Substituted heterocyclic compounds

Abstract

Disclosed are compounds of the following formula (I): # imgabs0 # or a stereoisomer or pharmaceutically acceptable salt thereof wherein all substituents are as defined herein, which are useful for modulating IL-12, IL-23 and / or IFN alpha by acting on Tyk-2 to induce inhibition of signal transduction. The compounds of the invention are useful in the treatment of neurodegenerative diseases or disorders.

Description

[0001] Cross - reference to related applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 382,747, filed on November 8, 2022, the disclosure of which is incorporated herein by reference in its entirety. Technical field

[0003] The present invention relates to compounds for modulating IL - 12, IL - 23, and / or IFNα by acting on Tyk - 2 to cause signal transduction inhibition. Substituted heterocyclic compounds, compositions containing such compounds, and methods of using the same are provided herein. The present invention also relates to pharmaceutical compositions containing at least one compound according to the present invention, which can be used to treat conditions in mammals related to the modulation of IL - 12, IL - 23, and / or IFNα. In particular, the present invention relates to compounds showing utility against neurodegenerative diseases. Background art

[0004] The heterodimeric cytokines interleukin (IL)-12 and IL-23, which share a common p40 subunit, are produced by activated antigen-presenting cells and are crucial in the differentiation and proliferation of Th1 and Th17 cells, two effector T cell lineages that play key roles in autoimmunity. IL-23 consists of the p40 subunit and a unique p19 subunit. IL-23 acts through a heterodimeric receptor composed of IL-23R and IL-12Rβ1 and is essential for the survival and expansion of Th17 cells that produce proinflammatory cytokines such as IL-17A, IL-17F, IL-6, and TNF-α (McGeachy, M.J. et al., "The link between IL-23 and Th17 cell-mediated immune pathologies", Semin. Immunol., 19:372-376 (2007)). These cytokines are key in mediating the pathophysiology of many autoimmune diseases, including rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, and lupus. In addition to the p40 subunit shared with IL-23, IL-12 also contains a p35 subunit and acts through a heterodimeric receptor composed of IL-12Rβ1 and IL-12Rβ2. IL-12 is essential for Th1 cell development and the secretion of IFNγ, a cytokine that plays a key role in immunity by stimulating MHC expression, class switching of B cells to IgG subclasses, and macrophage activation (Gracie, J.A. et al., "Interleukin-12 induces interferon-gamma-dependent switching of IgG alloantibody subclass", Eur. J. Immunol., 26:1217-1221 (1996); Schroder, K. et al., "Interferon-gamma: an overview of signals, mechanisms and functions", J. Leukoc. Biol., 75(2):163-189 (2004)).

[0005] In models of multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease, lupus, and psoriasis, mice deficient in p40, p19, or IL-23R are protected, demonstrating the importance of p40-containing cytokines in autoimmunity (Kyttaris, V.C. et al., "Cutting edge: IL-23 receptor deficiency prevents the development of lupus nephritis in C57BL / 6-lpr / lpr mice", J. Immunol., 184:4605-4609 (2010); Hong, K. et al., "IL-12, independently of IFN-gamma, plays a crucial role in the pathogenesis of a murine psoriasis like skin disorder", J. Immunol., 162:7480-7491 (1999); Hue, S. et al., "Interleukin-23 drives innate and T cell-mediated intestinal inflammation", J. Exp. Med., 203:2473-2483 (2006); Cua, D.J. et al., "Interleukin-23 rather than interleukin-12 is the critical cytokine for autoimmune inflammation of the brain", Nature, 421:744-748 (2003); Murphy, C.A. et al., "Divergent pro- and anti-inflammatory roles for IL-23 and IL-12 in joint autoimmune inflammation", J. Exp. Med., 198:1951-1957 (2003)).

[0006] In human diseases, high expression of p40 and p19 has been measured in psoriatic lesions, and Th17 cells have been identified in active lesions in the brains of MS patients and in the intestinal mucosa of patients with active Crohn's disease (Lee, E. et al., "Increased expression of interleukin 23p19 and p40 in lesional skin of patients with psoriasis vulgaris", J. Exp. Med., 199:125 - 130 (2004); Tzartos, J. S. et al., "Interleukin-17 production in central nervous system infiltrating T cells and glial cells is associated with active disease in multiple sclerosis", Am. J. Pathol., 172:146 - 155 (2008)). The mRNA levels of p19, p40, and p35 are also significantly higher in active SLE patients than in inactive SLE patients (Huang, X. et al., "Dysregulated expression of interleukin-23 and interleukin-12 subunits in systemic lupus erythematosus patients", Mod. Rheumatol., 17:220 - 223 (2007)), and T cells from lupus patients have a predominant Th1 phenotype (Tucci, M. et al., "Overexpression of interleukin-12 and T helper 1 predominance in lupus nephritis", Clin. Exp. Immunol., 154:247 - 254 (2008)).

[0007] In addition, genome-wide association studies have identified numerous loci associated with chronic inflammatory and autoimmune diseases that encode factors acting in the IL-23 and IL-12 pathways. These genes include IL23A, IL12A, IL12B, IL12RB1, IL12RB2, IL23R, JAK2, TYK2, STAT3, and STAT4 (Lees, C.W. et al., "New IBD genetics: common pathways with other diseases", Gut, 60:1739-1753 (2011); Tao, J.H. et al., "Meta-analysis of TYK2 gene polymorphisms association with susceptibility to autoimmune and inflammatory diseases", Mol. Biol. Rep., 38:4663-4672 (2011); Cho, J.H. et al., "Recent insights into the genetics of inflammatory bowel disease", Gastroenterology, 140:1704-1712 (2011)).

[0008] Indeed, anti-p40 therapy that inhibits both IL-12 and IL-23, as well as IL-23-specific anti-p19 therapy, have been shown to be effective in treating autoimmunity in diseases including psoriasis, Crohn's disease, and psoriatic arthritis (Leonardi, C.L. et al., "PHOENIX 1 study investigators. Efficacy and safety of ustekinumab, a human interleukin-12 / 23 monoclonal antibody, in patients with psoriasis: 76-week results from a randomized, double-blind, placebo-controlled trial (PHOENIX 1)", Lancet, 371:1665-1674 (2008); Sandborn, W.J. et al., "Ustekinumab Crohn's Disease Study Group. A randomized trial of Ustekinumab, a human interleukin-12 / 23 monoclonal antibody, in patients with moderate-to-severe Crohn's disease", Gastroenterology, 135:1130-1141 (2008); Gottlieb, A. et al., "Ustekinumab, a human interleukin 12 / 23 monoclonal antibody, for psoriatic arthritis: randomized, double-blind, placebo-controlled, crossover trial", Lancet, 373:633-640 (2009)). Accordingly, it is expected that agents that inhibit the actions of IL-12 and IL-23 would have therapeutic benefits in human autoimmune disorders.

[0009] The type I interferon group (IFN), which includes IFNα members as well as IFNβ, IFNε, IFNκ, and IFNω, acts through the heterodimeric IFNα / β receptor (IFNAR). Type I IFNs have multiple roles in the innate and adaptive immune systems, including activation of cellular and humoral immune responses and enhancement of the expression and release of self-antigens (Hall, J.C. et al., "Type I interferons: crucial participants in disease amplification in autoimmunity", Nat. Rev. Rheumatol., 6:40-49 (2010)).

[0010] In patients with systemic lupus erythematosus (SLE), a potentially fatal autoimmune disease, elevated serum levels of interferon (IFN) α, a type I interferon, or increased expression of type I IFN-regulated genes, a so-called IFNα signature, have been demonstrated in the peripheral blood monocytes and affected organs of most patients (Bennett, L. et al., "Interferon and granulopoiesis signatures in systemic lupus erythematosus blood", J. Exp. Med., 197:711-723 (2003); Peterson, K. S. et al., "Characterization of heterogeneity in the molecular pathogenesis of lupus nephritis from transcriptional profiles of laser-captured glomeruli", J. Clin. Invest., 113:1722-1733 (2004)), and some studies have shown that serum IFNα levels are correlated with disease activity and severity (Bengtsson, A. A. et al., "Activation of type I interferon system in systemic lupus erythematosus correlates with disease activity but not with antiretroviral antibodies", Lupus, 9:664-671 (2000)). The direct role of IFNα in the pathobiology of lupus is confirmed by the observation that administration of IFNα to patients with malignancies or viral diseases can induce a lupus-like syndrome.In addition, deletion of IFNAR in lupus-prone mice provides high protection against autoimmunity, disease severity, and mortality (Santiago-Raber, M. L. et al., "Type-I interferon receptor deficiency reduces lupus-like disease in NZB mice", J. Exp. Med., 197:777-788 (2003)), and genome-wide association studies have identified loci associated with lupus that encode factors acting in the type I interferon pathway, including IRF5, IKBKE, TYK2, and STAT4 (Deng, Y. et al., "Genetic susceptibility to systemic lupus erythematosus in the genomic era", Nat. Rev. Rheumatol., 6:683-692 (2010); Sandling, J. K. et al., "A candidate gene study of the type I interferon pathway implicates IKBKE and IL8 as risk loci for SLE", Eur. J. Hum. Genet., 19:479-484 (2011)). In addition to lupus, there is evidence that aberrant activation of the type I interferon-mediated pathway is important in the pathobiology of other autoimmune diseases such as Sjogren's syndrome and scleroderma ( U. et al., "Activation of the type I interferon system in primary syndrome: a possible etiopathogenic mechanism", Arthritis Rheum., 52:1185-1195 (2005); Kim, D. et al., "Induction of interferon-alpha by scleroderma sera containing autoantibodies to topoisomerase I: association of higher interferon-alpha activity with lung fibrosis", Arthritis Rheum., 58:2163-2173 (2008)). Thus, it is expected that agents that inhibit the type I interferon response will have therapeutic benefits in human autoimmune diseases.

