JP2010522753A - New imidazo heterocycle - Google Patents

Abstract

  The present invention relates to novel imidazopyrazine compounds that are useful as kinase inhibitors and are therefore useful in the treatment of certain conditions and diseases, particularly inflammatory conditions and diseases, and proliferative disorders and conditions such as cancer.

Description

  Protein phosphorylation at specific amino acid residues is important for the regulation of numerous cellular processes including cell cycle progression and division, signal transduction and apoptosis. The phosphorylation is usually a transfer reaction of a terminal phosphate group from ATP to a protein substrate. The specific structure in the target substrate to which the phosphate (phosphate) is transferred is a tyrosine, serine or threonine residue. Because these amino acid residues are the target structure for the phosphoryl transfer, these protein kinase enzymes are commonly referred to as tyrosine kinases or serine / threonine (S / T) kinases. Phosphorylation and counteracting phosphatase reactions on tyrosine, serine and threonine residues are involved in a myriad of cellular processes that result in responses to diverse intracellular signals, regulation of cellular functions and activation or inactivation of cellular processes Yes. A cascade of protein kinases (protein kinases) is often involved in intracellular signaling and is necessary for the realization of cellular processes. Because of their ubiquitous nature in these processes, the protein kinases can be considered as integral parts of the cell membrane or cytoplasmic enzymes and are often localized in the nucleus as components of enzyme complexes there is a possibility. In many cases, these protein kinases are essential elements of enzyme and structural protein complexes that determine where and when cellular processes occur in the cell. Given the importance and diversity of protein kinase function, it is not surprising that altered phosphorylation is associated with a number of diseases such as cancer, diabetes, inflammation and hypertension.

  Accordingly, it is desirable to identify effective small molecules that specifically inhibit protein kinases involved in abnormal or inappropriate cell proliferation, signaling, differentiation, protein production or metabolism. In particular, identification of methods and compounds that specifically inhibit the function of kinases involved in immune regulation or proliferation disorders is desirable.

  The present invention provides novel compounds that inhibit one or more S / T kinases or receptor or non-receptor tyrosine kinases. The compounds of the present invention affect cytokine inhibitory activity.

  Cytokine-mediated diseases and cytokine inhibition, suppression and antagonism are used in the context of diseases or conditions in which excessive or unregulated production or activity of one or more cytokines occurs. Specific examples of such cytokines include tumor necrosis factor alpha (TNFα), interleukin-1 (IL-1), interleukin-6 (IL-6) and interleukin-8 (IL-8). . There remains a need for compounds that are useful in the treatment of cytokine-mediated diseases and thus inhibit, suppress or antagonize the production or activity of cytokines such as TNF, IL-1, IL-6 and IL-8.

  The p38 MAP kinase (p38, also known as CSBP or SAPK) signaling pathway is an inflammatory cytokine (eg, TNF, IL-1, IL-6, IL-8) that is elevated in many inflammatory and autoimmune diseases. (See JC Lee, Nature Reviews Drug Discovery 2003, 2, 717-726 and references cited therein). This pathway is activated by cell stressors such as osmotic shock, UV light, free radicals, bacterial toxins, viruses, cytokines, chemokines and in response, TNF, IL-1, IL-6 and IL-8 ( It has been shown to mediate the expression of several cytokines, including but not limited to these. In myeloid lineage cells such as macrophages and monocytes, both IL-1 and TNFα are transcribed in response to p38 activation. Subsequent translation and secretion of these and other cytokines initiates local or systemic inflammatory responses in adjacent tissues and via leukocyte infiltration. While this response is a normal part of the physiological response to cellular stress, acute or chronic cellular stress results in excessive or unregulated expression of inflammatory cytokines. And this leads to tissue damage and often causes pain and weakness (G. Panayi, N Engl J Med 2001, 344 (12), 907; J. Smolen Nature Reviews Drug Discovery 2003, 2, 473 and cited therein). See references). The four known isoforms (p38 α, β, γ, δ) of p38 MAP kinase, each showing different expression levels, tissue distribution and regulation, support the view that they are involved in the causes of numerous diseases. ing.

  Many solid tumors increase in mass due to the proliferation of malignant cells and matrix cells (including endothelial cells). In order for a tumor to grow to a diameter of 2-3 mm or more, it must form vasculature, a process known as angiogenesis. Selective p38 inhibitors have been shown to suppress angiogenesis (JR Jackson, J. Pharmacol Exp. Therapeutics, 1998, 284, 687). Since angiogenesis is a critical component of solid tumor mass expansion, the development of novel p38 kinase inhibitors to inhibit this process represents a promising approach for anti-tumor therapy . The compounds of the present invention are also useful in inhibiting the growth of sensitive neoplasms (RM Schultz, Potential of p38 MAP Kinase Inhibitors in the therapy of cancer. , 60, 59-92). The term “sensitive neoplasm” as used in this application includes human cancers such as malignant melanoma, colorectal cancer, gastric cancer, breast cancer and non-small cell lung cancer.

  In addition, suppression of p38 kinase may be achieved in certain viral states such as influenza (J. Immunology, 2000, 164, 3222), rhinovirus (J. Immunology, 2000, 165, 5211) and HIV (Proc. Nat. Acad. Sci, 1998, 95, 7422).

  In summary, a number of inhibitors of p38 kinase are being actively studied for the treatment of various disorders (Boehm, Adams Exp. Opin. Ther. Patents 2000, 10 (1), 25-37). There remains a need for treatment in this field for compounds that are cytokine suppressive, ie compounds that can inhibit p38 kinase.

Nature Reviews Drug Discovery 2003, 2, 717-726 N Engl J Med 2001, 344 (12), 907 Nature Reviews Drug Discovery 2003, 2,473 J. et al. Pharmacol Exp. Therapeutics, 1998, 284, 687 Potential of p38 MAP Kinase Inhibitors in the treatment of cancer. In: E. Jucker (eds.), Progress in Drug Research 2003, 60, 59-92 J. et al. Immunology, 2000, 164, 3222 J. et al. Immunology, 2000, 165, 5211 Proc. Nat. Acad. Sci, 1998, 95, 7422 Exp. Opin. Ther. Patents 2000, 10 (1), 25-37

SUMMARY OF THE INVENTION In a first embodiment, the present invention provides a formula (1)

の化合物ならびにその医薬上許容される塩、プロドラッグおよび医薬上活性な代謝産物である。

And pharmaceutically acceptable salts, prodrugs and pharmaceutically active metabolites thereof.

  In the second embodiment, the present invention provides a formula (2)

の化合物ならびにその医薬上許容される塩、プロドラッグおよび医薬上活性な代謝産物である。

And pharmaceutically acceptable salts, prodrugs and pharmaceutically active metabolites thereof.

  In a third embodiment, the present invention provides a formula (3)

の化合物ならびにその医薬上許容される塩、プロドラッグおよび医薬上活性な代謝産物である。

And pharmaceutically acceptable salts, prodrugs and pharmaceutically active metabolites thereof.

Detailed Description of the Invention Protein kinases are a broad and diverse class of over 500 enzymes, including cancer, growth factors, receptors, signal transduction, intermediates, apoptosis-related kinases and cyclin dependent kinases. They cause the transfer of phosphate groups to specific tyrosine, serine or threonine amino acid residues and are broadly classified as tyrosine and serine / threonine kinases based on their substrate specificity. Serine / threonine kinases (S / T kinases) are a large subfamily of protein kinases that specifically transfer phosphate groups to the terminal hydroxyl portion of certain serine or threonine residues (Hanks et al., (1988) Science, 241: 42-52). A number of S / T kinase family members are involved in inflammatory signaling, tumor growth or cell transformation. For example, mitogen-activated protein kinase (MAPK) is an intermediate in the signaling cascade of Toll-like receptors (TLR) such as TLR4, growth / survival factors such as EGF and death receptors such as TNF receptor. S / T kinase that acts. Activation of MAPKs such as extracellular signal-regulated kinase (ERK1-2), p38α, c-Jun N-terminal kinase (JNK) or MAPKAP-K2 (MK2) transduces signals in cells such as monocytes / macrophages Has been shown to result in the extracellular production of inflammatory cytokines such as TNF.

  The p38 MAP kinase (p38, also known as CSBP or SAPK) signaling pathway is an inflammatory cytokine (eg, TNF, IL-1, IL-6, IL-8) that is elevated in many inflammatory and autoimmune diseases. (See JC Lee, Nature Reviews Drug Discovery 2003, 2, 717-726 and references cited therein). This pathway is activated by cell stressors such as osmotic shock, UV light, free radicals, bacterial toxins, viruses, cytokines, chemokines and in response, TNF, IL-1, IL-6 and IL-8 ( It has been shown to mediate the expression of several cytokines, including but not limited to these. In myeloid lineage cells such as macrophages and monocytes, both IL-1 and TNFα are transcribed in response to p38 activation. Subsequent translation and secretion of these and other cytokines initiates local or systemic inflammatory responses in adjacent tissues and via leukocyte infiltration. While this response is a normal part of the physiological response to cellular stress, acute or chronic cellular stress results in excessive or unregulated expression of inflammatory cytokines. And this leads to tissue damage and often causes pain and weakness (G. Panayi, N Engl J Med 2001, 344 (12), 907; J. Smolen Nature Reviews Drug Discovery 2003, 2, 473 and cited therein). See references). The four known isoforms (p38 α, β, γ, δ) of p38 MAP kinase, each showing different expression levels, tissue distribution and regulation, are involved in the causes of inflammation, autoimmunity and other diseases We support this view.

  In summary, a number of inhibitors of p38 kinase are being actively studied for the treatment of various disorders (Boehm, Adams Exp. Opin. Ther. Patents 2000, 10 (1), 25-37). There remains a need for treatment in this field for compounds that are cytokine suppressive, ie compounds that can inhibit p38 kinase.

  Protein tyrosine kinases (PTKs) are enzymes that catalyze the phosphorylation of specific tyrosine residues in cellular proteins. This post-translational modification of these substrate proteins (often the enzyme itself) acts as a molecular switch that regulates cell growth, activation or differentiation (for review see Schlessinger and Ulrich, 1992, Neuron 9: 383-391). See). Abnormal or excessive PTK activity is observed in a number of conditions, including benign and malignant proliferative disorders and diseases resulting from inappropriate activation of the immune system (eg, autoimmune disorders), allograft rejection and graft-versus-host disease Has been. In addition, endothelial cell-specific receptors PTK, such as KDR and Tie-2, mediate the angiogenic process, and thus other diseases involving cancer and inappropriate angiogenesis (eg, diabetic retinopathy, age-related Involved in supporting the progression of macular degeneration choroidal neovascularization, psoriasis, arthritis, retinopathy of prematurity and infantile hemangioma).

  Tyrosine kinases can be of the receptor type (having extracellular, transmembrane and intracellular domains) or non-receptor type (generally intracellular).

  Receptor tyrosine kinases (RTKs) comprise a large family of transmembrane receptors with diverse biological activities. Currently, at least 19 different RTK subfamilies have been identified. The receptor tyrosine kinase (RTK) family contains critical receptors critical to the growth and differentiation of various cell types (Yarden and Ulrich, Ann. Rev. Biochem. 57: 433-478, 1988; Ullrich and Schlessinger). , Cell 61: 243-254, 1990). The intrinsic function of RTKs is activated upon ligand binding, which causes phosphorylation of the receptor and multiple cellular substrates, followed by various cellular responses (Ullrich & Schlessinger, 1990, Cell 61: 203-212). Thus, receptor tyrosine kinase-mediated signal transduction is initiated by extracellular interactions with specific growth factors (ligands), typically followed by receptor dimerization, stimulation of endogenous protein tyrosine kinase activity. And receptor transphosphorylation occurs. Thereby, binding sites for intracellular signaling molecules are formed and appropriate cellular responses (eg, cell division, differentiation, metabolism, and changes in the extracellular microenvironment; Schlessinger and Ulrich, 1992, Neuron 9: 1-20). Which leads to the formation of a complex of a range of cytoplasmic signaling molecules that promotes.

  Non-receptor tyrosine kinases are a group of cellular enzymes that lack extracellular and transmembrane sequences. Over 24 non-receptor tyrosine kinases have been identified, including 11 subfamilies (Src, Frk, Btk, Csk, Abl, Zap70, Fes / Fps, Fak, Jak, Ack and LIMK). The Src subfamily of non-receptor tyrosine kinases is composed of the largest number of PTKs and includes Src, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk. The Src subfamily of enzymes has been linked to carcinogenesis and immune responses. A more detailed discussion of non-receptor tyrosine kinases is described in Bohlen, 1993, Oncogene 8: 2025-2031, which is incorporated herein by reference.

  Many of the kinases are involved in cellular signaling pathways involved in many pathologies including immune regulation, inflammation or proliferative disorders such as cancer, whether they are receptor or non-receptor tyrosine kinases tyrosine or S / T kinase It has been found that

  In a related aspect, the invention is a method for inhibiting p38 in a human patient suffering from a disorder in which P38 activity is detrimental, such that p38 activity is suppressed in the human patient and treatment is achieved. There is provided a method comprising administering a compound of formula 1, 2 or 3 to the human patient.

  A number of autoimmune diseases and diseases associated with chronic inflammation and acute response have been linked to p38 MAP kinase activation and overexpression or dysregulation of inflammatory cytokines. The compound is present in rheumatoid arthritis, osteoarthritis, asthma, chronic obstructive pulmonary disease (COPD), sepsis, psoriasis, psoriatic arthritis, inflammatory bowel disease, Crohn's disease, lupus, multiple sclerosis, juvenile chronic arthritis, Useful in the treatment of inflammatory disorders including but not limited to Lyme arthritis, reactive arthritis, septic arthritis, spondyloarthropathy and systemic lupus erythematosus.

  The compounds of the present invention are also useful in the treatment of cardiovascular disorders such as acute myocardial infarction, acute coronary syndrome, chronic heart failure, myocardial infarction, atherosclerosis, viral myocarditis, cardiac allograft rejection and sepsis related heart failure. Useful. Furthermore, the compounds of the present invention are also useful for the treatment of central nervous system disorders such as meningitis, Alzheimer's disease and Parkinson's disease.

