooled to room temperature and the resulting precipitate was filtered and washed with more ethanol to give the title compound as a golden yellow solid (295mg, 62%). MS (ES)⁺)289，(ES^-)287.δH(DMSO-d₆)7.16(1H，dd)，7.21(2H，m)，7.84(1H，m)，7.89(2H，m)，8.57(2H，br m)，8.65(2H，d)，8.98(1H，d)，9.44(1H，d)。

Various other compounds were prepared by methods substantially similar to those described in example 3. The characterization data for some of these compounds, including HPLC, LC/MS (Observation) and¹h NMR data.

Is summarized in Table 6 below¹H NMR data, wherein¹H NMR data were obtained at 400MHz in deuterated DMSO, with the exception ofIn addition, it was found to be consistent with the structure. The compound numbers correspond to the compound numbers listed in table 1.

Table 6: characterization data for selected compounds

Compound # (I-)	M+1(obs)	Rt (minutes)	1H-NMR
				2	318	9.68	3.81(3H，s)，7.07(4H，m)，7.12(1H，m)，7.75(2H，d)，7.84(1H，t)，8.54(1H，m)，8.58(1H，m)8.82(1H，d)9.15(1H，d)

3	366	/	(500MHz)7.14(m，3H)，7.46(t，1H)，7.60(dd，1H)，7.85(m，2H)，8.06(m，1H)，8.56(m，2H)，8.86(d，1H)，9.27(d，1H)
				4	318	/	(500MHz)3.86(s，3H)，6.97(m，1H)，7.09(s，2H)，7.13(m，1H)，7.39(m，3H)，7.84(m，1H)，8.55(m，1H)，8.58(d，1H)，8.86(d，1H)，9.23(d，1H)
5	302	10.12	2.40(3H，s)，7.15(3H，m)，7.35(2H，d)，7.75(2H，d)，7.90(1H，t)，8.60(2H，m)，8.90(1H，s)，9.20(1H，d)
				6	411	8.74	3.45(3H，s)，7.20(1H，m)，7.25(2H，s)，7.90(1H，t)，8.15(1H，d)，8.35(1H，d)，8.50(1H，s)，8.55(2H，m)，8.70(1H，s)，9.20(1H，d)
7	289	8.76	7.20(3H，m)，7.45(1H，m)，7.90(1H，t)，8.00(1H，t)，8.15(1H，d)，8.60(2H，m)，8.70(1H，m)，9.20(1H，s)，9.45(1H，s)
				8	290	8.29	7.20(1H，m)，7.35(2H，s)，7.90(1H，t)，8.20(1H，d)，8.60(2H，m)，8.90(1H，d)，9.25(2H，dd)，9.65(1H，s)
9	290	8.44	7.20(1H，m)，7.30(2H，s)，7.95(1H，t)，8.70(3H，m)，8.80(1H，s)，9.30(1H，s)，9.45(1H，s)，9.60(1H，s)

Compound # (I-)	M+1(obs)	Ry (minute)	1H-NMR
				10	333	6.64	7.15(1H，m)，7.25(2H，s)，7.95(1H，t)，8.05(1H，d)，8.15(1H，t)，8.40(1H，d)，8.60(2H，m)，9.30(1H，s)，9.80(1H，s)
11	333	6.65	7.15(1H，m)，7.25(2H，d)，7.90(1H，t)，8.25(1H，d)，8.35(1H，d)，8.60(2H，m)，9.15(1H，s)，9.25(1H，s)，9.55(1H，s)

12	360	8.34	3.10(6H，s)，7.15(1H，m)，7.20(1H，d)，7.55(1H，d)，7.90(1H，t)，8.05(1H，t)，8.20(1H，d)，8.60(2H，m)，9.20(1H，s)，9.45(1H，s)
				13	402	8.26	3.60(4H，m)，3.75(4H，m)，7.15(1H，m)，7.20(1H，d)，7.60(1H，d)，7.90(1H，t)，8.00(1H，t)，8.20(1H，d)，8.60(2H，m)，9.20(1H，s)，9.45(1H，s)
14	360	8.08	3.05(6H，s)，7.15(1H，m)，7.25(2H，s)，7.55(1H，d)，7.90(1H，t)，8.00(1H，d)，8.20(1H，d)，8.60(2H，m)，8.75(1H，s)，9.25(1H，s)，9.50(1H，s)
				15	402	8.06	3.65(8H，m)，7.15(1H，m)，7.25(2H，s)，7.90(1H，t)，8.00(1H，d)，8.20(1H，d)，8.60(2H，m)，8.75(1H，s)，9.20(1H，s)，9.50(1H，s)
16	403	7.78	2.20(6H，s)，2.45(2H，m)，3.50(2H，m)，7.15(1H，m)，7.25(2H，s)，7.50(1H，t)，8.00(1H，d)，8.10(1H，t)，8.30(1H，d)，8.60(2H，m)，9.15(1H，s)，9.45(1H，s)，9.90(1H，s)
				17	401	7.27	2.80(4H，m)，3.40(2H，m)，3.60(2H，m)，7.15(1H，m)，7.20(2H，s)，7.50(1H，d)，7.85(1H，t)，8.05(1H，t)，8.20(1H，d)，8.60(2H，m)，9.20(1H，s)，9.45(1H，s)
18	401	7.22	3.30(8H，m)，7.15(1H，m)，7.20(2H，brs)，7.90(1H，t)，8.10(1H，dd)，8.25(1H，d)，8.65(2H，m)，8.80(1H，s)，9.30(1H，s)，9.55(1H，s)

19	417	7.62	2.00(3H，s)，2.20(3H，s)，2.40(2H，m)，3.00(3H，s)，3.60(2H，m)，7.15(1H，m)，7.25(2H，s)，7.90(2H，m)，8.15(1H，m)，8.55(2H，m)，8.70(1H，s)，9.20(1H，s)，9.50(1H，s)
				20	322	/	(500MHz)7.11(s，2H)，7.14(m，1H)，7.56(d，2H)，7.83(m，1H)，7.86(d，2H)，8.55(m，1H)，8.57(d，1H)，8.85(d，1H)，9.24(d，1H)

Compound # (I-)	M+1(obs)	Rt (minutes)	1H-NMR
				21	340	/	(500MHz)7.18(m，1H)，7.28(s，2H)，7.85(m，1H)，7.89(m，2H)，8.15(m，2H)，8.59(m，1H)，8.63(d，1H)，9.40(d，1H)，9.69(s，1H)，9.78(s，1H)
22	417	/	(500MHz)3.25(m，2H)，3.55(m，2H)，3.62(m，2H)，3.8(br，2H)，4.05(be，2H)，4.43(m，2H)，7.16(m，3H)，7.80(d，2H)，7.85(t，1H)，8.56(m，2H)，8.83(d，1H)，9.16(d，1H)
				23	331	9.14	3.79(3H，s)，3.97(2H，s)，5.72(2H，s)，7.02-7.04(2H，m)，7.13(1H，m)，7.21-7.25(2H，m)，7.30-7.32(2H，m)，7.66(2H，d)，8.55(1H，s)，8.95(1H，s)
24	302	8.17	3.96(2H，s)，5.89(2H，s)，7.15(1H，m)，7.22-7.25(2H，m)，7.31-7.32(3H，m)，7.90(1H，t)，8.00(1H，m)，8.65(1H，d)，8.97(1H，s)，9.25(1H，s)
				25	302	7.87	3.95(2H，s)，5.94(2H，s)，7.13(1H，m)，7.21-7.25(2H，m)，7.30-7.32(2H，m)，7.81(2H，d)，8.60(2H，d)，8.73(1H，s)，9.23(1H，s)

Example 4

6-bromo-3-pyridin-2-yl-pyrazolo [1, 5-a ] pyrimidin-2-ylamine II-2

4- (pyridin-2-yl) -1H-pyrazole-3, 5-diamine (2.54g, 14.51mmol) and 2-bromo-malondialdehyde (2.41g, 15.97mmol) were suspended in anhydrous ethanol (35 mL). A small amount of glacial acetic acid (30 drops) was added and the mixture was heated at reflux for 3 hours. After allowing the reaction to cool to room temperature, the mixture was concentrated under reduced pressure. The solid obtained is purified by chromatography on silica gel, using a column containing 2% NH_3(aq)EtOAc of the solution eluted to give the title compound as a yellow solid (0.82g, 19% yield). MS (ES)⁺)290/292.δH(DMSO-d₆)7.2(3H，m)，7.9(1H，t)，8.5(1H，d)，8.6(2H，m)，9.3(1H，s)。

Example 5

4- (2-amino-3-pyridin-2-yl-pyrazolo [1, 5-a ] pyrimidin-6-yl) -phenol II-6

Reacting 6-bromo-3-pyridin-2-yl-pyrazolo [1, 5-a)]Pyrimidin-2-ylamine (50mg, 0.17mmol) and 4-hydroxyphenylboronic acid (48mg, 0.35mmol) were suspended in anhydrous DMF (1.4 ml). 2M Na was added₂CO₃Aqueous solution (0.346mL, 0.69mmol) and the reaction mixture was degassed. Adding [1, 1-bis (diphenylphosphino) ferrocene]Palladium (II) dichloride complex/dichloromethane (1: 1, 8mg, 0.010mmol) and the mixture was heated in a microwave at 120 ℃ for 20 minutes. After allowing the reaction to cool to room temperature, the mixture was concentrated under reduced pressure. The resulting solid was purified by silica gel chromatography eluting with EtOAc to give the title compound as a yellow solid (35mg, 67% yield). MS (ES)⁺)304.δH(DMSO-d₆)6.8(1H，d)，7.1(3H，m)，7.2(1H，d)，7.3(1H，t)，7.9(1H，t)，8.6(2H，m)，8.8(1H，s)，9.2(1H，s)，9.7(1H，s)。

Various other compounds were prepared by methods substantially similar to those described in example 5. The characterization data for some of these compounds, including HPLC, LC/MS (Observation) and¹h NMR data.

Is summarized in Table 7 below¹H NMR data, wherein¹H NMR data were obtained at 400MHz in deuterated DMSO, unless otherwise indicated, and were found to be consistent with the structure. The compound numbers correspond to the compound numbers listed in table 2.

Table 7: characterization data for selected compounds

Compound # (II-)	M+1(obs)	Rt (minutes)	1H-NMR
				1	288	9.72	7.15(3H，m)，7.90(2H，m)，7.40(1H，t)，7.50(2H，m)7.85(3H，m)，8.55(2H，m)，8.90(1H，s)，9.25(1H，s)

Chemical combinationSubstance # (II-)	M+1(obs)	Rt (minutes)	1H-NMR
				3	322	10.14	7.15(3H，m)，7.50(1H，m)，7.65(1H，m)7.90(1H，t)，8.60(3H，m)，9.05(1H，s)
4	359	8.63	3.05(6H，m)，7.15(3H，m)，7.55(1H，d)，7.90(3H，m)，8.60(2H，m)，8.90(1H，s)，9.30(1H，s)
				5	401	8.06	2.40(4H，m)，3.20(4H，m)，7.15(2H，t)，7.70(1H，d)，7.90(3H，m)，8.60(2H，m)，8.85(2H，m)，8.90(1H，s)，9.30(1H，s)
7	303	8.75	5.25(2H，s)，6.60(1H，d)，6.90(2H，m)，7.10(4H，m)，7.85(1H，t)，8.55(2H，m)，8.75(1H，s)，9.05(1H，s)
				8	402	8.60	2.20(6H，s)，2.45(2H，m)，3.40(2H，m)，7.15(3H，m)，7.85(1H，t)，7.95(4H，m)，8.60(3H，m)，8.95(1H，s)，9.30(1H，s)
9	416	8.71	2.00(3H，s)，2.25(3H，s)，2.40(2H，m)，3.00(4H，m)，3.55(1H，m)，7.15(3H，m)，7.50(2H，d)，7.90(3H，m)，8.60(3H，m)，8.90(1H，s)，9.30(1H，s)

Example 6

5-phenyl-3-pyridin-2-yl-6, 7-dihydro-pyrazolo [1, 5-a ] pyrimidin-2-ylamine

Reacting 6-bromo-3-pyridin-2-yl-pyrazolo [1, 5-a)]Pyrimidin-2-ylamine (0.52g, 3.01mmol) and 3-dimethylamino-1-phenyl-propan-1-one hydrochloride (0.64g, 3.01mmol) were suspended in anhydrous DMF (10 mL). The mixture was heated at 160 ℃ for 3 hours. While stirring. After allowing the reaction to cool to room temperature, the mixture was poured into cold water. The resulting precipitate was filtered and dried to give the title compound as a yellow solid (0.79g, 93% yield). MS (ES)⁺)290.δH(DMSO-d₆)3.3(2H，m)，4.1(2H，m)，6.1(2H，s)，7.2(1H，m)，7.7(4H，m)，7.9(1H，t)，8.2(1H，m)，8.4(1H，d)，8.6(1H，d)。

Example 7

5-phenyl-3-pyridin-2-yl-pyrazolo [1, 5-a ] pyrimidin-2-ylamine III-1

Reacting 5-phenyl-3-pyridin-2-yl-6, 7-dihydro-pyrazolo [1, 5-a ]]Pyrimidin-2-ylamine (0.297g, 1.03mmol) and 2, 3-dichloro-5, 6-dicyano-1, 4-benzoquinone (0.256g, 1.13mmol) were suspended in dry 1, 4-dioxane (10 mL). The mixture was heated at 110 ℃ for 90 minutes. After allowing the reaction to cool to room temperature, the mixture was concentrated under reduced pressure. The resulting solid was dissolved in ethyl acetate and 2M Na₂CO₃The aqueous solution was partitioned. The organic layer was treated with additional 2M Na₂CO₃Aqueous solution (2X 20mL), H₂O (1X 20mL) over Na₂SO₄Drying and filtering. Purification by silica gel chromatography eluting with EtOAc/petroleum ether (1/1) afforded the title compound as a yellow solid (0.15g, 51% yield). MS (ES)⁺)288.δH(DMSO-d₆)7.1(3H，m)，7.6(4H，m)，8.0(1H，s)，8.4(2H，d)，8.6(1H，m)，8.8(1H，d)，9.0(1H，d)。

Various other compounds were prepared by methods substantially similar to those described in example 7. The characterization data for some of these compounds, including HPLC, LC/MS (Observation) and¹h NMR data.