[0011] Tyrosine kinase 2 (TYK2) is a member of the Janus kinase (JAK) family of non-receptor tyrosine kinases and has been shown to play a key role in the signal transduction cascades downstream of the IL-12, IL-23, and type I interferon receptors in both mice (Ishizaki, M. et al., "Involvement of Tyrosine Kinase-2 in Both the IL-12 / Th1 and IL-23 / Th17 Axes In vivo", J. Immunol., 187:181-189 (2011); Prchal-Murphy, M. et al., "TYK2 kinase activity is required for functional type I interferon responses in vivo", PLoS One, 7:e39141 (2012)) and humans (Minegishi, Y. et al., "Human tyrosine kinase 2 deficiency reveals its requisite roles in multiple cytokine signals involved in innate and acquired immunity", Immunity, 25:745-755 (2006)). Tyk2 mediates receptor-induced phosphorylation of members of the STAT family of transcription factors, which is an essential signal leading to STAT protein dimerization and STAT-dependent pro-inflammatory gene transcription. Tyk2-deficient mice are resistant to experimental models of colitis, psoriasis, and multiple sclerosis, demonstrating the importance of Tyk2-mediated signaling in autoimmunity and related disorders (Ishizaki, M. et al., "Involvement of Tyrosine Kinase-2 in Both the IL-12 / Th1 and IL-23 / Th17 Axes In vivo", J. Immunol., 187:181-189 (2011); Oyamada, A. et al., "Tyrosine kinase 2 plays critical roles in the pathogenic CD4 T cell responses for the development of experimental autoimmune encephalomyelitis", J. Immunol., 183:7539-7546 (2009)).

[0012] In humans, individuals expressing an inactivating variant of Tyk2 are protected from multiple sclerosis and potentially other autoimmune disorders (Couturier, N. et al., "Tyrosine kinase 2 variant influences T lymphocyte polarization and multiple sclerosis susceptibility", Brain, 134:693-703 (2011)). Genome-wide association studies have shown other variants of Tyk2 to be associated with autoimmune diseases such as Crohn's disease, psoriasis, systemic lupus erythematosus, and rheumatoid arthritis, further demonstrating the importance of Tyk2 in autoimmunity (Ellinghaus, D. et al., "Combined Analysis of Genome-wide Association Studies for Crohn Disease and Psoriasis Identifies Seven Shared Susceptibility Loci", Am. J. Hum. Genet., 90:636-647 (2012); Graham, D. et al., "Association of polymorphisms across the tyrosine kinase gene, TYK2 in UK SLE families", Rheumatology (Oxford), 46:927-930 (2007); Eyre, S. et al., "High-density genetic mapping identifies new susceptibility loci for rheumatoid arthritis", Nat. Genet., 44:1336-1340 (2012)).

[0013] NPSLE and other neurodegenerative diseases are characterized by extensive involvement of inflammatory processes and CNS-specific cellular inflammation (e.g., CNS-resident microglia and the blood-brain barrier). Symptoms of these disorders can include cognitive dysfunction, a pathological process known to be associated with upregulation of TYK2 and downstream pathway mediators and effectors. Preclinical models of neurodegenerative diseases clearly demonstrate that only CNS-penetrant TYK2i molecules are able to inhibit central-mediated inflammatory and activation processes associated with the etiology and progression of the patient's disease. Therefore, the development of effective, selective, and CNS-penetrant TYK2 inhibitors is necessary for optimal therapeutic benefit.

[0014] TYK2 inhibition can also be used as a monotherapy and in combination with existing standards of care, including immunotherapy, for solid tumors and hematological malignancies.

[0015] In vitro studies of T-cell acute lymphoblastic leukemia (T-ALL) have shown that TYK2 is essential for T-ALL survival, suggesting a potential direct cancer-killing mechanism for TYK2 inhibitors in this indication. Sanda, T. et al., TYK2–STAT1–BCL2 Pathway Dependence in T-cell Acute Lymphoblastic Leukemia. Cancer Discov. 3, 564–577 (2013). Multiple activating mutations of TYK2 have been detected and characterized in T-ALL cell lines. The NPM1-TYK2 gene fusion has also been identified in a subset of cutaneous T-cell lymphoma (CTCL), and TYK2 has been shown to be a transforming oncogenic driver. Kuravi, S. et al., Functional characterization of NPM1–TYK2 fusion oncogene. Npj Precis. Oncol. 6, 3 (2022). Ablation of TYK2 signaling can inhibit this transforming potential.

[0016] There have been descriptions of effective tyrosine kinase 2 (TYK2) inhibitors; however, these compounds tend to be highly polar and have high efflux ratios in standard efflux models. Wrobleski, S.T. et al., Highly selective inhibition of Tyrosine Kinase 2 (TYK2) for the treatment of autoimmune diseases: Discovery of the allosteric inhibitor BMS-986165. J. Med. Chem. 62, 8973-8995 (2019). It is well established that one route to multidrug resistance is increased expression of efflux transporters. Gottesman, M.M. et al., Multidrug Resistance in Cancer: Role of ATP-Dependent Transporters. Nature Rev. Cancer 2, 48-58 (2002), Fletcher, J.I. et al., ABC transporters in cancer: more than just drug efflux pumps. Nature Rev. Cancer 10, 147-156 (2010).

[0017] Thus, compounds with a lower efflux ratio in an in vitro model may have a greater chance of effectively treating certain oncogenic indications.

[0018] Given the disorders that can benefit from treatments involving modulation of cytokines and / or interferons, novel compounds capable of modulating cytokines and / or interferons (such as IL-12, IL-23, and / or IFNα) and methods of using these compounds can provide substantial therapeutic benefits to a variety of patients in need. SUMMARY OF THE INVENTION

[0019] The present invention relates to compounds of formula I (see below), which can be used as modulators of IL-12, IL-23, and / or IFNα by inhibiting Tyk2-mediated signal transduction.

[0020] The present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and at least one compound of the present invention.

[0021] The present invention also provides a method of modulating IL-12, IL-23, and / or IFNα by inhibiting Tyk-2-mediated signal transduction, the method comprising administering to a host in need of such treatment a therapeutically effective amount of at least one compound of the present invention.

[0022] The present invention also provides a method of treating a neurodegenerative disease, the method comprising administering to a host in need of such treatment a therapeutically effective amount of at least one compound of the present invention.

[0023] The present invention also provides the compounds of the present invention for use in therapy.

[0024] As the disclosure continues, these and other features of the invention will be set forth in an expanded form.

[0025] DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

[0026] In a first aspect of the present invention, there is provided a compound of the following formula

[0027]

[0028] or a stereoisomer or a pharmaceutically acceptable salt thereof,

[0029] wherein

[0030] X is -N- or -CH-;

[0031] Y is -N- or -CH-;

[0032] R is

[0033] R 1 is CF3 , C 1-6 alkyl or C 3-6 cycloalkyl;

[0034] R 2 is H or C 1-6 alkyl;

[0035] Z is CHR 1 , CH 2 , CR 1 2 , O, NR 1 or C(O); and

[0036] n is 0, 1, 2 or 3.

[0037] In a second aspect of the present invention, there is provided a compound of the formula

[0038]

[0039] or a stereoisomer or a pharmaceutically acceptable salt thereof,

[0040] wherein

[0041] R is

[0042] R 1 is CF 3 , C 1-6 alkyl or C 3-6 cycloalkyl;

[0043] R 2 is H or C 1-6 alkyl;

[0044] Z is CHR 1 , CH 2 , CR 1 2 , O, NR 1 or C(O); and

[0045] n is 0, 1, 2 or 3.

[0046] In a third aspect of the present invention, there is provided a compound of the formula

[0047]

[0048] or a stereoisomer or a pharmaceutically acceptable salt thereof,

[0049] wherein

[0050] R is

[0051] R 1 is CF3 and C 1-6 alkyl or C 3-6 cycloalkyl;

[0052] Z is CHR 1 or CH 2 or CR 1 2 or O, NR 1 or C(O); and

[0053] n is 0, 1, 2 or 3.

[0054] In a fourth aspect of the present invention, there is provided a compound of the formula

[0055]

[0056] or a stereoisomer or a pharmaceutically acceptable salt thereof,

[0057] wherein

[0058] R is

[0059] R 1 is CF 3 or C 1-6 alkyl or C 3-6 cycloalkyl;

[0060] Z is CHR 1 or CH 2 or CR 1 2 or O, NR 1 or C(O); and

[0061] n is 0, 1, 2 or 3.