  The compounds of the present invention are also useful in the treatment of the following: eye conditions, cancer, solid tumors, sarcomas, fibrosarcomas, bone tumors, melanomas, retinoblastomas, rhabdomyosarcomas, glioblastomas Neuroblastoma, teratoblastoma, cancer such as lung cancer, breast cancer, stomach cancer, bladder cancer, colon cancer, pancreatic cancer, ovarian cancer, prostate cancer and rectal cancer and hematopoietic malignancies (leukemia and lymphoma), β-lipo Proteinemia, advanced cyanosis, acute and chronic parasitic or infectious processes, acute leukemia, acute lymphoblastic leukemia (ALL), acute myelocytic leukemia (AML), acute or chronic bacterial infection, acute pancreatitis, acute Renal failure, adenocarcinoma, air ectopic beat, AIDS dementia complications, alcohol-induced hepatitis, allergic conjunctivitis, allergic contact dermatitis, allergic rhinitis, alpha-1 antitrypsin deficiency, amyotrophic lateral cord Hard Keratosis, anemia, angina pectoris, anterior horn cell degeneration, anti-cd3 therapy, antiphospholipid syndrome, antireceptor hypersensitivity reaction, hypersensitivity reaction, hyperactivity disorder, hypersensitivity pneumonitis, hypertension, hypokinetic disorder, aorta Aneurysm and peripheral aneurysm, hypothalamic-pituitary-adrenal axis assessment, aortic dissection, arterial hypertension, arteriosclerosis, arteriovenous fistula, ataxia, streptococcal myositis, cerebellar structural lesion, subacute sclerosing panencephalitis , Syncope, cardiovascular syphilis, systemic anaphylaxis, systemic inflammatory response syndrome, systemic expression juvenile rheumatoid arthritis, T cells or FAB ALL, telangiectasia, obstructive thromboangitis, transplantation, trauma / bleeding, type III Hypersensitivity reaction, type IV hypersensitivity, unstable angina pectoris, uremia, urinary sepsis, hives, valvular heart disease, dilated meander vein, vasculitis, venous disease, venous thrombosis, ventricular fibrillation, virus and fungus Infection, virus Vital encephalitis / aseptic meningitis, virus-related hemophagocytic syndrome, Wernicke-Korsakov syndrome, Wilson disease, xenograft rejection of any organ or tissue, atrial fibrillation (persistent or seizure), Atrial flutter, atrioventricular block, B cell lymphoma, bone transplant rejection, bone marrow transplant (BMT) rejection, small intestine transplant rejection, spinal ataxia, leg block, Burkitt lymphoma, burn, cardiac arrhythmia, cardiac syncope syndrome, Heart tumor, cardiomyopathy, cardiopulmonary bypass inflammatory response, cartilage transplant rejection, cerebellar cortical degeneration, cerebellar disorder, chaotic or multifocal atrial tachycardia, chemotherapy related disorders, chronic myelogenous leukemia (CML), chronic alcoholism, Chronic inflammatory pathology, chronic lymphocytic leukemia, (CLL), chronic salicylic acid detoxification, colorectal cancer, congestive heart failure, conjunctivitis, pulmonary heart, coronary artery disease, Croy Tufert-Jakob disease, culture negative sepsis, cystic fibrosis, cytokine therapy-related disorders, fighting dementia, demyelinating disease, dengue hemorrhagic fever, dermatitis, skin disease, diabetic arteriosclerosis, diffuse Lewy body disease, dilated Congestive cardiomyopathy, brainstem ganglion disorder, middle-aged Down syndrome, drug-induced dyskinesia induced by drugs that block CNS dopamine receptors, drug sensitivity, eczema, encephalomyelitis, endocarditis, Endocrine disease, epiglottis, Epstein-Barr virus infection, erythema erythematosus, extrapyramidal and cerebellar disorders, familial hemophagocytic lymphohistiocytosis, fetal thymus transplant rejection, Friedreich ataxia, functional peripheral arterial disorder , Fungal sepsis, gas gangrene, gastric ulcer, glomerulonephritis, gram-negative sepsis, gram-positive sepsis, granulomas due to intracellular organisms, hair-like cell leukemia, Hallerfolden-si Patz disease, hay fever, heart transplant rejection, hemochromia, hemodialysis, hemolytic uremic syndrome / thrombolytic thrombocytopenic purpura, bleeding, idiopathic pulmonary fibrosis, antibody-mediated cytotoxicity, asthenia, infant spinal cord Muscular atrophy, aortic inflammation, influenza A, ionizing radiation exposure, iridocyclitis / uveitis / optic neuritis, juvenile rheumatoid arthritis, juvenile spinal muscular atrophy, renal transplant rejection, Legionella, leash Maniasis, steatosis, liver transplant rejection, lymphedema, malaria, malignant lymphoma, malignant histiocytosis, malignant melanoma, meningococcal disease, metabolic / idiopathic, migraine, mitochondrial multisystem disorder, single Clonal hypergammaglobulinemia, multiple myeloma, multiple systemic degeneration (Mencel DeJerine-Thomas Shi-Drager) Machado-Joseph, myasthenia gravis, intracellular mycobacterium avium, mycobacterium tuberculosis, myelodysplastic syndrome, myocardial ischemia disorder, nasopharyngeal cancer, newborn Chronic lung disease, nephritis, nephrosis, neurodegenerative disease, neurogenic I muscle atrophy, neutropenic fever, non-Hodgkin lymphoma, obstruction of the abdominal aorta and its branches, obstructive arterial injury, okt3 therapy, testitis / Epididymis, testicularitis / vasectomy reversal, organ hypertrophy, osteoporosis, pancreas transplant rejection, pancreatic cancer, paraneoplastic syndrome / hypercalcemia (malignant disease), parathyroid transplant rejection, pelvis Internal infection, perennial rhinitis, pericardial disease, Kaposi's sarcoma, Hodgkin's disease, lymphoma, myeloma, leukemia Malignant ascites, hematopoietic cancer, Crow-Fukase (POEMS) syndrome (polyneuropathy, organ hypertrophy, endocrine disease, monoclonal hypergammaglobulinemia and skin change syndrome), diabetic conditions such as insulin-dependent diabetes glaucoma , Diabetic retinopathy or microangiopathy, sickle cell anemia, chronic inflammation, synovitis, glomerulonephritis, graft rejection, Lyme disease, von Hippel-Lindau disease, pemphigoid, Paget's disease, fibrosis, Sarcoidosis, cirrhosis, thyroiditis, hyperviscosity syndrome, Osler-Weber-Langue disease, chronic obstructive pulmonary disease, post-burn edema or asthma, trauma, radiation, stroke, hypoxia, ischemia, ovarian hyperstimulation syndrome Post-perfusion syndrome, post-pump syndrome, MI post-cardiac incision syndrome, preeclampsia, functional uterine bleeding, endometriosis, pulmonary hypertension, infantile hemangioma, or herpes simplex Virus, herpes zoster virus, human immunodeficiency virus, parapoxvirus infection, protozoa or toxoplasmosis, progressive supranuclear paralysis, primary pulmonary hypertension, radiation therapy, Raynaud's phenomenon and Raynaud's disease, Raynaud's disease , Lefsum disease, regular narrow QRS tachycardia, renovascular hypertension, restrictive cardiomyopathy, sarcoma, senile chorea, senile dementia of Lewy bodies, shock, skin allograft, skin change syndrome, eye or macular edema, Ocular neovascular disease, scleritis, radial keratotomy, uveitis, vitreitis, myopia, papillary pit, chronic retinal detachment, complications after laser treatment, conjunctivitis, Stargardt disease, Eels disease, retinopathy, macular Degeneration, recurrent stenosis, ischemia / reperfusion injury, ischemic stroke, vascular occlusion, carotid artery occlusion disease, ulcerative colitis, inflammatory bowel disease, diabetes, true Urinary disease, insulin-dependent diabetes, allergic disease, dermatitis scleroderma, graft-versus-host disease, organ transplant rejection (including but not limited to bone marrow and solid organ rejection), acute or related to organ transplant Chronic inflammatory disease, sarcoidosis, disseminated intravascular coagulation, Kawasaki disease, nephrotic syndrome, chronic fatigue syndrome, Wegener's granulomatosis, Henoho-Schönlein purpura, renal microvasculitis, chronic active hepatitis, septic shock , Toxic shock syndrome, sepsis syndrome, cachexia, infection, parasitic diseases, acquired immune deficiency syndrome, acute transverse myelitis, Huntington's chorea, stroke, primary biliary cirrhosis, hemolytic anemia, malignant disease , Addison's disease, idiopathic Addison's disease, sporadic, type I multi-gland deficiency and type II multi-gland deficiency, Schmidt syndrome, adult (acute) respiratory distress symptoms , Alopecia, alopecia areata, seronegative arthropathy, arthropathy, Reiter's disease, psoriatic arthropathy, ulcerative colitis arthritis, enteritis synovitis, chlamydia, Yersinia and Salmonella-related arthropathy, atheromatous Disease / arteriosclerosis, atopic allergy, autoimmune bullous disease, pemphigus vulgaris, lobular pemphigus, pemphigoid, linear IgA disease, autoimmune hemolytic anemia, Coombs-positive hemolytic anemia, acquired Pernicious anemia, juvenile pernicious anemia, peripheral angiopathy, peritonitis, pernicious anemia, myalgia encephalitis / royal free disease, chronic mucocutaneous candidiasis, giant cell arteritis, primary sclerosing hepatitis, idiopathic autoimmune hepatitis , Acquired immune deficiency syndrome, acquired immune deficiency related disease, hepatitis A, hepatitis B, hepatitis C, his bundle arrhythmia, HIV infection / HIV neuropathy, vulgaris immunodeficiency Hypofibrinogenemia), dilated cardiomyopathy, female infertility, ovarian failure, immature ovarian failure, fibrotic lung disease, chronic wound healing, idiopathic fibrotic alveolitis, post-inflammatory interstitial lung disease, interstitial Pneumonia, Pneumocystis carini pneumonia, pneumonia, connective tissue disease-related interstitial lung disease, mixed connective tissue disease, mixed lung disease, systemic sclerosis-related interstitial lung disease, rheumatoid arthritis-related interstitial disease Lung disease, systemic lupus erythematosus related lung disease, dermatomyositis / polymyositis related lung disease, Sjogren's disease related lung disease, ankylosing spondylitis related lung disease, vasculitis diffuse lung disease, hemosiderin deposition related lung disease, drug induction Interstitial lung disease, radiation fibrosis, obstructive bronchiolitis, chronic eosinophilic pneumonia, lymphocyte infiltrating lung disease, post-infectious interstitial lung disease, urate arthritis, autoimmune hepatitis, type 1 Autoimmune hepatitis (classical self Immunity or lupoid hepatitis), type 2 autoimmune hepatitis (anti-LKM antibody hepatitis), autoimmune-mediated hypoglycemia, type B insulin resistance with black epidermopathy, hypoparathyroidism, acute immunity related to organ transplantation Disease, chronic immune disease related to organ transplantation, osteoarthritis, primary sclerosing cholangitis, type 1 psoriasis, type 2 psoriasis, idiopathic leukopenia, autoimmune neutropenia, kidney disease NOS, thread For spherical nephritis, renal microvasculitis, Lyme disease, discoid lupus erythematosus, idiopathic or NOS male infertility, sperm autoimmunity, multiple sclerosis (all subtypes), sympathetic ophthalmitis, connective tissue disease Secondary pulmonary hypertension, acute and chronic pain (various forms of pain), Goodpasture syndrome, pulmonary manifestations of polyarteritis nodosa, acute rheumatic fever, rheumatoid spondylitis, Still's disease, systemic sclerosis, Sjogren Disease Group, Takayasu disease / arteritis, autoimmune thrombocytopenia, toxicity, transplantation, idiopathic thrombocytopenia, autoimmune thyroid disease, hyperthyroidism, goiter autoimmune hypothyroidism (Hashimoto disease) , Atrophic autoimmune hypothyroidism, primary myxedema, phacogenic uveitis, primary vasculitis, vitiligo, acute liver disease, chronic liver disease, alcoholic cirrhosis, alcohol-induced liver Injury, cholestasis, idiosyncratic liver disease, drug-induced hepatitis, nonalcoholic steatohepatitis, allergy and asthma, group B streptococcal (GBS) infection, mental disorders (eg depression and schizophrenia), Th2 And Th1-type mediated diseases, as well as diseases involving inappropriate angiogenesis, such as diabetic retinopathy, retinopathy of prematurity, choroidal neovascularization and breast with age-related macular degeneration Infantile hemangioma (in humans). Such compounds also include, for example, ascites, exudates and exudates such as macular edema, brain edema, acute lung injury, adult respiratory distress syndrome (ARDS), proliferative disorders such as recurrent stenosis, fibrotic disorders such as cirrhosis. And atherosclerosis, mesangial cell proliferative disorders such as diabetic retinopathy, malignant nephrosclerosis, thrombotic microangiopathy syndrome, and glomerulopathy, myocardial neovascularization, coronary and cerebral side branches, ischemic limb neovascularization , Ischemia / reperfusion injury, peptic ulcer Helicobacter-related disease, virus-induced angiogenesis disorder, preeclampsia, functional uterine bleeding, cat scratching fever, rubeosis, neovascular glaucoma, and retinopathy such as diabetic retinopathy, It may be useful in the treatment of disorders such as those associated with retinopathy of prematurity or age-related macular degeneration. These compounds also have hyperproliferative disorders such as thyroid hyperplasia (especially Graves disease) and cysts such as ovarian stroma hyperangiogenesis and cystic kidney disease characteristic of polycystic ovary syndrome (Stein-Levantal syndrome). It can be used as an active substance for. This is because such diseases require the proliferation of vascular cells for proliferation and / or metastasis.

  The compounds of formula 1, 2 or 3 of the present invention may be used alone or in combination with other therapeutic substances for treating such diseases. It will be understood that the compounds of the present invention may be used alone or in combination with additional substances such as therapeutic substances, such other substances being selected by those skilled in the art for their intended purpose. Should be. For example, the additional agent can be a therapeutic agent recognized in the art as useful for treating a disease or condition being treated by the compounds of the present invention. The additional material may also be a material that imparts a beneficial attribute to the therapeutic composition, such as a material that affects the viscosity of the composition.

  Further, the combinations to be included in the present invention should be understood as being useful combinations for their intended purpose. The following materials are shown for illustrative purposes and are not intended to be limiting. The combination that is part of the invention may be a compound of the invention and at least one additional substance selected from those listed below. The combination can also include two or more additional substances, such as two or three additional substances, provided that the composition formed can perform its intended function.

  A preferred combination is a non-steroidal anti-inflammatory drug, also called NSAID, which includes drugs such as ibuprofen. Another preferred combination is a corticosteroid containing prednisone, a well-known side effect of steroid use is that in combination with the p38 inhibitors of the present invention, gradually reduce the steroid dose required in treating a patient. Can be reduced or even eliminated. Non-limiting examples of therapeutic agents for rheumatoid arthritis to which the compounds of Formula 1, 2, or 3 of the present invention may be combined may include the following: cytokine inhibitory anti-inflammatory drugs (CSAIDs) Other human cytokines or growth factors such as TNF, LT, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-12, Antibodies or antagonists to IL-15, IL-16, IL-21, IL-23, interferon, EMAP-II, GM-CSF, FGF and PDGF. S / T kinase inhibitors of the present invention are cell surface receptors such as CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69, CD80 (B7.1), CD86 (B7.2), It can be combined with antibodies to CD90, CTLA or their ligands, eg CD154 (gp39 or CD40L).

  Preferred combinations of therapeutic agents can prevent various points in the autoimmunity and subsequent inflammatory cascades, and preferred embodiments include TNF antagonists such as chimeric, humanized or human TNF antibodies, HUMIRA®. (US Pat. No. 6,090,382), CA2 (REMICADE ™), CDP 571, and possible p55 or p75 TNF receptors, derivatives thereof (p75TNFR1gG (ENBREL ™ or p55TNFR1gG (Lenercept)), Also included are TNFα converting enzyme (TACE) inhibitors, as well as IL-1 inhibitors (interleukin-1 converting enzyme inhibitors, IL-1RA, etc.) may be effective for the same reason. Is Leukin 11. Still other preferred combinations include other key functional factors of the autoimmune response that can act in a dependent or concerted manner, corresponding to IL-18 function, and are particularly preferred. IL-12 antagonists, including IL-12 antibodies or soluble IL-12 receptors, or IL-12 binding proteins IL-12 and IL-18 have been shown to have overlapping but distinct functions A combination of antagonists to both may be most effective, yet another preferred combination is a non-depleting anti-CD-4 inhibitor, yet another preferred combination comprises an antibody, a soluble receptor or an antagonistic ligand. An antagonist of the costimulatory pathway CD80 (B.71) or CD86 (B7.2) is included.

  The compounds of formula 1, 2 or 3 of the present invention can also be combined with, for example, the following substances: methotrexate, 6-MP, azathioprine sulfasalazine, mesalazine, olsalazine chloroquine (olsalazine) chloroquinine / hydroxychloroquine, penicillamine, aurothiomalate (intramuscular and oral), azathioprine, oral corticosteroid (oral, inhalation and topical injection), beta Body agonist (salbutamol (salbuta mol), terbutaline, salmeteral, xanthine (theophylline), aminophylline, chromoglycate, nedocromil, etiotropium, ketotrofen Tropium, cyclosporin, FK506, rapamycin, mycophenolate mofetil, leflunomide, NSAID such as ibuprofen, ibuprofen Substances that interfere with signaling by inflammatory cytokines such as luticosteroids such as prednisolone, phosphodiesterase inhibitors, adenosine inhibitors, antithrombotic agents, complement inhibitors, adrenergic agents, TNFα or IL-1 (eg , IRAK, NIK or IKK), IL-1β converting enzyme inhibitors, T cell signaling inhibitors such as kinase inhibitors, metalloproteinase inhibitors, sulfasalazine, 6-mercaptopurine, angiotensin converting enzyme inhibitors, soluble cytokine receptors and their Derivatives (eg soluble p55 or p75 TNF receptor and derivatives p75TNFRIgG (Enb el ™ and p55TNFRIgG (Lenercept)), sEL-1RI, sIL-1RII, sIL-6R), anti-inflammatory cytokines (eg, IL-4, IL-10, IL-11, IL-13 and TGFβ), Celecoxib, folic acid, hydroxychloroquine sulfate, rofecoxib, etanercept, infliximab, infliximab, naproxen (naproxen), valdecoxib, ulfasalazine, ulfasalazine ), Methylprednisolone acetate, gold sodium thiomalate, Aspirin, triamcinolone acetonide, propoxyphene naphthylate / apap, folate, nabumetone, diclofenac, diclofenac, diclofenac, diclofenac diclofenac sodium, oxaprozin, oxycodone HCl, hydrocodone bitartrate / apap, diclofenac sodium / misoprostol, fentanyl ), Anakinra, tramadol HCl, salsalate, sulindac, cyanocobalamin / fa / pyridoxine, adolaminophene sodium, alenaminophen, sodium , Prednisolone, morphine sulfate, lidocaine hydrochloride, indomethacin, glucosamine sulfo / chondroitin (chondroitin) amitriptyline HCl, sulfadiazine, oxycodone HCl / acetaminophen, olopatadine HCl misoprostol, naproxen sodium, naproxen sodium, naproxen sodium, naproxen sodium, naproxen sodium (Cyclophosphamide), rituximab, IL-1 TRAP, MRA, CTLA4-IG, IL-18BP, anti-IL-12, anti-IL15, BIRB-796, SCI-469, VX-702, AMG-548, VX -740, Roflumilast, IC 485, CDC-801 and mesopram (Mesopram). Preferred combinations include methotrexate or leflunomide and, in the case of moderate or severe rheumatoid arthritis, cyclosporine and anti-TNF antibodies (above).

  Non-limiting examples of therapeutic agents for inflammatory bowel disease that can be combined with the compounds of Formula 1, 2, or 3 of the present invention include: budesonide; epidermal growth factor; Corticosteroids; cyclosporine, sulfasalazine; aminosalicylate; 6-mercaptopurine; azathioprine; metronidazole; lipoxygenase inhibitor; mesalamine; olsalazine; balsalazide; Thromboxane inhibitor; IL-1 receptor antagonist; anti-IL-1β monoclonal antibody; anti-IL-6 monoclonal antibody; growth factor; elastase inhibitor Pyridinyl-imidazole compounds; other human cytokines or growth factors such as TNF, LT, IL-1, IL-2, IL-6, IL-7, IL-8, IL-12, IL-15, IL- 16, antibodies or antagonists to EMAP-II, GM-CSF, FGF and PDGF; cell surface molecules such as CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69, CD90 or their ligands; Methotrexate; cyclosporine; FK506; rapamycin; mycophenolate mofetil; leflunomide; NSAID, eg ibuprofen Corticosteroids such as prednisolone; phosphodiesterase inhibitors; adenosine agonists; antithrombotic substances; complement inhibitors; adrenergic substances; substances that interfere with signaling by inflammatory cytokines such as TNFα or IL-1 (eg NIK or IKK); IL-1β converting enzyme inhibitor; TNFα converting enzyme inhibitor; T cell signaling inhibitor such as kinase inhibitor; metalloproteinase inhibitor; sulfasalazine; azathioprine; 6-mercaptopurine; Soluble cytokine receptors and derivatives thereof (eg soluble p 55 or p75 TNF receptor, sIL-1RI, sIL-1RII, sIL-6R) and anti-inflammatory cytokines (eg, IL-4, IL-10, IL-11, IL-13 and TGFβ). Preferred specific examples of therapeutic agents for Crohn's disease that can be combined with compounds of Formula 1, 2, or 3 include: TNF antagonists, such as anti-TNF antibodies, HUMIRA® (US Pat. 6,090,382), CA2 (REMICADE ™), CDP 571, TNFR-Ig constructs (p75TNFRIgG (ENBREL ™) and p55TNFRIgG (LENERCEPT ™)) inhibitors and PDE4 inhibitors. Compounds of formula 1, 2 or 3 may be combined with: corticosteroids such as budenoside and dexamethasone; sulfasalazine, 5-aminosalicylic acid; olsalazine; and inflammatory Substances that interfere with the synthesis or action of cytokines such as IL-1, such as IL-1β converting enzyme inhibitors and IL-1ra; T cell signaling inhibitors such as tyrosine kinase inhibitors 6-mercaptopurine; IL-11; mesalamine; prednisone (Prednisone); azathioprine; mercaptopurine; infliximab; methyl succinate Sodium prednisolone (diphenoxylate); atropsulfate; loperamide hydrochloride; methotrexol; ) / Dextrose-water; hydrocodone bitartrate / apap; tetracycline hydrochloride; fluocinonide; metronidazole; thimerosal Boric acid; cholestyramine / sucrose; ciprofloxacin hydrochloride; hyoscyamine sulfate; meperidine hydrochloride; Acetaminophen; promethazine hydrochloride; sodium phosphate; sulfamethoxazole / trimethoprim; celecoxib Polycarbophil; propoxyphene naphthylate; hydrocortisone; multivitamins; ); Methylprednisolone; natalizumab and interferon-gamma.

  Non-limiting specific examples of therapeutic agents for multiple sclerosis that can be combined with compounds of Formula 1, 2, or 3 include: corticosteroids; prednisolone; methylprednisolone ( methylprednisolone; azathioprine; cyclophosphamide; cyclosporine; methotrexate (Bio); 4-aminopyridine; -Β1b (BETASERON®; Chiron / Berlex); interferon α-n3 (Interferon Sciences / Fujimoto), Interferon-α (Alfa Wassermann / J & J), Interferon β1A-IF (Serono / Inhale Therapeutics), Peginterferon (p) interferon (p) interferon (p) interferon (p-interferon) 1; COPAXONE®; Teva Pharmaceutical Industries, Inc .; hyperbaric oxygen; intravenous immunoglobulin; clavribine; other human cytokines or growth factors and their receptors such as TNF, LT, IL- 1, IL-2, IL-6, IL-7, IL-8, IL- 2, IL-23, IL-15, IL-16, EMAP-II, GM-CSF, antibody or antagonist to FGF and PDGF. Compounds of formula 1, 2 or 3 are antibodies against cell surface molecules such as CD2, CD3, CD4, CD8, CD19, CD20, CD25, CD28, CD30, CD40, CD45, CD69, CD80, CD86, CD90 or their ligands Can be combined with Compounds of formula 1, 2 or 3 can also be combined with the following substances: methotrexate, cyclosporine, FK506, rapamycin, mycophenolate mofetyl, leflunomide NSAIDs such as ibuprofen, corticosteroids such as prednisolone, phosphoesterase inhibitors, adensocine agonists, antithrombotic agents, complement inhibitors, adrenergic agents, inflammatory cytokines such as TNFα or IL Substances that interfere with signaling by -1 (eg IR K, NIK, IKK), IL-1β converting enzyme inhibitor, TACE inhibitor, T cell signaling inhibitor such as kinase inhibitor, metalloproteinase inhibitor, sulfasalazine, azathioprine, 6-mercaptopurine, angiotensin converting enzyme inhibitor, soluble cytokine receptor And derivatives thereof (eg, soluble p55 or p75 TNF receptor, sIL-1RI, sIL-1RJI, sIL-6R) and anti-inflammatory cytokines (eg, IL-4, IL-10, IL-13 and TGFβ).

  Preferred specific examples of therapeutic agents for multiple sclerosis that can be combined with compounds of formula 1, 2 or 3 include interferon-β, such as IFNβ1a and IFNβ1b; copaxone, corticosteroids, caspase inhibitors, such as Inhibitors of caspase-1, IL-1 inhibitors, TNF inhibitors, and antibodies to CD40 ligand and CD80 are included.

  Compounds of formula 1, 2 or 3 may also be combined with, for example, the following substances: alemtuzumab, dronabinol, daclizumab, mitoxantrone, xaliproden hydrochloride, xaliproden hydrochloride (Fampridine), glatiramer acetate, natalizumab, sinnabidol, a-immunokine NNS03, ABR-215062, Anergix. MS, chemokine receptor antagonist, BBR-2778, carraguarine, CPI-1189, LEM (liposome-encapsulated mitoxantrone), THC. CBD (cannabinoid agonist), MBP-8298, mesopram (PDE4 inhibitor), MNA-715, anti-IL-6 receptor antibody, neurovax, pirfenidone allotrap 1258 (RDP-1258), sTNF -R1, Talampanel, teriflunomide, TGF-beta2, tiplimotide, VLA-4 antagonists (eg TR-14035, VLA4 Ultrahaler, Antegran-ELAN / Biogen and IL-Gamma antagonists) 4 agonists.