Is summarized in Table 8 below¹H NMR data, wherein¹H NMR data were obtained at 400MHz in deuterated DMSO, unless otherwise indicated, and were found to be consistent with the structure. The compound numbers correspond to the compound numbers listed in table 3.

Table 8: characterization data for selected compounds

Compound # (III-)	M+1(obs)	Rt (minutes)	1H-NMR
				2	318	9.82	3.90(3H，s)，7.05(2H，m)，7.20(3H，m)，7.50(1H，d)，7.95(1H，t)，8.30(2H，d)，8.60(1H，m)，8.80(1H，d)，8.90(1H，d)

Example 8

2-amino-3-pyridin-2-yl-pyrazolo [1, 5-a ] pyrimidine-6-carboxylic acid ethyl ester IV-1

A mixture of 4- (pyridin-2-yl) -1H-pyrazole-3, 5-diamine (0.10g, 0.57mmol), (ethoxycarbonyl) malondialdehyde (0.084g, 0.57mmol) and 2 drops of acetic acid in 1-propanol (3mL) was heated in a microwave at 180 ℃ for 15min with stirring. The reaction was cooled to room temperature, filtered, washed with ethanol, and dried under vacuum at 60 ℃ for 3 days to give the title compound as a brown solid (0.098g, 61% yield). MS (ES)⁺)284.δH(500Mhz，DMSO-d₆)1.35(3H，t)，4.36(2H，q)，7.19(1H，m)，7.36(2H，s)，7.87(1H，dt)，8.58(2H，m)，8.82(1H，d)，9.18(1H，d)。

Example 9

2-amino-3-pyridin-2-yl-pyrazolo [1, 5-a ] pyrimidine-6-carboxylic acid IV-3

2-amino-3-pyridin-2-yl-pyrazolo [1, 5-a]Ethyl pyrimidine-6-carboxylate (1.58g, 5.58mmol) was refluxed with a solution of potassium hydroxide (0.95g, 16.7mmol) in ethanol (30mL) for 3h and then cooled to room temperature. The precipitate is filtered off, washed with diethyl ether, suspended in diethyl ether and washed with 2M HCl/Et₂Treatment with O (15mL, 30mmol), stirring for 0.5h, filtration, and washing with diethyl ether gave the HCl salt of the title compound as a brown solid (0.88g, 54% yield). MS (ES)⁺)256.δH(500Mhz，DMSO-d₆)7.25(1H，m)，7.94(1H，t)，8.56(1H，d)，8.60(1H，d)，8.83(1H，d)，9.16(1H，d)。

Example 10

(2-amino-3-pyridin-2-yl-pyrazolo [1, 5-a ] pyrimidin-6-yl) morpholin-4-yl-methanone (IV-12)

2-amino-3-pyridin-2-yl-pyrazolo [1, 5-a]A mixture of pyrimidine-6-carboxylic acid (0.06g, 0.20mmol), morpholine (0.02mL, 0.25mmol) and HBTU (0.09g, 0.25mmol) in tetrahydrofuran (5mL) was stirred at room temperature under a nitrogen atmosphere for 24 h. Filtering off the precipitate, purifying by chromatography on silica gel using NH₄OH: methanol: dichloromethane (0.5: 5: 95) to give the title compound (0.02g, 35% yield). MS (ES)⁺)325.δH(500Mhz，DMSO-d6)3.63(8H，m)，7.17(3H，m)，7.85(1H，m)，8.52(1H，s)，8.56(2H，m)，8.99(1H，s)。

A variety of other compounds are prepared by methods substantially similar to those described herein. Characterization data for some of these compounds, including HPLC, LC/MS (observed) and 1HNMR data, are summarized in table 7 below.

Is summarized in Table 9 below¹H NMR data, wherein¹H NMR data were obtained at 500MHz in deuterated DMSO, unless otherwise indicated, and were found to be consistent with the structure. The compound numbers correspond to the compound numbers listed in table 5.

Table 9: characterization data for selected compounds

Compound # (IV-)	M+1(obs)	Rt (minutes)	1H-NMR
				2	366	/	(500MHz)1.70(6H，m)，2.50(6H，m)，3.34(2H，m)，7.16(1H，m)，7.23(2H，s)，7.86(1H，dt)，8.56(2H，m)，8.61(1H，t)，8.84(1H，d)，9.19(1H，d)

Compound # (IV-)	M+1(obs)	Ry (minute)	1H-NMR
				4	382	/	(500MHz)1.71(2H，m)，2.35(6H，m)，3.33(2H，m)，3.58(4H，m)，7.16(1H，m)，7.23(2H，s)，7.86(1H，dt)，8.56(3H，m)，8.85(1H，d)，9.20(1H，d)
5	366	/	(500MHz)1.49(2H，m)，1.67(2H，m)，1.92(1H，m)，1.98(1H，m)，2.15(2H，m)，2.29(3H，m)，3.00(1H，m)，3.34(2H，m)，7.16(1H，m)，7.23(2H，s)，7.86(1H，dt)，8.57(3H，m)，8.84(1H，d)，9.19(1H，d)
				6	368	/	(500MHz)1.2(6H，m)，2.50(4H，m)，3.12(5H，m)，3.75(2H，m)，7.18(3H，m)，7.85(1H，m)，8.56(3H，m)，9.05(1H，m)
7	326	/	(500MHz)2.26(3H，br s)，2.70(2H，m)，3.05(3H，br s)，3.49(2H，m)，7.16(3H，m)，7.84(1H，dt)，8.54(4H，m)，9.10(1H，m)

8	352	/	(500MHz)1.07(2H，m)，1.65(3H，m)，2.46(2H，m)，2.97(2H，m)，3.17(2H，m)，7.16，(1H，m)，7.23(2H，s)，7.86(1H，dt)，8.56(3H，m)，8.86(1H，d)，9.23(1H，d)
				9	340	/	(500MHz)1.72(2H，m)，2.24(6H，s)，2.41(2H，t)，3.33(2H，m)，7.17(1H，m)，7.23(2H，s)，7.86(1H，dt)，8.56(2H，m)，8.60(1H，t)，8.84(1H，d)，9.20(1H，d)
10	395	/	(500MHz)1.70(2H，m)，2.17(3H，s)，2.36(10H，m)，3.33(2H，m)，7.16(1H，m)，7.23(2H，s)，7.56(1H，dt)，8.56(3H，m)，8.84(1H，d)，9.20(1H，d)
				11	338	/	(500MHz)2.21(3H，s)，2.36(4H，m)，3.59(4H，m)，7.16(3H，m)，7.84(1H，dt)，8.50(1H，d)，8.55(2H，m)，8.96(1H，d)
13	295	/	(500MHz，)0.60(2H，m)，0.74(2H，m)，2.86(1H，m)，7.16(1H，m)，7.23(2H，s)，7.85(1H，dt)，8.55(3H，m)，8.82(1H，d)，9.18(1H，d)

Example 11: synthesis of Compounds according to scheme VII

Reagents and conditions: (a) DMF-DMA, 85 ℃, 16 hours; (b) EtOH, AcOH, microwave irradiation, 120 ℃ for 30 minutes.

Scheme VII above shows another general synthetic route for the preparation of Compound D of the present invention, wherein Z, R, R³And R⁴As described herein. Intermediate B was prepared from compound a according to scheme VII, step (a). Preparation of pyrazolo [1, 5-a by microwave-assisted cyclization of diaminopyrazoles C with enaminones B]A pyrimidine (D).

Exemplary compounds prepared according to the general procedures described in schemes VI and VII are shown below:

example 12: 6-methyl-3- (6- (methylsulfinyl) pyrimidin-4-yl) pyrazolo[1，5-a]Synthesis of pyrimidin-2-amines

Synthesis of 4-chloro-6- (methylthio) pyrimidine

4, 6-dichloropyrimidine (15.4g, 0.10mol) was dissolved in THF (120mL) at room temperature, and NaSMe (8.5g, 0.12mol) was added as a solid. The reaction mixture was heated to 60 ℃ for 16 hours, then cooled to room temperature, and then diluted with ethyl acetate (300mL) and water (300 mL). The organic layer was washed with brine, dried over sodium sulfate, concentrated to an orange oil and recrystallized from hexane to give a light yellow solid (9.85g, 0.061mol, 61%). H NMR (500MHz, CDCl3)8.74(s, 1H), 7.23(s, 1H), 2.58(s, 3H).

Synthesis of 2- (6- (methylthio) pyrimidin-4-yl) malononitrile

Sodium hydride (10.8g, 0.27mol) was suspended in THF (100mL) at 0 deg.C. To this suspension was added dropwise malononitrile (8.0g, 0.122mmol) in 50mL THF. After the addition was complete, the reaction mixture was stirred for an additional 10 minutes, then a solution of 4-chloro-6- (methylthio) pyrimidine (9.85g, 0.61mol) in 50mL THF was added. Nitrogen was bubbled through the reaction mixture for 10min, Pd (PPh) was added₃)₄(3.3g, 3.0mmol) of solid and the reaction mixture was heated to 60 ℃. After heating for 4hr, the reaction mixture was cooled to room temperature and carefully quenched with water. The reaction mixture was then concentrated to the residue and 100mL of water was added followed by 6N HCl until pH 1. The resulting precipitate was filtered, washed with water and diethyl ether, suspended in ethyl acetate (100mL) and filtered again. The solid was dried (11.4g, 0.06mol, 98%). H NMR (500MHz, DMSO-d6)8.28(s, 1H), 6.47(s, 1H), 2.58(s, 3H).

Synthesis of 4- (6- (methylthio) pyrimidin-4-yl) -1H-pyrazole-3, 5-diamine

2- (6- (methylthio) pyrimidin-4-yl) malononitrile (11.4g, 0.06mol) was suspended in 150mL EtOH and hydrazine hydrate (3mL, 0.06mol) was added. The reaction mixture was heated to reflux for 3 days. The reaction mixture was concentrated to a solid, suspended in ethyl acetate (100mL) and filtered. The solid was dried to give the product (10.4g, 46.8mmol, 78%, ca.80% pure).

Synthesis of 6-methyl-3- (6- (methylsulfinyl) pyrimidin-4-yl) pyrazolo [1, 5-a ] pyrimidin-2-amine

4- (6- (methylthio) pyrimidin-4-yl) -1H-pyrazole-3, 5-diamine (1.0g, 4.5mmol) and 3-ethoxy-2-methylacrolein (0.63g, 5.3mmol) were combined in isopropanol (12 mL). Acetic acid (0.1mL) was added to the reaction mixture, which was then sealed in a microwave container and heated to 160 ℃ for 15 min. The reaction mixture was diluted with ethyl acetate (200mL) and 1N NaOH (60mL), the layers were separated, and the organic layer was concentrated to 60 mL. The solution was cooled to 0 ℃ and 77% m-chloroperbenzoic acid (1.9g, 8.5mmol) was added. After 1hr, a yellow precipitate formed, which was filtered off, washed with ethyl acetate and dried to give the product (0.39g, 30% yield). LCMS MH +289.29 HNMR (500MHz, DMSO-d6)9.02(s, 1H), 8.91(s, 1H), 8.88(s, 1H), 8.54(s, 1H), 7.14(s, 2H), 2.91(s, 3H), 2.34(s, 3H).

Synthesis of 1- (6- (2-amino-6-methylpyrazolo [1, 5-a ] pyrimidin-3-yl) pyrimidin-4-yl) piperidine-3-amide

Reacting 6-methyl-3- (6- (methylsulfinyl) pyrimidin-4-yl) pyrazolo [1, 5-a]Pyrimidin-2-amine (60mg, 0.21mmol) was dissolved in NMP containing piperidine carboxamide (100mg, 0.78mmol) and heated to 200 ℃ in a microwave for 20 min. The reaction mixture was subjected to RP HPLC (C18, CH)₃CN/H₂O0.1% TFA) and the pure fraction was poured into ethyl acetate/0.5N NaOH. The organic phase was dried over sodium sulfate, concentrated to a yellow solid, dissolved in 1N HCl in MeOH, and concentrated to a HCl salt as a yellow solid (20mg, 0.052mmol, 26%). LC/MS MH +353.4.¹H NMR(500MHz，DMSO-d6)8.93(s，1H)，8.70(s，1H)，8.54(d，J＝1.9Hz，1H)，7.48(s，1H)，7.11(s，1H)，6.96(s，1H)，3.32-3.25(m，2H)，2.47(m，2H)，2.36(s，3H)，2.00-1.96(m，1H)，1.88-1.83(m，2H)，1.72(m，1H)，1.52(s，1H)。

Is summarized in Table 10 below¹H NMR data, wherein¹H NMR data were obtained at 500MHz in deuterated DMSO, unless otherwise indicated, and were found to be consistent with the structure. Compound (I)The numbers correspond to the compound numbers listed in table 5.