[0062] In a fifth aspect of the present invention, there is provided a compound of the formula

[0063]

[0064] or a stereoisomer or a pharmaceutically acceptable salt thereof,

[0065] wherein

[0066] R is

[0067] R 1 is CF 3 or C 1-6 alkyl or C 3-6 cycloalkyl;

[0068] Z is CHR 1 or CH 2 or CR 12 , O, NR 1 or C(O); and

[0069] n is 0, 1, 2 or 3.

[0070] In another aspect, there is provided a compound selected from the exemplary instances within the scope of the first aspect or a pharmaceutically acceptable salt thereof.

[0071] In another aspect, there is provided a compound which is any subset list of compounds selected from within the scope of any one of the above aspects.

[0072] In another aspect, there is provided a compound (IUPAC naming rules) or a pharmaceutically acceptable salt thereof, which is selected from

[0073] 4-{[3-Methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl]amino}-N-(2H3)methyl-6-[2-oxo-3-(propan-2-yl)imidazolidin-1-yl]pyridazine-3-carboxamide;

[0074] 4-{[3-Methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl]amino}-N-(2H3)methyl-6-[2-oxo-3-(propan-2-yl)imidazolidin-1-yl]pyridine-3-carboxamide;

[0075] 4-{[2-Methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl]amino}-N-(2H3)methyl-6-[2-oxo-3-(propan-2-yl)imidazolidin-1-yl]pyridine-3-carboxamide;

[0076] 6-(3-Cyclopropyl-2-oxoimidazolidin-1-yl)-4-{[3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl]amino}-N-(2H3)methylpyridazine-3-carboxamide;

[0077] 6-(3-Cyclopropyl-2-oxoimidazolidin-1-yl)-4-{[2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl]amino}-N-(2H3)methylpyridazine-3-carboxamide;

[0078] 6-(3-Isopropyl-2-oxoimidazolidin-1-yl)-4-((3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d 3 )pyridazine-3-carboxamide;

[0079] 6-(3-Cyclopropyl-2-oxoimidazolidin-1-yl)-4-((3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d3)pyridazine-3-carboxamide;

[0080] 6-(3-Cyclobutyl-2-oxoimidazolidin-1-yl)-4-{[3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl]amino}-N-(2H3)methylpyridazine-3-carboxamide; and

[0081] 4-{[2-Methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl]amino}-N-(2H3)methyl-6-[2-oxo-3-(trifluoromethyl)imidazolidin-1-yl]pyridine-3-carboxamide.

[0082] In another embodiment, there is provided a pharmaceutical composition comprising one or more compounds of formula I and a pharmaceutically acceptable carrier or diluent.

[0083] The present invention also relates to a pharmaceutical composition useful for treating diseases associated with modulating IL-12, IL-23, and / or IFNα by acting on Tyk-2 to cause signal transduction inhibition, which comprises a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or diluent.

[0084] The present invention also relates to a method of treating diseases associated with the modulation of IL-12, IL-23, and / or IFNα, which method comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I.

[0085] The present invention also provides methods and intermediates for preparing the compounds of the present invention.

[0086] The present invention also provides a method for treating proliferative, metabolic, allergic, autoimmune, and inflammatory diseases (or the use of a compound of the present invention in the manufacture of a medicament for treating these diseases), which comprises administering to a host in need of such treatment a therapeutically effective amount of at least one compound of the present invention.

[0087] The present invention also provides a method of treating inflammatory or autoimmune diseases (or the use of a compound of the present invention in the manufacture of a medicament for treating these diseases), which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I.

[0088] The present invention also provides a method for treating diseases (or the use of the compounds of the present invention in the preparation of a medicament for treating these diseases), which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I, wherein the diseases are rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus, inflammatory bowel disease, psoriasis, Crohn's disease, psoriatic arthritis, Sjogren's syndrome, systemic sclerosis, ulcerative colitis, Graves' disease, discoid lupus erythematosus, adult onset Still's disease, systemic onset juvenile idiopathic arthritis, gout, gouty arthritis, type 1 diabetes, insulin-dependent diabetes, sepsis, septic shock, Shigellosis, pancreatitis (acute or chronic), glomerulonephritis, autoimmune myasthenia gravis, diabetes, autoimmune hemolytic anemia, autoimmune neutropenia, thrombocytopenia, atopic dermatitis, myasthenia gravis, pancreatitis (acute or chronic), ankylosing spondylitis, pemphigus vulgaris, Goodpasture's disease, antiphospholipid syndrome, idiopathic thrombocytopenia, ANCA-associated vasculitis, pemphigus, Kawasaki disease, chronic inflammatory demyelinating polyneuropathy (CIDP), dermatomyositis, polymyositis, uveitis, Guillain-Barre syndrome, autoimmune pneumonia, autoimmune thyroiditis, autoimmune inflammatory eye diseases and chronic demyelinating polyneuropathy.

[0089] The present invention also provides a method for treating neurodegenerative diseases (or the use of the compounds of the present invention in the preparation of a medicament for treating said diseases), which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I, wherein the diseases are selected from Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis (RMS and / or progressive MS, including clinically isolated syndrome (CIS)), optic neuritis or neuromyelitis optica.

[0090] The present invention also provides a method for treating rheumatoid arthritis (or the use of the compounds of the present invention in the preparation of a medicament for treating rheumatoid arthritis), which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I.

[0091] In addition, the present invention also provides a method for treating a disorder (or the use of a compound of the present invention in the preparation of a medicament for treating these disorders), which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I, wherein the disorder is selected from acute myeloid leukemia, chronic myeloid leukemia, metastatic melanoma, Kaposi's sarcoma, multiple myeloma, solid tumors, ocular neovascularization and infantile hemangioma, B-cell lymphoma, systemic lupus erythematosus (SLE), rheumatoid arthritis, psoriatic arthritis, polyangiitis, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, allergic rhinitis, multiple sclerosis (MS), transplant rejection, type I diabetes, membranous nephritis, inflammatory bowel disease, autoimmune hemolytic anemia, autoimmune thyroiditis, cold agglutinin disease, Evans syndrome, hemolytic uremic syndrome / thrombotic thrombocytopenic purpura (HUS / TTP), sarcoidosis, Sjogren's syndrome, peripheral neuropathy, pemphigus vulgaris and asthma.

[0092] The present invention also provides a method for treating an IL-12, IL-23 and / or IFNα-mediated disease (or the use of a compound of the present invention in the preparation of a medicament for treating these diseases), which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I.

[0093] The present invention also provides a method for treating an IL-12, IL-23 and / or IFNα-mediated disease (or the use of a compound of the present invention in the preparation of a medicament for treating these diseases), which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I, wherein the IL-12, IL-23 and / or IFNα-mediated disease is a disease regulated by IL-12, IL-23 and / or IFNα.

[0094] The present invention also provides a method for treating a disease, which comprises administering to a patient in need of such treatment a therapeutically effective amount of a combination of a compound of formula I and other therapeutic agents.

[0095] The present invention also provides a compound of the present invention for use in therapy.

[0096] In another embodiment, the compound of formula I is selected from the exemplary compounds or a combination of exemplary compounds or other embodiments herein.

[0097] In another embodiment is a compound having an IC 50 <1000 nM in at least one of the following assays.

[0098] The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present invention encompasses all combinations of the preferred aspects and / or embodiments of the present invention described herein. It should be understood that any and all embodiments of the present invention may be used in combination with any other embodiment or embodiments to describe additional, more preferred embodiments. It should also be understood that each individual element of a preferred embodiment is a preferred embodiment in its own right. In addition, any element of an embodiment is intended to be combined with any and all other elements from any embodiment to describe additional embodiments. Detailed Description

[0099] The following are definitions of terms that may be used in this specification and the appended claims. Unless otherwise indicated, the initial definitions provided herein for a group or term apply to that group or term, either individually or as part of another group, throughout the specification and claims.

[0100] The compounds of the present invention may have one or more asymmetric centers. Unless otherwise indicated, all chiral (enantiomers and diastereomers) and racemic forms of the compounds of the present invention are included in the present invention. Many geometric isomers of alkenes, C=N double bonds, etc. may also be present in the compounds, and all such stable isomers are encompassed by the present invention. The cis and trans geometric isomers of the compounds of the present invention are described and may be separated as a mixture of isomers or as isolated isomeric forms. The compounds of the present invention may be optically active and isolated or in racemic form. It is well known in the art how to prepare optically active forms, for example, by resolution of racemic forms or by synthesis from optically active starting materials. All chiral (enantiomers and diastereomers) and racemic forms of the structure, as well as all geometric isomeric forms, are contemplated unless a specific stereochemistry or isomeric form is specifically indicated.

[0101] When any variable (e.g., R 3 ) appears more than once in any part of a compound or formula, its definition at each occurrence is independent of its definition at every other occurrence. Thus, for example, if a group is shown to be substituted with 0 - 2 R 3 substituents, then said group may optionally be substituted with up to two R 3 substituents, and R 3 is independently selected from the definition of R 3 at each occurrence. In addition, combinations of substituents and / or variables are permitted only if such combinations result in stable compounds.