Non-limiting examples of therapeutic agents for angina that can be combined with the compounds of Formula 1, 2, or 3 of the present invention include: aspirin, nitroglycerin, isosorbide dinitrate mononitrate, metoprolol succinate, atenolol, metoprolol tartrate, amodipine disulfate, diltiazemate hydrochloride clopidogrel bisulfate), nifedipine (nifedipi) ne), atorvastatin calcium, potassium chloride, furosemide, simvastatin, verapamil HCl, digoxin, propranolol hydrochloride, propranolol hydrochloride. , Spironolactone, hydrochlorothiazide, enalapril maleate, nadolol, ramipril, enoxaparin sodium (enoxaline) arin sodium, heparin sodium, valsartan, sotalol hydrochloride, fenofibrate, ezetimibe, rosartyl, rosartyl, o hydrochlorothiazide, felodipine, captopril, bisoprolol fumarate, ibuprofen, diclofenac and diisofenstol prostol, naproxen, meloxicam, indomethacin, diclofenac, celecoxib, celecoxib, rofecoxib, sulfasalazine (sulfathine). Prednisone, enteraccept, infliximab, albuterol, salmeterol / fluticasone, montelukast sodium (m) ntelukast sodium, fluticasone propionate, budesonide, prednisone, salmeterol xinafoate, valterol Prednisolone sodium phosphate, tracinolone acetonide, beclomethasone dipropionate, ip bromide Ratropium bromide, azithromycin, pyrbuterol acetic acid, theophylline anhydrous, succinic acid methylprednisolone sodium Formolol fumarate (formerol fumarate), influenza virus vaccine, methylprednisolone sodium succate, amoxicillin trihydrate (amoxicillin t ihydrate, flunisolide / menthol, allergic injection, cromolyn sodium, fexofenadine hydrochloride, exon, levofloxacin, levofloxacin, guanifine, guanofine, sodium phosphate, moxifloxacin HCl, doxycycline hydrate, fuaifenesin / d-methorphan, p-ephedrine (ephedrine) / Cod / chlorphenyl, gatifloxacin, cetirizine hydrochloride, mometasone furoate, benzonate, phexoncoyl (Chlophenir), cetirizine HCl / pseudoephed, phenphrine / code / promethazine, codeine / promethazine, ceprozil, ceprozil, ceprozil Dexamethasone, guaifenesin / pseudoephedrine, chlorophenylamine, hydrocodone, terephthalate, nedocromil sulfate Proterenol sulfate, mesalamine, azathioprine, mercaptopurine, diphenoxylate / atrop sulf, loperamide hydrochloride ride), omeprazole, folate, ciprofloxacin / dextrose-water, hydrocodone bitartrate fluoridone / apap, tetracycline hydrochloride (tetracrine hydrochloride) ), Thimerosal / boric acid, cholestyramine / sucrose, ciprofloxacin hydrochloride, hyoscyamine sulfate, meperidine hydrochloride ridine hydrochloride, midazolam hydrochloride, oxycodone hcl / acetaminophen, phthamethine hydrochloride, disulfide, sulfahydrate trimethoprim, polycarbophil, propoxyphene naphthylate, hydrocortisone, multivitamin, balsalazide disodium (balsalazide) isodium, codeine phosphate / apap, colesevelam HCl, cyanocobalamin, folic acid, levofloxacin, natalizumag, interferon-gamma, montelukast sodium ), Tiliacinolone acetonide, levofloxacin, guaifenesin, levalbuterol HCl, flunisolide, ceftriaxide. Sodium (ceftriaxone sodium), gatifloxacin, amoxicillin / clavulanate, flunisolide / menthol, chlorpheniramine / hydrocodone metaproterenol sulfate (on methyl) , Mometasone furoate, p-ephedrine / cod / chlorphenyl, p-ephedrine / loratadine, terbutaline sulfate line sulfate, tiotropium bromide, (R, R) -formoterol, TgAAT, cilomilast, roflumilast, interferon-alpha-2α, interferon-alpha-2α, interferon-alpha-2α, interferon conl, interferon-alpha-nl, pegylated interferon-alpha-2β, pegylated interferon-alpha-2β, ribavirin, peginterferon alpha-2β and ribavirin (ribavirin) Deoxycholic acid (Ursode oxycholic Acid), glycyrrhizic acid, thymalfasin, maxamine, VX-497, any compound used to treat HCV by intervention in the following targets: HCV polymerase, HCV protease , HCV helicase, HCV IRES (internal ribosome entry site), azathioprine, colchicine, albuterol sulfate, gamma interferon, lorazepam, furosemide, lisinopril (lisinoprirmid) cyclophosphamide), actinomycin (acti) nomycin) d, alteplase, levofloxacin, metaproterenol sulfate (metaprote)
renol sulfate, morphine sulfate, oxycodone HCl, triamcinolone acetonide, tacrolimus anhydrus, calcium, interferon-alpha -1b, clopidogrel hydrogensulfate (clopidogrel bisulfate), atenolol (atenolol), morphine sulfate (metoprolol succinate), warfarin sodium (warfarin sod) um), isosorbide mononitrate, simvastatin, tenecteplase, torsemide, retabase, losartanpitalase, ), Enalapril rat, aminodarone hydrochloride, tirofiban HCl m-hydrate, diltiazem hydrochloride, captopril, captopril irbesartan, propranolol hydrochloride, fosinopril sodium, fosinopril sodium salt, fosinopril sodium salt, fosinopril sodium salt Sotalol hydrochloride, docusate sodium, dobutamine HCl, alprazolam, pravastatin sora ium), atorvastatin calcium (atorvastatin calcium), midazolam hydrochloride (midazolam hydrochloride), meperidine hydrochloride (meperidine hydrochloride), isosorbide dinitrate (isosorbide dinitrate), epinephrine (epinephrine), dopamine hydrochloride (dopamine hydrochloride), bivalirudin (bivalirudin), rosuvastatin (Rosuvastatin), ezetimibe / simvastatin, avasimibe, cariporide, calcipotriene, propion Clobetasole propionate, triamcinolone acetonide, halobetasol propionate, fluotenone, fluocinone, fluocinone, fluocinone Acitretin, tar shampoo, betamethasone valerate (betamethasone valerate), mometasone furoate, ketoconazole, pramoxin (pr moxine / fluocinolone, hydrocortisone valerate, flulandrenolide, urea, betamethasone hydrode, clobetasol propionate, desonide), coal tar, diflorazone diacetate, etanercept, lactic acid, methoxsalen, HCl / bismuth, subgal / znox / resor, methyprednisolone acetate dnisolone acetate, sunscreen, halcinonide, salicylic acid, anthralin, crocortolone pivalate, coal extract, coal tar / salicylic acid, coal tar / salicylic acid / sulfur, dexamethasone Softeners, fluocinonides / softeners, mineral oil / castor oil / na lact, mineral oil / peanut oil, petroleum / isopropyl myristate, psoralen, soap / tribromosaran, thimerosal / boric acid, cyclosporine, alefacept aleceptpt, efalizumab, pimecrolimus (pime) crolimus), PUVA, UVB, naproxen, leflunomide, hydroxydequinone sulfate, prednisone, sulindac, sulindac Dimethyl sulfoxide, piroxicam, diclofenac sodium, ketoprofen, nabumetone, tolmetin sodium, calcipochi En (calcipotriene), cyclosporine, sodium / misoprostol, fluocinonide, glucosamine sulfate, sodium gold thiomalate, hydrocodone / saptaric acid disulfate ), Thioguanine, valdecoxib, hydroxychloroquine sulfate, leflunomide, valdecoxib, methylprednisothiolone, methylprednisothiolone , Triamcinolone acetonide, propoxyphene naphthylate / apap, nabumetone, piroxicam, oxydoxycoate, etodolac , Fentanyl, human recombinant anakinra, tramadol HCl, salsalate, sulindac, yanocobalamin / fa / pyridoxine, acetaminophen, alle Sodium dronate (alendronate sodium), prednisolone, morphine sulfate, lidocaine hydrochloride, dizine sulfate, glucosamine sulfate, glucosamine sulfate (Oxycodone) HC1 / acetaminophen, olopatadine HCl, misoprostol, omeprazole, mycophenolate mofetil (mycophe) olate mofetil), rituximab, IL-1 TRAP, MRA, CTLA4-IG, IL-18BP, ABT-874, anti-EL18 antibody, anti-IL15, BIRB-796, SCIO-469, X-702, AMG- 548, VX-740, Roflumilast, IC-485, CDC-801, Mesopram, Sirolimus, paclitaxel, Everolimus, Acro-T Hydrocodone / apap, cyclobenzaprine HCl, oxycodone HCl / acetaminophen (Acetaminophen), valdecoxib, codeine phosphate / apap, tramadol HCl / acetaminophen, metaxalone, methocarbamol, lidocaine hydrochloride, lidocaine (lidocine hydrochloride) dilofenac sodium, gabapentin, dexamethasone, carisoprodol, ketorolac tromethamine, diazepam, nabetone e), oxycodone HCl, tizanidine HCl, propoxyphene naphthylate / apap, asa / oxycod / oxycodone ter, ibuprofen (ibuprodone) ), Propoxyphene HCl, amitriptyline HCl, carisoprodol / codeine phos / asa, morphine sulfate, orphenadrine citrate, orphenadeline citrate epim), epidermal growth factor, corcosteroid, cyclosporine, aminosalicylate, 6-mercaptopurine, azathioprine, metronidazole, lipoxygenase inhibitor, mesalamine, olsalazine, balsalazide, balsalazide , Thromboxane inhibitor, IL-1 receptor antagonist, anti-IL-1β monoclonal antibody, anti-IL-6 monoclonal antibody, growth factor, elastase inhibitor, pyrimidinyl-imidazole compound, TNF, LT, IL-1, IL-2, IL -6, IL-7, IL-8, IL-15, IL-16, IL-18, EMAP-II, GM-CSF, FGF and PDGF antibodies Other agonists, CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69, CD90, or antibodies to their ligands, Sik
Rosporine, FK506, rapamycin, mycophenolate mofetil, leflunomide, NSAID, corticosteroid, prednisolone, phosphodiesterase inhibitor, adenosine agonist, anti-thrombotic agent, complement inhibitor agonist , IRAK, NIK, IKK, IL-1β converting enzyme inhibitor, TNFα converting enzyme inhibitor, T cell signaling inhibitor, metalloproteinase inhibitor, 6-mercaptopurine, angiotensin converting enzyme inhibitor, soluble cytokine receptor, soluble p55 TNF receptor, soluble p75 TNF receptor, sIL-1RI, sIL-1RII, s IL-6R, anti-inflammatory cytokines, IL-4, IL-10, IL-11, IL-13 and TGFβ.

  Non-limiting examples of therapeutic agents for ankylosing spondylitis that can be combined with compounds of Formula 1, 2, or 3 include: ibuprofen, diclofenac, misopro Styrofoam, naproxen, meloxicam, indomethacin, diclofenac, celecoxib, rofecoxib, ulfaxazib, ulfasamicin ), Prednisone, etanel Script (etanercept) and infliximab (infliximab).

  Non-limiting examples of therapeutic agents for asthma that can be combined with compounds of Formula 1, 2, or 3 include: albuterol, salmeterol / fluticasone, Montelukast sodium, fluticasone propionate, budesonide, prednisone, salmeterol xinafoate, salmeterol xinafoate, salmeterol xinafoate, salmeterol xinafoate, salmeterol xinafoate ), Prednisolone sodium phosphate, triamcinolone acetonide, beclomethasone dipropionate, ipratropium bromide (Prednisolone), anhydrous theophylline, methylprednisolone sodium succinate, clarithromycin clarithromycin, zafirlukast, formoterol fumarate (formoterol fumarate), influenza virus vaccine, amoxicillin trihydrate, flunisolidine hydrochloride allergy (Fexofenadine hydrochloride), flunisolide / methanol, amoxicillin / clavulanate, levofloxacin, inhalation aids, guaifenesin, dexamethasone sodium phosphate (dexamesone) thathone sodium phosphate, moxifloxacin HCl, doxycycline hydrate, guaifenesin / d-methorphan, methotrephine / phlephrine (Gatifloxacin), cetirizine hydrochloride, mometasone furoate, salmeterol xinafoate, benzonate, phenexate, ph exonce Codon / chlorphenyl, cetirizine HCl / pseudoephed, phenylephrine / code / promethazine, codeine, prometazine, promethazine, promethazine , Guanifenesin / pseudoephedrine, chlorpheniramine / hydrocodone, nedocromil sodium, terbutaline sulfate te), epinephrine (epinephrine), methylprednisolone (methylprednisolone), and metaproterenol sulfate (metaproterenol sulfate).

  Non-limiting examples of therapeutic agents for COPD that can be combined with compounds of Formula 1, 2, or 3 include: albuterol sulfate / ipratropium, ipratropium bromide (Ipratropium bromide), salmeterol (flumeterasone), albuterol (albutyrool), salmeterol xinafoate (salmeterol xinafoate), fluticasone propionate (fluticasone propionate) Methylpred succinate Two pyrazolone sodium (methylprednisolone sodium succinate), montelukast sodium (montelukast sodium), budesonide (budesonide), formoterol fumarate (formoterol fumarate), triamcinolone acetonide (triamcinolone acetonide), levofloxacin (levofloxacin), guaifenesin (guaifenesin), azithromycin (azithromycin ), Beclomethasone dipropionate dipropionate, levalbuterol HCl, flunisolide, ceftriaxo Sodium (ceftriaxone sodium), amoxicillin trihydrate, gatifloxacin, zafirlukast, amoxicillin, amoxicillin, amoxicillin Hydrocodone, metaproterenol sulfate, methylprednisolone, mometasone furoate, p-ephedrine / cod / chlorphenyl, pyrbuterol acetate Pirbuterol acetate, p-ephedrine / loratadine, terbutaline sulfate, tiotropium bromide, (R, R) -formoterol, lomastol, rogadine (Roflumilast).

  Non-limiting specific examples of therapeutic agents for HCV that can be combined with compounds of Formula 1, 2, or 3 include: interferon-alpha-2a, interferon-alpha-2b, interferon-alpha. conl, interferon-alpha-nl, pegylated interferon-alpha-2a, pegylated interferon-alpha-2b, ribavirin, peginterferon alpha-2b + ribavirin acid, ursodeoxylic acid Treat HCV with glycyrrhizic acid, thymalfasin, maxamine, VX-497, and the following target interventions Any compound used to: HCV polymerase, HCV protease, HCV helicase, and HCV IRES (internal ribosome entry site).

  Non-limiting examples of therapeutic agents for idiopathic pulmonary fibrosis that may be combined with compounds of Formula 1, 2, or 3 include: prednisone, azathioprine, albuterol , Colchicine, albuterol sulfate, digoxin, gamma interferon, methylprednisolone sod succ, lorazepam, furosemide, lisinopyrimidyl phosphide, lisinopyrimido Ipratropium bromide, actinomycin d, alteplase, fluticasone propionate ( luticasone propionate, levofloxacin, metaproterenol sulfate, morphine sulfate, oxycodone HCl, potassium chloride, triamcinolone acetonide, tricalcine acetolide methotrexate), mycophenolate mofetil and interferon-gamma-1β.

  Non-limiting examples of therapeutic agents for myocardial infarction that can be combined with compounds of Formula 1, 2, or 3 include the following: aspirin, nitroglycerin, metroprolol tartrate, Enoxaparin sodium, heparin sodium, clopidogrel hydrogensulfate (clopidogrel bisulfate), carvedilol, atenolol, morphine sulfate, metoprolol sodium succinate Isosorbide mononitrate (isosorbide monitit) ate), digoxin, furosemide, simvastatin, ramipril, tenecteplase, enalapril maleate, torsemide, torsemide, torsemide (Quinapril) HCl / mag carb, bumetanide, alteplase, enalapril, amiodarone hydrochloride, trifiban dihydrate HCl, tirofibane hydrate HCl ltiazem hydrochloride), captopril (captopril), irbesartan (irbesartan), valsartan (valsartan), propranolol hydrochloride (propranolol HYDROCHLORIDE), fosinopril sodium (fosinopril sodium), lidocaine hydrochloride (lidocaine HYDROCHLORIDE), eptifibatide (eptifibatide), cefazolin sodium ( cefazolin sodium), atropine sulfate, aminocaproic acid, spironolactone, interferon, sotalol hydrochloride, potassium chloride, doxate sodium (Docusate sodium), dobutamine HCl, alprazolam, pravastatin sodium, atorvastatin in sorbide, iodamine hydrochloride, hydrate Epinephrine, dopamine hydrochloride, bivalirudin, rosuvastatin, ezetimibe / simvastatin, avasimibe and cariporide (ca) iporide).

  Non-limiting examples of therapeutic agents for psoriasis that can be combined with compounds of Formula 1, 2, or 3 include: calcipotriene, clobetasol propionate, triamcinolone acetonide (triamcinolone acetonide), halobetasol propionic acid (halobetasol propionate), tazarotene (tazarotene), methotrexate (methotrexate), fluocinonide (fluocinonide), strengthening diprop betamethasone (betamethasone), fluocinolone acetonide (fluocinolone acetonide), acitretin ( actr tin), tar shampoo, betamethasone valerate, mometasone furoate, ketoconazole, pramoxine, fluocinolol, fluocinolol, fluocinolol , Betamethasone, clobetasol propionate / emol, fluticasone propionate, azithromycin, hydrocortisone, moisturizing agent, desoni ), Pimecrolimus, coal tar, diflorazone diacetate, etanercept folate, lactic acid, methoxallen (hmu / sulzomethrezo) ), Prednisone, sunscreen, halcinonide, salicylic acid, anthralin, crocortolone pivalate, coal extract, coal tar / salicylic acid, coal tar / salicylic acid / sulfur, Tazon (desoximetasone), diazepam, softener, fluocinonide / softener, mineral oil / castor oil / na lact, mineral oil / peanut oil, petroleum / isopropyl myristate, psoralen, salicylic acid, soap / t-ribran ), Thimerosal / boric acid, celecoxib, infliximab, cyclosporine, alefaccept, efalizumab, tacrolimus, UV, pumecrolimis (ulB), pimecrolimus (ul)

  Non-limiting examples of therapeutic agents for psoriatic arthritis that can be combined with compounds of Formula 1, 2, or 3 include the following: methotrexate, etanercept, rofecoxib ), Celecoxib, folic acid, sulfasalazine, naproxen, leflunomide, methylprednisolone acetate, indomethacin (indomethine) Sulindac (su lindac), reinforced Diprop betamethasone (betamethasone diprop augmented), infliximab (infliximab), methotrexate (methotrexate), Horato (folate), triamcinolone acetonide (triamcinolone acetonide), diclofenac (diclofenac), dimethyl sulfoxide (dimethylsulfoxide), piroxicam (piroxicam) , Diclofenac sodium, ketoprofen, meloxicam, methylprednisolone, nabumetone one), tolmetin sodium, calcipotriene, cyclosporine, diclofenac sodium / misoprostocol, fluocinonide acid, fluocinonide acid, fluocinonide acid / Apap, ibuprofen, risedronate sodium, sulfadiazine, thioguanine, valdecoxib, alefaccept and efalizumab.

  Non-limiting examples of therapeutic agents for recurrent stenosis that can be combined with compounds of Formula 1, 2, or 3 include sirolimus, paclitaxel, everolimus, tacrolimus, ABT-578 and acetaminophen are included.

  Non-limiting examples of therapeutic agents for sciatica that can be combined with compounds of Formula 1, 2, or 3 include the following: hydrocodone bitartrate / apap, rofecoxib, cyclobenzaprine (Cyclobenzaprine) HCl, methylprednisolone, naproxen, ibuprofen, oxycodone HCl / acetaminophen (b), celcoxib (b) ), Prednisone, Codeine acid / apap, tramadol hcl / acetaminophen, metaxalone, meloxicam, methocalamol, lidocaine hydrochloride, lidocaine hydrochloride Gabapentin, dexamethasone, carisoprodol, ketorolac tromethamine, indomethacin, acetaminophen, di Azepam, nabumetone, oxycodone HCl, tizanidine HCl, diclofenac sodium / misoprostoyl, propoxyphenopyphene napsilate / Oxycodone ter, ibuprofen / hydrocodone bit, tramadol HCl, etodolac, propoxyphene HCl, amitriptyline Cl, carisoprodol / codeine phos / asa, morphine sulfate, multivitamins, naproxen sodium, orphenadrine citrate and temazepam.