Table 10: characterization data for selected compounds

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				1	225.10	2.60	(500MHz，dmso-d6)8.59(d，1H)，7.81(d，2H)，7.40(t，2H)，7.19(t，1H)，6.65(d，1H)，2.45(s，3H)ppm
2	265.90	2.62	1H NMR(500MHz，DMSO-d6)d 9.20(1H，s)，8.81(1H，s)，7.72(2H，d)，7.05(2H，d)，6.70(2H，br s)，3.80(3H，s)ppm.
				3	348.20	2.10

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				4	316.20	2.30
5	356.10	2.30

6	330.20	1.90
				7	302.20	2.20
8	304.20	1.90
				9	316.20	2.30
10	313.10	2.00
				11	332.10	1.80
12	316.20	2.30
				13	306.10	2.10
14	318.20	2.10
				15	364.10	2.50
16	381.20	1.80
				17	372.10	2.20
18	380.10	1.90
				19	372.10	2.40
20	303.20	1.70
				21	333.10	2.00
22	319.10	1.80
				23	345.20	1.50
24	317.00	3.00	1H NMR(500MHz，DMSO-d6)d 9.22(1H，s)，8.90(1H，s)，7.85(2H，d)，7.59(2H，d)，6.50(2H，br s)，4.30(2H，q)，1.30(3H，t)ppm.

25	284.00	2.10	1H NMR(500MHz，DMSO-d6)d 9.40(1H，s)，9.22(1H，s)，8.70(1H，d)，8.10(1H，d)，7.91(1H，dd)，7.40(1H，dd)，6.55(2H，brs)，4.30(2H，q)，1.30(3H，t)ppm.
				26	285.00	2.00	1H MR(500MHz，DMSO-d6)d 9.50(1H，s)，9.35(1H，s)，9.22(1H，s)，8.85(1H，d)，8.65(1H，d)，6.60(2H，br s)，4.30(2H，q)，1.30(3H，t)ppm.
27	285.00	1.90	1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.30(1H，s)，9.25(1H，s)，8.90(1H，d)，8.21(1H，d)，6.62(2H，br s)，4.30(2H，q)，1.30(3H，t)ppm.
				28	257.00	1.50	1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.20(1H，s)，9.00(1H，s)，8.80(1H，d)，8.15(1H，d)，6.80(2H，br s)ppm.
29	330.60	3.41	H NMR(500MHz，DMSO-d6)9.03(d，J＝2.3Hz，1H)，8.67(d，J＝2.3Hz，1H)，7.76(d，J＝8.8Hz，2H)，7.66(d，J＝8.1Hz，2H)，7.30(d，J＝8.0Hz，2H)，7.01(d，J＝8.8Hz，2H)，5.71(s，2H)，3.79(s，3H)，2.36(s，3H)

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				30	378.70	3.90	H NMR(500MHz，DMSO-d6)9.09(d，J＝2.3Hz，1H)，8.73(d，J＝2.3Hz，1H)，7.85(d，J＝8.6Hz，2H)，7.67(d，J＝8.1Hz，2H)，7.60(d，J＝8.6Hz，2H)，7.30(d，J＝7.9Hz，2H)，5.90(s，2H)，2.36(s，3H)

31	378.60	3.41	H NMR(500MHz，DMSO-d6)8.03(d，J＝2.2Hz，1H)，7.75(d，J＝2.2Hz，1H)，6.97(d，J＝8.0Hz，1H)，6.76(d，J＝8.1Hz，2H)，6.67(m，2H)，6.52(m，3H)，1.60(s，3H)
				32	382.30	2.50	1H NMR(500MHz，DMSO-d6)d 9.65(1H，s)，9.25(2H，m)，8.9(1H，d)，8.20(2H，m)，7.45(1H，m)，7.20(1H，m)，7.05(1H，m)，6.70(2H，m)，4.60(2H，s)ppm.
33	325.20	1.90	1H NMR(500MHz，DMSO-d6)d 9.50(1H，s)，9.20(1H，m)，9.15(1H，s)，8.85(1H，d)，8.20(1H，m)，7.45(1H，m)，3.4(3H，m)，1.20(6H，d)，1.1(3H，m)ppm.
				34	393.40	1.40	1H NMR(500MHz，DMSO-d6)d 9.60(1H，s)，9.25(1H，m)，9.20(1H，s)，8.85(1H，d)，8.20(1H，m)，4.5(1H，m)，3.90(1H，m)，3.80(1H，m)，3.60(2H，m)，3.45(1H，m)，3.30(1H，m)，3.15(1H，m)，3.05(1H，m)，2.15(1H，m)，1.95(2H，m)，1.90(4H，m)，1.75(2H，m)ppm.
35	417.40	1.60	1H NMR(500MHz，DMSO-d6)d 9.60(1H，s)，9.21(2H，m)，8.89(1H，d)，8.20(1H，m)，7.35*3H，m)，7.29(2H，m)，4.85(2H，s)，3.74(2H，m)，3.35(2H，m)，2.80(6H，s)ppm.
				36	375.40	1.30	1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.21(1H，m)，8.89(1H，d)，8.80(2H，m)，8.20(1H，m)，7.80(2H，m)，4.97(2H，s)，3.55(2H，m)，1.15(3H，t)ppm.
37	390.40	1.90	1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.21(2H，m)，8.85(1H，d)，8.20(1H，m)，7.30(5H，m)，4.80(2H，s)，4.0(2H，m)，3.50(2H，m)ppm.

38

399.40

2.20

1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.21(2H，m)，8.85(1H，d)，8.20(1H，m)，7.25(5H，m)，4.80(2H，s)，3.70(2H，m)，2.85(2H，m)ppm.

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				39	340.30	1.50	1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.21(1H，s)，9.20(1H，s)，8.85(1H，d)，8.20(1H，m)，3.55(2H，m)，3.35(1H，m)，1.95(2H，m)，1.90(2H，m)，1.70(2H，m)ppm.
40	323.30	1.50	1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.25(2H，m)，8.89(1H，d)，8.20(1H，m)，3.70(2H，m)，3.20(3H，s)，2.95(2H，m)ppm.
				41	360.40	2.10	1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.25(2H，m)，8.89(1H，d)，8.20(1H，m)，7.30(5H，m)，4.70(2H，s)，3.0(3H，s)ppm.
42	374.40	2.30	1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.25(2H，m)，8.89(1H，d)，8.20(1H，m)，7.30(5H，m)，4.75(2H，s)，3.40(2H，m)，1.10(3H，t)ppm.
				43	389.40	1.50	1H NMR(500MHz，DMSO-d6)d 9.60(1H，s)，9.22(2H，m)，8.89(1H，d)，8.20(1H，m)，8.0(1H，m)，7.20(2H，m)，6.75(3H，m)，4.45(2H，s)，2.90(6H，s)ppm.
44	326.20	1.30	1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.22(2H，m)，8.89(1H，d)，8.20(1H，m)，3.90(2H，m)，3.40(1H，m)，2.20(1H，m)，1.95(2H，m)ppm.

45	310.30	1.70	1H NMR(500MHz，DMSO-d6)d 9.55(1H，s)，9.22(2H，m)，8.89(1H，d)，8.20(1H，m)，3.70(2H，m)，3.55(2H，m)，1.85(4H，m)ppm.
				46	376.40	2.00	1H NMR(500MHz，DMSO-d6)d 9.60(1H，s)，9.30(1H，s)，9.25(1H，s)，8.89(1H，d)，8.40(1H，m)，8.20(1H，m)，7.30(5H，m)，5.20(1H，m)，3.75(2H，m)ppm.
47	413.10	3.40	1H NMR(500MHz，DMSO-d6)d 9.50(1H，s)，9.20(1H，s)，8.70(1H，t)，8.30(1H，t)，8.10(1H，d)，7.91(1H，t)，7.60(2H，m)，7.40(1H，d)，7.35(1H，d)，6.61(2H，s)，4.55(2H，m)ppm.
				48	307.10	2.15	1H NMR(500MHz，DMSO-d6)d 8.79(1H，m)，8.46(1H，m)，6.32(2H，s)，4.30(2H，m)，4.05(2H，m)，2.69(2H，m)，2.0(3H，s)，1.91(2H，m)，1.29(3H，t)ppm.
49	373.90	3.90
				50	319.20	1.70
51	414.20	1.50
				52	290.00	1.30

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				53	306.20	1.30
54	402.40	1.40
				55	414.40	1.30
56	394.10	2.50
				57	296.70	2.62

58	280.90	3.22
				59	294.00	3.36
60	451.40	0.80
				61	443.50	1.30
62	403.40；401.20	1.30；1.90
				63	429.00	1.30
64	458.40	1.80
				65	403.40	1.30
66	444.40	1.70
				67	388.40	1.70
68	459.40	1.30
				69	415.50	1.30
70	429.40	1.30
				71	430.40	1.50
72	384.40	1.70
				73	378.40	1.70
74	430.20	1.50
				75	404.40	1.60
76	472.50；473.50	1.90；1.30
				77	388.40	1.90
78	445.40	1.30
				79	416.40	1.50

80	431.50	1.50
				81	401.40	1.20
82	444.40	1.60
				83	444.40	1.60
84	469.50	1.30
				85	431.40	1.40
86	372.30	1.60
				87	418.40	1.80
88	445.50	1.40
				89	374.40	1.60
90	413.40	1.70
				91	400.40	1.80
92	418.40	1.60
				93	415.20	1.50
94	389.40	0.80
				95	432.40	1.60

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				96	402.40	1.50
97	402.40	1.90
				98	388.40	1.70
99	404.40	1.50
				100	451.40	1.40
101	443.50	1.30

102	429.40	1.30
				103	388.40	1.78
104	415.40	1.30
				105	429.40	1.30
106	384.40	1.80
				107	378.30	1.80
108	404.40	1.80
				109	388.40	2.10
110	431.50	1.60
				111	418.40	1.60
112	416.40	1.60
				113	431.50	1.60
114	399.40	1.60
				115	373.20	1.70
116	413.40	1.70
				117	473.50	1.40
118	345.90	2.09
				119	401.40	1.30
120	389.40	1.40
				121	469.50	1.40
122	431.50	1.50
				123	372.40	1.70
124	445.50	1.40
				125	402.40	2.00

126	402.40	2.00
				127	388.40	1.80
128	404.40	1.50
				129	473.50	1.30
130	403.40	1.30
				131	315.70	1.61	HNMR(500MHz，DMSO-d6)9.61(d，J＝2.3Hz，1H)，8.95(d，J＝2.3Hz，1H)，8.78(dd，J＝5.1，1.4Hz，2H)，8.13(d，J＝6.3Hz，2H)，7.76(d，J＝7.0Hz，2H)，7.57(m，1H)，7.47(t，J＝7.6Hz，2H)，7.09(br s，2H).
132	302.80	1.62	H NMR(500MHz，DMSO-d6)9.56(d，J＝2.3Hz，1H)，9.04(d，J＝2.3Hz，1H)，8.79(m，2H)，8.49(d，J＝8.3Hz，1H)，8.43(d，J＝2.0Hz，1H)，8.19(d，J＝6.1Hz，2H)，7.72(dd，J＝8.4，2.1Hz，1H)，2.33(s，3H).

Compound # (V-)	M+1(obs)	Rt (minute)	1H-NMR
				133	302.80	1.90	H NMR(500MHz，DMSO-d6)9.63(d，J＝2.1Hz，1H)，9.13(d，J＝2.1Hz，1H)，8.85(d，J＝6.4Hz，2H)，8.46(d，J＝5.1Hz，1H)，8.42(s，1H)，8.30(d，J＝6.1Hz，2H)，7.08(d，J＝4.5Hz，1H)，2.42(s，3H).
134	395.00	1.67	H NMR(500MHz，DMSO-d6)9.60(d，J＝2.2Hz，1H)，9.00(d，J＝2.2Hz，1H)，8.85(d，J＝6.6Hz，2H)，8.41(d，J＝4.8Hz，1H)，8.30(d，J＝6.5Hz，2H)，8.00(d，J＝8.4Hz，1H)，7.67(dd，J＝8.3，5.3Hz，1H)，7.49(d，J＝7.2Hz，2H)，7.33(m，3H)，5.30(s，2H).
				135	302.80	0.37	HNMR(500MHz，DMSO-d6)9.62(d，J＝2.0Hz，1H)，9.00(d，J＝2.0Hz，1H)，8.80(d，J＝6.2Hz，2H)，8.70(d，J＝5.1Hz，1H)，8.30(d，J＝6.9Hz，1H)，8.18(d，J＝5.1Hz，2H)，7.74(m，1H)，2.43(s，3H).