[0102] When the bond showing the substituent crosses the bond connecting two atoms in the ring, the substituent may be bonded to any atom in the ring. When a substituent is listed without indicating the atom through which the substituent is bonded to the remainder of the compound of a given formula, the substituent may be bonded through any atom in the substituent. Combinations of substituents and / or variables are permitted only if such combinations result in stable compounds.

[0103] In the case where there are nitrogen atoms (such as amines) on the compounds of the present invention, these can be converted into N-oxides by treatment with an oxidizing agent (such as MCPBA and / or hydrogen peroxide) to obtain other compounds of the present invention. Accordingly, all shown and claimed nitrogen atoms are considered to cover the shown nitrogen and its N-oxide (N→O) derivatives.

[0104] According to the convention used in the art, in the structural formulas herein, a bond is used to describe the point of attachment of a moiety or substituent to the core or backbone structure.

[0105] A dash "-" that is not between two letters or symbols is used to indicate the point of attachment of a substituent. For example, -CONH 2 is attached through a carbon atom.

[0106] The term "optionally substituted" with respect to a particular moiety of the compound of formula I (e.g., an optionally substituted heteroaryl) means a moiety having 0, 1, 2 or more substituents. For example, "optionally substituted alkyl" encompasses both "alkyl" and "substituted alkyl" as defined below. Those skilled in the art will understand that for any group containing one or more substituents, such groups are not intended to introduce any substitutions or substitution patterns that are sterically impracticable, synthetically infeasible and / or inherently unstable.

[0107] As used herein, the term "at least one chemical entity" may be used interchangeably with the term "compound".

[0108] As used herein, the term "alkyl" or "alkylene" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the designated number of carbon atoms. For example, "C 1-10 alkyl" (or alkylene) is intended to include C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 and C 10 alkyl. Additionally, for example, "C 1 -C 6"Alkyl" means an alkyl group having 1 to 6 carbon atoms. The alkyl group can be unsubstituted or substituted such that one or more of its hydrogens are replaced by another chemical group. Exemplary alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, tert-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), etc.

[0109] Those skilled in the art will understand that when the designation "CO 2 " is used herein, this refers to the group

[0110] When the term "alkyl" is used in conjunction with another group, such as in "arylalkyl", the linkage more specifically defines at least one substituent that the substituted alkyl will contain. For example, "arylalkyl" refers to a substituted alkyl as defined above, wherein at least one substituent is an aryl group, such as benzyl. Thus, the term aryl(C 0-4 )alkyl includes substituted lower alkyls having at least one aryl substituent, and also includes an aryl group directly bonded to another group, i.e., aryl(C 0 )alkyl. The term "heteroarylalkyl" refers to a substituted alkyl as defined above, wherein at least one substituent is a heteroaryl group.

[0111] The term "alkoxy" refers to an oxygen atom substituted by an alkyl or substituted alkyl group as defined herein. For example, the term "alkoxy" includes groups -O-C 1-6 alkyl, such as methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentyloxy, 2-pentyloxy, isopentyloxy, neopentyloxy, hexyloxy, 2-hexyloxy, 3-hexyloxy, 3-methylpentyloxy, etc. "Lower alkoxy" refers to an alkoxy group having 1 to 4 carbons.

[0112] It should be understood that those skilled in the art can select all groups, including, for example, alkoxy, thioalkyl, and aminoalkyl, to provide stable compounds.

[0113] As used herein, the term "substituted" means that any one or more hydrogens on a specified atom or group are replaced by a selection from a specified group, provided that the normal valence of the specified atom is not exceeded. When the substituent is oxo or a keto group (i.e., =O), 2 hydrogens on the atom are replaced. There are no keto group substituents on aromatic moieties. Unless otherwise stated, substituents should be named into the core structure. For example, it should be understood that when (cycloalkyl)alkyl is listed as a possible substituent, the point of attachment of the substituent to the core structure is in the alkyl portion. As used herein, a ring double bond is a double bond formed between two adjacent ring atoms (e.g., C=C, C=N, or N=N).

[0114] Combinations of substituents and / or variables are permitted only if such combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure means a compound that is sufficiently robust to withstand isolation to a useful degree of purity from a reaction mixture and subsequent formulation into an effective therapeutic agent. Preferably, the compounds of the present invention do not contain N-halo, S(O) 2 H or S(O)H groups.

[0115] The term "cycloalkyl" refers to a cyclic alkyl group, including monocyclic, bicyclic or polycyclic systems. C 3-7 Cycloalkyl is intended to include C 3 、C 4 、C 5 、C 6 and C 7 cycloalkyl. Exemplary cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl, etc. As used herein, "carbocyclic" or "carbocyclic residue" is intended to mean any stable 3-, 4-, 5-, 6- or 7-membered monocyclic or bicyclic or 7-, 8-, 9-, 10-, 11-, 12- or 13-membered bicyclic or tricyclic ring, any of which may be saturated, partially unsaturated, unsaturated or aromatic. Examples of such carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, adamantyl, cyclooctyl, cyclooctenyl, cyclooctadienyl, [3.3.0]bicyclooctane, [4.3.0]bicyclononane, [4.4.0]bicyclodecane, [2.2.2]bicyclooctane, fluorenyl, phenyl, naphthyl, indanyl, adamantyl, anthracenyl and tetrahydronaphthyl (tetralin). As shown above, bridged rings are also included in the definition of carbocycles (e.g., [2.2.2]bicyclooctane). Unless otherwise specified, preferred carbocycles are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and phenyl. When the term "carbocyclic" is used, it is intended to include "aryl". When one or more carbon atoms connect two non-adjacent carbon atoms, a bridged ring is formed. Preferred bridges are one or two carbon atoms. It should be noted that a bridge always converts a monocyclic ring into a bicyclic ring. When a ring is bridged, substituents of the ring may also be present on the bridge.

[0116] The term "aryl" is a monocyclic or bicyclic aromatic hydrocarbon group having 6 to 12 carbon atoms in the ring portion, such as phenyl and naphthyl, each of which may be substituted.

[0117] Thus, in the compounds of formula I, the term "cycloalkyl" includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclooctyl, etc., and the following ring systems:

[0118]

[0119] etc., which may optionally be substituted at any available atom of the ring(s).

[0120] Preferred cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0121] The term "halo" or "halogen" refers to chlorine, bromine, fluorine and iodine.

[0122] The term "haloalkyl" refers to a substituted alkyl group having one or more halo substituents. For example, "haloalkyl" includes mono-, di- and trifluoromethyl.

[0123] The term "haloalkoxy" refers to an alkoxy group having one or more halo substituents. For example, "haloalkoxy" includes OCF 3 .

[0124] Unless otherwise specified, when referring to a specifically named aryl (e.g., phenyl), cycloalkyl (e.g., cyclohexyl), heterocyclic (e.g., pyrrolidinyl, piperidinyl and morpholinyl) or heteroaryl (e.g., tetrazolyl, imidazolyl, pyrazolyl, triazolyl, thiazolyl and furyl) group, the reference is intended to include, where appropriate, rings having 0 to 3, preferably 0 to 2 substituents selected from the above aryl, cycloalkyl, heterocyclic and / or heteroaryl groups.

[0125] The term "carbocyclic" or "carbocyclic ring" refers to a saturated or unsaturated monocyclic or bicyclic ring in which all atoms of all rings are carbon. Thus, the term includes cycloalkyl and aryl rings. Monocyclic carbocycles have 3 to 6 ring atoms, more typically 5 or 6 ring atoms. Bicyclic carbocycles have 7 to 12 ring atoms, for example arranged in a bicyclo[4,5], [5,5], [5,6] or [6,6] system, or 9 or 10 ring atoms arranged in a bicyclo[5,6] or [6,6] system. Examples of monocyclic and bicyclic carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, phenyl and naphthyl. The carbocycle may be substituted, in which case the substituents are selected from those listed above for cycloalkyl and aryl.

[0126] When the term "unsaturated" is used herein to refer to a ring or group, the ring or group may be fully unsaturated or partially unsaturated.

[0127] Throughout the specification, those skilled in the art can select their groups and substituents to provide stable moieties and compounds and compounds useful as pharmaceutically acceptable compounds and / or intermediate compounds for preparing pharmaceutically acceptable compounds.

[0128] The compounds of formula I may exist in free form (non-ionized) or may form salts, which are also within the scope of the present invention. Unless otherwise indicated, reference to a compound of the present invention is to be understood as including reference to the free form and its salts. The term "salt" denotes acidic and / or basic salts formed with inorganic and / or organic acids and bases. In addition, the term "salt" may include zwitterions (inner salts), for example, when a compound of formula I contains a basic moiety (such as an amine or pyridine or imidazole ring) and an acidic moiety (such as a carboxylic acid). Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, such as acceptable metal and amine salts, where the cation does not significantly contribute to the toxicity or biological activity of the salt. However, other salts may be useful, for example, in separation or purification steps that may be employed during preparation and are therefore contemplated within the scope of the present invention. The salts of the compounds of formula I may be formed, for example, by reacting a compound of formula I with a quantity of an acid or base (e.g., an equivalent amount) in a medium such as salt precipitation or in an aqueous medium and then lyophilizing.