  Preferred specific examples of therapeutic agents for SLE (Lupus) that can be combined with compounds of Formula 1, 2, or 3 include the following: NSAIDs such as diclofenac, naproxen, ibuprofen ( ibuprofen, piroxicam, indomethacin; COX2 inhibitors such as celecoxib, rofecoxib, valdecoxib; antimalarial substances such as hydroxychloroquine , Prednisolone, budenosi (Budenoside), dexamethasone (dexamethasone); cytotoxic agent, eg azathioprine, cyclophosphamide, mycophenolate mofetil (mycophenolate mofetil), methotrexate (methotrexate); PDE4 inhibitors or purine synthesis inhibitor, for example Cellcept (R). Compounds of formula 1, 2 or 3 can also be combined with, for example, the following substances: sulfasalazine, 5-aminosalicylic acid, olsalazine, Imuran®, and inflammatory cytokines such as IL-1 Substances that interfere with the synthesis, production or action of, for example, caspase inhibitors, such as IL-1β converting enzyme inhibitors and IL-1ra. Also, the compound of Formula 1, 2, or 3 is a T cell signaling inhibitor, such as a tyrosine kinase inhibitor; or a molecule that targets a T cell activation molecule, such as CTLA-4-IgG or an anti-B7 family antibody, anti-PD-1 Can be used with family antibodies. Compounds of formula 1, 2 or 3 are IL-11 or anti-cytokine antibodies, such as fonotolizumab (anti-IFNg antibody) or anti-receptor receptor antibodies, such as anti-IL-6 receptor antibodies and antibodies to B cell surface molecules Can be combined with Compounds of formula 1, 2 or 3 are also LJP394 (abetimus), a substance that depletes or inactivates B cells, such as rituximab (anti-CD20 antibody), lymphoblast-B (anti-antibody). BlyS antibody), TNF antagonists such as anti-TNF antibodies, HUMIRA® (US Pat. No. 6,090,382), CA2 (REMICADE ™), CDP 571, TNFR-Ig construct (p75TNFRIgG (ENBREL ™) )) And p55TNFRIgG (LENERCEPT ™)).

  In the present invention, the following definitions apply.

  A “therapeutically effective amount” is a compound of formula 1, 2, or 3 or two or more such compounds that completely or partially inhibits the progression of the disease state or at least partially ameliorates one or more of the symptoms of the disease state The amount of combination. A therapeutically effective amount can also be an amount that is prophylactically effective. The amount that is therapeutically effective depends on the size and sex of the patient, the condition being treated, the severity of the condition and the outcome sought. For a given patient, the therapeutically effective amount can be determined by methods known to those skilled in the art.

  “Physiologically acceptable salts” retain the biological effectiveness and properties of the free base and are inorganic acids or organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid. Acids such as sulfonic acid, carboxylic acid, organophosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, citric acid, fumaric acid, maleic acid, succinic acid, benzoic acid, salicylic acid, lactic acid, tartaric acid (e.g. (+) Or (−)-tartaric acid or a mixture thereof), a salt obtained by reaction with an amino acid (for example, (+) or (−)-amino acid or a mixture thereof) and the like. These salts can be prepared by methods known to those skilled in the art.

  Certain compounds of formula 1, 2 or 3 having acidic substituents may exist as salts with pharmaceutically acceptable bases. The present invention includes such salts. Specific examples of such salts include sodium salt, potassium salt, lysine salt and arginine salt. These salts can be prepared by methods known to those skilled in the art.

  Certain compounds of Formula 1, 2, or 3 and their salts can exist in more than one crystal form, and the present invention includes each crystal form and mixtures thereof.

  Certain compounds of Formula 1, 2, or 3 and their salts may exist in the form of solvates, for example hydrates, and the present invention includes each solvate and mixtures thereof.

  Certain compounds of Formula 1, 2, or 3 can exist as various tautomers or various geometric isomers, and the present invention provides each tautomer of Formula 1, 2, or 3 and / or Including geometric isomers as well as mixtures thereof.

  Certain compounds of Formula 1, 2, or 3 can exist in various stable conformational forms that can be separated. Torsional asymmetry due to restricted rotation around an asymmetric single bond (eg, due to steric hindrance or ring distortion) may allow separation of different conformers. The present invention includes each conformational isomer of compounds of formula 1, 2 or 3 and mixtures thereof.

  Certain compounds of formula 1, 2 or 3 can exist in zwitterionic form and the present invention includes each zwitterionic form of compounds of formula 1, 2 or 3 as well as mixtures thereof.

  As used herein, the term “prodrug” refers to a substance that is converted into the parent drug in vivo by some physiochemical process (eg, a prodrug is exposed to physiological pH to form the desired drug form). Converted to). In some situations, prodrugs are often useful because they can be administered more easily than their parent drug. They are bioavailable, for example by oral administration, but the parent drug cannot. A prodrug may have improved solubility in a pharmacological composition compared to its parent drug. In the compounds of the present invention, which is given as a non-limiting example of a prodrug, it is administered as an ester (“prodrug”) to facilitate permeation through the cell membrane where water solubility does not favor. Once entered into the cell, where water solubility favors, it will be metabolically hydrolyzed to the carboxylic acid.

  Prodrugs can have a number of useful properties. For example, prodrugs can be more water soluble than the final drug, thereby facilitating intravenous administration of the drug. Prodrugs can also have higher levels of oral bioavailability than the final drug. After administration, the prodrug is cleaved enzymatically or chemically so that the final drug is carried into the blood or tissue.

Typical prodrugs are cleaved to release the corresponding free acid, and such hydrolyzable ester-forming residues of the compounds of the invention include carboxylic acid substituents (eg, — (CH ₂ ) C (O) OH, or a moiety containing a carboxylic acid), in which case the free hydrogen is (C ₁ -C ₄ ) alkyl, (C ₂ -C ₁₂ ) Alkanoyloxymethyl, (C ₄ -C ₉ ) 1- (alkanoyloxy) ethyl, 1-methyl-1- (alkanoyloxy) -ethyl having 5-10 carbon atoms, 3-6 carbon atoms Having alkoxycarbonyloxymethyl, 1- (alkoxycarbonyloxy) ethyl having 4-7 carbon atoms, 1-methyl-1- (alkoxycarbonyl) having 5-8 carbon atoms Xyl) ethyl, N- (alkoxycarbonyl) aminomethyl having 3 to 9 carbon atoms, 1- (N- (alkoxycarbonyl) amino) ethyl having 4 to 10 carbon atoms, 3-phthalidyl, 4- Crotonolactonyl, gamma-butyrolactone-4-yl, di-N, N- (C ₁ -C ₂ ) alkylamino (C ₂ -C ₃ ) alkyl (eg β-dimethylaminoethyl), carbamoyl- (C Substituted by _1- C ₂ ) alkyl, N, N-di (C ₁ -C ₂ ) -alkylcarbamoyl- (C ₁ -C ₂ ) alkyl and piperidino-, pyrrolidino- or morpholino (C ₂ -C ₃ ) alkyl ing.

Other exemplary prodrugs release an alcohol of formula 1, 2 or 3, in which case the hydroxyl substituent (eg R ¹ contains hydroxyl) is (C ₁ -C ₆ ) alkanoyloxymethyl, 1-((C ₁ -C ₆ ) alkanoyloxy) ethyl, 1-methyl-1-((C ₁ -C ₆ ) alkanoyloxy) ethyl, (C ₁ -C ₆ ) alkoxycarbonyloxymethyl, N- (C _1- C ₆ ) alkoxycarbonylamino-methyl, succinyl, (C ₁ -C ₆ ) alkanoyl, α-amino (C ₁ -C ₄ ) alkanoyl, arylactyl and α-aminoacyl, or α-aminoacyl-α-aminoacyl (Wherein the α-aminoacyl moiety is independently any of the naturally occurring L-amino acids found in proteins. In _a), is replaced by P (O) (OH) 2 , -P (O) (O (C 1 -C 6) _{alkyl) 2} or glycosyl (the radical resulting from detachment of the hydroxyl of the hemiacetal of a carbohydrate) Yes.

  As used herein, the term “heterocycle” or “heterocyclyl” includes non-aromatic ring systems such as, but not limited to, monocyclic, bicyclic and tricyclic rings, These can be fully saturated or can contain one or more units of unsaturation (for the avoidance of doubt, the degree of unsaturation does not produce an aromatic ring system). ), Having at least one heteroatom, for example 3 to 12 atoms including nitrogen, oxygen or sulfur. By way of illustration (which should not be construed as limiting the scope of the invention), specific examples of heterocycles are listed below: azepine, azetidinyl, morpholinyl, oxopiperidinyl, oxopyrrolidinyl, piperazinyl, Piperidinyl, pyrrolidinyl, quinicledinyl, thiomorpholinyl, tetrahydropyranyl and tetrahydrofuranyl.

  As used herein, the term “heteroaryl” includes ring systems including, but not limited to monocyclic, bicyclic and tricyclic rings, and includes at least one heteroatom, such as It has 3 to 12 atoms including nitrogen, oxygen or sulfur. Illustrative examples (which should not be construed as limiting the scope of the invention) include the following: azaindole, benzo (b) thienyl, benzimidazolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzo Thiadiazolyl, benzooxadiazolyl, furan, imidazole, imidazopyridine, indole, indolinyl, indazole, isoindolinyl, isoxazole, isothiazole, oxadiazole, oxazole, purine, pyran, pyrazine, pyrazole, pyridine, pyrimidine, pyrrole, Pyrrolo [2,3-d] pyrimidine, pyrazolo [3,4-d] pyrimidine, quinoline, quinazoline, triazole, thiazole, thiophenyl, tetrahydroindole, tetrazole, thiadiazole Thienyl, thiomorpholine or tropanyl.

When the term “substituted heterocycle” (or heterocyclyl) or “substituted heteroaryl” is used, it provides one or more substituents that can be prepared by one of ordinary skill in the art to provide molecules that are kinase inhibitors. Means that the heterocyclic group is substituted. Illustratively (which should not be construed as limiting the scope of the invention), preferred substituents of the heterocycles of the invention are each independently alkenyl, alkoxy, alkoxyalkoxy, alkoxyalkyl, alkoxy Carbonyl, alkoxycarbonylheterocycloalkoxy, alkyl, alkylcarbonyl, alkyl ester, alkyl-O-C (O)-, alkyl-heterocyclyl, alkyl-cycloalkyl, alkyl-nitrile, alkynyl, amide group, amino, aminoalkyl, amino carbonyl, carbonitrile, carbonylalkoxy, _{carboxamido, CF 3, CN, -C (} O) OH, -C (O) H, -C (O) -C (CH 3) 3, -OH, -C (O) O-alkyl, -C (O) O-cycloalkyl, -C ( ) O-heterocyclyl, -C (O) -alkyl, -C (O) -cycloalkyl, -C (O) -heterocyclyl, cycloalkyl, dialkylaminoalkoxy, dialkylaminocarbonylalkoxy, dialkylaminocarbonyl, halogen, heterocyclyl, heterocycloalkyl group, heterocyclyloxy, hydroxy, hydroxyalkyl, nitro, OCF _3, oxo, _{phenyl, -SO 2 CH 3, -SO 2} CR 3, tetrazolyl, thienyl alkoxy, trifluoromethyl carbonyl amino, trifluoromethyl sulfonamide , Heterocyclylalkoxy, heterocyclyl-S (O) _p , cycloalkyl-S (O) _p , alkyl-S-, heterocyclyl-S, heterocycloalkyl, cycloalkylalkyl, hetero Shikurochio, ^{_{cycloalkylthio, -Z 105 -C (O) N}} (R) 2, -Z 105 -N (R) -C (O) -Z 200, -Z 105 -N (R) -S (O) 2 ^{-Z 200, -Z 105 -N (R} ) -C (O) -N (R) -Z 200, -N (R) -C (O) R, -N (R) -C (O) OR, Selected from optionally substituted substituents consisting of OR-C (O) -heterocyclyl-OR, R _c and —CH ₂ OR _c , wherein
R ₃ is C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl or phenyl;
p is 0, 1 or 2;
R _c is independently in each case hydrogen, optionally substituted alkyl, optionally substituted aryl, — (C ₁ -C ₆ ) —NR _d R _e , —E _{- (CH 2) t -NR d} R e, -E- (CH 2) t -O- alkyl, -E- _{(CH 2)} t -S- alkyl, or _{-E- (CH} 2) t -OH ( Where t is an integer from about 1 to about 6.
Z ¹⁰⁵ is independently in each case a covalent bond, alkyl, alkenyl or alkynyl and Z ²⁰⁰ is in each case independently alkyl, alkenyl, alkynyl, phenyl, alkyl-phenyl, alkenyl-phenyl. Or an optionally substituted substituent selected from the group consisting of alkynyl-phenyl,
E is a direct bond, O, S, S (O), S (O) ₂ or NR _f , where R _f is H or alkyl, and R _d and R _e are independently H, Alkyl, alkanoyl or SO ₂ -alkyl, R _d , R _e and the nitrogen to which they are attached together form a 5- or 6-membered heterocycle.

  As used herein, a “heterocycloalkyl” group is a heterocyclic group that is linked to a compound by an aliphatic group having from 1 to about 8 carbon atoms. For example, a preferred heterocycloalkyl group is a morpholinomethyl group.

As used herein, a designation such as “aliphatic” or “aliphatic group” or “(C ₀ -C ₈ )” is a straight-chain that is fully saturated or contains one or more unsaturated units. Includes chain or branched hydrocarbons and thus includes hydrocarbons containing alkyl, alkenyl, alkynyl, and mixtures of single, double, and triple bonds. When the group is C ₀ it means that the moiety is not present, in other words it is a bond. “Alkyl” as used herein means C ₁ -C ₈ and includes straight or branched hydrocarbons, which are fully saturated. Preferred alkyl are methyl, ethyl, propyl, butyl, pentyl, hexyl and isomers thereof. As used herein, “alkenyl” and “alkynyl” mean C ₂ -C ₈ and include one or more unsaturated units, one or more double bonds for alkenyl and one or more triple bonds for alkynyl. Including linear or branched hydrocarbons.

  As used herein, aromatic groups (or aryl groups) include aromatic carbocyclic ring systems (eg, phenyl and cyclopentyldienyl) and fused polycyclic aromatic ring systems (eg, naphthyl, biphenylenyl, and 1, 2,3,4-tetrahydronaphthyl).

Cycloalkyl as used herein, fully saturated from or one or more C ₃ -C ₁₂ monocyclic or polycyclic have an unsaturated bond does not reach the aromatic group (e.g., two (Cyclic, tricyclic, etc.) means hydrocarbon. Preferred specific examples of the cycloalkyl group include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl and cyclohexenyl.

  As used herein, an amide group means —NHC (═O) —.

  As used herein, an acyloxy group is —OC (O) R.

A number of moieties or substituents as used herein are referred to as “substituted” or “optionally substituted”. When a moiety is modified by one of these terms, it means that any part of the moiety known to those skilled in the art to be available for substitution can be substituted unless otherwise indicated. Where it contains one or more substituents and when two or more substituents are present, each substituent is independently selected. Such means for substitution are well known in the art and / or taught by the present disclosure. By way of illustration (which should not be construed as limiting the scope of the invention), several examples of substituent groups are given below: alkenyl groups, alkoxy groups (—O—C ₁ —C ₆ -alkyl-OR, —O—C ₁ -C ₆ -alkyl-N (R) ₂ , and OCF ₃ itself can be substituted), alkoxyalkoxy, alkoxycarbonyl, alkoxycarbonylpiperidinyl- alkoxy, alkyl group _(-C 1 _{-C 6} - alkyl -OR, _-C 1 _{-C 6} - alkyl -N _{(R) 2,} and as in -CF _3, which itself may be substituted), alkylamino, alkylcarbonyl, alkyl ester, alkyl nitrile, alkylsulfonyl, amino, _aminoalkoxy, CF 3, COH, COOH, CN, cycloalkyl, di Alkylamino, dialkylaminoalkoxy, dialkylaminocarbonyl, dialkylaminocarbonylalkoxy, dialkylaminosulfonyl, ester (—C (O) —OR, wherein R is alkyl, heterocycloalkyl (which can be substituted), heterocyclyl, etc. (They can be substituted), halogen or halo groups (F, Cl, Br, I), hydroxy, morpholinoalkoxy, morpholinoalkyl, nitro, oxo, OCF ₃ , optionally substituted May be phenyl, S (O) ₂ CH ₃ , S (O) ₂ CF ₃ , and sulfonyl, N-alkylamino or N, N-dialkylamino, where the alkyl group may also be substituted.

  One or more compounds of the invention may treat or treat a disease or condition described herein, either by itself or as a pharmaceutical composition in admixture with a biologically suitable carrier or excipient. It can be administered to human patients at a dose to improve. Also, mixtures of these compounds can be administered as a simple mixture or as a suitable formulated pharmaceutical composition. A therapeutically effective amount refers to that amount of the compound sufficient to result in prevention or alleviation of the diseases or conditions described herein. Techniques for formulation and administration of the compounds of this application are described in “Remington's Pharmaceutical Sciences,” Mack Publishing Co. , Easton, PA (latest edition) and can be found in references well known to those skilled in the art.

  Suitable routes of administration include, for example, oral, ophthalmic, rectal, transmucosal, topical or enteral administration; parenteral delivery such as intramuscular, subcutaneous, intrathecal injection, and intrathecal, directly intraventricular, intravenous, Intraperitoneal, intranasal or intraocular injection may be included.

  Alternatively, the compound can be administered locally rather than systemically, for example, by injection of the compound directly into the edema site, often as a depot or sustained release agent.

  Furthermore, the drug can be administered by targeted drug delivery systems, for example, in liposomes coated with endothelial cell specific antibodies.

  The pharmaceutical composition of the present invention can be produced by a method known per se, for example, a usual mixing, dissolution, granulation, dragee production, syrup, emulsification, encapsulation, capture or lyophilization method.

  Accordingly, the pharmaceutical compositions used in the present invention employ one or more physiologically acceptable carriers containing excipients and adjuvants that facilitate the processing of the active compound into pharmaceutically usable formulations. It can be produced by conventional methods. Proper formulation is dependent upon the route of administration chosen.

  For injection, the substances of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiologically buffered saline. For transmucosal administration, permeation enhancers suitable for the barrier to be permeated are used in the formulation. Such permeation enhancers are generally known in the art.

  For oral administration, the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers are formulated with the compounds of the invention as tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions, etc. for ingestion by the patient to be treated. Make it possible. Oral pharmaceutical preparations may be prepared by combining the active compound with a solid excipient and, optionally, crushing the resulting mixture and optionally adding appropriate adjuvants, then processing the granule mixture to produce tablets. Alternatively, it can be obtained by obtaining a sugar-coated core. Suitable excipients include in particular fillers such as sugars such as lactose, sucrose, mannitol or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methylcellulose, hydroxypropyl Mention may be made of methyl-cellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar or alginic acid, or a salt thereof such as sodium alginate.

  The sugar-coated core is provided with an appropriate coating. For this purpose, concentrated sugar solutions can be used, which in some cases are gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol and / or titanium dioxide, lacquer solutions, and suitable organic solvents or It may contain a solvent mixture. Dyestuffs or pigments may be added to the tablets or dragees for identification or to characterize different combinations of active compound doses.

  Pharmaceutical formulations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules contain the active ingredient together with fillers such as lactose, binders such as starch, and / or lubricants such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, the active compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. A stabilizer may also be added. All formulations for oral administration should be in dosage forms suitable for such administration.

  For buccal administration, the composition may take the form of tablets or lozenges formulated in conventional manner.

  For administration by inhalation, the compounds used in the present invention can be prepared in pressurized packs or in suitable propellants such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases. It is conveniently transported from the nebulizer in the form of an aerosol spray. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a certain amount. For example, gelatin capsules and cartridges for use in an inhaler or ventilator may be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch.

  The compounds may be formulated for parenteral administration by injection, eg, bolus injection or continuous infusion. Formulations for injection may be provided in unit dosage forms such as ampoules or multi-dose containers with a preservative added. The compositions can take the form of suppositories, solutions or emulsions in oily or aqueous vehicles and contain formulation agents such as suspending, stabilizing and / or dispersing agents. Yes.

  Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. The active compound suspensions may also be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. In some cases, the suspension may also contain suitable stabilizers or substances that increase the solubility of the compound to allow the production of highly concentrated solutions.

  Alternatively, the active ingredient may be in powder form for reduction prior to use with a suitable vehicle, such as sterile pyrogen-free water.

  The compounds can be formulated as rectal compositions, suppositories, or retention enemas (eg, containing conventional suppository bases such as cocoa butter or other glycerides).

  In addition to the formulations already described, the compounds can also be formulated as a depot. Such long acting formulations may be administered by implantation, for example subcutaneously or intramuscularly or by intramuscular injection. Thus, for example, the compound can be formulated with a suitable polymerizable or hydrophobic material or ion exchange resin (eg, as an acceptable emulsion in oil) or as a poorly soluble derivative (eg, a poorly soluble salt). .