136	438.10	2.46	1H NMR(500MHz，CDCl3)d 9.25(2H，m)，8.99(1H，s)，8.83(1H，d)，8.20(1H，s)，7.38(1H，d)，6.95(1H，d)，5.91(2H，s)ppm.
				137	367.20	2.00	500MHz，MeOD，7.82(s，1H)，7.49(s，1H)，7.25(d，2H)，7.11(d，1H)，7.78(d，2H)，6.18(t，1H)，5.80(d，1H)
138	412.00	1.82
				139	444.40	1.80
140	416.40	1.50
				141	444.50	1.60
142	432.40	1.70
				143	358.40	1.40
144	374.40	1.30
				145	387.50	1.30
146	415.50	1.30
				147	371.20	1.50
148	417.50	1.20
				149	415.50	1.30
150	461.50	1.30
				151	373.40	1.20
152	441.50	1.40
				153	469.50	1.40
154	469.50	1.40
				155	403.40	1.40
156	362.00	1.30

157	375.40	1.10
				158	389.20	1.40
159	375.40	1.20

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				160	401.50	1.20
161	412.50	1.10
				162	451.50	1.50
163	464.20	1.20
				164	441.50	1.20
165	387.40	1.10
				166	401.50	1.30
167	444.20	1.60
				168	423.40	1.30
169	415.50	1.10
				170	390.40	1.50
171	346.30	1.30
				172	374.40	1.50
173	417.50	1.40
				174	445.50	1.60
175	469.50	1.50
				176	441.50	1.40
177	469.50	1.40

178	302.80	1.21	H NMR(500MHz，DMSO-d6)9.70(d，J＝2.3Hz，1H)，9.13(d，J＝2.3Hz，1H)，8.93(d，J＝6.8Hz，2H)，8.50(d，J＝6.8Hz，2H)，8.37(d，J＝8.0Hz，1H)，7.83(t，J＝7.8Hz，1H)，7.12(d，J＝7.5Hz，1H)，2.56(s，3H)
				179	373.00	1.46	H NMR(500Hz，DMSO-d6)9.64(s，1H)，9.16(s，1H)，8.84(d，J＝5.1Hz，2H)，8.58(s，1H)，8.27(d，J＝5.1Hz，2H)，7.59(s，1H)，3.73(m，4H)，1.65(m，6H)
180	361.00	1.35	500MHz，MeOD，8.12(d，1H)，7.77(d，1H)，7.51(d，2H)，7.28(s，1H)，6.88(d，2H)，5.89(s，1H)，2.47(m，4H)，1.01(t，6H)
				181	319.30	1.19	DMSO，500MHz，9.61(s，1H)，9.052(s，1H)，8.80(d，2H)，8.55(s，1H)，8.14(d，2H)，7.47(s，1H)，2.93(s，3H)
182	347.30	1.37	DMSO，500MH，9.58(s，1H)，9.01(s，1H)，8.79(d，2H)，8.52(s，1H)，8.08(d，2H)，7.46(s，1H)，4.15(m，1H)，1.19(d，6H)
				183	387.40	1.64	DMSO，500MHz，9.56(s，1H)，9.01(s，1H)，8.78(d，2H)，8.51(s，1H)，8.10(d，2H)，7.47(s，1H)，1.91(m，2H)，1.86(m，2H)，1.62(m，1H)，1.30(m，6H)
184	401.40	1.76	DMOS，500MHz，9.58(s，1H)，9.01(s，1H)，8.77(d，2H)，8.50(s，1H)，8.11(d，2H)，7.48(s，1H)，3.20(s，2H)，1.69(m，6H)，1.18(m，3H)，0.96(q，2H)

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				185	361.40	1.50	DMSO，500MHz，9.59(s，1H)，9.13(s，1H)，8.81(d，2H)，8.56(s，1H)，8.18(d，2H)，7.50(s，1H)，3.21(s，2H)，1.89(m，1H)，0.94(d，6H)

186			(400MHz，DMSO-d6)3.81(3H，s)，5.83(2H，br s)，7.05(2H，d)，7.21(1H，m)，7.43(2H，t)，7.71(2H，d)，7.86(2H，d)，8.69(1H，s)and9.05(1H，s).
				187			(DMSO)6.05(2H，m，NH2)，7.25(1H，m，ArH)，7.50(2H，m，4x ArH)，7.90(4H，m，4x ArH)，8.70(2H，d，ArHx 2)，8.85(1H，s，ArH)，9.40(1H，s，ArH).
188			(DMSO)2.65(1H，m)，2.91(6H，br s)，2.97(3H，s)，3.63(1H，m)，3.80(2H，m)，7.15(1H，t)，7.60(2H，d)，7.87(1H，t)，7.92(2H，d)，8.53-8.60(2H，m)，8.92(1H，s)，9.32(1H，s)，9.40(2H，br s).
				189			(DMSO)2.69(3H，s)，2.97(3H，s)，3.54-3.80(4H，m)，7.20(1H，m)，7.48-7.61(2H，m)，7.80-7.94(3H，m)，8.45(2H，br s)，8.54-8.58(2H，m)，8.88(1H，s)，9.28(1H，s)
190	416.60	3.73
				191	416.61	3.79	(DMSO)1.91(2H，m)，2.61(2H，m)，2.96(6H，m)，3.52(2H，m)，7.15(1H，t)，7.55(2H，d)，7.86(1H，t)，7.90(2H，d)，8.39(2H，br s)，8.57(2H，m)，8.91(1H，s)，9.32(1H，s).(DMSO)1.91-2.00(2H，m)，2.58-2.64(2H，m)，2.95-2.99(3H，m)，3.49-3.54(3H，br s)，3.64-3.74(2H，m)，7.15(1H，t)，7.55(2H，d)，7.86(1H，t)，7.90(2H，d)，8.39(2H，br s)，8.57(2H，m)，8.91(1H，s)，9.32(1H，s).；(DMSO)1.91(2H，m)，2.61(2H，m)，2.96(6H，m)，3.52(2H，m)，7.15(1H，t)，7.55(2H，d)，7.86(1H，t)，7.90(2H，d)，8.39(2H，br s)，8.57(2H，m)，8.91(1H，s)，9.32(1H，s)
192			(DMSO)1.38(9H，s)，4.17(2H，m)，7.08-7.15(3H，m)，7.34(2H，d)，7.48(1H，m)，7.78(2H，d)，7.84(1H，t)，8.52-8.58(2H，m)，8.85(1H，s)，9.21(1H，s)

193

316.40

4.42

(DMSO)7.16(1H，t)，7.21(2H，br s)，7.88(1H，t)，8.01(2H，d)，8.10(2H，d)，8.56-8.60(2H，m)，8.98(1H，s)，9.40(1H，s)，10.09(1H，s).

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				194		4.14	(DMSO)4.55(2H，d)，5.27(1H，t)，7.10-7.18(3H，m)，7.45(2H，d)，7.79(2H，d)，7.86(1H，t)，8.54-8.60(2H，m)，8.89(1H，s)，9.23(1H，s)
195	317.44	3.72	(DMSO)4.09(2H，m)，7.19(1H，t)，7.60(2H，d)，7.88-7.92(3H，m)，8.28(2H，br s)，8.55-8.59(2H，m)，8.92(1H，s)，9.32(1H，s).
				196			DMSO 7.15(1H，t，ArH)，7.20(2H，br s，NH2)，7.70(1H，t，ArH)，7.90(2H，d，ArHx2)，8.20(2H，d，ArHx 2)，8.60(2H，m，ArHx 2)，8.95((1H，s，ArH)，9.40(1H，s，ArH).
197			DMSO 7.15(1H，t，ArH)，7.70(1H，t，ArH)，7.90(2H，d，ArHx 2)，8.20(1H，d，ArH)，8.35(1H，s，ArH)，8.60(2H，m，ArHx 2)，8.95((1H，s，ArH)，9.40(1H，s，ArH).
				198	359.80	4.42	(DMSO)2.9(2H，t)，3.6(2H，t)，7.1-7.3(6H，m)，7.9(1H，t)，8.5(2H，m)，8.8(1H，m)，8.9(1H，s)，9.2(1H，s)
199	360.80	3.80	(DMSO)4.3(2H，d)，5.0(NH2)，6.5(2H，d)，7.1(2H，d)，7.2(1H，m)，7.3(NH2)，7.9(1H，m)，8.5(2H，m)，8.9(NH)，9.0(1H，m)，9.3(1H，s)
				200	346.70	3.75	(DMSO)4.6(2H，m)，7.2(1H，m)，7.3(NH2)，7.4(1H，m)，7.8(1H，m)，7.9(1H，m)，8.4-8.7(4H，m)，8.9(1H，s)，9.2(1H，m)，9.3(NH)

201	452.90	4.65	DMSO)1.1(2H，m)，1.4(9H，m)，1.7(3H，m)，2.7(2H，m)，4.0(2H，m)，7.1(1H，m)，7.3(NH2)，7.9(1H，m)，8.4-8.6(3H，m)，8.9(1H，s0，9.3(1H，s)
				202	317.44	3.82	(DMSO)4.09-4.17(2H，m)，7.20(1H，t)，7.50(1H，d)，7.58(1H，t)，7.88(1H，d)，7.91(1H，t)，8.02(1H，s)，8.40(2H，br s)，8.58-8.62(2H，m)，8.92(1H，s)，9.29(1H，s).
203	336.44	4.85	(DMSO)4.84(2H，s)，7.11-7.17(3H，m)，7.58(2H，d)，7.82-7.86(3H，m)，8.53-8.56(1H，m)，8.58(1H，d)，8.89(1H，s)，9.27(1H，s)
				204	269.70	3.50	(DMSO)2.8(3H，d)，7.1(1H，m)，7.3(2H，brs)，7.9(1H，m)，8.5-8.7(3H，m)，8.9(1H，s)，9.2(1H，s)
205	311.74	4.24	(DMSO)0.9(6H，m)，1.9(1H，m)，3.1(2H，m)，7.1(1H，m)，7.2(NH2)，7.9(1H，t)，8.5-8.6(3H，m)，8.9(1H，s)，9.3(1H，s)

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				206	338.75	3.30	(DMSO)0.8(2H，m)，1.6(1H，m)，2.0(2H，m)，2.9(2H，m)，3.1-3.3(2H，m)，7.2(1H，m)，8.0(1H，m)，8.4(1H，m)，8.6(1H，m)，8.7(NH2)，8.9(1H，s)，9.2(1H，s)
207	345.75	4.30	(DMSO)4.6(2H，d)，7.2(1H，m)，(7.3，2H，s)，7.4(3H，m)，7.9(1H，m)，8.5(2H，m)，8.9(1H，s)，9.2(1H，m)，9.3(NH，s)
				208	387.54	4.59	(DMSO)2.40(4H，m)，3.52(2H，s)，3.59(4H，t)，7.09-7.15(3H，m)，7.45(2H，d)，7.78(2H，d)，7.84(1H，t)，8.55(1H，d)，8.59(1H，d)，8.86(1H，s)，9.22(1H，s).

209			(DMSO)7.10(3H，m，ARH，NH2)，7.25(2H，m，ArHx 2)，7.40(3H，m，ArH x 3)，7.60(2H，m，ArHx 2)，7.85(1H，t，ArH)，8.55(2H，m，ArHx 2)，8.85(1H，s，ArH)，9.10(1H，s，ArH).
				210	336.42	4.85	(DMSO)4.84(2H，s)，7.12-7.17(3H，m)，7.47-7.56(2H，m)，7.81(1H，d)，7.86(1H，t)，7.91(1H，s)，8.56(1H，d)，8.59(1H，d)，8.86(1H，s)，9.24(1H，s).
211			1H NMR(DMSO)2.55(1H，m)，2.89(4H，m)，3.02(6H，br s)，3.82(2H，br s)，3.96(2H，s)，5.98(2H，s)，7.14(1H，m)，7.22-7.26(2H，m)，7.31-7.32(2H，m)，7.98(1H，m)，8.14(1H，m)，8.73(1H，m)，9.01(1H，m)，9.32(1H，s)1H NMR(DMSO)2.55(1H，m)，2.89(4H，m)，3.02(6H，br s)，3.82(2H，br s)，3.96(2H，s)，5.98(2H，s)，7.14(1H，m)，7.22-7.26(2H，m)，7.31-7.32(2H，m)，7.98(1H，m)，8.14(1H，m)，8.73(1H，m)，9.01(1H，m)，9.32(1H，s)
				212			DMSO 6.50(1H，s，ArH)，7.10(2H，s，NH2)，7.15(1H，t，ArH)，7.40(1H，d，ArH)，7.55(2H，dd，ArHx 2)，7.85(1H，t，ArH)，8.00(1H，s，ArH)，8.50(1H，m，ArH)，8.60(1H，d，ArH)，8.90(1H，s，ArH)，9.15(1H，s，ArH).
213			(DMSO)3.00(2H，s，CH2)，3.30(2H，d，CH2)，6.95(2H，s，NH2)，7.10(1H，t，ArH)，7.30(5H，m，ArHx 5)，7.80(1H，t，ArH)，8.40(1H，s，ArH)，8.50(2H，m，ArHx 2)，8.70(1H，s，ArH).
				214	345.52	4.30	(DMSO)2.75(6H，s)，4.32(2H，s)，7.18(1H，t)，7.68(2H，d)，7.87-7.98(3H，m)，8.55-8.60(2H，m)，8.93(1H，s)，9.34(1H，s)，10.40(2H，m).