[0129] Exemplary acid addition salts include acetates (such as those formed with acetic acid or trihaloacetic acids, e.g., trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecyl sulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides (formed with hydrochloric acid), hydrobromides (formed with hydrogen bromide), hydroiodides, 2-hydroxyethanesulfonates, lactates, maleates (formed with maleic acid), methanesulfonates (formed with methanesulfonic acid), 2-naphthalenesulfonates, nicotinates, nitrates, oxalates, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (such as those formed with sulfuric acid), sulfonates (such as those mentioned herein), tartrates, thiocyanates, toluenesulfonates such as tosylates, undecanoates, and the like.

[0130] Exemplary basic salts include ammonium salts, alkali metal salts such as sodium salts, lithium salts and potassium salts; alkaline earth metal salts such as calcium salts and magnesium salts; barium, zinc and aluminum salts; salts with organic bases (such as organic amines) such as trialkylamines such as triethylamine, procaine, dibenzylamine, N-benzyl-β-phenethylamine, 1-phenylpropylamine, N,N′-dibenzylethylenediamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, dicyclohexylamine or similar pharmaceutically acceptable amines and salts with amino acids such as arginine, lysine and the like. The basic nitrogen-containing groups can be quaternized with reagents such as lower alkyl halides (such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides), dialkyl sulfates (such as dimethyl, diethyl, dibutyl and dipentyl sulfates), long-chain halides (such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (such as benzyl and phenethyl bromides) and the like. Preferred salts include the monohydrochloride, bisulfate, mesylate, phosphate or nitrate.

[0131] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response or other problems or complications and are commensurate with a reasonable benefit / risk ratio.

[0132] As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds in which the parent compound is modified by making its acid or base salts. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic groups such as amines; and alkali metal salts or organic salts of acidic groups such as carboxylic acids. Pharmaceutically acceptable salts include, for example, conventional non-toxic salts or quaternary ammonium salts of the parent compound formed from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid; and salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid and isethionic acid and the like.

[0133] The pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing basic or acidic moieties. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of both; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. A list of suitable salts is given in Remington’s Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, Easton, PA (1990), the disclosure of which is incorporated herein by reference.

[0134] All stereoisomers of the compounds of the present invention are encompassed, whether in the form of mixtures or in pure or substantially pure form. Stereoisomers can include compounds that are optical isomers by virtue of having one or more chiral atoms, and compounds that are optical isomers by virtue of restricted rotation about one or more bonds (atropisomers). The definition of the compounds according to the present invention includes all possible stereoisomers and mixtures thereof. It particularly includes racemic forms and isolated optical isomers having a specific activity. Racemic forms can be resolved by physical methods, such as fractional crystallization, separation or crystallization of diastereomeric derivatives or by chiral column chromatography. The individual optical isomers can be obtained from the racemate by conventional methods, for example, by forming salts with an optically active acid and then crystallizing.

[0135] The present invention is intended to include all isotopes of the atoms present in the compounds of the present invention. Isotopes include those atoms having the same atomic number but different mass numbers. As general examples and not by way of limitation, isotopes of hydrogen include deuterium and tritium. Isotopes of carbon include 13 C and 14 C. The isotopically labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described herein, using appropriate isotopically labeled reagents in place of the unlabeled reagents otherwise employed.

[0136] Prodrugs and solvates of the compounds of the present invention are also contemplated. The term "prodrug" refers to a compound that, when administered to a subject, is chemically transformed by a metabolic or chemical process to produce a compound of formula I and / or its salts and / or solvates. Any compound that is transformed in vivo to provide a bioactive agent (i.e., a compound of formula I) is a prodrug within the scope and spirit of the present invention. For example, a compound containing a carboxyl group can form a physiologically hydrolysable ester, which is used as a prodrug by hydrolyzing in vivo to produce the compound of formula I itself. Such prodrugs are preferably administered orally, since hydrolysis occurs mainly under the influence of digestive enzymes in many cases. In the case where the ester itself is active, or where hydrolysis occurs in the blood, parenteral administration can be used. Examples of physiologically hydrolysable esters of the compounds of formula I include C 1-6 alkylbenzyl, 4-methoxybenzyl, indanyl, phthalyl, methoxymethyl, C 1-6 alkanoyloxy-C 1-6 alkyl, such as acetoxymethyl, pivaloyloxymethyl or propionyloxymethyl, C 1-6 alkoxycarbonyloxy-C 1-6 alkyl, such as methoxycarbonyloxy methyl or ethoxycarbonyloxy methyl, glycyl-oxy methyl, phenylglycyl-oxy methyl, (5-methyl-2-oxo-1,3-dioxol-4-yl)-methyl, and other well-known physiologically hydrolysable esters, for example in the field of penicillins and cephalosporins. Such esters can be prepared by conventional techniques known in the art.

[0137] Various forms of prodrugs are well known in the art and are described in Rautio, J. et al., Nature Review Drug Discovery, 17, 559-587 (2018).

[0138] The compounds of formula I and their salts can exist in their tautomeric forms, in which a hydrogen atom is transposed to other parts of the molecule and the chemical bonds between the atoms of the molecule are thus rearranged. It should be understood that all tautomeric forms, as long as they may exist, are included in the present invention. Additionally, the compounds of the present invention can have trans- and cis-isomers.

[0139] It should also be understood that solvates (e.g., hydrates) of the compounds of formula I are also within the scope of the present invention. Methods of solvation are generally known in the art.

[0140] Practical

[0141] The compounds of the present invention modulate IL-23-stimulated and IFNα-stimulated cellular functions, including gene transcription. Other types of cellular functions that can be modulated by the compounds of the present invention include, but are not limited to, IL-12-stimulated responses.

[0142] Thus, the compounds of formula I act on Tyk2 to mediate signal transduction and can be used to treat conditions related to modulating the functions of IL-23 and / or IFNα, particularly selectively inhibiting the functions of IL-23, IL-12, and / or IFNα. Such conditions include IL-23-, IL-12-, or IFNα-related diseases, in which the pathogenic mechanism is mediated by these cytokines, subsequently activating the Tyk2 pathway, and subsequently a pro-inflammatory response may occur in the peripheral and / or central compartments.

[0143] As used herein, the terms "treating" or "treatment" cover treating a disease state in a mammal, particularly a human, and include: (a) preventing or delaying the occurrence of the disease state in a mammal, particularly when the mammal is predisposed to the disease state but has not been diagnosed with the disease state; (b) inhibiting the disease state, i.e., preventing or slowing its development; and / or (c) achieving complete or partial alleviation of symptoms or the disease state, and / or alleviating, improving, reducing, or curing the disease or disorder and / or its symptoms.

[0144] In view of their activity as regulators of IL-23-, IL-12-, and / or IFNα-stimulated cellular responses, the compounds of formula I can be used to treat IL-23-, IL-12-, and / or IFNα-related diseases, which include but are not limited to inflammatory diseases such as Crohn's disease, ulcerative colitis, asthma, graft-versus-host disease, allograft rejection, chronic obstructive pulmonary disease; autoimmune diseases such as Graves' disease, rheumatoid arthritis, systemic lupus erythematosus (SLE), cutaneous lupus, lupus nephritis, discoid lupus erythematosus, neuropsychiatric SLE, psoriasis; autoinflammatory diseases including CAPS, TRAPS, FMF, adult-onset Still's disease, systemic-onset juvenile idiopathic arthritis, gout, gouty arthritis; metabolic diseases including type 1 diabetes, type 2 diabetes, atherosclerosis, myocardial infarction; destructive bone diseases such as bone resorption diseases, osteoarthritis, osteoporosis, bone diseases associated with multiple myeloma; proliferative diseases such as acute myeloid leukemia, chronic myeloid leukemia; angiogenic disorders, e.g., angiogenic disorders including solid tumors, ocular neovascularization, and infantile hemangioma; infectious diseases such as sepsis, septic shock, and shigellosis; neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, ALS, multiple sclerosis (RMS and / or progressive MS, including clinically isolated syndrome (CIS)), optic neuritis, neuromyelitis optica, cerebral ischemia or neurodegenerative diseases caused by traumatic injury, tumors, and viral diseases such as metastatic melanoma, Kaposi's sarcoma, multiple myeloma, as well as HIV infection and CMV retinitis, AIDS.