  An example of a pharmaceutical carrier for the hydrophobic compounds of the present invention includes a cosolvent system comprising benzyl alcohol, a nonpolar surfactant, a hydratable organic polymer, and an aqueous phase. The co-solvent system can be a VPD co-solvent system. VPD is a solution of 3% w / v benzyl alcohol, 8% w / v nonpolar surfactant polysorbate 80 and 65% w / v polyethylene glycol 300 adjusted to the appropriate volume in absolute ethanol. The VPD co-solvent system (VPD: 5W) consists of VPD diluted 1: 1 with 5% dextrose in aqueous solution. This co-solvent system dissolves hydrophobic compounds well and itself exhibits low toxicity upon systemic administration. In essence, the ratio of a co-solvent system can vary considerably without compromising its solubility and toxicity characteristics. Furthermore, the entity of the co-solvent component can vary. For example, other low-toxic nonpolar surfactants can be used in place of polysorbate 80, the fraction size of polyethylene glycol can vary, and other biocompatible polymers can be used instead of polyethylene glycol. For example, polyvinylpyrrolidone can be used, and other sugars or polysaccharides can be used instead of dextrose.

  Alternatively, other delivery systems for hydrophobic pharmaceutical compounds can be used. Liposomes and emulsions are well known examples of delivery vehicles or carriers for hydrophobic drugs. Some organic solvents such as dimethyl sulfoxide can also be used, but usually have the disadvantage of greater toxicity. The compounds can also be delivered using sustained release systems, such as a semi-permeable matrix of solid hydrophobic polymers containing the therapeutic substance. Various sustained-release materials have been established and are well known by those skilled in the art. Sustained release capsules can release the compound over several weeks to over 100 days depending on their chemical properties. Depending on the chemical nature and biological stability of the therapeutic agent, additional methods for protein stabilization may be used.

  The pharmaceutical composition may also include a suitable solid or gel phase carrier or excipient. Examples of such carriers or excipients include, but are not limited to, polymers such as calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polyethylene glycol. Absent.

  Many of the compounds of the present invention can be provided as salts with pharmaceutically compatible counterions. Pharmaceutically compatible salts can be formed with numerous acids, including but not limited to hydrochloric acid, sulfuric acid, acetic acid, lactic acid, tartaric acid, malic acid, succinic acid, and the like. Salts tend to be more stable than the corresponding free base form in aqueous or other protic solvents.

  Pharmaceutical compositions suitable for use in the present invention include compositions containing the active ingredient in an effective amount to achieve its intended purpose. More specifically, a therapeutically effective amount means an amount effective to prevent the occurrence of symptoms in the subject being treated or to reduce existing symptoms in the subject being treated. The determination of an effective amount is well within the capability of those skilled in the art.

For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cellular assays. For example, doses that achieve a circulating concentration range that includes an IC ₅₀ (ie, the concentration of a test compound that achieves half-maximal inhibition of a given protein kinase activity) determined in a cellular assay can be determined in cell and animal models. . In some cases it is appropriate to determine the IC _{50 in} the presence of 3-5% serum albumin. This is because such a determination estimates the binding effect of plasma proteins on the compound. Such information can be used to more accurately determine useful doses in humans. Furthermore, the most preferred compounds for systemic administration effectively inhibit protein kinase signaling in intact cells at levels that can be safely achieved in plasma.

A therapeutically effective amount refers to that amount of the compound that results in amelioration of symptoms in a patient. Toxicity and therapeutic efficacy of such compounds is determined by standard pharmaceutical in cell cultures or experimental animals, eg, to determine maximum tolerated dose (MTD) and ED ₅₀ (effective amount for 50% maximal response). It can be determined by the method. The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio between MTD and ED _50. Compounds that exhibit high therapeutic indices are preferred. Data obtained from these cell culture assays and animal studies can be used in determining dosage ranges for human use. The dosage of such compounds is preferably within a range of circulating concentrations that include the ED ₅₀ with little or no toxicity. The dosage may vary within this range depending on the dosage form used and the route of administration used. The exact formulation (formulation), route of administration and dosage can be selected by the individual physician in view of the patient's condition (eg, Fingl et al., 1975, in “The Pharmaceutical Basis of Therapeutics”, Ch. 1 pl. See). In crisis treatment, an acute bolus or infusion close to the MTD may be required for rapid response.

  Dosage amount and interval may be adjusted individually to provide plasma levels or minimal effective concentrations (MECs) of the active moiety which are sufficient to maintain the kinase modulating effects. The MEC will vary for each compound, but can be estimated from in vitro data, eg, the concentration required to achieve 50-90% inhibition of protein kinase using the assays described herein. The dosage required to achieve MEC will depend on individual characteristics and route of administration. On the other hand, HPLC assays or bioassays can be used to determine plasma concentrations.

  In addition, the administration interval can be determined using the MEC value. The compound is administered using a regimen that maintains plasma levels above the MEC for 10-90%, preferably 30-90%, most preferably 50-90% of the time until the desired improvement in symptoms is achieved. Should be. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.

  The amount of composition administered will, of course, be dependent on the subject being treated (patient), the subject's weight, the severity of the affliction, the manner of administration and the judgment of the prescribing physician.

  The composition may be provided by a pack or dispensing device that may contain one or more unit dosage forms containing the active ingredients. The pack can include, for example, a metal or plastic foil, such as a blister pack. The pack or dispenser device can be accompanied by instructions for administration. A composition comprising a compound of the invention formulated in a compatible pharmaceutical carrier is manufactured, placed in a suitable container, and an indication is made regarding the treatment of the indicated condition.

  In some formulations, it may be beneficial to use the compounds of the present invention in the form of very small sized particles obtained, for example, by fluid energy milling.

  The use of the compounds of the invention in the manufacture of a pharmaceutical composition is illustrated by the following description. In this description, the term “active compound” refers to any compound of the invention, in particular any compound that is one end product of the previous example.

a) Capsules In the production of capsules, 10 parts by weight of active compound and 240 parts by weight of lactose are deagglomerated and mixed. The mixture is filled into hard gelatin capsules such that each capsule contains a unit dose or part of a unit dose of active compound.

b) Tablets Tablets can be produced, for example, from the following ingredients.

Parts by weight of active compound 10
Lactose 190
Corn starch 22
Polyvinylpyrrolidone 10
Magnesium stearate 3
The active compound, the lactose, and some of the starch can be deagglomerated and mixed, and the resulting mixture can be granulated with a solution of polyvinylpyrrolidone in ethanol. It is possible to mix the dry granules with the magnesium stearate and the rest of the starch. The mixture is then compressed in a tablet machine to give tablets, each containing a unit dose or part of a unit dose of active compound.

c) Enteric coated tablets Tablets can be produced by the method described in (b) above. The tablets are enteric coated by conventional methods using a solution of 20% cellulose acetate phthalate and 3% diethyl phthalate in ethanol: dichloromethane (1: 1).

d) Suppositories In the manufacture of suppositories, for example, 100 parts by weight of the active compound are added to 1300 parts by weight of a triglyceride suppository base and the mixture is added to a suppository each containing a therapeutically effective amount of the active compound. Mold into agent.

  In the compositions of the invention, the active compound can be mixed with other compatible pharmacologically active ingredients, if desired. For example, the compounds of the present invention can be administered in combination with other therapeutic agents known to treat the diseases or conditions described herein. For example, inhibiting or preventing the production of VEGF or angiopoietin, reducing the intracellular response to VEGF or angiopoietin, blocking intracellular signaling, inhibiting vascular permeability, reducing inflammation, or forming edema or Combined with one or more additional pharmaceutical substances that inhibit or prevent angiogenesis. The compounds of the invention can be administered before, after or simultaneously with the additional drug substance, whatever the course of administration. The additional pharmaceutical substances include anti-edema steroids, NSAIDs, ras inhibitors, anti-TNF substances, anti-IL-1 substances, antihistamines, PAF-antagonists, COX-1 inhibitors, COX-2 inhibitors, NO synthase inhibitors, Akt / These include but are not limited to PTB inhibitors, IGF-1R inhibitors, PKC inhibitors, PI3 kinase inhibitors, calcineurin inhibitors and immunosuppressive substances. The compounds of the invention and the additional pharmaceutical substance act additively or synergistically. Therefore, administration of such a combination of substances that suppress angiogenesis, increased vascular permeability, and suppress edema formation may result in a greater reduction in the adverse effects of hyperproliferative disorders than administration of either substance alone. . In the treatment of malignant disorders, combinations with antiproliferative or cytotoxic chemotherapy or radiation are included within the scope of the present invention.

  The invention also includes the use of a compound of formula 1, 2 or 3 as a medicament.

  Another aspect of the present invention is the use of Formulas 1, 2 in the manufacture of a medicament for treating increased vascular permeability, angiogenesis-dependent disorders, proliferative diseases and / or immune system disorders in mammals, particularly humans. Or use of a compound of 3 or a salt thereof.

  The present invention also provides a method of treating increased vascular permeability, inappropriate angiogenesis, proliferative diseases and / or disorders of the immune system, comprising the formula 1, 2 or 3 for mammals, particularly humans, in need thereof. Methods are provided that include administration of a therapeutically effective amount.

Enzyme assays The in vitro potency of Formulas 1, 2, or 3 in the inhibition of one or more of the protein kinases described herein or described in the art can be determined by the methods detailed below. .

  The potency of the compound of formula 1, 2 or 3 is the inhibition of phosphorylation of the exogenous substrate (eg, synthetic peptide (Z. Songyang et al., Nature. 373: 536-539)) by the test compound as compared to the control. It can be determined by quantity.

p38 Kinase Assay Materials: Active p38α enzyme is available from Upstate Biotechnology Inc. (UBI) is available for purchase. Anti-phospho MBP specific antibodies are commercially available from UBI and can be labeled with Europium (Eu) cryptate by Cis-Bio International. SAXL (streptavidin-linked XL) is available for Prozyme. Biotin-MBP-peptide (Biot-Ahx-VHFFKNIVTPRTPPPSQGKGAEGQR-OH) can be manufactured by New England Peptide. The HTRF reader RUBYstar is available from BMG Labtech. The kinase assay is performed using the homogeneous time-resolved fluorescence (HTRF) method (Mabile, 1991; Mathis, 1993). The assay mixture is in a buffer containing 20 mM MOPS (pH 7.2), 10 mM MgCl ₂ , 5 mM EGTA, 5 mM β-phosphoglycerol, 1 mM Na ₃ VO ₄ , 0.01% Triton-X-100, 1 mM DTT, Contains 7.8 nM p38α, 0.5 μM biotin-MBP-peptide, 0.1 mM ATP and compound (up to final 5% DMSO). The reaction is performed at room temperature in 96 half-well black plates (Corning). At the indicated times, EDTA (up to 0.1M final) is added to quench the reaction. The product is detected by addition of developer reagents (final 11 ng anti-phospho-MBP-Eu antibody and 0.34 μg SAXL). The plate is incubated overnight at 4 ° C. in the dark and read in the HTRF reader RUBYstar. The IC ₅₀ is calculated using the ratio between the signals at 620 nm and 665 nm at various inhibitor concentrations.
References:
(1) M.M. Mabil, G.M. Mathis, E .; J. et al. P. Jolu, D .; Pouyat, C.I. Dumont, Patent WO92: 13264, 1991
(2) G. Mathis, Clin. Chem. 39 (1993) 1953-1959

Method Kinase Assay: The kinase assay was performed using the homogeneous time-resolved fluorescence (HTRF) method (Mabile et al .; Mathis et al.). IKKα and IKKβ (in-house prepared) assays consisted of buffer (20 mM MOPS pH 7, 10 mM MgCl ₂ , 5 mM EGTA, 5 mM β-phosphoglycerol, 1 mM Na ₃ VO ₄ , 0.01% Triton-X-100, 1 mM DTT, 5 % DMSO) containing 6.7 nM IKKα or 1.7 nM IKKβ, 0.5 μM biotin-IκBα-peptide (Cell Signaling), 0.01 mM ATP and compound. The p38α and CDK2 (UBI) assays contained 7.8 nM p38α or 2.7 nM CDK2 / cyclin A and 0.5 μM biotin-MBP-peptide, 0.1 mM ATP and compounds in KK buffer. The p38β assay contained 0.3 nM p38β and 0.1 μM biotin-MBP-protein (UBI), 0.1 mM ATP and compounds in IKK buffer. The JNK, JNK2, and JNK3 assays were those containing 11.1 nM JNK1, 7.6 nM JNK2, or 2.4 nM JNK3, 1 μM biotin-ATF2-peptide (Cell Signaling), 0.01 mM ATP and compounds in IKK buffer. It was. KDR (in-house prepared) assay is in a buffer containing 50 mM HEPES, pH 7.1, 10 mM MgCl ₂ , 2 mM MnCl ₂ , 2.5 mM DTT, 0.01% BSA, 0.1 mM Na ₃ VO ₄ and 5% DMSO It contained 4.0 nM KDR, 2 μM biotin-FGFR-peptide, 0.1 mM ATP and compound. The JAK1 (in-house prepared) assay is in a buffer containing 50 mM MOPSO, pH 6.5, 10 mM MgCl ₂ , 2 mM MnCl ₂ , 2.5 mM DTT, 0.01% BSA, 0.1 mM Na ₃ VO ₄ and 5% DMSO. It contained 3.6 nM JAK1, 2 μM biotin-FGFR-peptide, 0.001 mM ATP and compound. All assays were performed at room temperature for 60 minutes and stopped by the addition of EDTA. The product was detected by the addition of a color reagent containing a europium labeled phospho-specific antibody and SAXL. The plates were incubated overnight at 4 ° C. in the dark and read in the HTRF reader RUBYstar (BMG).
References:
(3) M.M. Mabil, G.M. Mathis, E .; J. et al. P. Jolu, D .; Pouyat, C.I. Dumont, Patent WO 92/13264, 1991
(4) G. Mathis, Clin. Chem. 39 (1993) 1953-1959

Cell assay THP-1 cells from ATCC (TIB-202) were serum starved and seeded at a density of 2 × 10 ⁵ / well in 100 μL of low serum RPMI medium (0.5% FBS). A 50 μl sample of the compound in the appropriate serial dilution is added to the well. Compound stock and dilutions (in 100% DMSO) are prepared so that the final concentration of DMSO in RPMI medium is 0.5%. Cells and compounds or controls are preincubated for 1 hour in a 37 ° C. incubator.

Cytokine release and P-Hsp27 induction are stimulated by LPS treatment. LPS (Sigma, L-4516) was reduced (reconstituted) in endotoxin-free dIH ₂ O to a concentration of 1 mg / ml, diluted in RPMI medium, and 50 μl / well to a final concentration of 1 μg / ml. To each well (except negative control wells). Plates containing cells, compounds and LPS are incubated at 37 ° C. for 45 minutes. If new THP-1 is thawed, recalibration is necessary at this point.

  For analysis of P-Hsp27 (phosphorylated Hsp27 protein), the plates are vacuum filtered to remove media and compounds. Cells are washed twice with buffer (UBI, Assay Buffer # 1, 43-010) using vacuum filtration. Then 100 μl of cell lysis buffer (Biorad, 171-304011) per well is added, the plate is covered and shaken at 4 ° C. for 20 minutes to lyse the cells. The lysate is transferred directly to a flat bottom 96 well plate for analysis or stored at −20 ° C. until analysis. Lysates are diluted 1: 2 with assay buffer # 1 and analyzed by the Luminex method on a Bio-Plex instrument according to the manufacturer's instructions (UBI, Phospho-HSP27 Beadmates kit, 46-607).

  For analysis of cytokine release, after incubation with LPS, the plate is centrifuged at 1000 rpm for 5 minutes and 100 μl of supernatant medium is transferred directly to a second 96 well plate. The test plate containing the cells is returned to the incubator O / N assayed for toxicity the next day (see below). The supernatant is stored at −20 ° C. until analysis. Supernatant media sample plates are analyzed in standard ELISA format according to manufacturer's instructions (R & D, huTNFα ELISA assay kit). Toxicity analysis is performed after overnight incubation with compounds. Add 50 μl of a 2.5 mg / ml solution of MTT (Sigma, M2128) to the cells. Incubate the plate at 37 ° C. for 3 hours. Then 50 μl of 20% SDS is added to solubilize the formazen dye. Plates are incubated for an additional 3 hours at 37 ° C. and OD570 is measured with a spectrophotometer.

material:
Blood donors are in-house volunteers. The tube used for blood collection is 3.2% buffered sodium citrate from Monoject, Mansfield, MA, catalog number 340486. Dilution plates and assay plates were from Corning, COSTAR catalog numbers 3365 and 3599, respectively. Dimethyl sulfoxide (DMSO) is available from Sigma, St. From Louis, MO, catalog number D2650. RPMI medium 1640 and HEPES buffer solution (1M) are from Invitrogen GIBCO Cell Culture Systems, Carlsbad, CA, catalog numbers 11875 and 15630. Escherichia coli 0127: B8 derived lipopolysaccharide (LPS) was from Sigma, catalog number L4516. Tumor necrosis factor alpha (TNF-α / TNFSF1A) ELISA kit is available from R & D Systems, Inc. Minneapolis, Minn., Catalog number PDTA00C.

Method:
Blood is collected from healthy donors into sodium citrate tubes within 1 hour of the assay. Drugs were prepared in dimethyl sulfoxide (DMSO) and serially diluted (1: 3) with DMSO in a dilution plate to give 8 dilution points for each test compound. Further drug dilution (1: 100) was performed in RPMI medium 1640, 20 mM HEPES. Apply 100 μL / well of diluted drug or control (RPMI medium 1640, 1% DMSO in 20 mM HEPES) and 80 μL of blood into the wells of a 96-well assay plate and pre-mix for 30 minutes in an incubator set at 37 ° C. Incubate. Tumor necrosis factor alpha (TNF-α) is then stimulated for 3.5 hours at 37 ° C. by the addition of lipopolysaccharide (LPS, 50 ng / ml) from Escherichia coli 0127: B8. Centrifuge the plate for 10 minutes at 183 g (1000 rpm in a Beckman / Coulter Allegra 6KR centrifuge). Cell-free supernatant (75 μL / well) is collected and TNF-α is measured with a commercial ELISA kit according to the manufacturer's protocol. The potency of a drug that inhibits TNF-α in vitro is determined as the percent reduction in measured TNF-α in wells containing the drug compared to control wells containing no drug. Results are expressed as IC ₅₀ values.
Reference: Current Protocols in Immunology (2005) 7.18B-7.18B12

LPS-induced TNF production material:
Escherichia coli serotype 0111: Lipopolysaccharide (LPS) from B4 (Sigma, cat # L-4130, lot # 095K4056).

  Phosphate buffered saline pH 7.2 (Gibco).

  PEG200 (Sigma, cat # P3015).

  Methylcellulose (Sigma, cat # M7027).

  Male Lewis rats, 200-300 g (Charles River Laboratories).

  Rat tumor necrosis factor α (TNFα) ELISA kit (R & D Systems cat # RTA00).

Method:
Test compounds are prepared in vehicle (in 5% PEG200, 0.5% methylcellulose) at the desired concentration for administration (1, 3, 10, 30, 100 mg / kg). The compound is pre-administered intraperitoneally (ip) or orally (po) to Lewis rats at 0.002 ml / gram body weight 1-2 hours prior to LPS challenge. Negative controls include rats treated only with vehicle (5% PEG200 in 0.5% methylcellulose). LPS is dissolved in phosphate buffered saline, sonicated and the rats are injected intravenously (iv) with 1 mg / kg at 0.001 ml / gram body weight. One hour after the LPS challenge, blood is drawn from the rat's heart and serum is analyzed by ELISA for TNFα. Also determine the compound concentration in serum.

The average concentration of TNFα in the vehicle treated group is considered the maximum (100%) response. The mean TNFα level in the compound treatment group is expressed as a ratio to the maximum response. The ratio of the compound to maximal TNFα response at various doses or serum concentrations is further analyzed using a 4-parameter curve fit of the log-transformed data (Graphpad Prism 4 software) to obtain ED ₅₀ and EC ₅₀ .

Related references:
Azab A et al. (1998). Life Sci. 63: 323-327.

  Martinez EF et al. (2004) Biochem. Pharma. 68: 1321-1329.

  The entire teachings of all references, including journal articles, patents and published patent applications, are incorporated herein by reference.

  The compounds of the present invention can be prepared using the synthetic scheme illustrated in Scheme 1. Starting materials are commercially available or can be prepared by methods described herein or by methods well known to those skilled in organic chemistry. The variables in the scheme are as defined herein or as defined in the claims.