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				215	345.52	4.32	(DMSO)2.75(6H，d)，4.36(2H，d)，7.25(1H，t)，7.59(1H，t)，7.61(1H，t)，7.92-8.00(2H，m)，8.13(1H，s)，8.57(1H，d)，8.60(1H，d)，8.99(1H，s)，9.38(1H，s)，10.83(2H，m).
216			(DMSO)2.65(3H，s，CH3)，3.85(3H，s，CH3)，7.10(5H，m，ArHx 3，NH2)，7.50(2H，d，ArH x 2)，7.85(1H，t，ArH)，8.40(1H，s，ArH)，8.55(1H，d，ArH)，8.65(1H，d，ArH).
				217	386.56	2.97	(DMSO)2.56-2.65(4H，m)，3.09-3.14(4H，m)，3.61(2H，s)，7.10-7.18(2H，m)，7.43(2H，d)，7.81(2H，d)，7.85(1H，t)，8.53-8.61(3H，m)，8.88(1H，s)，9.22(1H，s).
218			(DMSO)3.85(3H，s，CH3)，7.10(2H，d，ArHx 2)，7.20(1H，t，ArH)，7.40(4H，m，ArHx2，NH2)，7.90(1H，t，ArH)，8.50(1H，s，ArH)，8.60(2H，m，ArH x 2).
				219			1H NMR(DMSO)3.81(3H，s)，6.14(2H，brs)，7.07(2H，d)，7.74(2H，d)，7.96(2H，d)，8.53(2H，d)，8.81(1H，s)，9.14(1H，s)
220	332.73	3.73	(DMSO)2.3(3H，s)，3.9(3H，s)，7.0(NH2)，7.1(2H，m)，7.6-7.8(3H，m)，8.3(1H，s)，8.4(1H，m)，8.8(1H，s)，9.2(1H，s)
				221	318.42	3.22	4.60(2H，s)，7.10-7.16(3H，m)，7.38(1H，d)，7.49(1H，t)，7.68(1H，d)，7.71(1H，s)，7.85(1H，t)，8.53-8.57(1H，m)，8.59(1H，d)，8.85(1H，s)，9.19(1H，s)

222			(DMSO)3.30(4H，s，CH2x 2)，3.50(4H，s，CH2x 2)，7.05(1H，d，ArH)，7.15(1H，d，ArH)，7.30(1H，d，ArH)，7.40(2H，m，ArH x2)，7.90(1H，t，ArH)，8.55(2H，m，ArHx 2)，8.75(2H，s，NH2)，8.90(1H，s，ArH)，9.25(1H，s，ArH).
				223	386.52	3.05	(DMSO)2.31-2.37(4H，m)，2.70(4H，t)，3.50(2H，s)，7.08-7.13(3H，m)，7.33(1H，d)，7.46(1H，t)，7.68-7.71(2H，m)，7.83(1H，t)，8.54-8.60(2H，m)，8.83(1H，s)，9.20(1H，s).
224	400.54	3.15	(DMSO)1.68(2H，m)，2.58(2H，m)，2.64(2H，m)，2.75(2H，m)，2.80(2H，m)，3.68(2H，s)，7.10(3H，m)，7.46(2H，d)，7.76(2H，d)，7.84(1H，t)，8.54(1H，d)，8.57(1H，d)，8.86(1H，s)，9.20(1H，s)
				225	387.50	3.50	(DMSO)2.39-2.43(4H，m)，3.54(2H，s)，3.59(4H，t)，7.09-7.16(3H，m)，7.37(1H，d)，7.48(1H，t)，7.68-7.73(2H，m)，7.85(1H，t)，8.56-8.60(2H，m)，8.87(1H，s)，9.22(1H，s).

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				226			(DMSO)1.3-1.4(1H，m)，1.9-2.3(3H，m)，2.5-2.6(1H，m)，2.6-2.7(1H，m)，3.6-3.7(2H，m)，7.0-7.2(2.5H，m)，7.3-7.4(1H，m)，7.4-7.5(1H，m)，7.6-7.7(2H，m)，7.8-7.9(1H，m)，8.5-8.6(2H，m)，8.8-8.9(1H，s)，9.2(1H，s)
227			(DMSO)1.3-1.4(1H，m)，1.6-2.2(3H，m)，2.5-2.6(1H，m)，2.6-2.7(1H，m)，3.6-3.7(2H，m)，7.0-7.2(2.5H，m)，7.3-7.4(1H，m)，7.4-7.5(1H，m)，7.6-7.7(2H，m)，7.8-7.9(1H，m)，8.5-8.6(2H，m)，8.8-8.9(1H，s)，9.2(1H，s)
				228			(DMSO)1.6-1.7(2H，m)，2.5-2.9(8H，m)，3.7(2H，m)，7.0-7.2(2.8H，m)，7.3-7.4(1H，m)，7.4-7.5(1H，m)，7.7(2H，m)，7.8-7.9(1H，m)，8.5-8.6(2H，m)，8.8-8.9(1H，s)，9.2(1H，s)

229			(DMSO)1.2-1.3(2H，m)，1.6-1.7(2H，m)，1.9-2.0(2H，m)，2.4-2.6(1H，m)，2.7-2.9(2H，m)，3.5-3.6(2H，m)，7.0-7.2(3H，m)，7.3(1H，m)，7.4-7.5(1H，m)，7.7-7.8(2H，m)，7.8-7.9(1H，m)，8.5-8.6(2H，m)，8.8(1H，m)，9.2(1H，m)
				230			(DMSO)1.60(2H，quin)，2.12(3H，s)，2.27(2H，br s)，2.38(2H，t)，2.50(3H，m)，3.49(2H，s)，7.08-7.14(3H，m)，7.40(2H，d)，7.77(2H，d)，7.85(1H，t)，8.54-8.59(2H，m)，8.86(1H，s)，9.20(1H，s).
231			(DMSO)1.40-1.48(1H，m)，2.00-2.10(1H，m)，2.22(dd)，2.41-2.49(2H，m)，2.58-2.69(2H，m)，3.37-3.43(2H，m)，3.58(2H，d)，3.64(2H，d)，7.09-7.15(3H，m)，7.42(2H，d)，7.75(2H，d)，7.84(1H，t)，8.53-8.59(2H，m)，8.86(1H，s)，9.21(1H，s).
				232			(DMSO)1.74-1.79(2H，m)，2.22-2.28(2H，m)，2.95(2H，t)，3.36-3.40(2H，m)，4.18-4.24(2H，m)，7.09-7.20(2H，m)，7.14(1H，t)，7.62(2H，d)，7.86(1H，t)，8.93(2H，d)，8.53-8.58(2H，m)，8.89(1H，s)，9.29(1H，s).
233			(DMSO)1.40-1.48(1H，m)，2.00-2.10(1H，m)，2.22(dd)，2.41-2.49(2H，m)，2.58-2.69(2H，m)，3.37-3.43(2H，m)，3.58(2H，d)，3.64(2H，d)，7.09-7.15(3H，m)，7.42(2H，d)，7.75(2H，d)，7.84(1H，t)，8.53-8.59(2H，m)，8.86(1H，s)，9.21(1H，s).

Compound # (V-)

M+1(obs)

Rt (minutes)

1H-NMR

234			(DMSO)1.48-1.60(2H，m)，1.81-1.92(2H，m)，1.97-2.08(2H，m)，2.81-2.90(1H，m)，2.95-3.09(1H，m)，3.48-3.59(2H，m)，7.10-7.18(3H，m)，7.38-7.44(2H，m)，7.75-7.89(3H，m)，8.54-8.60(2H，m)，8.88(1H，br s)，9.22(1H，br s).
				235	331.80	3.23	(DMSO)7.0(NH2)，7.1(1H，m)，7.5-7.7(3H，m)，7.9(H，m)，8.0(2H，m)，8.5(2H，m)，8.7(1H，s)，9.4(1H，s)，10.7(NH)
236	331.84	2.96	(DMSO)1.37(1H，s)，2.29(3H，s)，3.70(2H，s)，7.08-7.14(3H，m)，7.43(2H，d)，7.75(2H，d)，7.85(1H，t)，8.55(1H，d)，8.58(1H，d)，8.88(1H，s)，9.20(1H，s).
				237	294.73	3.22	(DMSO)1.49(6H，s)，5.54(1H，s)，7.12-7.19(3H，m)，7.85(1H，t)，8.44(1H，s)，8.50(1H，d)，8.56(1H，d)，9.01(1H，s).
238	266.63	2.95	(DMSO)4.33(2H，d)，5.43(1H，t)，7.10-7.20(3H，m)，7.82(1H，t)，8.47-8.60(3H，m)，9.09(1H，s).
				239	286.69	2.69	(DMSO)3.83(3H，s)，7.08(2H，d)，7.34(2H，br s)，7.80(2H，d)，9.08(1H，s)，9.37(1H，s)
240	256.68	2.54	(DMSO)3.79(3H，s)，7.02(2H，d)，7.64(2H，d)，8.49(1H，s)，8.84(1H，s)
				241	381.40	1.59	DMSO，500MHz，9.73(s，1H)，9.45(s，1H)，9.101(s，1h)，8.82(d，2H)，8.66(s，1H)，8.15(d，2H)，7.93(s，1H)，7.73(d，2H)，7.38(t，2H)，7.14(t，1H)
242	411.40	1.56	DMSO，500MHz，9.61(s，1H)，9.02(s，1H)，8.84(d，2H)，8.58(s，1H)，8.24(d，2H)，7.81(s，1H)，7.77(d，1H)，7.19(t，1H)，7.14(d，1H)，7.00(t，1H)，3.86(s，3H)

243	375.40	1.56	DMSO，500MHz 9.61(s，1H)，9.12(s，1H)，8.80(d，2H)，8.55(s，1H)，8.19(d，2H)，7.58(s，1H)，3.51(m，2H)，3.18(s，3H)，2.11(m，1H)，0.96(d，6H)
				244	375.40	1.52	DMSO，500MHz，9.63(s，1H)，9.12(s，1H)，8.79(d，2H)，8.57(s，1H)，8.21(d，2H)，7.58(s，1H)，3.66(m，2H)，3.50(m，2H)，1.68(q，2H)，1.21(t，3H)，0.96(t，3H)
245	362.40	1.40
				246	375.40	1.20
247	463.20	1.30
				248	417.00	1.20
249	461.50	1.20
				250	412.50	1.30
251	388.40	1.40

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				252	409.40	1.20
253	444.50	1.60
				254	401.50	1.10
255	451.50	1.40
				256	441.50	1.20
257	374.40	1.30
				258	358.40	1.30
259	372.20	1.40
				260	430.50	1.40

261	441.50	1.50
				262	388.40	0.70
263	423.20	1.30
				264	464.20	1.30
265	441.50	1.20
				266	417.20	1.30
267	401.50	1.20
				268	386.20	1.30
269	415.50	1.30
				270	402.50	1.30
271	416.50	1.30
				272	364.50	1.30
273	284.30；284.40	1.61；1.53	H NMR(500MHz，DMSO-d6)8.71(s，1H)，8.37(d，J＝1.6Hz，2H)，7.45(s，1H)，7.20(s，1H)，6.89(s，2H)，4.10-4.07(m，1H)，2.30(s，3H)，1.16(d，J＝6.4Hz，3H).
				274	347.80	2.55
275	399.70	2.86
				276	310.30；310.44	1.80	H NMR(500MHz，DMSO-d6)8.90(m，2H)，8.65(s，1H)，8.52(d，J＝1.7Hz，1H)，3.81(m，4H)，2.35(s，3H)，1.69(m，2H)，1.63(m，4H).
277	296.30	1.80
				278	326.30	1.60
279	318.30	1.90

280	245.70	2.20
				281	273.40	2.73	DMSO，500MHz，8.70(s，2H)，8.48(d，1H)，8.31(s，1H)，2.59(s，3H)，2.31(s，3H)
282	298.50	1.95	H NMR(500MHz，CDCl3)9.93(brs，1H)，8.45(s，1H)，8.34(s，1H)，8.29(s，1H)，7.86(s，1H)，3.69-3.67(m，1H)，2.43(s，3H)，1.85-1.68(m，2H)，1.40(d，J＝6.5Hz，3H)，1.05(t，J＝7.4Hz，3H)，

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				283	300.28	1.41	H NMR(500MHz，MeOD)8.65(s，1H)，8.55(s，1H)，8.51(s，1H)，8.48(s，1H)，7.15(brs，1H)，4.29(brs，1H)，3.67-3.60(m，2H)，2.40(s，3H)，1.29(d，J＝6.7Hz，3H)
284	339.36	1.19
				285	415.44	1.51	H NMR(500MHz，MeOD)8.67(s，1H)，8.63(s，H)，8.52(s，1H)，)7.54-7.50(m，5H)，7.07(s，1H)，4.34(s，2H)，3.61(m，2H)，3.33-3.30(m，2H)，2.40(s，3H)，2.33(m，2H)，2.31(m，1H)，1.91(m，2H)
286	401.44	1.48	H NMR(500MHz，MeOD)8.67(s，1H)，8.63(s，H)，8.52(s，1H)，8.51(s，1H)，7.54-7.50(m，5H)，7.07(s，1H)，3.61(m，2H)，3.33-3.30(m，2H)，2.40(s，3H)，2.33(m，2H)，2.31(m，1H)，1.91(m，2H)
				287	314.13	1.52	H NMR(500MHz，MeOD)8.66(s，1H)，8.55(s，1H)，8.52(s，1H)，8.51(1H)，7.27(s，1H)，4.38(m，1H)，3.71-3.62(，m，2H)，2.65(s，3H)，1.62-1.59(m，2H)，1.02(t，J＝7.4Hz，3H)

288	314.40	1.69	H NMR(500MHz，MeOD)8.65(s，1H)，8.54(s，1H)，8.50(s，1H)，7.19(s，1H)，3.72-3.62(m，2H)，2.40(s，3H)，2.2.02(d，J＝10.5Hz，1H)，1.80-1.75(m，1H)，1.64-1.58(m，1H)，1.02(t，J＝7.4Hz，3H)
				289	282.30	1.60
290	296.30	1.70
				291	324.30	2.00
292	314.30	1.60
				293	300.30	1.40
294	298.30	1.80
				295	300.30	1.40
296	312.30	2.00	H NMR(500MHz，DMSO-d6)8.84(s，2H)，8.53(s，1H)，8.47(s，1H)，2.34(m，4H)，1.81(d，J＝6.6Hz，1H)，1.14-1.11(m，3H)，0.90(m，6H).
				297	352.40	2.30
298	342.30	1.80
				299	310.40	1.73
300	346.40	2.17
				301	346.40	2.21
302	340.30	2.70
				303	372.40	3.30
304	400.40	3.70
				305	379.40	2.70
306	326.30	2.20