[0145] More specifically, specific conditions or diseases treatable with the compounds of the present invention include, but are not limited to, pancreatitis (acute or chronic), asthma, allergy, adult respiratory distress syndrome, chronic obstructive pulmonary disease, glomerulonephritis, rheumatoid arthritis, systemic lupus erythematosus, cutaneous lupus, lupus nephritis, discoid lupus erythematosus, scleroderma, chronic thyroiditis, Graves' disease, autoimmune gastritis, diabetes, autoimmune hemolytic anemia, autoimmune neutropenia, thrombocytopenia, atopic dermatitis, chronic active hepatitis, myasthenia gravis, multiple sclerosis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, psoriasis, graft-versus-host disease, endotoxin-induced inflammatory response, tuberculosis, atherosclerosis, muscle degeneration, cachexia, psoriatic arthritis, Reiter's syndrome, gout, traumatic arthritis, rubella arthritis, acute synovitis, pancreatic beta cell disease; diseases characterized by massive neutrophil infiltration; rheumatoid spondylitis, gouty arthritis and other arthritic conditions, cerebral malaria, chronic pulmonary inflammatory diseases, silicosis, pulmonary sarcoidosis, bone resorption diseases, allograft rejection, infection-induced fever and myalgia, cachexia secondary to infection, keloid formation, scar tissue formation, ulcerative colitis, fever, influenza, osteoporosis, osteoarthritis, acute myelogenous leukemia, chronic myelogenous leukemia, metastatic melanoma, Kaposi's sarcoma, multiple myeloma, sepsis, septic shock and Shigellosis; Alzheimer's disease, Parkinson's disease, multiple sclerosis (RMS and / or progressive MS, including clinically isolated syndrome (CIS)), optic neuritis, neuromyelitis optica, cerebral ischemia or neurodegenerative diseases caused by traumatic injury; angiogenesis disorders, including solid tumors, ocular neovascularization and infantile hemangioma; viral diseases, including acute hepatitis infections (including hepatitis A, hepatitis B and hepatitis C), HIV infection and CMV retinitis, AIDS, ARC or malignancies and herpes; stroke, myocardial ischemia, ischemia in stroke heart attack, organ hypoxia, angiogenesis, cardiac and renal reperfusion injury, thrombosis, cardiac hypertrophy, thrombin-induced platelet aggregation, endotoxemia and / or toxic shock syndrome, conditions associated with prostaglandin endoperoxide synthase-2 and pemphigus vulgaris. Preferred methods of treatment are those in which the condition is selected from Alzheimer's disease, Parkinson's disease, ALS, multiple sclerosis (RMS and / or progressive MS, including clinically isolated syndrome (CIS)), optic neuritis or neuromyelitis optica.

[0146] When the terms "IL-23-, IL-12- and / or IFNα-related conditions" or "IL-23-, IL-12- and / or IFNα-related diseases or disorders" are used herein, each is intended to encompass all of the conditions identified above as if recited in detail, as well as any other conditions affected by IL-23, IL-12 and / or IFNα.

[0147] Accordingly, the present invention provides methods for treating such conditions, which comprise administering to a subject in need a therapeutically effective amount of at least one compound of formula I or a salt thereof. "Therapeutically effective amount" is intended to include the amount of the compounds of the present invention that are effective to inhibit IL-23, IL-12, and / or IFNα function and / or treat the disease when administered alone or in combination.

[0148] The methods for treating IL-23-, IL-12-, and / or IFNα-related conditions may comprise administering the compound of formula I alone or in combination with each other and / or with other suitable therapeutic agents useful for treating such conditions. Accordingly, "therapeutically effective amount" is also intended to include the amount of a combination of the claimed compounds that is effective to inhibit IL-23, IL-12, and / or IFNα function and / or treat diseases related to IL-23, IL-12, and / or IFNα.

[0149] Exemplary of such other therapeutic agents include corticosteroids, rolipram, calcipotriene, cytokine inhibitory anti-inflammatory drugs (CSAIDs), interleukin-10, glucocorticoids, salicylates, nitric oxide, and other immunosuppressants; nuclear translocation inhibitors such as deoxyspergualin (DSG); non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen, celecoxib, and rofecoxib; steroids such as prednisone or dexamethasone; antiviral agents such as abacavir; antiproliferative agents such as methotrexate, leflunomide, FK506 (tacrolimus, ); antimalarial drugs such as hydroxychloroquine; cytotoxic drugs such as azathioprine and cyclophosphamide; TNF-α inhibitors such as tenidap, anti-TNF antibodies or soluble TNF receptors, and rapamycin (sirolimus or ), or derivatives thereof.

[0150] When used in combination with the compounds of the present invention, the above other therapeutic agents may be used, for example, in the amounts indicated in the Physician's Desk Reference (PDR) or as otherwise determined by one of ordinary skill in the art. In the methods of the present invention, such other therapeutic agents may be administered before, simultaneously with, or after the administration of the compounds of the present invention. The present invention also provides pharmaceutical compositions capable of treating IL-23-, IL-12-, or IFNα-related conditions by inhibiting Tyk2-mediated signal transduction, including the IL-23-, IL-12-, and / or IFNα-mediated diseases as described above.

[0151] The compositions of the present invention may contain other therapeutic agents as described above and may be formulated, for example, by using conventional solid or liquid vehicles or diluents and pharmaceutical additives of the type suitable for the desired mode of administration (e.g., excipients, binders, preservatives, stabilizers, flavoring agents, etc.) according to techniques well known in the art of pharmaceutical formulations.

[0152] Accordingly, the present invention also includes a composition comprising one or more compounds of Formula I and a pharmaceutically acceptable carrier.

[0153] "Pharmaceutically acceptable carrier" refers to a medium generally accepted in the art for delivering a bioactive agent to an animal, particularly a mammal. Pharmaceutically acceptable carriers are formulated based on a number of factors within the purview of one of ordinary skill in the art. These include, but are not limited to, the type and nature of the active agent being formulated; the subject to which the reagent-containing composition will be administered; the intended route of administration of the composition; and the targeted therapeutic indication. Pharmaceutically acceptable carriers include aqueous and non-aqueous liquid media, as well as a variety of solid and semi-solid dosage forms. In addition to the active agent, such carriers may include a number of different ingredients and additives, which are included in the formulation for various reasons, such as stabilization of the active agent, binders, etc., which are well known to one of ordinary skill in the art. Descriptions of suitable pharmaceutically acceptable carriers and the factors involved in their selection can be found in a variety of readily available sources, such as Remington’s Pharmaceutical Sciences, 17th Edition (1985), which is incorporated herein by reference in its entirety.

[0154] The compounds of Formula I can be administered by any means suitable for the condition to be treated, which may depend on the need for site-specific therapy or the amount of drug to be delivered. Topical administration is generally preferred for skin-related diseases, and systemic therapy is preferred for cancerous or pre-cancerous conditions, although other delivery modes are also contemplated. For example, the compounds can be delivered orally, such as in the form of tablets, capsules, granules, powders or liquid preparations (including syrups); topically, such as in the form of solutions, suspensions, gels or ointments; sublingually; orally; parenterally, such as by subcutaneous, intravenous, intramuscular or intrasternal injection or infusion techniques (e.g., as a sterile injectable aqueous or non-aqueous solution or suspension); nasally, such as by inhalation spray; topically, such as in the form of creams or ointments; rectally, such as in the form of suppositories; or in liposomes. Dosage unit formulations containing non-toxic, pharmaceutically acceptable vehicles or diluents can be administered. The compounds can be administered in a form suitable for immediate release or extended release. Immediate release or extended release can be achieved with suitable pharmaceutical compositions, or particularly in the case of extended release, with devices such as subcutaneous implants or osmotic pumps.

[0155] Exemplary compositions for topical administration include topical carriers such as (mineral oil gelled with polyethylene).

[0156] Exemplary compositions for oral administration include suspensions which may contain, for example, microcrystalline cellulose as a filler, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweeteners or flavoring agents such as those known in the art; and immediate-release tablets which may contain, for example, microcrystalline cellulose, dibasic calcium phosphate, starch, magnesium stearate, and / or lactose and / or other excipients, binders, extenders, disintegrants, diluents, and lubricants such as those known in the art. The compounds of the invention may also be orally delivered by sublingual and / or buccal administration, for example, with molded, compressed, or lyophilized tablets. Exemplary compositions may include instant diluents such as mannitol, lactose, sucrose, and / or cyclodextrin. High molecular weight excipients such as cellulose or polyethylene glycol (PEG); excipients that assist in mucoadhesion such as hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), sodium carboxymethylcellulose (SCMC), and / or maleic anhydride copolymers (e.g., ); and controlled release agents such as polyacrylic acid copolymers (e.g., CARBOPOL ). Lubricants, glidants, flavoring agents, coloring agents, and stabilizers may also be added to facilitate manufacture and use.

[0157] Exemplary compositions for nasal aerosol or inhalation administration include solutions which may contain, for example, benzyl alcohol or other suitable preservatives, absorption promoters that enhance absorption and / or bioavailability, and / or other solubilizing or dispersing agents such as those known in the art.

[0158] Exemplary compositions for parenteral administration include injectable solutions or suspensions which may contain, for example, suitable non-toxic parenterally acceptable diluents or solvents such as mannitol, 1,3-butanediol, water, Ringer's solution, isotonic sodium chloride solution, or other suitable dispersing or wetting agents and suspending agents including synthetic monoglycerides or diglycerides of glycerol, and fatty acids including oleic acid.

[0159] Exemplary compositions for rectal administration include suppositories which may contain, for example, suitable non-irritating excipients such as cocoa butter, synthetic glycerides, or polyethylene glycol which are solid at room temperature but liquefy and / or dissolve in the rectal cavity to release the drug.