A method for producing the imidazopyridine compound of the present invention is illustrated in Scheme 1. In Scheme 1, step i, an appropriately substituted α-bromoketone 1 is reacted with an optionally substituted 2-aminoheterocycle 2. These types of cyclization reactions are well established in the literature (see, for example, Spitzer et al., J Med Chem 1988, 31, 1590-1595). This reaction is typically performed in an organic solvent (eg, AC, EtOH, or DMF) below the reflux temperature (eg, 80 ° C.). Product 3 is typically isolated from the reaction mixture as a solid by concentrating the mixture and then after extraction work-up with a suitable organic solvent (e.g. IPA, DCM or EtOAc) Use as product or purify by crystallization or flash silica gel chromatography with trituration in an organic solvent (eg DCM, EtOH or EtOAc). Compound 3 is directly usable, or first, to methods known to those skilled in the art (e.g., Larock, R.C.Comprehensive Organic Transformations: A Guide to Functional Group Preparations, 2 n edition, 1999, Wiley-VCH Publishers, (See New York). In one non-limiting example where R1 = Cl, the alkyl group is introduced by iron-mediated addition of a Grignard reagent using conditions such as those described in Furstner et al., J Am Chem Soc, 2002, 124, 13856-13863. It is possible. In another non-limiting example where R 1 = Cl, alkyl groups are introduced by reaction with alkylidene phosphoranes (eg, Taylor, EC and Martin, SF, J Am Chem Soc, 1974). 96, 8095-8102). Coupling of compound 3 with a substituted pyrimidine such as heterocycle 4 to give compound 5 as shown in step ii (Scheme 1) is often Pd (OAc) ₂ / PPh ₃ or PdCl ₂ ( This is done by palladium-mediated arylation using a catalyst / ligand system such as PPh ₃ ) ₂ (see, eg, Pivsa-Art et al., Bull Chem Soc Japan, 1998, 71, 467-473). This reaction is typically performed using a base (eg, Cs ₂ CO ₃ , CsOAc or KOAc) in a solvent such as DMF or NMP at an elevated temperature (eg, 80-100 ° C.). Oxidation as shown in step iii (Scheme 1) typically involves a solution of 5 in an organic solvent (eg, DCM and / or MeOH) and an oxidant (eg, Oxone® or an aqueous solution of m-CBPA). ) To give product 6 (eg Kennedy, RJ. And Stock, AM J Org Chem, 1960, 25, 1901-1906 or Zanatta et al., Synthesis 2003, (6), 894-898). Replacement of the 6 sulfone leaving group with a primary amine to obtain 7 shown in step iv (Scheme 1) can be accomplished by various methods known to those skilled in the art. For example, compound 6 can be reacted with the desired primary amine in an organic solvent (eg, dioxane, toluene or DMSO) at elevated temperatures in the presence or absence of a bulky organic base (eg, TEA). (See, for example, Clark et al., J Med Chem, 2004, 47, 2724-2727). Compound 7 can then be isolated and purified using standard techniques (eg, crystallization, flash column chromatography or reverse phase liquid chromatography).

Abbreviations ACN Acetonitrile bp Boiling point CsOAc Cesium acetate DCM Dichloromethane (methylene chloride)
DME 1,2-dimethoxyethane DMF N, N-dimethylformamide EtMgBr ethylmagnesium bromide EtOAc ethyl acetate Et ₂ O diethyl ether IPA isopropyl alcohol KOAc potassium acetate MeMgBr methyl magnesium bromide MeOH methyl alcohol NMP N-methylpyrrolidone Pd (OAc) ₂ acetic acid Palladium (II)
PPh ₃ Triphenylphosphine THF Tetrahydrofuran TLC Thin layer chromatography

  Example

  4- {4- [2- (2,4-difluorophenyl) -8-methylimidazo [1,2-a] pyrazin-3-yl] -pyrimidin-2-ylamino} -2-methyl-butane-2- Oar

工程Ａ：８−クロロ−２−（２，４−ジフルオロフェニル）−イミダゾ［ｌ，２−α］ピラジン

Step A: 8-Chloro-2- (2,4-difluorophenyl) -imidazo [l, 2-α] pyrazine

ＡＣＮ（８００ｍＬ）中の２−ブロモ−１−（２，４−ジフルオロフェニル）エタノン（１０７．３１ｇ，４４２．８９ｍｍｏｌ）および３−クロロピラジン−２−アミン（９８．００ｇ，７５６．５ｍｍｏｌ）の混合物を還流温度で約２０時間攪拌した。該反応混合物を約２５℃で冷却した後、得られた固体を集めた。濾液溶媒を真空中で除去して褐色固体を得た。この濾過は、該ＡＣＮの除去に伴う突沸を無効にするために行った。ついで、合わせた固体を水（７５０ｍＬ）に懸濁させ、攪拌しながら２Ｎ ＮａＯＨ（７５０ｍＬ）で塩基性化した。約３０分後、該生成物をＤＣＭ（９×１０００ｍＬ）間に分配させ、不溶性物質から濾過して抽出過程を助けた。該有機抽出物を合わせ、２．５Ｎ ＨＣｌ（４×７５０ｍＬ）と共に攪拌した。最終的に該有機層を２．０Ｎ ＮａＯＨ（５００ｍＬ）および水（２×５００）で洗浄し、ＭｇＳＯ_４で乾燥させ、Ｆｌｏｒｉｓｉｌ（登録商標）パッド（直径３インチ×厚さ３インチ）で濾過して元の物質を除去した。生成物がＴＬＣにより検出されなくなるまで、該Ｆｌｏｒｉｓｉｌ（登録商標）パッドを反復量の溶媒で洗浄した。該有機溶媒を真空中で除去して黄色固体を得た。該固体をＩＰＡ（２００ｍＬ）に約８０℃で約１５分間懸濁させ、ついで約２０℃に冷却した。該固体を集め、氷冷ＩＰＡ（２×４０ｍＬ）で洗浄し、ついで石油エーテル［ｂｐ３０〜６０℃］（３×８０ｍＬ）で洗浄して不純物を除去した。該固体を真空中、約７０℃で一晩乾燥させて、表題化合物を黄色粉末固体として得た（６９．７５ｇ，５７％）：ＬＣ／ＭＳ（表１，方法ａ）Ｒ_ｔ＝２．７０分；ＭＳ ｍ／ｚ：２６６．１（Ｍ＋Ｈ）^＋。

Mixture of 2-bromo-1- (2,4-difluorophenyl) ethanone (107.31 g, 442.89 mmol) and 3-chloropyrazin-2-amine (98.00 g, 756.5 mmol) in ACN (800 mL) Was stirred at reflux temperature for about 20 hours. After cooling the reaction mixture at about 25 ° C., the resulting solid was collected. The filtrate solvent was removed in vacuo to give a brown solid. This filtration was performed to negate bumping associated with the removal of the ACN. The combined solids were then suspended in water (750 mL) and basified with 2N NaOH (750 mL) with stirring. After about 30 minutes, the product was partitioned between DCM (9 × 1000 mL) and filtered from insoluble material to aid the extraction process. The organic extracts were combined and stirred with 2.5N HCl (4 × 750 mL). Finally, the organic layer was washed with 2.0N NaOH (500 mL) and water (2 × 500), dried over MgSO ₄ and filtered through a Florisil® pad (3 inches diameter × 3 inches thick). The original material was removed. The Florisil® pad was washed with repeated amounts of solvent until no product was detected by TLC. The organic solvent was removed in vacuo to give a yellow solid. The solid was suspended in IPA (200 mL) at about 80 ° C. for about 15 minutes and then cooled to about 20 ° C. The solid was collected and washed with ice cold IPA (2 × 40 mL) and then with petroleum ether [bp 30-60 ° C.] (3 × 80 mL) to remove impurities. The solid was dried in vacuo at about 70 ° C. overnight to give the title compound as a yellow powdered solid (69.75 g, 57%): LC / MS (Table 1, Method a) R _t = 2.70. Min; MS m / z: 266.1 (M + H) ⁺ .

  Step B: 2- (2,4-Difluorophenyl) -8-methylimidazo [1,2-α] pyrazine

機械攪拌器を備えた三頚反応フラスコ内に８−クロロ−２−（２，４−ジフルオロフェニル）イミダゾ［１，２−ａ］ピラジン（４０．００ｇ，１５０．６ｍｍｏｌ）、鉄（ＩＩＩ）アセチルアセトナート（２．６６ｇ，７．５３ｍｍｏｌ）、ＴＨＦ（９７０ｍＬ）およびＮＭＰ（８６ｍＬ）を加えた。該フラスコに窒素を充填し、それを約−５〜０℃に冷却した後、Ｅｔ_２Ｏ（１５０ｍＬ）中の３Ｍ ＭｅＭｇＢｒを約２０分かけて滴下した。該添加が完了した後、該反応を約１５分間攪拌し、ついで冷却浴を取り外した。該反応混合物を外界温度に約１時間加温し、ついで一晩攪拌した。該反応液を濃縮し、残渣を水（１０００ｍＬ）およびＥｔＯＡｃ（１０００ｍＬ）と共に約１５分間攪拌した。該混合物をＣｅｌｉｔｅ（登録商標）パッドで濾過して塩を除去した。該Ｃｅｌｉｔｅ（登録商標）パッドをこすり、ＥｔＯＡｃ（３×２５０ｍＬ）と共に攪拌した。塩基性水相を分離し、ＥｔＯＡｃ（３×２５０ｍＬ）で抽出した。有機層を合わせ、水（３×３５０ｍＬ）で洗浄し、ＭｇＳＯ_４で乾燥させ、Ｆｌｏｒｉｓｉｌ（登録商標）パッドで濾過して元の物質を除去した。溶媒を真空中で除去して黄色固体を得、これを沸騰ＭｅＯＨ（１２５ｍＬ）で処理し、約１５℃に冷却し、攪拌しながら石油エーテル［ｂｐ３０〜６０℃］（２５０ｍＬ）で処理した。得られた固体を濾過し、石油エーテル［ｂｐ３０〜６０℃Ｃ］（３×５０ｍＬ）で洗浄し、真空中、７０℃で乾燥させて表題化合物を淡黄色粉末固体（２３．２５ｇ，６４％）として得た：ＬＣ／ＭＳ（表１，方法ａ）Ｒ_ｔ＝２．４２分；ＭＳ ｍ／ｚ：２４６．１（Ｍ＋Ｈ）^＋。

In a three-necked reaction flask equipped with a mechanical stirrer, 8-chloro-2- (2,4-difluorophenyl) imidazo [1,2-a] pyrazine (40.00 g, 150.6 mmol), iron (III) acetyl Acetonate (2.66 g, 7.53 mmol), THF (970 mL) and NMP (86 mL) were added. The flask was filled with nitrogen and cooled to about −5 to 0 ° C., then 3M MeMgBr in Et ₂ O (150 mL) was added dropwise over about 20 minutes. After the addition was complete, the reaction was stirred for about 15 minutes and then the cooling bath was removed. The reaction mixture was warmed to ambient temperature for about 1 hour and then stirred overnight. The reaction was concentrated and the residue was stirred with water (1000 mL) and EtOAc (1000 mL) for about 15 minutes. The mixture was filtered through a Celite® pad to remove salts. The Celite® pad was rubbed and stirred with EtOAc (3 × 250 mL). The basic aqueous phase was separated and extracted with EtOAc (3 × 250 mL). The organic layers were combined, washed with water (3 × 350 mL), dried over MgSO ₄ and filtered through a Florisil® pad to remove the original material. The solvent was removed in vacuo to give a yellow solid which was treated with boiling MeOH (125 mL), cooled to about 15 ° C., and treated with petroleum ether [bp 30-60 ° C.] (250 mL) with stirring. The resulting solid was filtered, washed with petroleum ether [bp 30-60 ° C.] (3 × 50 mL) and dried in vacuo at 70 ° C. to give the title compound as a pale yellow powdered solid (23.25 g, 64%) Obtained as: LC / MS (Table 1, Method a) R _t = 2.42 min; MS m / z: 246.1 (M + H) ⁺ .

  Step C: 2- (2,4-Difluorophenyl) -8-methyl-3- (2-methylsulfanylpyrimidin-4-yl) -imidazo [1,2-α] pyrazine

ＰＰｈ_３（４．２８ｇ，１６．３ｍｍｏｌ）およびＰｄ（ＯＡｃ）_２（１．８３ｇ，８．１６ｍｍｏｌ）の混合物をＤＭＦ（９０ｍＬ）中で攪拌した。該混合物を窒素で脱気し、ついで、それが暗赤色溶液になるまで約１００℃で約１０分間加熱した。該反応を加熱から離し、ついで２−（２，４−ジフルオロフェニル）−８−メチルイミダゾ［１，２−ａ］ピラジン（１０．０ｇ，４０．８ｍｍｏｌ）およびＣｓＯＡｃ（１５．７ｇ，８１．８ｍｍｏｌ）を加えた。該反応を約１００℃の加熱に戻し、ＤＭＦ（４０ｍＬ）中の４−ヨード−２−（メチルチオ）ピリミジン（２０．５６ｇ，８１．６ｍｍｏｌ）の溶液を添加漏斗を介して約４時間にわたって加えた。該添加の終了後、該反応を約１００℃で約１６時間加熱し、ついで外界温度に冷却し、減圧下で濃縮した。得られた固体をＤＣＭ（５００ｍＬ）に溶解し、１Ｎ ＨＣｌ（５×２００ｍＬ）で洗浄し、ついで０．５Ｎ ＮａＯＨ（２００ｍＬ）で洗浄し、Ｃｅｌｉｔｅ（登録商標）で濾過して、生じたエマルションを破壊した。層を分離し、有機層をＭｇＳＯ_４で乾燥させ、濾過し、減圧下で濃縮した。得られた残渣を、ＥｔＯＡｃを使用するシリカゲル栓で濾過し、ついで減圧下で濃縮し、ＤＣＭ（２００ｍＬ）に溶解した。該ＤＣＭ溶液を、ＤＣＭ：ＡＣＮの勾配（１：０で４分間、４：１に下げ、４０分間維持し、２０分間かけて１：１への勾配とし、生成物の溶出が終了するまで維持する）を用いるシリカゲルクロマトグラフィー（３３０ｇカラム）により精製した。全ての生成物含有画分（トリフェニルホスフィンオキシドを含有するものを含む）を合わせ、暗色液中に大量の沈殿物が生じるまで、減圧下で濃縮した。ついでＩＰＡを加え、減圧下で更に濃縮してＤＣＭを除去した。得られた懸濁液を濾過し、ＩＰＡで洗浄し、ついで石油エーテル（ｂ．ｐ．３０〜６０℃）で洗浄し、真空オーブン中で約７０℃で乾燥させて表題化合物（５．７ｇ，３８％）を得た：ＬＣ／ＭＳ（表１，方法ａ）Ｒ_ｔ＝３．３９分；ＭＳ ｍ／ｚ：３７０．３（Ｍ＋Ｈ）^＋。

A mixture of PPh ₃ (4.28 g, 16.3 mmol) and Pd (OAc) ₂ (1.83 g, 8.16 mmol) was stirred in DMF (90 mL). The mixture was degassed with nitrogen and then heated at about 100 ° C. for about 10 minutes until it became a dark red solution. The reaction was removed from heating and then 2- (2,4-difluorophenyl) -8-methylimidazo [1,2-a] pyrazine (10.0 g, 40.8 mmol) and CsOAc (15.7 g, 81.8 mmol). ) Was added. The reaction was returned to about 100 ° C. heating and a solution of 4-iodo-2- (methylthio) pyrimidine (20.56 g, 81.6 mmol) in DMF (40 mL) was added via addition funnel over about 4 hours. . After completion of the addition, the reaction was heated at about 100 ° C. for about 16 hours, then cooled to ambient temperature and concentrated under reduced pressure. The resulting solid was dissolved in DCM (500 mL), washed with 1N HCl (5 × 200 mL), then with 0.5N NaOH (200 mL), filtered through Celite®, and the resulting emulsion was Destroyed. The layers were separated and the organic layer was dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was filtered through a plug of silica gel using EtOAc, then concentrated under reduced pressure and dissolved in DCM (200 mL). The DCM solution was ramped to a DCM: ACN gradient (1: 0 at 4 min, 4: 1, maintained for 40 min, ramped to 1: 1 over 20 min, maintained until product elution was complete Purified by silica gel chromatography (330 g column). All product-containing fractions (including those containing triphenylphosphine oxide) were combined and concentrated under reduced pressure until a large amount of precipitate formed in the dark liquid. IPA was then added and further concentrated under reduced pressure to remove DCM. The resulting suspension was filtered, washed with IPA, then with petroleum ether (bp 30-60 ° C.) and dried in a vacuum oven at about 70 ° C. to give the title compound (5.7 g, 38%): LC / MS (Table 1, Method a) R _t = 3.39 min; MS m / z: 370.3 (M + H) ⁺ .

  Step D: 2- (2,4-Difluorophenyl) -3- (2-methanesulfonylpyrimidin-4-yl) -8-methylimidazo [1,2-α] pyrazine

ＤＣＭ（３７３ｍＬ）およびＭｅＯＨ（３７３ｍＬ）中の２−（２，４−ジフルオロフェニル）−８−メチル−３−（２−メチルスルファニルピリミジン−４−イル）−イミダゾ［１，２−α］ピラジン（１７．２ｇ，４６．６ｍｍｏｌ）の高速攪拌溶液に水（１８７ｍＬ）中のＯｘｏｎｅ（登録商標）（５７．４ｇ，９３．４ｍｍｏｌ）の溶液を外界温度で加えた。約１６時間後、該混合物を水（９００ｍＬ）で希釈し、ＤＣＭ（３×３００ｍＬ）で抽出した。合わせた有機層をブライン（３００ｍＬ）で洗浄し、ＭｇＳＯ_４で乾燥させ、濾過し、減圧下で濃縮した。粗物質を、ＤＣＭ：ＥｔＯＡｃ（１；１で１０分間、２０分かけて０：１の勾配とし、さらに２０分間維持する）の勾配を用いるシリカゲルクロマトグラフィー（３３０ｇカラム）により精製した。生成物に富む画分を熱ＡＣＮから結晶化した。生じた固体を濾過し、追加的なＡＣＮで洗浄し、真空オーブン中、約７０℃で約５時間乾燥させて、１５．９ｇ（８５％）の表題化合物を得た。濾液の更なる精製から追加的な生成物が得られうる：ＬＣ／ＭＳ（表１，方法ｂ）Ｒ_ｔ＝１．７７分；ＭＳ ｍ／ｚ：４０２．１（Ｍ＋Ｈ）^＋。

2- (2,4-Difluorophenyl) -8-methyl-3- (2-methylsulfanylpyrimidin-4-yl) -imidazo [1,2-α] pyrazine in DCM (373 mL) and MeOH (373 mL) To a rapidly stirred solution of 17.2 g, 46.6 mmol) was added a solution of Oxone® (57.4 g, 93.4 mmol) in water (187 mL) at ambient temperature. After about 16 hours, the mixture was diluted with water (900 mL) and extracted with DCM (3 × 300 mL). The combined organic layers were washed with brine (300 mL), dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (330 g column) using a gradient of DCM: EtOAc (1: 1, 10 minutes, 0: 1 gradient over 20 minutes and maintained for an additional 20 minutes). The product rich fraction was crystallized from hot ACN. The resulting solid was filtered, washed with additional ACN, and dried in a vacuum oven at about 70 ° C. for about 5 hours to give 15.9 g (85%) of the title compound. Additional product can be obtained from further purification of the filtrate: LC / MS (Table 1, Method b) R _t = 1.77 min; MS m / z: 402.1 (M + H) ⁺ .

  Step E: 4- {4- [2- (2,4-difluorophenyl) -8-methylimidazo [1,2-a] pyrazin-3-yl] -pyrimidin-2-ylamino} -2-methyl-butane -2-ol

ＡＣＮ（２００ｍＬ）中の２−（２，４−ジフルオロフェニル）−３−（２−メタンスルホニルピリミジン−４−イル）−８−メチル−イミダゾ［１，２−ａ］ピラジン（２０．０ｇ，４９．８ｍｍｏｌ）の混合物に４−アミノ−２−メチルブタン−２−オール（ＷＯ ２００３１０１９６８ Ａ１、１９．５ｇ，１８９ｍｍｏｌ）を加えた。該混合物を約８０℃で一晩加熱した。外界温度に冷却した後、該反応混合物を濾過し、該固体を熱ＡＣＮから連続的に３回、再結晶した。得られた固体を真空中、約７０℃で乾燥させて、表題化合物（１４．８ｇ，７０％）を得た：ＬＣ／ＭＳ（表１１，方法ａ）Ｒ_ｔ＝２．９３分；ＭＳ ｍ／ｚ：４２５．４（Ｍ＋Ｈ）^＋，ｍｐ １７５℃。

2- (2,4-Difluorophenyl) -3- (2-methanesulfonylpyrimidin-4-yl) -8-methyl-imidazo [1,2-a] pyrazine (20.0 g, 49 in ACN (200 mL). .8 mmol) was added to 4-amino-2-methylbutan-2-ol (WO 2003011968 A1, 19.5 g, 189 mmol). The mixture was heated at about 80 ° C. overnight. After cooling to ambient temperature, the reaction mixture was filtered and the solid was recrystallized three times consecutively from hot ACN. The resulting solid was dried in vacuo at about 70 ° C. to give the title compound (14.8 g, 70%): LC / MS (Table 11, Method a) R _t = 2.93 min; MS m / Z: 425.4 (M ⁺ H) ⁺ , mp 175 ° C.