Compound # (V-)	M+1(obs).	Rt (minutes)	1H-NMR
				307	390.40	3.20
308	374.30	1.90
				309	353.40	1.90；1.40	H NMR(500MHz，DMSO-d6)8.93(s，1H)，8.70(s，1H)，8.54(d，J＝1.9Hz，1H)，7.48(s，1H)，7.11(s，1H)，6.96(s，1H)，3.32-3.25(m，2H)，2.47(m，2H)，2.36(s，3H)，2.00-1.96(m，1H)，1.88-1.83(m，2H)，1.72(m，1H)，1.52(s，1H).
310	270.10	1.30
				311	323.40	1.12
312	337.20	1.20
				313	339.40	0.90
314	337.42	0.90
				315	366.50	1.00
316	297.30	1.00
				317	339.40	1.30
318	311.40	1.10
				319	325.40	1.20
320	353.50	1.30
				321	382.50	1.10
322	309.20	1.20
				323	351.50	1.40
324	323.20	1.30
				325	339.40	1.00
326	327.40	1.10

327	311.40	1.90
				328	355.40	1.20
329	353.40	1.20
				330	409.50	1.40
331	380.50	1.10
				332	310.30	1.90
333	336.40	2.10
				334	416.50	2.20
335	312.30	1.50
				336	416.50	2.20
337	311.43	1.48	H NMR(500MHz，DMSO-d6)8.80(s，1H)，8.55(s，1h)，8.44(d，J＝1.8Hz，1H)，8.09(m，3H)，7.35(s，1H)，3.99(m，1H)，3.81(m，1H)，3.79-3.58(m，2H)，2.32(s，3H)，2.19-2.13(m，1H)，1.93-1.87(m，1H).
				338	409.50	1.80
339	324.30	2.00
				340	340.30	1.60
341	353.40	1.40
				342	326.30	1.50

Compound # (V-)	M+1(obs)	Rt (minutes)	1H-NMR
				343	407.40	1.68
344	353.40	1.50
				345	270.30	1.50
346	339.30	1.30

347	312.30	1.40
				348	395.10	1.70
349	296.30	1.70
				350	373.30	1.50
351	346.20	1.50
				352	429.40	1.90
353	373.30	1.50

Example 13: ITK inhibition assay (radio assay)

Compounds were screened for their ability to inhibit Itk using a radioactive phosphate binding assay. The assay was at 100mM HEPES (pH 7.5), 10mM MgCl₂25mM NaCl, 0.01% BSA in a mixture with 1mM DTT. The final substrate concentration was 15. mu.M [ gamma-³³P]ATP(400mCi³³P ATP/mmol ATP, Amersham Pharmacia Biotech/Sigma Chemicals) and 2. mu.M peptide (SAM68 protein D332-443). The assay was performed at 25 ℃ in the presence of 30nM Itk. Assay stock buffer solutions were prepared containing all the above reagents, with the exception of ATP and related test compounds. 50 μ l of stock solution was placed in 96-well plates, followed by addition of 1.5 μ l of DMSO stock solution containing serial dilutions of test compound (usually starting at a final concentration of 15 μ M, 2-fold serial dilutions), in duplicate (final DMSO concentration 1.5%). The plate was preincubated at 25 ℃ for 10 minutes, and 50. mu.l of [ gamma-³³P]ATP initiated the reaction (final concentration 15. mu.M).

After 10min the reaction was stopped by adding 50. mu.l of TCA/ATP mixture (20% TCA, 0.4mM ATP). Unifilter GF/C96 well plates (Perkin Elmer Life Sciences, Cat No.6005174) were pretreated with 50. mu.l Milli Q water and the entire reaction mixture (150. mu.l) was added. Plates were washed with 200. mu.l Milli Q water, followed by 200ml of TCA/ATP mix (5% TCA, 1mM ATP). This washing cycle was repeated 2 additional times. After drying, 30 μ l Optiphase 'SuperMix' Liquid scintillation cocktail reagent (PerkinElmer) was added to the wells, followed by scintillation counting (1450Microbeta Liquid scintillation counter, Wallac).

Using the Prism Software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego California, USA), the number of initial rates was countedCalculating IC according to nonlinear regression analysis₅₀And (4) data.

The assay was at 20mM MOPS (pH 7.0), 10mM MgCl₂0.1% BSA in a mixture with 1mM DTT. The final substrate concentration was 7.5. mu.M [ gamma. ]³³P]ATP(400mCi³³PATP/mmol ATP, Amersham Pharmacia Biotech/Sigma Chemicals) and 3. mu.M peptide (SAM68 protein D332-443). The assay was performed at 25 ℃ in the presence of 50nM Itk. Assay stock buffer solutions were prepared containing all the above reagents, with the exception of ATP and related test compounds. 50 μ l of stock solution was placed in 96-well plates, followed by addition of 2 μ l of DMSO stock solution containing serial dilutions of test compound (usually starting at a final concentration of 50 μ M, 2-fold serial dilutions), in duplicate (final DMSO concentration 2%). The plates were preincubated at 25 ℃ for 10 minutes and 50. mu.l of [ gamma. ] was added³³P]ATP initiated the reaction (final concentration 7.5. mu.M).

After 10min, 100mL of 0.2M phosphoric acid + 0.01% Tween 20 was added to stop the reaction. Multiscreen phosphocellulose 96-well filter plates (Millipore, Cat No. maphn0b50) were pretreated with 100 μ L of 0.2M phosphoric acid + 0.01% tween 20, and then 170mL of a stop assay mixture was added. Plates were washed with 4 × 200 μ L0.2M phosphate + 0.01% tween 20. After drying, 30 μ L Optiphase 'Supermix' liquid Scintillation cocktail reagent (Perkin Elmer) was added to the wells, followed by Scintillation counting (1450Microbetaliquid Scintillation Counter, Wallac).

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

Example 14: ITK inhibition assay (UV)

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

The assay was at 20mM MOPS (pH 7.0), 10mM MgCl₂0.1% BSA, 1mM DTT, 2.5mM phosphorusAcid enol pyruvate, 300. mu.M NADH, 30. mu.g/ml pyruvate kinase and 10. mu.g/ml lactate dehydrogenase. The final substrate concentrations in the assay were 100. mu.M ATP (Sigma Chemicals) and 3. mu.M peptide (biotinylated SAM68D 332 89332-443). The assay was performed at 25 ℃ in the presence of 100nM Itk.

Assay stock buffer solutions were prepared containing all the above reagents, with the exception of ATP and related test compounds. 60 μ l of stock solution was placed in each well of a 96-well plate, followed by the addition of 2 μ l DMSO (usually starting at a final concentration of 15 μ M) containing serial dilutions of the test compound. The plates were preincubated at 25 ℃ for 10 minutes and 5. mu.l ATP was added to initiate the reaction. The initial reaction rate was determined for 10 minutes using a Molecular Devices SpectraMax Plus plate reader. IC was calculated from nonlinear regression analysis using the Prism Software package (GraphPad Prism version 3.0cxfor Macintosh, GraphPad Software, San Diego California, USA)₅₀And Ki data.

In general, the compounds of the present invention, including the compounds of table 1, are effective in the inhibition of ITK.

Example 15: assay for BTK inhibition

Compounds were screened for their ability to inhibit Btk using the radioactive phosphate binding assay in Vertex Pharmaceuticals. The assay was at 20mM MOPS (pH 7.0), 10mM MgCl₂0.1% BSA in a mixture with 1mM DTT. The final substrate concentration was 50. mu.M [ gamma-³³P]ATP(200mCi³³PATP/mmol ATP, Amersham Pharmacia Biotech, Amersham, UK/Sigma Chemicals) and 2. mu.M peptide (SAM68D 332-443). The assay was performed in the presence of 25nM Btk at 25 ℃. Assay stock buffer solutions were prepared containing all the above reagents, except peptides and related test compounds. 75 μ l of stock solution was placed in 96-well plates, followed by addition of 2 μ l of DMSO stock solution (usually starting at a final concentration of 15 μ M) containing serial dilutions of the test compound in duplicate (final DMSO concentration 2%). The plates were preincubated at 25 ℃ for 15 minutes and 25. mu.l peptide was added to initiate the reaction (finalConcentration 2. mu.M). Prior to priming with peptide, 100mL of 0.2M phosphate + 0.01% tween was added to control wells containing assay stock buffer and DMSO, and background counts were determined.

After 10min, 100mL of 0.2M phosphoric acid + 0.01% Tween was added to stop the reaction. Multiscreen phosphocellulose 96-well filter plates (Millipore, Cat No. maphn0b50) were pretreated with 100 μ L of 0.2M phosphoric acid + 0.01% tween 20, and then 170mL of a stop assay mixture was added. Plates were washed with 4 × 200 μ L0.2M phosphate + 0.01% tween 20. After drying, 30 μ L Optiphase 'Supermix' liquid Scintillation cocktail reagent (Perkin Elmer) was added to the wells, followed by Scintillation counting (1450Microbetaliquid Scintillation Counter, Wallac).

After removing the average background value for all data points, ki (app) data was calculated from nonlinear regression analysis using the Prism Software package (GraphPadPrism version 3.0cx for Macintosh, GraphPad Software, San diego california, USA).

AlphaScreen was used in Vertex Pharmaceuticals^TMCompounds are screened for their ability to inhibit Btk. The assay was at 20mM MOPS (pH 7.0), 10mM MgCl₂0.1% BSA in a mixture with 1mM DTT. The final substrate concentrations in the assay were 50. mu.M ATP (Sigma Chemicals) and 2. mu.M peptide (biotinylated SAM68D 332-443). The assay was performed in the presence of 25nM Btk at 25 ℃. Assay stock buffer solutions were prepared containing all the above reagents, except peptides and related test compounds. 37.5 μ l of stock solution was placed in each well of a 96-well plate, followed by the addition of 1 μ l DMSO containing serial dilutions of test compounds (usually starting at a final concentration of 15 μ M), in duplicate (final DMSO concentration 2%). The plates were preincubated at 25 ℃ for 15 minutes and the reaction was initiated by the addition of 12.5. mu.l peptide (final concentration 2. mu.M). Prior to priming with biotin-SAM 68, 5 μ l of 500mM EDTA was added to control wells containing assay stock buffer and DMSO, and background counts were determined.

After 30 minutes, the 225-fold reaction mixture was diluted to MOPS buffer (20mM MOPS (pH 7.0), 1mM DTT, 10mM MgCl) containing 50mM EDTA₂，0.1% BSA) to give a final peptide concentration of 9 nM.

Alphascreen was prepared according to the manufacturer's instructions^TMReagent (AlphaScreen)^TMPhosphotyrosine (P-Tyr-100) assay kit, Perkinelmer catalog number 6760620C). Under soft light, 20. mu.l Alphascreen^TMReagents were placed in each well of a white half-zone 96-well plate (Corning Inc. -COSTAR 3693) containing 30. mu.l of the stopped diluted kinase reaction. The plates were incubated in the dark for 60 minutes and then read on a Fusion Alpha plate reader (PerkinElmer).

After removing the average background value for all data points, ki (app) data was calculated from nonlinear regression analysis using the Prism Software package (GraphPadPrism version 3.0cx for Macintosh, GraphPad Software, San diego california, USA).

In general, compounds of the invention, including the compounds of table 1, are effective in the inhibition of Btk.

Example 16: RLK inhibition assay

Compounds were screened for their ability to inhibit Rlk using a standard coupled enzyme assay (Fox et al, Protein Sci., (1998)7, 2249). The assay was at 20mM MOPS (pH 7.0), 10mM MgCl₂0.1% BSA in a mixture with 1mM DTT. The final substrate concentrations in the assay were 100. mu.M ATP (Sigma Chemicals) and 10. mu.M peptide (Poly Glu: Tyr 4: 1). The assay was performed at 30 ℃ in the presence of 40nM Rlk. The final concentrations of the components of the coupled enzyme system were 2.5mM phosphoenolpyruvate, 300. mu.M NADH, 30. mu.g/ml pyruvate kinase and 10. mu.g/ml lactate dehydrogenase.

Assay stock buffer solutions were prepared containing all the above reagents, with the exception of ATP and related test compounds. 60 μ l of stock solution was placed in each well of a 96-well plate, followed by the addition of 2 μ l DMSO (usually starting at a final concentration of 7.5 μ M) containing serial dilutions of the test compound. The plates were preincubated at 30 ℃ for 10 minutes and 5. mu.l ATP was added to initiate the reaction. Utilizing Molecular Devices SpectraMax Plus PinpinThe plate reader measures the initial reaction rate for 10 minutes. IC was calculated from nonlinear regression analysis using the Prism Software package (GraphPad Prism version 3.0cxfor Macintosh, GraphPad Software, San Diego California, USA)₅₀And Ki data.

In general, the compounds of the present invention, including the compounds of table 1, are effective in the inhibition of Rlk.

Example 17: assay for inhibition of JAK3

JAK inhibition of compounds was determined by the method described in g.r.brown et al, bioorg.med.chem.lett.2000, vol.10, pp 575-. To Maxisorb plates previously coated with Poly (Glu, Ala, Tyr) 6: 3: 1 at 4 ℃ and washed with phosphate buffered saline 0.05% and Tween (PBST) was added 2. mu.M ATP, 5mM MgCl₂And DMSO solutions of compounds. The reaction was started with JAK enzyme and the plates were incubated at 30 ℃ for 60 minutes. The plates were then washed with PBST, 100. mu.l of HRP-conjugated 4G10 antibody was added, and the plates were incubated at 30 ℃ for 90 minutes. The plate was washed again with PBST, 100. mu.l TMB solution was added, and the plate was incubated at 30 ℃ for another 30 minutes. The reaction was stopped by addition of sulfuric acid (100. mu.l, 1M), and the plates were read at 450nm to obtain optical densities for analytical determination of K_iThe value is obtained.