[0160] The therapeutically effective amount of the compounds of the present invention can be determined by those of ordinary skill in the art and includes exemplary dosages for mammals of about 0.05 to 1000 mg / kg per day; 1 - 1000 mg / kg; 1 - 50 mg / kg; 5 - 250 mg / kg; 250 - 1000 mg / kg body weight of the active compound, which can be administered as a single dose or in separate divided doses, for example, 1 to 4 times a day. It should be understood that the specific dosage levels and dosage frequencies for any particular subject may vary and will depend on a variety of factors, including the activity of the particular compound employed, the metabolic stability and duration of action of the compound, the species, age, body weight, general health, sex and diet of the subject, the mode and time of administration, the rate of excretion, drug combinations and the severity of the particular disorder. Preferred subjects for treatment include animals, most preferably mammalian species such as humans and domestic animals such as dogs, cats, horses, etc. Thus, when the term "patient" is used herein, the term is intended to include all subjects affected by the regulation of functions mediated by IL-23, IL-12 and / or IFNα, most preferably mammalian species.

[0161] Preparation method

[0162] The compounds of the present invention can be prepared in a variety of ways well known to those skilled in the art of organic synthesis. The compounds of the present invention can be synthesized using the following methods as well as synthetic methods known in the field of synthetic organic chemistry or variations thereof understood by those skilled in the art. Preferred methods include, but are not limited to, those described below. All references cited herein are incorporated herein by reference in their entirety.

[0163] The compounds of the present invention can be prepared using the reactions and techniques described in this section. The reactions are carried out in solvents suitable for the reagents and materials used and are suitable for the transformations effected. In addition, in the description of the synthetic methods described below, it should be understood that all proposed reaction conditions, including the choice of solvent, reaction atmosphere, reaction temperature, duration of the experiment, and work-up procedures, are selected as standard conditions for the reaction, which should be readily recognized by those skilled in the art. Those skilled in the art of organic synthesis will understand that the functional groups present on the various parts of the molecule must be compatible with the proposed reagents and reactions. Such limitations on substituents compatible with the reaction conditions will be apparent to those skilled in the art, and alternative methods must therefore be used. This sometimes requires judgment to change the order of synthetic steps or to choose one particular process scheme over another to obtain the desired compounds of the present invention. It should also be recognized that another major consideration in planning any synthetic route in the art is the wise choice of protecting groups for protecting the reactive functional groups present in the compounds described in the present invention. Greene and Wuts (Protective Groups In Organic Synthesis, Third Edition, Wiley and Sons, 1999) is an authoritative description of many alternatives for the trained practitioner.

[0164] The key intermediate shown in Figure 1 can be assembled in a variety of ways known to those skilled in the art of synthetic organic chemistry using the building blocks shown in Figure 1 or others to give Compound I.

[0165] Figure 1

[0166]

[0167] Scheme 1

[0168] Scheme 1 shows how those skilled in the art can treat Compound Ib with a strong base, particularly n-BuLi / TMEDA, and then quench with a trialkyl borate, particularly triisopropyl borate, to give Compound Ic. The resulting Compound Ic can be treated with Compound Ia under palladium-catalyzed conditions, particularly under Suzuki conditions, to give Compound Id. Id can be reacted with many reagents known to those skilled in the art, particularly 2,4-dimethoxybenzylamine, to give a protected amine, such as Compound Ie. These protected amines can be deprotected using appropriate methods known to those skilled in the art, particularly in the case of Ie, trifluoroacetic acid, to give the key intermediate IIa.

[0169]

[0170] Scheme 2

[0171] Using many conditions known to those skilled in the art, especially ferric chloride and ammonium chloride in ethanol and water at elevated temperatures, the commercially available compound If can be reduced to the corresponding amine Ig. Compound Ig can be treated with bis(pinacolato)diboron under palladium-catalyzed conditions to obtain the pinacol borate 1g. Other reagents known to those skilled in the art can replace the shown system, such as tin reagents and other boron-based reagents, to obtain compounds analogous to compound Ig in utility. Ig can be coupled with Ia under various palladium-catalyzed conditions known to those skilled in the art to obtain the key compound IIa.

[0172]

[0173] Scheme 3

[0174] Scheme 3 shows how a person skilled in the art of organic synthesis couples compound III (see Moslin et al., J. Med. Chem 2019, 62, 8953 - 8972 or US 9,505,748) and a compound of general formula II to provide an intermediate of general formula IIIa. The reaction involves mixing the two reagents in a suitable aprotic solvent (especially THF or 2-methyl-THF), depending on the circumstances, between 0 °C and 50 °C, and adding a suitable base, especially lithium hexamethyldisilazide, sodium hexamethyldisilazide, potassium hexamethyldisilazide or sodium hydride.

[0175]

[0176] Scheme 4

[0177] Compounds of general formula IV are commercially available or can be prepared by a person skilled in the art of organic synthesis by the method described in Scheme 4 (or by other known methods). 2-Chloroethyl isocyanate can be reacted with a primary amine and then treated with various bases in various aprotic solvents, especially sodium hydride in tetrahydrofuran, to obtain a compound of general formula IV. Additionally, in cases where the primary amine is not available or does not react, such as when R = CF 3 in the case of, the cyclic urea IVa can be monoprotected with a labile group, especially 4-methoxybenzyl, to obtain compound IVb. In the presence of silver triflate, cesium fluoride, 2-fluoropyridine and , IVb is treated with trimethylsilyl trifluoromethane in dichloromethane and chlorobenzene to obtain compound IVc. IVc can then be deprotected under many conditions known to those skilled in the art (especially using trifluoroacetic acid at elevated temperatures) to obtain compound IVd.

[0178]

[0179] Scheme 5

[0180] Scheme 5 shows how a person skilled in the art of organic synthesis couples a compound of general formula III to a suitable substrate IV to prepare a compound of general formula I in one step. This one-step method involves the coupling of a compound of general formula III with a urea of general formula IV under transition metal catalysis conditions. In particular, the favorable conditions for this reaction include the use of Pd 2 (dba) 3 as the catalyst, 1,1′-bis(dicyclohexylphosphino)ferrocene as the ligand, and Cs 2 CO 3 as the base at an elevated temperature, in particular 110 °C, in a Buchwald-type coupling in 1,4-dioxane as the solvent. This catalyst / ligand / base system can be varied in a manner known to a person skilled in the art of organic synthesis.

[0181]

[0182] Specify the method (LCMS, etc.) at the beginning, for example:

[0183] LCMS - Method A:

[0184] A linear gradient of 2% to 40% solvent B in 4 minutes, held at 100% B for 0.6 minutes, and

[0185] then gradient to 20% B for 0.1 minute and held at 20% B for 0.3 minutes.

[0186] Solvent A: 5 mM ammonium formate pH 3.3: ACN (98:02)

[0187] Solvent: B: ACN: buffer (98:02)

[0188] Flow rate: 1.0 ml / min

[0189] Column: Kinetex XB - C18 (75x 3.0) mm, 2.6 μm

[0190] UV visualization at 220 nanometers (“nm”).

[0191] LCMS - Method B:

[0192] A linear gradient of 20% to 100% solvent B in 4 minutes, held at 100% B for 0.6 minutes, and

[0193] then gradient to 20% B for 0.1 minute and held at 20% B for 0.3 minutes.

[0194] Solvent A: 5 mM ammonium formate pH 3.3: ACN (98:02)

[0195] Solvent: B: ACN: buffer (98:02)

[0196] Flow rate: 1.0 ml / min

[0197] Column: Kinetex XB-C18 (75x 3.0) mm, 2.6 μm

[0198] Ultraviolet (“UV”) visualization at 220 nanometers (“nm”).

[0199] LCMS-Method C:

[0200] Linear gradient of 5% to 95% solvent B in 2.5 minutes, held at 95% B for 1.5 minutes, and

[0201] then a 0.5-minute gradient to 5% B and held at 5% B for 1 minute

[0202] Solvent A: 0.1% TFA, in H 2 O

[0203] Solvent: B: 0.1% TFA, in ACN

[0204] Flow rate: 1.5 ml / min

[0205] Column: XBridge C18 (50x 4.6) mm, 3.5 μm Ultraviolet (“UV”) visualization at 220 nanometers (“nm”).

[0206] GCMS-Method D:

[0207] Column: HP-5 (30m x 320μm x 0.25μm)

[0208] Column length 30m, inner diameter 0.32mm, thickness 0.25μm

[0209] Inlet temperature: 250 °C; Carrier gas: He

[0210] Detector temperature: 300 °C; Column flow rate 2 mL / min;

[0211] Gas flow 400 ml / min; H 2 Flow rate 40 mL / min.

[0212] Heating schedule: Held at 120 °C for 3 minutes; then heated to 300 °C at a rate of 40 °C / minute and held for 2 minutes. Source temperature: 230 °C.

[0213] Recorded on: 7890B GC, with 5977B MSD from Agilent technologies.