  1- {4- [2- (2,4-difluorophenyl) -8-ethylimidazo [1,2-a] pyrazin-3-yl] -pyrimidin-2-ylamino} -2-methylpropan-2-ol

工程Ａ：８−クロロ−２−（２，４−ジフルオロフェニル）−イミダゾ［１，２−α］ピラジン

Step A: 8-Chloro-2- (2,4-difluorophenyl) -imidazo [1,2-α] pyrazine

ＡＣＮ（８００ｍＬ）中の２−ブロモ−１−（２，４−ジフルオロフェニル）エタノン（１０７．３１ｇ，４４２．８９ｍｍｏｌ）および３−クロロピラジン−２−アミン（９８．００ｇ，７５６．５ｍｍｏｌ）を還流温度で約２０時間攪拌した。該反応混合物を約２５℃に冷却した後、生じた固体を集めた。濾液溶媒を真空中で除去して褐色固体を得た。この濾過は、該ＡＣＮの除去に伴う突沸を無効にするために行った。ついで、合わせた固体を水（７５０ｍＬ）に懸濁させ、攪拌しながら２Ｎ ＮａＯＨ（７５０ｍＬ）で塩基性化した。約３０分後、該生成物をＤＣＭ（９×１０００ｍＬ）間に分配させ、不溶性物質から濾過して抽出過程を助けた。該有機抽出物を合わせ、２．５Ｎ ＨＣｌ（４×７５０ｍＬ）と共に攪拌した。最終的に該有機層を２．０Ｎ ＮａＯＨ（５００ｍＬ）および水（２×５００）で洗浄し、ＭｇＳＯ_４で乾燥させ、Ｆｌｏｒｉｓｉｌ（登録商標）パッド（直径３インチ×厚さ３インチ）で濾過して元の物質を除去した。生成物がＴＬＣにより検出されなくなるまで、該Ｆｌｏｒｉｓｉｌ（登録商標）パッドを反復量の溶媒で洗浄した。該有機溶媒を真空中で除去して黄色固体を得た。該固体をＩＰＡ（２００ｍＬ）に約８０℃で約１５分間懸濁させ、ついで約２０℃に冷却した。該固体を集め、氷冷ＩＰＡ（２×４０ｍＬ）で洗浄し、ついで石油エーテル［ｂｐ３０〜６０℃］（３×８０ｍＬ）で洗浄して不純物を除去した。該固体を真空中、約７０℃で一晩乾燥させて、表題化合物を黄色粉末固体として得た（６９．７５ｇ，５７％）：ＬＣ／ＭＳ（表１，方法ａ）Ｒ_ｔ＝２．７０分；ＭＳ ｍ／ｚ：２６６．１（Ｍ＋Ｈ）^＋。

Reflux 2-bromo-1- (2,4-difluorophenyl) ethanone (107.31 g, 442.89 mmol) and 3-chloropyrazin-2-amine (98.00 g, 756.5 mmol) in ACN (800 mL). Stir at temperature for about 20 hours. After the reaction mixture was cooled to about 25 ° C., the resulting solid was collected. The filtrate solvent was removed in vacuo to give a brown solid. This filtration was performed to negate bumping associated with the removal of the ACN. The combined solids were then suspended in water (750 mL) and basified with 2N NaOH (750 mL) with stirring. After about 30 minutes, the product was partitioned between DCM (9 × 1000 mL) and filtered from insoluble material to aid the extraction process. The organic extracts were combined and stirred with 2.5N HCl (4 × 750 mL). Finally, the organic layer was washed with 2.0N NaOH (500 mL) and water (2 × 500), dried over MgSO ₄ and filtered through a Florisil® pad (3 inches diameter × 3 inches thick). The original material was removed. The Florisil® pad was washed with repeated amounts of solvent until no product was detected by TLC. The organic solvent was removed in vacuo to give a yellow solid. The solid was suspended in IPA (200 mL) at about 80 ° C. for about 15 minutes and then cooled to about 20 ° C. The solid was collected and washed with ice cold IPA (2 × 40 mL) and then with petroleum ether [bp 30-60 ° C.] (3 × 80 mL) to remove impurities. The solid was dried in vacuo at about 70 ° C. overnight to give the title compound as a yellow powdered solid (69.75 g, 57%): LC / MS (Table 1, Method a) R _t = 2.70. Min; MS m / z: 266.1 (M + H) ⁺ .

  Step B: 2- (2,4-Difluorophenyl) -8-ethylimidazo [1,2-α] pyrazine

無水Ｅｔ_２Ｏ（５０ｍＬ）中のブロモエタン（４２．１ｍＬ，５６５ｍｍｏｌ）の溶液をＥｔ_２Ｏ（１５０ｍＬ）中のマグネシウムの削りくず（１３．７ｇ，５６５ｍｍｏｌ）の攪拌懸濁液に約１時間にわたって滴下し、内部温度を約２５〜３０℃に維持した。ついで該反応混合物を外界温度で一晩攪拌した。別のフラスコ内で、８−クロロ−２−（２，４−ジフルオロフェニル）イミダゾ［１，２−ａ］ピラジン（５０．０ｇ，１８８ｍｍｏｌ）、鉄（ＩＩＩ）アセチルアセトナート（３．３２ｇ，９．４１ｍｍｏｌ）、ＮＭＰ（１０７ｍＬ，１１１１ｍＬ）およびＴＨＦ（１０００ｍＬ）の懸濁液を約−５〜０℃で約５分間攪拌した後、Ｅｔ_２Ｏ中のＥｔＭｇＢｒの新たに調製した溶液（前記からのもの）を約１時間にわたって滴下した。該添加が完了した後、該反応を約−５〜０℃で約３０分間攪拌し、ついで冷却浴を取り外し、該反応液を外界温度に一晩加温した。溶媒を真空中で除去し、残渣を水（１０００ｍＬ）およびＥｔＯＡｃ（１０００ｍＬ）で希釈し、ついで約１５分間攪拌し、Ｃｅｌｉｔｅ（登録商標）パッドで濾過した。集めた褐色ペースト（生成物およびＭｇ塩）を該パッドから取り出し、ＥｔＯＡｃ（３００ｍＬ）と共に攪拌した。この手順を２回繰返した。元の水層をこれらの洗液で抽出した。合わせた有機層を水（４×５００ｍＬ）で洗浄し、ＭｇＳＯ_４で乾燥させ、濾過し、減圧下で濃縮して褐色固体を得た。該固体を沸騰ＭｅＯＨ（６０ｍＬ）でトリチュレーションし、ついで約１５〜２０℃に冷却し、該固体を集め、氷冷ＭｅＯＨ（２×２０ｍＬ）で洗浄し、ついで石油エーテル［ｂｐ３０〜６０℃］（３×２５ｍＬ）で洗浄し、乾燥させて暗淡黄褐色粉末状固体（２９．４ｇ，５８％）を得た：ＬＣ／ＭＳ（表１，方法ｂ）Ｒ_ｔ＝２．２０分；ＭＳ ｍ／ｚ：２６０．１（Ｍ＋Ｈ）^＋。

A solution of bromoethane (42.1 mL, 565 mmol) in anhydrous Et ₂ O (50 mL) was added dropwise over about 1 hour to a stirred suspension of magnesium shavings (13.7 g, 565 mmol) in Et ₂ O (150 mL). The internal temperature was maintained at about 25-30 ° C. The reaction mixture was then stirred overnight at ambient temperature. In a separate flask, 8-chloro-2- (2,4-difluorophenyl) imidazo [1,2-a] pyrazine (50.0 g, 188 mmol), iron (III) acetylacetonate (3.32 g, 9 .41 mmol), a suspension of NMP (107 mL, 1111 mL) and THF (1000 mL) was stirred at about −5 to 0 ° C. for about 5 minutes, and then a freshly prepared solution of EtMgBr in Et ₂ O (from Were added dropwise over about 1 hour. After the addition was complete, the reaction was stirred at about −5 to 0 ° C. for about 30 minutes, then the cooling bath was removed and the reaction was allowed to warm to ambient temperature overnight. The solvent was removed in vacuo and the residue was diluted with water (1000 mL) and EtOAc (1000 mL), then stirred for about 15 minutes and filtered through a Celite® pad. The collected brown paste (product and Mg salt) was removed from the pad and stirred with EtOAc (300 mL). This procedure was repeated twice. The original aqueous layer was extracted with these washings. The combined organic layers were washed with water (4 × 500 mL), dried over MgSO ₄ , filtered and concentrated under reduced pressure to give a brown solid. The solid was triturated with boiling MeOH (60 mL), then cooled to about 15-20 ° C., the solid collected, washed with ice-cold MeOH (2 × 20 mL), then petroleum ether [bp 30-60 ° C.] (3 × 25 mL) and dried to give a dark tan powdered solid (29.4 g, 58%): LC / MS (Table 1, Method b) R _t = 2.20 min; MS m / Z: 260.1 (M ⁺ H) ⁺ .

  Step C: 2- (2,4-Difluorophenyl) -8-ethyl-3- (2-methylsulfanylpyrimidin-4-yl) -imidazo [1,2-α] pyrazine

ＰＰｈ_３（６．１２ｇ，２３．３ｍｍｏｌ）およびＰｄ（ＯＡｃ）_２（２．６２ｇ，１１．７ｍｍｏｌ）の混合物をＤＭＦ（１３６ｍＬ）中で攪拌した。該混合物を窒素で脱気し、ついで、それが暗赤色溶液になるまで約１００℃で約１５分間加熱した。該反応を加熱から離し、ついで２−（２，４−ジフルオロフェニル）−８−エチルイミダゾ［１，２−ａ］ピラジン（１５．１ｇ，５８．３ｍｍｏｌ）およびＣｓＯＡｃ（１５．７ｇ，８１．８ｍｍｏｌ）を加えた。該反応を加熱に戻し、ＤＭＦ（６０ｍＬ）中の４−ヨード−２−（メチルチオ）ピリミジン（２９．４ｇ，１１７ｍｍｏｌ）の溶液をシリンジポンプを介して約８時間にわたって加えた。該添加の終了後、該反応を約１５時間加熱し、ついで減圧下で濃縮した。得られた粗物質をＤＣＭ（６００ｍＬ）に溶解し、１Ｎ ＨＣｌ（５×３００ｍＬ）で洗浄し、ついで０．５Ｎ ＮａＯＨ（３００ｍＬ）で洗浄し、生じたエマルションをＣｅｌｉｔｅ（登録商標）で濾過した。該濾液の層を分離し、有機層をブライン（２×３００ｍＬ）で洗浄し、ＭｇＳＯ_４で乾燥させ、濾過し、減圧下で濃縮した。粗製物質を、ＤＣＭ：ＡＣＮの勾配（１：０で４分間、１分間かけて４：１への勾配とし、３５分間維持し、２０分間かけて１：１への勾配とし、さらに５分間維持する）を用いるシリカゲルクロマトグラフィー（３３０ｇカラム）により精製した。生成物に富むカラム画分を合わせ、減圧下で濃縮し、ＩＰＡ（７５ｍＬ）でトリチュレーションし、５分間音波処理し、ついで濾過し、追加的なＩＰＡ（１０ｍＬ）で洗浄し、ついで石油エーテル（ｂ．ｐ．３０〜６０℃）で洗浄し、真空オーブン中で約７０℃で乾燥させて表題化合物を淡褐色固体（１２．３ｇ，５１％）を得た：ＬＣ／ＭＳ（表１，方法ｂ）Ｒ_ｔ＝２．２５分；ＭＳ ｍ／ｚ：３８４．２（Ｍ＋Ｈ）^＋。

A mixture of PPh ₃ (6.12 g, 23.3 mmol) and Pd (OAc) ₂ (2.62 g, 11.7 mmol) was stirred in DMF (136 mL). The mixture was degassed with nitrogen and then heated at about 100 ° C. for about 15 minutes until it became a dark red solution. The reaction was removed from heating and then 2- (2,4-difluorophenyl) -8-ethylimidazo [1,2-a] pyrazine (15.1 g, 58.3 mmol) and CsOAc (15.7 g, 81.8 mmol). ) Was added. The reaction was brought back to heat and a solution of 4-iodo-2- (methylthio) pyrimidine (29.4 g, 117 mmol) in DMF (60 mL) was added via syringe pump over about 8 hours. After the addition was complete, the reaction was heated for about 15 hours and then concentrated under reduced pressure. The resulting crude material was dissolved in DCM (600 mL), washed with 1N HCl (5 × 300 mL), then with 0.5N NaOH (300 mL), and the resulting emulsion was filtered through Celite®. The filtrate layers were separated and the organic layer was washed with brine (2 × 300 mL), dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude material was DCM: ACN gradient (1: 0 at 4 minutes, 4: 1 gradient over 1 minute, maintained for 35 minutes, 1: 1 gradient over 20 minutes, maintained for an additional 5 minutes) Purified by silica gel chromatography (330 g column). The product rich column fractions are combined, concentrated under reduced pressure, triturated with IPA (75 mL), sonicated for 5 minutes, then filtered, washed with additional IPA (10 mL), and then petroleum ether. (B.p. 30-60 ° C.) and dried in a vacuum oven at about 70 ° C. to give the title compound as a light brown solid (12.3 g, 51%): LC / MS (Table 1, method _b) R t = 2.25 min; MS m / z: 384.2 ( M + H) +.

  Step D: 2- (2,4-Difluorophenyl) -8-ethyl-3- (2-methanesulfonylpyrimidin-4-yl) -imidazo [1,2-α] pyrazine

ＤＣＭ（４８６ｍＬ）およびＭｅＯＨ（４８６ｍＬ）中の２−（２，４−ジフルオロフェニル）−８−エチル−３−（２−メチルスルファニルピリミジン−４−イル）−イミダゾ［１，２−α］ピラジン（純度９３％，２３．３ｇ，５６．５ｍｍｏｌ）の高速攪拌溶液に水（２４３ｍＬ）中のＯｘｏｎｅ（登録商標）（７４．７ｇ，１２２ｍｍｏｌ）の溶液を外界温度で加えた。約１７時間後、該混合物を水（１．２Ｌ）で希釈し、ＤＣＭ（３×３００ｍＬ）で抽出した。合わせた有機層をブライン（３００ｍＬ）で洗浄し、ＭｇＳＯ_４で乾燥させ、濾過し、減圧下で濃縮した。粗物質を、ＤＣＭ：ＥｔＯＡｃ（１；１で１０分間、３０分かけて０：１の勾配とし、さらに２０分間維持する）の勾配を用いるシリカゲルクロマトグラフィー（３３０ｇカラム）により精製した。生成物に富む画分を熱ＡＣＮから結晶化した。生じた固体を濾過し、追加的なＡＣＮで洗浄し、真空オーブン中、約７０℃で一晩乾燥させて、９．８５ｇ（４２％）の表題化合物を得た。濾液の更なる精製から追加的な生成物が得られうる：ＬＣ／ＭＳ（表１，方法ｂ）Ｒ_ｔ＝１．８３分；ＭＳ ｍ／ｚ：４１６．１（Ｍ＋Ｈ）^＋。

2- (2,4-Difluorophenyl) -8-ethyl-3- (2-methylsulfanylpyrimidin-4-yl) -imidazo [1,2-α] pyrazine (DCM in DCM (486 mL) and MeOH (486 mL) To a rapidly stirring solution of 93% purity, 23.3 g, 56.5 mmol) was added a solution of Oxone® (74.7 g, 122 mmol) in water (243 mL) at ambient temperature. After about 17 hours, the mixture was diluted with water (1.2 L) and extracted with DCM (3 × 300 mL). The combined organic layers were washed with brine (300 mL), dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (330 g column) using a gradient of DCM: EtOAc (1: 1, 10 min, 0: 1 gradient over 30 min and maintained for an additional 20 min). The product rich fraction was crystallized from hot ACN. The resulting solid was filtered, washed with additional ACN, and dried in a vacuum oven at about 70 ° C. overnight to give 9.85 g (42%) of the title compound. Additional product can be obtained from further purification of the filtrate: LC / MS (Table 1, Method b) R _t = 1.83 min; MS m / z: 416.1 (M + H) ⁺ .

  Step E: 4- {4- [2- (2,4-difluorophenyl) -8-ethylimidazo [1,2-a] pyrazin-3-yl] -pyrimidin-2-ylamino} -2-methylpropane- 2-ol

ＡＣＮ（５０ｍＬ）中の２−（２，４−ジフルオロフェニル）−８−エチル−３−（２−メタンスルホニルピリミジン−４−イル）−イミダゾ［１，２−ａ］ピラジン（２．５３ｇ，６．０６ｍｍｏｌ）および１−アミノ−２−メチルプロパン−２−オール（Ｔｙｇｅｒ，６．３４ｇ，７１．１ｍｍｏｌ）の混合物を約８５℃で一晩加熱した。該反応混合物を減圧下で濃縮し、溶離液としてＥｔＯＡｃを使用するシリカゲルクロマトグラフィーにより精製して、表題化合物（１．９７ｇ，７６％）を得た：ＬＣ／ＭＳ（表１，方法ａ）Ｒ_ｔ＝２．１１分；ＭＳ ｍ／ｚ：４２５．２（Ｍ＋Ｈ）^＋，ｍｐ １６４〜１６５℃。

2- (2,4-Difluorophenyl) -8-ethyl-3- (2-methanesulfonylpyrimidin-4-yl) -imidazo [1,2-a] pyrazine (2.53 g, 6 in ACN (50 mL) .06 mmol) and 1-amino-2-methylpropan-2-ol (Tyger, 6.34 g, 71.1 mmol) were heated at about 85 ° C. overnight. The reaction mixture was concentrated under reduced pressure and purified by silica gel chromatography using EtOAc as eluent to give the title compound (1.97 g, 76%): LC / MS (Table 1, Method a) R _t = 2.11 min; MS m / z: 425.2 (M + H) ^<+> , mp 164-165 [deg.] C.