Compounds may also be assayed for JAK inhibition in the following manner: compounds were screened for their ability to inhibit JAK3 using the assays shown below. The reaction was carried out in kinase buffer containing 100mM HEPES (pH 7.4), 1mM DTT, 10mM MgCl₂25mM NaCl and 0.01% BSA. The substrate concentrations in the assay were 5. mu.M ATP (200. mu. Ci/. mu.mol ATP) and 1. mu.M poly (Glu)₄Tyr. The reaction was carried out at 25 ℃ and 1nM JAK 3. To each well of a 96-well polycarbonate plate, 1.5. mu.l of candidate JAK3 inhibitor and 50. mu.l of kinase buffer containing 2. mu.M poly (Glu)₄Tyr and 10. mu.M ATP. Then mixed, and the reaction was started by adding 50. mu.l of kinase buffer containing 2nM JAK3 enzyme. After 20 minutes at room temperature (25 ℃), the reaction was stopped with 50. mu.l of 20% trichloroacetic acid (TCA) also containing 0.4mM ATP.The entire contents of each well were then transferred to a 96-well glass fiber filter plate using a TomTek cell harvester. After washing, 60. mu.l scintillation fluid was added and the assay was performed on a Perkin Elmer TopCount³³And (3) binding of P.

In general, the compounds of the invention, including the compounds of table 1, are effective for the inhibition of JAKs (e.g., JAK-3).

Example 18: aurora B (Aurora-1) inhibition assay

An assay buffer solution was prepared having a composition of 25mM HEPES (pH 7.5), 10mM MgCl₂0.1% BSA and 10% glycerol. A22 nM Aurora-B solution was prepared in assay buffer, also containing 1.7mM DTT and 1.5mM Kemptide (LRRASLG). In 96-well plates, 2. mu.l of DMSO stock solution of the compound was added to 22. mu.l of Aurora-B solution, and the mixture was allowed to equilibrate for 10 minutes at 25 ℃. Add 16. mu.l of [ gamma-³³P]ATP stock solution (. about.20 nCi/. mu.l) to a final assay concentration of 800. mu.M, initiated the enzymatic reaction. After 3 hours the reaction was stopped by adding 16. mu.l of 500mM phosphoric acid, which was determined by the following method³³Level of P binding to peptide substrate.

Phosphocellulose 96-well plates (Millipore, Cat No. MAPNNEOB50) were pretreated with 100. mu.l of 100mM phosphoric acid, followed by addition of the enzyme reaction mixture (40. mu.l). The solution was soaked on a phosphocellulose membrane for 30 minutes, followed by washing the plate four times with 200. mu.l of 100mM phosphoric acid. To each well of the dry plate was added 30 μ l Optiphase 'Supermix' liquid scintillation cocktail reagent (Perkin Elmer) followed by scintillation counting (1450Microbeta Liquid science Counter, Wallac). The background radioactivity level without enzyme catalysis was determined by adding 16. mu.l of 500mM phosphoric acid containing all assay components (which serve to denature the enzyme) to a control well followed by [ gamma-³³P]-an ATP solution. Calculating enzyme catalyzed by subtracting the average background counts from the counts measured at each inhibitor concentration³³P binding level. At each time K_iFor the assay, 8 data points were obtained, usually covering the concentration range of 0-10 μ M compound, in duplicate (fromStock solutions of 10mM starting compound were serially diluted 1: 2.5 to prepare DMSO stock solutions). K was calculated from the initial data by means of non-linear regression using the Prism Software package (Prism 3.0, Graphpad Software, San Diego, Calif.)_iThe value is obtained.

Example 19: Aurora-A (Aurora-2) inhibition assay

Compounds were screened for their ability to inhibit Aurora-2 using a standard coupled enzyme assay (Fox et al, Protein ScL., (1998)7, 2249). The assay was at 100mM HEPES (pH 7.5), 10mM MgCl₂1mM DTT, 25mM NaCl, 2.5mM phosphoenolpyruvate, 300. mu.M NADH, 30. mu.g/ml pyruvate kinase and 10. mu.g/ml lactate dehydrogenase. The final substrate concentrations were 400. mu.M ATP (Sigma Chemicals) and 570. mu.M Peptide (Kemptide, American Peptide, Sunnyvale, Calif.). The measurement was carried out at 30 ℃ in the presence of 40nMA aurora-2.

Assay stock buffers were prepared containing all the above reagents, except Aurora-2 and the relevant test compounds. Place 55 μ l of stock solution into a 96-well plate, followed by addition of 2 μ l of DMSO stock solution containing serial dilutions of test compound (usually starting at a final concentration of 7.5 μ M). The plates were preincubated at 30 ℃ for 10 minutes and 10. mu.l of Aurora-2 was added to initiate the reaction. Initial reaction rates were determined using a Molecular Devices SpectraMax Plus plate reader over a 10 minute time course. IC was calculated from nonlinear regression analysis using the Prism Software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, San Diego California, USA)₅₀And K_iAnd (4) data.

Example 20: c-Met inhibition assay

Compounds were screened for their ability to inhibit c-Met kinase activity using a standard coupled enzyme assay (Fox et al, Protein Sci, 19987, 2249). The reaction was carried out in the presence of 100mM HEPES (pH 7.5), 10mM MgCl₂25mM NaCl, 300. mu.M NADH, 1mM DTT and 1.5% DMSO. The final substrate concentration in the assay was 20mu.M ATP (SigmaChemicals, St Louis, MO) and 10. mu.M polyGluTyr (Sigma chemical company, St. Louis). The assay was performed in the presence of 80nM c-Met at 30 ℃. The final concentrations of the components of the coupled enzyme system were 2.5mM phosphoenolpyruvate, 300. mu. MNADH, 30. mu.g/ml pyruvate kinase and 10. mu.g/ml lactate dehydrogenase.

An assay stock buffer solution was prepared containing all of the above reagents, with the exception of ATP and the test compound of the invention. Assay stock buffer (175. mu.l) was incubated with 5. mu.l of test compound of the invention in 96-well plates at 30 ℃ for 10 minutes to a final concentration of 0.006. mu.M to 12.5. mu.M. Typically, 12-point titrations are performed by preparing serial dilutions of test compounds of the invention (starting from a 10mM stock of compound) in DMSO in daughter plates. The reaction was initiated by the addition of 20. mu.l ATP (final concentration 200. mu.M). The reaction rate was obtained over 10min at 30 ℃ using a molecular devices Spectramax plate reader (Sunnyvale, Calif.). Determination of K from rate data_iValue as a function of inhibitor concentration.

Table 11 shows the enzyme inhibition data (K) for certain exemplary compounds_i). The compound numbers in table 11 correspond to those given in table 5. In Table 11, "A" represents K_iLess than 0.5. mu.M, "B" represents K_iBetween 0.5 and 5.0. mu.M, "C" represents K_iGreater than or equal to 5.0. mu.M for the indicated enzymes.

Table 11: biological characterization data for selected compounds

Compound # (V-)

AurA

AurB

ITK

JAK3

Met

RLK

1

C

2

B

B

C

B

3

B

C

B

4

B

C

B

5

B

C

B

6

B

B

B

7

B

C

B

8

B

C

B

9

B

C

B

10

B

C

B

11

B

C

B

12

B

C

B

13

B

C

B

14

B

B

C

B

15

B

C

B

16

B

C

B

17

B

C

B

18

A

A

C

A

19

B

C

B

20

B

C

B

21

B

C

B

22

B

B

C

A

23

B

C

B

24

B

B

C

B

25

B

C

B

26

B

C

B

27

B

C

B

28

B

C

B

29

B

C

B

30

B

C

B

31

B

C

B

32

B

C

B

33

B

B

C

B

34

B

C

B

35

B

C

B

36

B

C

B

37

B

C

B

38

B

C

B

39

B

C

B

40

B

C

B

Compound # (V-)

AurA

AurB

ITK

JAK3

Met

RLK

41

B

C

B

42

B

C

B

43

B

C

B

44

B

C

B

45

B

C

B

46

B

C

B

47

B

C

B

48

B

C

B

49

B

C

B

50

B

C

A

51

B

A

B

A

52

B

C

B

53

B

C

A

54

B

B

C

A

55

B

B

C

A

56

B

C

B

57

B

B

C

A

58

B

C

A

59

B

C

B

60

B

B

A

61

A

B

A

62

B

A

B

A

63

A

A

B

A

64

A

A

B

A

65

B

A

B

A

66

B

B

A

67

A

B

B

A

68

A

A

B

A

69

A

B

A

70

A

A

B

A

71

A

A

B

A

72

A

A

B

A

73

B

B

C

A

74

B

A

B

A

75

A

B

C

A

76

A

A

B

A

77

B

C

B

78

A

A

B

A

79

A

A

B

A

80

A

C

A

81

A

A

B

A

82

B

A

C

A

83

A

A

B

A

84

A

B

A

85

B

A

B

A

Compound # (V-)

AurA

AurB

ITK

JAK3

Met

RLK

86

B

B

C

A

87

A

A

B

A

88

A

B

A

89

A

A

B

A

90

A

A

B

A

91

A

A

A

92

A

A

B

A

93

A

A

B

A

94

B

A

C

A

95

A

B

A

96

B

B

C

A

97

A

A

B

A

98

A

A

B

A

99

A

B

A

100

B

C

A

101

B

B

C

A

102

B

B

C

A

103

B

C

A

104

B

B

C

A

105

B

C

A

106

B

B

C

A

107

B

B

A

108

B

C

A

109

B

C

A

110

A

B

A

111

B

C

A

112

B

B

C

A

113

A

B

A

114

B

B

C

A

115

B

B

C

A

116

B

B

C

A

117

B

C

A

118

B

C

A

119

B

B

A

120

A

B

A

121

B

C

A7

122

A

B

A

123

B

C

A

124

B

B

C

A

125

B

C

A

126

B

C

A

127

B

B

C

A

128

B

C

A

129

A

A

B

A

130

B

A

C

A

Compound # (V-)

AurA

AurB

ITK

JAK3

Met

RLK

131

B

B

B

132

B

B

B

C

A

133

A

B

B

B

A

134

B

B

B

C

B

135

B

B

B

C

B

136

B

C

B

137

B

C

B

138

B

C

B

139

B

B

C

A

140

B

C

A

141

B

C

A

142

B

B

A

143

B

C

144

B

C

A

145

A

C

A

146

B

A

C

A

147

B

C

B

148

A

B

A

149

B

B

A

150

A

C

A

151

B

A

C

A

152

B

A

C

A

153

A

B

A

154

B

B

C

A

155

B

A

B

A

156

B

B

C

A

157

A

C

A

158

B

C

A

159

B

A

B

A

160

B

B

C

A

161

B

C

A

162

B

C

B

163

B

C

A

164

A

C

A

165

A

A

C

A

166

B

A

C

A

167

B

C

A

168

B

C

B

169

A

C

A

170

B

C

B

171

B

C

A

172

B

C

B

173

B

A

B

A

174

B

C

B

175

B

B

B

A

Compound # (V-)

AurA

AurB

ITK

JAK3

Met

RLK

176

B

A

B

A

177

A

A

B

A

178

A

B

A

C

B

179

A

A

A

A

A

180

A

A

A

A

181

A

A

A

A

182

A

A

A

A

183

A

A

B

A

184

A

A

B

A

185

A

A

A

A

186

A

C

B

187

B

C

A

188

A

A

B

A

A

189

B

A

B

A

190

B

A

B

A

191

A

A

B

A

A

192

A

A

193

A

194

A

A

B

A

195

A

A

B

A

B

196

A

197

A

C

A

198

B

C

B

199

B

C

B

200

B

C

B

201

B

C

B

202

B

A

C

A

B

203

A

204

B

B

C

B

205

B

C

B

206

B

B

C

B

207

B

C

B

208

A

A

B

B

209

A

C

B

210

A

211

B

C

B

212

A

A

C

A

213

B

C

B

214

A

A

B

A

B

215

B

A

C

A

B

216

B

B

C

B

217

A

A

B

A

A

218

B

219

B

B

C

B

220

B

Compound # (V-)

AurA

AurB

ITK

JAK3

Met

RLK

221

B

B

B

A

222

A

A

B

A

B

223

B

A

B

A

B

224

A

A

N

A

225

B

B

C

A

226

B

A

C

A

227

B

A

C

A

B

228

B

A

C

A

B

229

B

A

C

A

B

230

A

A

B

A

A

231

A

A

B

A

B

232

B

A

C

A

B

233

A

A

B

A

B

234

A

A

B

A

B

235

B

C

B

236

A

A

B

A

B

237

B

A

B

A

238

B

B

B

A

239

B

C

B

240

B

C

B

241

A

A

A

A

242

A

A

A

A

243

A

A

A

A

244

A

A

A

A

245

B

C

A

246

B

C

A

247

B

C

A

248

B

B

A

249

B

B

A

250

A

B

A

251

B

C

A

252

B

C

B

253

B

C

B

254

B

C

B

255

B

C

B

256

B

C

B

257

B

C

B

258

B

C

B

259

B

C

B

260

B

C

B

261

B

C

B

262

B

C

B

263

B

C

B

264

B

C

B

265

B

C

B

Compound # (V-)

AurA

AurB

ITK

JAK3

Met

RLK

266

B

C

B

267

B

C

B

268

B

C

B

269

B

C

B

270

B

C

B

271

B

C

B

272

B

C

B

273

B

B

B

B

A

274

B

B

A

B

A

275

B

C

B

276

B

A

B

A

277

B

B

B

A

278

B

B

B

A

279

A

A

A

A

280

A

B

C

B

281

B

B

B

A

282

B

B

A

283

B

B

B

284

B

B

A

285

B

B

B

A

286

B

B

B

A

287

B

B

A

288

A

B

A

289

B

B

B

A

290

B

B

B

A

291

B

B

B

A

292

B

B

A

293

B

B

B

A

294

B

B

B

A

295

B

B

B

A

296

A

B

A

297

B

B

A

298

A

B

A

299

A

A

A

300

B

A

A

301

B

C

B

302

A

B

A

303

B

B

B

304

B

C

B

305

B

C

B

306

B

B

A

307

B

B

A

308

A

A

A

309

B

A

A

310

B

C

B

Compound # (V-)

AurA

AurB

ITK

JAK3

Met

RLK

311

B

C

A

312

B

C

B

313

B

C

B

314

B

C

B

315

B

C

B

316

B

C

B

317

B

C

B

318

B

C

B

319

B

C

A

320

B

C

B

321

B

C

B

322

B

C

B

323

B

C

B

324

B

C

A

325

B

C

B

326

B

C

B

327

B

C

B

328

B

C

B

329

B

C

A

330

B

C

B

331

B

C

B

332

A

B

A

333

A

B

A

334

B

B

A

335

B

B

A

336

B

B

A

337

A

A

A

338

B

B

A

339

A

A

A

340

B

B

A

341

B

B

B

342

B

B

A

343

B

B

B

While we have described some embodiments of this invention, it is apparent that our basic examples can be modified to provide other embodiments that employ the compounds and methods of this invention. It will therefore be appreciated that the scope of the invention is defined by the claims rather than the specific embodiments represented by the examples above.