  1- {4- [8-cyclopropylmethyl-2- (2,4-difluorophenyl) -imidazo [1,2-a] pyrazin-3-yl] -pyrimidin-2-ylamino} -2-methylpropane- 2-ol

工程Ａ：８−クロロ−２−（２，４−ジフルオロフェニル）−イミダゾ［１，２−α］ピラジン

Step A: 8-Chloro-2- (2,4-difluorophenyl) -imidazo [1,2-α] pyrazine

ＡＣＮ（８００ｍＬ）中の２−ブロモ−１−（２，４−ジフルオロフェニル）エタノン（１０７．３１ｇ，４４２．８９ｍｍｏｌ）および３−クロロピラジン−２−アミン（９８．００ｇ，７５６．５ｍｍｏｌ）を還流温度で約２０時間攪拌した。該反応混合物を約２５℃に冷却した後、生じた固体を集めた。濾液溶媒を真空中で除去して褐色固体を得た。この濾過は、該ＡＣＮの除去に伴う突沸を無効にするために行った。ついで、合わせた固体を水（７５０ｍＬ）に懸濁させ、攪拌しながら２Ｎ ＮａＯＨ（７５０ｍＬ）で塩基性化した。約３０分後、該生成物をＤＣＭ（９×１０００ｍＬ）間に分配させ、不溶性物質から濾過して抽出過程を助けた。該有機抽出物を合わせ、２．５Ｎ ＨＣｌ（４×７５０ｍＬ）と共に攪拌した。最終的に該有機層を２．０Ｎ ＮａＯＨ（５００ｍＬ）および水（２×５００）で洗浄し、ＭｇＳＯ_４で乾燥させ、Ｆｌｏｒｉｓｉｌ（登録商標）パッド（直径３インチ×厚さ３インチ）で濾過して元の物質を除去した。生成物がＴＬＣにより検出されなくなるまで、該Ｆｌｏｒｉｓｉｌ（登録商標）パッドを反復量の溶媒で洗浄した。該有機溶媒を真空中で除去して黄色固体を得た。該固体をＩＰＡ（２００ｍＬ）に約８０℃で約１５分間懸濁させ、ついで約２０℃に冷却した。該固体を集め、氷冷ＩＰＡ（２×４０ｍＬ）で洗浄し、ついで石油エーテル［ｂｐ３０〜６０℃］（３×８０ｍＬ）で洗浄して不純物を除去した。該固体を真空中、約７０℃で一晩乾燥させて、表題化合物を黄色粉末固体として得た（６９．７５ｇ，５７％）：ＬＣ／ＭＳ（表１，方法ａ）Ｒ_ｔ＝２．７０分；ＭＳ ｍ／ｚ：２６６．１（Ｍ＋Ｈ）^＋。

Reflux 2-bromo-1- (2,4-difluorophenyl) ethanone (107.31 g, 442.89 mmol) and 3-chloropyrazin-2-amine (98.00 g, 756.5 mmol) in ACN (800 mL). Stir at temperature for about 20 hours. After the reaction mixture was cooled to about 25 ° C., the resulting solid was collected. The filtrate solvent was removed in vacuo to give a brown solid. This filtration was performed to negate bumping associated with the removal of the ACN. The combined solids were then suspended in water (750 mL) and basified with 2N NaOH (750 mL) with stirring. After about 30 minutes, the product was partitioned between DCM (9 × 1000 mL) and filtered from insoluble material to aid the extraction process. The organic extracts were combined and stirred with 2.5N HCl (4 × 750 mL). Finally, the organic layer was washed with 2.0N NaOH (500 mL) and water (2 × 500), dried over MgSO ₄ and filtered through a Florisil® pad (3 inches diameter × 3 inches thick). The original material was removed. The Florisil® pad was washed with repeated amounts of solvent until no product was detected by TLC. The organic solvent was removed in vacuo to give a yellow solid. The solid was suspended in IPA (200 mL) at about 80 ° C. for about 15 minutes and then cooled to about 20 ° C. The solid was collected and washed with ice cold IPA (2 × 40 mL) and then with petroleum ether [bp 30-60 ° C.] (3 × 80 mL) to remove impurities. The solid was dried in vacuo at about 70 ° C. overnight to give the title compound as a yellow powdered solid (69.75 g, 57%): LC / MS (Table 1, Method a) R _t = 2.70. Min; MS m / z: 266.1 (M + H) ⁺ .

  Step B: 8-cyclopropylmethyl-2- (2,4-difluorophenyl) imidazo [1,2-α] pyrazine

約−３０℃〜−４０℃に維持した無水ＤＭＥ（８００ｍＬ）中の（シクロプロピルメチル）トリフェニルホスホニウムブロミド（Ａｌｆａ Ａｅｓａｒ，１６５ｇ，４１４ｍｍｏｌ）の懸濁液にヘキサン中のｎ−ブチルリチウムの２．５Ｍ溶液（１６６ｍＬ，４１４ｍｍｏｌ）を加えた。−３０℃〜−４０℃で約１時間攪拌した後、８−クロロ−２−（２，４−ジフルオロフェニル）イミダゾ［１，２−ａ］ピラジン（５０．０ｇ，１８８ｍｍｏｌ）を加え、ＤＭＥ（１００ｍＬ）で洗浄した。該混合物を約１時間にわたって外界温度に加温し、ついで約８５℃で約２時間加熱し、この時点でＮａ_２ＣＯ_３（２１．９４ｇ，２０７ｍｍｏｌ）を加え、ついで水（２００ｍＬ）を注意深く加え、加熱を約１．５時間継続した。該揮発性溶媒を減圧下で除去し、生じた暗赤色反応混合物にＥｔＯＡｃ（７５０ｍＬ）およびＨ_２Ｏ（７５０ｍＬ）を加え、約１０分間攪拌し、ついで不溶性物質を濾過し、ＥｔＯＡｃ（３×１００ｍＬ）で洗浄した。水相を分離し、ＥｔＯＡｃ（３×１５０ｍＬ）で洗浄した。合わせた有機抽出物を水（３×２５０ｍＬ）で洗浄し、ＭｇＳＯ_４で乾燥させ、濾過し、減圧下で濃縮して暗赤色油／ガムを得た。残渣をエーテル（４×２５０ｍＬ）でトリチュレーションし、ＰＰｈ_３Ｏ不溶性物質が残り、上清液をＦｌｏｒｉｓｉｌ（登録商標）パッドで濾過して、同様にして元の物質を除去した。溶媒を減圧下で除去して暗オレンジ色油を得た。該油を約６０℃でヘプタン（３×２００ｍＬ）でトリチュレーションし、約２５〜３０℃に冷却し、ついで該上清液を該暗色油からデカントし、さらに約２０℃に冷却した。生じた黄色固体を集め、氷冷石油エーテル［ｂｐ３０〜６０℃］（２×１５ｍＬ）で洗浄し、乾燥させて、表題化合物の初期バッチ（７．３ｇ，２９％）を得た。該暗色油残渣をＥｔ_２Ｏ（２００ｍＬ）に溶解し、該ヘプタン濾液と合わせた。この溶液に５Ｎ ＨＣｌ溶液（４００ｍＬ）を加えた。該懸濁液を約１時間攪拌し、該ＨＣｌ塩を濾過して暗褐色固体を得た。この固体をＡＣＮ（２×６０ｍＬ）と共に攪拌し、濾過し、乾燥させて黄色固体を得、これを１Ｎ ＮａＯＨ（２００ｍＬ）、水（２００ｍＬ）と共に約１０分間攪拌した。該遊離塩基生成物をＥｔＯＡｃ（４×１５０ｍＬ）および該塩基性水相間に分配した。合わせた有機抽出物を水（３×２５０ｍｌ）で洗浄し、ＭｇＳＯ_４で乾燥させ、濾過し、減圧下で濃縮して追加的な表題化合物を淡黄色粉末（３０．２ｇ，５６％）として得た：ＬＣ／ＭＳ（表１，方法ｂ）Ｒ_ｔ＝２．２６分；ＭＳ ｍ／ｚ：２８６．１（Ｍ＋Ｈ）^＋。

A suspension of (cyclopropylmethyl) triphenylphosphonium bromide (Alfa Aesar, 165 g, 414 mmol) in anhydrous DME (800 mL) maintained at about −30 ° C. to −40 ° C. 2. A 5M solution (166 mL, 414 mmol) was added. After stirring at −30 ° C. to −40 ° C. for about 1 hour, 8-chloro-2- (2,4-difluorophenyl) imidazo [1,2-a] pyrazine (50.0 g, 188 mmol) was added, and DME ( 100 mL). The mixture was warmed to ambient temperature over about 1 hour, then heated at about 85 ° C. for about 2 hours, at which point Na ₂ CO ₃ (21.94 g, 207 mmol) was added followed by careful addition of water (200 mL). The heating was continued for about 1.5 hours. The volatile solvent was removed under reduced pressure and EtOAc (750 mL) and H ₂ O (750 mL) were added to the resulting dark red reaction mixture and stirred for about 10 minutes, then the insoluble material was filtered and EtOAc (3 × 100 mL) was added. ). The aqueous phase was separated and washed with EtOAc (3 × 150 mL). The combined organic extracts were washed with water (3 × 250 mL), dried over MgSO ₄ , filtered and concentrated under reduced pressure to give a dark red oil / gum. The residue was triturated with ether (4 × 250 mL), leaving PPh ₃ O insoluble material, and the supernatant was filtered through a Florisil® pad to remove the original material in the same manner. The solvent was removed under reduced pressure to give a dark orange oil. The oil was triturated with heptane (3 × 200 mL) at about 60 ° C. and cooled to about 25-30 ° C., then the supernatant was decanted from the dark oil and further cooled to about 20 ° C. The resulting yellow solid was collected, washed with ice-cold petroleum ether [bp 30-60 ° C.] (2 × 15 mL) and dried to give the initial batch of title compound (7.3 g, 29%). The dark oil residue was dissolved in Et ₂ O (200 mL) and combined with the heptane filtrate. To this solution was added 5N HCl solution (400 mL). The suspension was stirred for about 1 hour and the HCl salt was filtered to give a dark brown solid. The solid was stirred with ACN (2 × 60 mL), filtered and dried to give a yellow solid that was stirred with 1N NaOH (200 mL), water (200 mL) for about 10 minutes. The free base product was partitioned between EtOAc (4 × 150 mL) and the basic aqueous phase. The combined organic extracts were washed with water (3 × 250 ml), dried over MgSO ₄ , filtered and concentrated under reduced pressure to give additional title compound as a pale yellow powder (30.2 g, 56%). LC / MS (Table 1, Method b) R _t = 2.26 min; MS m / z: 286.1 (M + H) ⁺ .

  Step C: 8- (cyclopropylmethyl) -2- (2,4-difluorophenyl) -3- (2- (methylthio) pyrimidin-4-yl) imidazo [1,2-a] pyrazine

ゴム隔壁および窒素入口針を備えた５００ｍＬ丸底フラスコ内にＤＭＦ（１７５ｍＬ）中のＰＰｈ_３（８．０９ｇ，３０．８ｍｍｏｌ）およびＰｄ（ＯＡｃ）_２（３．４６ｇ，１５．４ｍｍｏｌ）を充填して黄色懸濁液を得た。該混合物を真空下で排気し、窒素ガスを再充填した（３回行った）。該反応混合物を約１００℃で約１０分間加熱した。ＣｓＯＡｃ（２９．６ｇ，１５４ｍｍｏｌ）および８−（シクロプロピルメチル）−２−（２，４−ジフルオロフェニル）イミダゾ［１，２−ａ］ピラジン（２２ｇ，７７ｍｍｏｌ）をそれぞれ連続的に１回で加えて紫色懸濁液を得た。該混合物を真空下で排気し、窒素ガスを再充填し（３回行った）、ついで約１００℃で約５分間加熱した。ＤＭＦ（７０ｍＬ）中の４−ヨード−２−（メチルチオ）ピリミジン（３８．９ｇ，１５４ｍｍｏｌ）をシリンジポンプを介して約８時間にわたって加えて黒色懸濁液を得た。該反応混合物を約１００℃で約２０時間加熱した。該反応混合物を真空中で濃縮し、ＤＣＭで希釈し、Ｈ_２Ｏ、飽和水性ＮａＣｌおよびＨ_２Ｏ（各２５０ｍＬ）で順次希釈した。有機層をＭｇＳＯ_４で乾燥させ、Ｃｅｌｉｔｅ（登録商標）で濾過し、真空中で濃縮した。粗製物質を、ＤＣＭ：ＥｔＯＡｃの勾配（１：０で６０分間、６０分間かけて１：１への勾配とし、５分間維持する）を用いるシリカゲルクロマトグラフィー（３３０ｇカラム）により精製した。生成物含有カラム画分を合わせ、減圧下で濃縮し、熱ＥｔＯＡｃから結晶化した。生じた針状結晶を濾過により単離し、石油エーテル（ｂ．ｐ．３０〜６０℃）で洗浄し、真空オーブン中で約７０℃で一晩乾燥させて表題化合物（１５．２ｇ，４２％）を得た。濾液の更なる精製から追加的な生成物が得られうる：ＬＣ／ＭＳ（表１，方法ｂ）Ｒ_ｔ＝２．４分；ＭＳ ｍ／ｚ：４１０．２（Ｍ＋Ｈ）^＋。

A 500 mL round bottom flask equipped with a rubber septum and a nitrogen inlet needle was charged with PPh ₃ (8.09 g, 30.8 mmol) and Pd (OAc) ₂ (3.46 g, 15.4 mmol) in DMF (175 mL). To obtain a yellow suspension. The mixture was evacuated under vacuum and refilled with nitrogen gas (performed 3 times). The reaction mixture was heated at about 100 ° C. for about 10 minutes. CsOAc (29.6 g, 154 mmol) and 8- (cyclopropylmethyl) -2- (2,4-difluorophenyl) imidazo [1,2-a] pyrazine (22 g, 77 mmol) were added in succession once each. A purple suspension was obtained. The mixture was evacuated under vacuum, refilled with nitrogen gas (performed 3 times) and then heated at about 100 ° C. for about 5 minutes. 4-iodo-2- (methylthio) pyrimidine (38.9 g, 154 mmol) in DMF (70 mL) was added via syringe pump over about 8 hours to give a black suspension. The reaction mixture was heated at about 100 ° C. for about 20 hours. The reaction mixture was concentrated in vacuo, diluted with DCM, and sequentially diluted with H ₂ O, saturated aqueous NaCl, and H ₂ O (250 mL each). The organic layer was dried over MgSO ₄ , filtered through Celite® and concentrated in vacuo. The crude material was purified by silica gel chromatography (330 g column) using a DCM: EtOAc gradient (1: 0 for 60 minutes, 1: 1 gradient over 60 minutes and maintained for 5 minutes). Product containing column fractions were combined, concentrated under reduced pressure and crystallized from hot EtOAc. The resulting needle crystals were isolated by filtration, washed with petroleum ether (bp 30-60 ° C.) and dried in a vacuum oven at about 70 ° C. overnight to give the title compound (15.2 g, 42%). Got. Additional product can be obtained from further purification of the filtrate: LC / MS (Table 1, Method b) R _t = 2.4 min; MS m / z: 410.2 (M + H) ⁺ .

  Step D: 8- (Cyclopropylmethyl) -2- (2,4-difluorophenyl) -3- (2- (methanesulfonyl) pyrimidin-4-yl) imidazo [1,2-a] pyrazine

ＤＣＭ（２００ｍＬ）およびＭｅＯＨ（２３４ｍＬ）中の８−（シクロプロピルメチル）−２−（２，４−ジフルオロフェニル）−３−（２−（メチルチオ）ピリミジン−４−イル）イミダゾ［１，２−ａ］ピラジン（１５．２ｇ，〜８７ｗｔ％，３２．２ｍｍｏｌ）の高速攪拌溶液に水（１２３ｍＬ）中のＯｘｏｎｅ（登録商標）（３９．６ｇ，６４．５ｍｍｏｌ）の溶液を外界温度で加えた。約１６時間後、該混合物を水（１５０ｍＬ）で希釈し、ＤＣＭ（２００ｍＬ）で抽出した。層を分離した。水層をＤＣＭ（３×１００ｍＬ）で抽出した。合わせた有機層をブライン（３００ｍＬ）で洗浄し、ＭｇＳＯ_４で乾燥させ、濾過し、減圧下で濃縮した。粗物質を、ＤＣＭ：ＥｔＯＡｃ（１；１で２０分間、２０分かけて０：１の勾配とし、さらに２０分間維持する）の勾配を用いるシリカゲルクロマトグラフィーにより精製した。生成物に富む画分を熱ＡＣＮから結晶化した。生じた固体を濾過し、ＡＣＮおよび石油エーテル（ｂ．ｐ．３０〜６０℃）で洗浄し、真空オーブン中、約７０℃で約５時間乾燥させて、１０．０ｇ（７０％）の表題化合物を得た。濾液の更なる精製から追加的な生成物が得られうる：ＬＣ／ＭＳ（表１，方法ｂ）Ｒ_ｔ＝２．０分；ＭＳ ｍ／ｚ：４４２．２（Ｍ＋Ｈ）^＋。

8- (cyclopropylmethyl) -2- (2,4-difluorophenyl) -3- (2- (methylthio) pyrimidin-4-yl) imidazo [1,2- in DCM (200 mL) and MeOH (234 mL) a] To a rapidly stirring solution of pyrazine (15.2 g, ˜87 wt%, 32.2 mmol) was added a solution of Oxone® (39.6 g, 64.5 mmol) in water (123 mL) at ambient temperature. After about 16 hours, the mixture was diluted with water (150 mL) and extracted with DCM (200 mL). The layers were separated. The aqueous layer was extracted with DCM (3 × 100 mL). The combined organic layers were washed with brine (300 mL), dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by silica gel chromatography using a gradient of DCM: EtOAc (1: 1, 20 minutes, 0: 1 gradient over 20 minutes and maintained for an additional 20 minutes). The product rich fraction was crystallized from hot ACN. The resulting solid was filtered, washed with ACN and petroleum ether (bp 30-60 ° C.) and dried in a vacuum oven at about 70 ° C. for about 5 hours to give 10.0 g (70%) of the title compound. Got. Additional product may be obtained from further purification of the filtrate: LC / MS (Table 1, Method b) R _t = 2.0 min; MS m / z: 442.2 (M + H) ⁺ .

  Step E: 1- {4- [8-cyclopropylmethyl-2- (2,4-difluorophenyl) -imidazo [1,2-a] pyrazin-3-yl] -pyrimidin-2-ylamino} -2- Methylpropan-2-ol

ＡＣＮ（１６２ｍＬ）中の８−シクロプロピルメチル−２−（２，４−ジフルオロフェニル）−３−（２−メタンスルホニルピリミジン−４−イル）イミダゾ［１，２−ａ］ピラジン（１０．０ｇ，２２．６ｍｍｏｌ）および１−アミノ−２−メチルプロパン−２−オール（Ｔｙｇｅｒ，６．０６ｇ，６８．０ｍｍｏｌ）の混合物を約８０℃で加熱した。約１６時間後、該反応混合物を減圧下で濃縮し、ＤＣＭ：ＭｅＯＨ（１；０で５分間、５分かけて９：１の勾配とし、さらに２０分間維持する）の勾配を用いるシリカゲルクロマトグラフィー（１２０ｇカラム）により精製した。生成物含有画分を減圧下で濃縮して白色固体を得た。また、生成物および出発物質の混合物（〜３ｇ）を回収し、ＡＣＮ（１６２ｍＬ）中の１−アミノ−２−メチルプロパン−２−オール（Ｔｙｇｅｒ，０．５０５ｇ，５．６６ｍｍｏｌ）で処理した。該反応混合物を約８０℃で約５時間加熱し、ついで減圧下で濃縮して黄色固体を得た。該黄色固体を、ＤＣＭ：ＭｅＯＨ（１；０で１０分間、１０分かけて４：１の勾配とし、さらに１０分間維持する）の勾配を用いるシリカゲルクロマトグラフィー（１２０ｇカラム）により精製した。生成物含有画分を減圧下で濃縮し、ついで、ＭｅＯＨに溶解することにより第１カラムからの白色固体と一緒にした。該溶液を、沈殿物が形成し始めるまで、減圧下で濃縮した。該懸濁液を、均一な固体が全体に形成するまで、音波処理し、ついで減圧下で濃縮し、真空下、約１００℃で乾燥させて、表題化合物を白色固体（９．５ｇ，９３％）を得た：ＬＣ／ＭＳ（表１，方法ｂ）Ｒ_ｔ＝２．１分；ＭＳ ｍ／ｚ：４５１．２（Ｍ＋Ｈ）^＋，ｍｐ １７４〜１７６℃。

8-cyclopropylmethyl-2- (2,4-difluorophenyl) -3- (2-methanesulfonylpyrimidin-4-yl) imidazo [1,2-a] pyrazine (10.0 g, ACN (162 mL). 22.6 mmol) and 1-amino-2-methylpropan-2-ol (Tyger, 6.06 g, 68.0 mmol) were heated at about 80 ° C. After about 16 hours, the reaction mixture is concentrated under reduced pressure and silica gel chromatography using a gradient of DCM: MeOH (1: 0 at 5: 1, 5: 1 gradient over 5 minutes and maintained for an additional 20 minutes). (120 g column). Product containing fractions were concentrated under reduced pressure to give a white solid. Also, the product and starting material mixture (˜3 g) was collected and treated with 1-amino-2-methylpropan-2-ol (Tyger, 0.505 g, 5.66 mmol) in ACN (162 mL). The reaction mixture was heated at about 80 ° C. for about 5 hours and then concentrated under reduced pressure to give a yellow solid. The yellow solid was purified by silica gel chromatography (120 g column) using a gradient of DCM: MeOH (1: 0 at 10: 1 for a 4: 1 gradient over 10 minutes and maintained for an additional 10 minutes). The product containing fractions were concentrated under reduced pressure and then combined with the white solid from the first column by dissolving in MeOH. The solution was concentrated under reduced pressure until a precipitate began to form. The suspension was sonicated until a uniform solid formed entirely, then concentrated under reduced pressure and dried at about 100 ° C. under vacuum to give the title compound as a white solid (9.5 g, 93% ) Obtained: LC / MS (Table 1, Method b) R _t = 2.1 min; MS m / z: 451.2 (M + H) ⁺ , mp 174-176 ° C.

Claims (3)

Formula (1)

And pharmaceutically acceptable salts, prodrugs and pharmaceutically active metabolites thereof. Formula (2)

And pharmaceutically acceptable salts, prodrugs and pharmaceutically active metabolites thereof. Formula (3)

And pharmaceutically acceptable salts, prodrugs and pharmaceutically active metabolites thereof.