Claims (16)

1. Compounds of formula I

Of formula I'

Or a pharmaceutically acceptable salt thereof, wherein

R is- (C ═ Q) R^2aCN or Y; wherein

Y is a 5-6 membered aryl or heteroaryl ring, wherein the heteroaryl ring is selected from

Each Y is independently and optionally substituted with 0-4J^YSubstitution;

q is O, NH, NR' or S;

r' is C_1-6Alkyl, optionally halogenated with 0 to 4 occurrences, C_1-6Aliphatic radical, NO₂、NH₂、-N(C_1-6Alkyl), -N (C)_1-6Alkyl radical)₂、SH、-S(C_1-6Alkyl), OH, -O (C)_1-6Alkyl), -C (O) (C)_1-6Alkyl), -C (O) NH₂、-C(O)N(C_1-6Alkyl) or-C (O) N (C)_1-6Alkyl radical)₂Substitution;

R^2ais C_1-6Aliphatic radical, C_6-10Aryl, 5-10 membered heteroaryl, 5-10 membered heterocyclyl, OR⁵Or N (R)⁵)₂(ii) a Each R^2aIndependently and optionally by 0-5J^2aSubstitution;

R¹is H, -C (O) (C)_1-6Alkyl), -C (O) O (C)_1-6Alkyl), -C (O) NH₂、-C(O)N(C_1-6Alkyl), -C (O) N (C)_1-6Alkyl radical)₂Or C_1-6An aliphatic group; each R¹Optionally halogenated groups present 0 to 4 times, C_1-6Haloalkyl, C_1-6Aliphatic radical, NO₂、NH₂、-N(C_1-6Alkyl), -N (C)_1-6Alkyl radical)₂、SH、-S(C_1-6Alkyl), OH or-O (C)_1-6Alkyl) substitution;

z is a bond;

R³and R⁴Each independently of the others being H, halogen, C_1-6Alkoxy, N (R)⁵)₂、CN、NO₂Or U_m-V, wherein m is 0 or 1;

v is H, C_6-10Aryl, 5-10 membered heteroaryl, C_3-10Cycloaliphatic, 5-to 10-membered heterocyclic radical or C_1-12An aliphatic radical, wherein up to two methylene units of the alkylene chain are optionally and independentlyIs replaced by a heteroatom selected from O, N or S in a chemically stable arrangement; v is optionally substituted by 0-4R⁸Substitution;

u is C_1-12An alkylene chain wherein up to two methylene units of the chain are optionally and independently replaced by-NH-, -NR⁵-、-O-、-S-、-CO₂-、-OC(O)-、-C(O)CO-、-C(O)-、-C(O)NH-、-C(O)NR⁵-、-C(＝N-CN)、-NHCO-、-NR⁵CO-、-NHC(O)O-、-NR⁵C(O)O-、-SO₂NH-、-SO₂NR⁵-、-NHSO₂-、-NR⁵SO₂-、-NHC(O)NH-、-NR⁵C(O)NH-、-NHC(O)NR⁵-、-NR⁵C(O)NR⁵-、-OC(O)NH-、-OC(O)NR⁵-、-NHNH-、-NHNR⁵-、-NR⁵NR⁵-、-NR⁵NH-、-NHSO₂NH-、-NR⁵SO₂NH-、-NHSO₂NR⁵-、-NR⁵SO₂NR⁵-、-SO-、-SO₂-、-PO-、-PO₂-or-POR⁵-substituted; u is optionally substituted by 0-6J^USubstitution;

R⁵is C_1-4Haloalkyl, -C (O) COR⁶、-C(O)R⁶、-C(O)OR⁶、-C(O)N(R⁶)₂、-SO₂R⁶、C_0-6Alkyl-heterocyclyl radical, C_0-6Alkyl-heteroaryl, C_0-6Alkyl-aryl, C_0-6Alkyl-cycloaliphatic radicals or C_1-6An aliphatic radical wherein up to three methylene units of the aliphatic chain are optionally and independently replaced by-NR' -, -O-, -S-, -CO₂-、-OC(O)-、-C(O)CO-、-C(O)-、-C(O)NR″-、-NR″CO-、-NR″C(O)O-、-SO₂NR″-、-NR″SO₂-、-C(O)NR″NR″-、-NR″C(O)NR″-、-OC(O)NR″-、-NR″NR″-、-NR″SO₂NR″-、-SO-、-SO₂-、-PO-、-PO₂-or-POR "-is replaced by a chemically stable arrangement; each R⁵Independently and optionally by 0-5J^R5Substitution; or two R⁵The groups together with the atoms to which they are attached are optionally joined to form a 5-10 membered carbocyclic or heterocyclic ring; wherein the ring is optionallySubstituted with 0-4J';

R⁶is H, C_1-6Alkoxy radical, C_1-4Haloalkyl, C_0-6Alkyl-heterocyclyl radical, C_0-6Alkyl-heteroaryl, C_0-6Alkyl-aryl, C_0-6Alkyl-cycloaliphatic radicals or C_1-6An aliphatic group wherein up to two methylene units of the aliphatic chain are optionally and independently replaced in a chemically stable arrangement with a heteroatom selected from O, N or S; each R⁶Independently and optionally by 0-5J^R6Substitution; or two R⁶The groups together with the atoms to which they are attached are optionally joined to form a 5-10 membered carbocyclic or heterocyclic ring; wherein said ring is optionally substituted with 0-4J';

R⁸is halogen, C_1-4Haloalkyl, phenyl, 5-8 membered heterocyclyl, 5-6 membered heteroaryl, -OR⁶、-N(R⁶)₂、-SR⁶、NO₂、CN、-COOR⁶、-C(O)N(R⁶)₂、-SO₂R⁶、-SO₂N(R⁶)₂、-NR⁶C(O)R⁶、-C(O)R⁶、-OC(O)R⁶、-NR⁶C(O)O-R⁶、-NR⁶SO₂-R⁶、-C(O)NR⁶N(R⁶)₂、-NR⁶C(O)N(R⁶)₂、-OC(O)N(R⁶)₂、-NR⁶N(R⁶)₂、-NR⁶SO₂N(R⁶)₂Or C_1-12An aliphatic radical, wherein up to three methylene units of the aliphatic chain may optionally be replaced by-C (O) R⁶、-C(O)O-、-OC(O)-、-C(O)-、-C(O)N(R⁶)-、-NR⁶CO(R⁶)-、-O-、-NR⁶-or-S-interrupted; each R⁸Independently and optionally by 0-5J^R8Substitution;

each J^Y、J^2a、J^U、J^R5、J^R6、J^R8J 'and J' are independently selected from N (R)⁹)₂、SR⁹、OR⁹Halo, CN, NO₂、COOR⁹、C(O)R⁹、SO₂R⁹、SOR⁹、-X-CF₃、-X-SH、-X-OH、C_1-4Haloalkyl, C_6-10Aryl, -X- (C)_6-10Aryl), 5-10 membered heteroaryl, -X- (5-10 membered heteroaryl), C_3-10Cycloaliphatic radical, -X- (C)_3-10Cycloaliphatic), 5-10 membered heterocyclyl, -X- (5-10 membered heterocyclyl), or X;

x is C_1-12An aliphatic radical in which up to two methylene units of the alkylene chain are optionally and independently replaced by-NH-, -NR' -, -O-, -S-, -CO₂-、-OC(O)-、-C(O)CO-、-C(O)-、-C(O)NH-、-C(O)NR″-、-C(＝N-CN)、-NHCO-、-NR″CO-、-NHC(O)O-、-NR″C(O)O-、-SO₂NH-、-SO₂NR″-、-NHSO₂-、-NR″SO₂-、-NHC(O)NH-、-NR″C(O)NH-、-NHC(O)NR″-、-NR″C(O)NR″、-OC(O)NH-、-OC(O)NR″-、-NHNH-、-NHNR″-、-NR″NR″-、-NR″NH-、-NHSO₂NH-、-NR″SO₂NH-、-NHSO₂NR″-、-NR″SO₂NR″-、-SO-、-SO₂-、-PO-、-PO₂-or-POR "-is replaced by a chemically stable arrangement; wherein R' is H or C_1-6An aliphatic group;

each J^Y、J^2a、J^U、J^R5、J^R6N (R) optionally and independently substituted with 0-4 occurrences of J' and J ″⁹)₂、SR⁹、OR⁹Halo, CN, NO₂、COOR⁹、C(O)R⁹、SO₂R⁹、SOR⁹、-X-CF₃、-X-SH、-X-OH、C_1-4Haloalkyl, C_6-10Aryl, -X- (C)_6-10Aryl), 5-10 membered heteroaryl, -X- (5-10 membered heteroaryl), C_3-10Cycloaliphatic radical, -X- (C)_3-10Cycloaliphatic), 5-10 membered heterocyclyl, -X- (5-10 membered heterocyclyl), or X; and

R⁹is H, C_1-6Aliphatic radical, C_1-4Haloalkyl, C_6-10Aryl, -X- (C)_6-10Aryl), 5-10 membered heteroaryl, -X- (5-10 membered heteroaryl), C_3-10Cycloaliphatic radical, -X- (C)_3-10Cycloaliphatic), 5-10 membered heterocyclyl, -X- (5-10 membered)Heterocyclyl) or X, or wherein two R are⁹Together with the atoms to which they are attached form a 5-10 membered heterocyclyl, wherein said heterocyclyl is optionally substituted with 0-4 occurrences of halo, CN, NO₂、-COOH、-COO(C_1-6Alkyl), -C (O) H, SO₂H、SO₂(C_1-6Alkyl group), C_1-6Halogenated aliphatic radical, NH₂、-NH(C_1-6Alkyl), -N (C)_1-6Alkyl radical)₂、SH、-S(C_1-6Alkyl), OH, -O (C)_1-6Alkyl), -C (O) (C)_1-6Alkyl), -C (O) NH₂、-C(O)NH(C_1-6Alkyl), -C (O) N (C)_1-6Alkyl radical)₂、-C(O)NH₂、-C(O)NH(C_1-6Alkyl), -C (O) N (C)_1-6Alkyl radical)₂、C_1-4Haloalkyl, C_6-10Aryl, -X- (C)_6-10Aryl), 5-10 membered heteroaryl, -X- (5-10 membered heteroaryl), C_3-10Cycloaliphatic radical, -X- (C)_3-10Cycloaliphatic), 5-10 membered heterocyclyl, -X- (5-10 membered heterocyclyl), or X,

wherein said heteroaryl, heterocyclyl or heterocyclic ring system each contains one or more heteroatoms selected from nitrogen, oxygen or sulfur.

2. A compound according to claim 1, wherein R⁴Is H, R³Is not H.

3. A compound according to claim 1, wherein R¹Is H.

4. A compound according to claim 1, wherein R is

5. A compound according to claim 4, wherein R^2aIs OR⁵、N(R⁵)₂Or a 5-8 membered heterocyclic group.

6. The compound according to claim 1, wherein R is Y.

7. A compound according to claim 1, wherein R³And R⁴Each independently is U_m-V。

8. A compound according to claim 1, wherein Z is a bond and R is selected from

9. A compound according to claim 1, wherein Z is a bond and R is

10. The compound according to claim 1, wherein said compound is of formula II:

or a pharmaceutically acceptable salt thereof, wherein ring a is Y.

11. The compound according to claim 10, wherein said compound is of formula III:

or a pharmaceutically acceptable salt thereof, wherein

R³Is halogen, C_1-6Aliphatic radical, C_1-6Alkoxy, N (R)⁵)₂、CN、NO₂Or U_m-V。

12. The compound according to claim 11, wherein said compound is of formula IV:

wherein each Z¹And Z²Is CH or N.

13. A compound according to claim 1 selected from.

14. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

15. A method of inhibiting Tec family kinase activity in a biological sample selected from saliva, urine, feces, semen or tears, or an extract thereof, the method comprising contacting the biological sample with a compound or composition comprising the compound according to claim 1.

16. Use of a pharmaceutical composition according to claim 14 in the manufacture of a medicament for the treatment of a cancer selected from colon, breast, stomach, ovary, prostate or pancreas.