[0214]

[0215] Intermediate 1

[0216]

[0217] Step 1:

[0218] At -78 °C, n-butyllithium (37.8 mL, 94 mmol) was added to a stirred solution of 2-fluoro-3-methoxypyridine (10 g, 79 mmol) and TMEDA (23.74 mL, 157 mmol) in THF (120 mL). The mixture was then stirred at the same temperature for 2 h. At -78 °C, triisopropyl borate (27.4 mL, 118 mmol) was added to the reaction mixture and stirred for 2 h. The reaction mixture was quenched with water (50 mL) at -78 °C and then extracted with ether (2 × 100 mL). The aqueous layer was acidified with acetic acid (pH ~4) and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na 2 SO 4 and filtered and concentrated. The desired product, (2-fluoro-3-methoxypyridin-4-yl)boronic acid (11 g, 63.7 mmol, 81% yield), was isolated as an off-white solid.

[0219] MS (M+1) m / z: 172.2 [M+1] + . LC retention time 0.63 Min [Method A].

[0220] Step 2:

[0221] At ambient temperature, K 3 PO 4 (12.74 g, 73.1 mmol) was added to a stirred solution of (2-fluoro-3-methoxypyridin-4-yl)boronic acid (5 g, 29.3 mmol) in water (6 mL) and 1,4-dioxane (60 mL), and the mixture was then degassed for 10 min under N 2 atmosphere. 4-Bromo-2-methyl-2H-1,2,3-triazole (4.74 g, 29.3 mmol) and PdCl 2 (dppf)[DCM adduct (2.389 g, 2.93 mmol) were added to the reaction mixture and degassed for 5 min. The reaction mixture was stirred in a sealed tube at 80 °C for 6 h. The reaction mixture was filtered through a pad and washed with ethyl acetate (2 × 100 mL). The combined organic layers were washed with brine solution (50 mL) and dried over anhydrous Na 2 SO4 , filtered, and concentrated. The crude product was purified by silica gel (230 - 400 mesh) column chromatography, eluted with 10 - 15% ethyl acetate / petroleum ether, to give 2 - fluoro - 3 - methoxy - 4-(2 - methyl - 2H - 1,2,3 - triazol - 4 - yl)pyridine (5.2 g, 22.98 mmol, 79% yield), as an off - white solid.

[0222] MS (M + 1) m / z: 209.0 [M + 1] + . LC retention time 1.49 Min [Method B].

[0223] 1 H - NMR (400 MHz, DMSO - d 6 ): δ8.29 (s, 1H), 7.98 (dd, J = 1.20, 5.00 Hz, 1H), 7.78 (dd, J = 0.40, 5.00 Hz, 1H), 4.27 (s, 3H), 3.98 (d, J = 2.40 Hz, 3H).

[0224] Step 3:

[0225] 2 - Fluoro - 3 - methoxy - 4-(2 - methyl - 2H - 1,2,3 - triazol - 4 - yl)pyridine (5 g, 24.02 mmol) and (2,4 - dimethoxyphenyl)methanamine (25 g, 150 mmol) were stirred in a sealed tube at 100 °C for 16 h. The reaction mixture was partitioned between ethyl acetate (100 mL) and water (100 ml). The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (2 x 150 mL). The combined organic layers were washed with brine solution (50 mL), dried over anhydrous Na 2 SO 4 and filtered and concentrated. The crude product was purified by silica gel (230 - 400 mesh) column chromatography, eluted with 20 - 25% ethyl acetate / petroleum ether, to give the off - white solid N-(2,4 - dimethoxybenzyl)-3 - methoxy - 4-(2 - methyl - 2H - 1,2,3 - triazol - 4 - yl)pyridin - 2 - amine (6.2 g, 14.31 mmol, 59.6% yield).

[0226] MS (M + 1) m / z: 356.2 [M + 1] + . LC retention time 1.45 Min [Method B].

[0227] Step 4:

[0228] At 0 °C, to the stirred N-(2,4 - dimethoxybenzyl)-3 - methoxy - 4-(2 - methyl - 2H - 1,2,3 - triazol - 4 - yl)pyridin - 2 - amine (6 g, 16.88 mmol) in CH 2Cl 2 (60 mL) of solution was added to TFA (13.01 mL, 169 mmol). The reaction mixture was stirred at ambient temperature for 16 h. The reaction mixture was quenched with 10% aqueous NaHCO 3 solution (100 mL), and then extracted with ethyl acetate (2 × 200 mL). The combined organic layers were washed with brine solution (50 mL), dried over anhydrous Na 2 SO 4 and filtered and concentrated. The crude product was washed with 5% EtOAc / petroleum ether to give the desired 3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-amine (3.80 g, 14.81 mmol, 88% yield) as an off-white solid.

[0229] MS (M+1) m / z: 206.2 [M+1] + . LC retention time 0.89 Min [Method A].

[0230] 1 1H-NMR (400 MHz, DMSO-d 6 ): δ 8.17 (s, 1H), 7.74 (d, J = 5.60 Hz, 1H), 6.98 (d, J = 5.60 Hz, 1H), 6.17 (s, 2H), 4.24 (s, 3H), 3.65 (s, 3H).

[0231] Intermediate 2

[0232]

[0233] Step 1:

[0234] To a solution of 1-bromo-2-methoxy-3-nitrobenzene (2.0 g, 8.62 mmol) in ethanol (20 mL) and water (5 mL) was added iron (3.37 g, 60.3 mmol) and ammonium chloride (2.3 g, 43.3 mmol). The reaction mixture was stirred at 60 °C for 3 h, diluted with ethanol (50 mL) and filtered through a pad of diatomaceous earth. The filtrate was concentrated under reduced pressure to give the crude product. The crude residue was diluted with EtOAc (100 mL) and washed with water (2 × 20 mL) and brine (2 × 20 mL). The collected organic layer was dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure to give 3-bromo-2-methoxyaniline (1.8 g, 8.55 mmol, 99% yield) as a brown liquid.

[0235] MS (M+1) m / z: 202.0 (M+H) + . LC retention time 1.84 [A].

[0236] Step 2:

[0237] To a stirred solution of 3-bromo-2-methoxyaniline (1.80 g, 8.91 mmol) in 1,4-dioxane (15 mL) in a sealed tube was added bis(pinacolato)diboron (3.39 g, 13.36 mmol) and potassium acetate (2.62 g, 26.7 mmol). The reaction mixture was purged with nitrogen for 5 minutes and then PdCl 2 (dppf)[DCM adduct] (0.73 g, 0.89 mmol) was added. The reaction mixture was stirred at 90 °C for 5 h and then cooled to room temperature and diluted with EtOAc (100 mL). The reaction mixture was passed through filtration, and the filtrate was washed with water (50 mL) and brine (50 mL). The collected organic extract was dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (25% ethyl acetate / petroleum ether) to give 2-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (1.8 g, 6.88 mmol, 77% yield) as a light brown solid.

[0238] MS (M+1) m / z: 250.4 (M+H) + . LC retention time 2.11 [A].

[0239] Step 3:

[0240] To a stirred solution of 2-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (2.31 g, 9.26 mmol) and 4-bromo-2-methyl-2H-1,2,3-triazole (1.50 g, 9.26 mmol) in DME (15 mL) and water (5 mL) was added sodium carbonate (2.45 mg, 23.15 mmol). The reaction mixture was purged with nitrogen for 5 minutes and Pd(Ph 3 P) 4 (1.07 g, 0.93 mmol) was added. The reaction mixture was stirred at 90 °C for 6 h. The reaction mixture was filtered through a pad and washed with methanol (50 mL). The filtrate was concentrated under reduced pressure, and the crude residue was partitioned between ethyl acetate (150 mL) and water (150 mL). The collected organic layer was dried over anhydrous Na 2 SO 4Dry, filter, and concentrate. The crude residue was purified by silica gel column chromatography (50% ethyl acetate / petroleum ether) to afford the desired 2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)aniline (1.70 g, 7.67 mmol, 83% yield) as a brown crystalline solid.

[0241] MS (M+1) m / z: 205.2 (M+H) + 。LC retention time 1.20 [A]

[0242] Intermediate 3

[0243]

[0244] Step 1:

[0245] At 0 °C, oxalyl chloride (9.07 mL, 104.0 mmol) and 1 mL of DMF were added to a solution of lithium 4,6-dichloropyridazine-3-carboxylate (10.0 g, 51.8 mmol) in DCM (200 mL). The reaction mixture was warmed to room temperature over 1 h. The solvent was removed under reduced pressure, and DCM (200 mL) was added to the reaction mixture. The resulting solution was cooled to -30 °C, and deuterated methylamine hydrochloride (4.02 g, 57.0 mmol) was added, followed by DIPEA (18.1 mL, 104.0 mmol). The reaction mixture was stirred at -30 °C for 1 h, then warmed to room temperature and continued for 1 h. The reaction mixture was concentrated under reduced pressure, and the crude residue was dissolved in ethyl acetate (200 mL) and washed with water (100 mL) and brine (100 mL). The collected organic layer was dried over anhydrous Na 2 SO 4 and then concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography (10 - 15% ethyl acetate / petroleum ether) to give the desired product 4,6-dichloro-N-(methyl-d 3 )pyridazine-3-carboxamide (4.1 g, 19.42 mmol, 37.5% yield) as a pale yellow solid.

[0246] MS (M+1) m / z: 208.6 (M+H) + 。LC retention time 1.18 min [B].

[0247] Step 2:

[0248] At ambient temperature, 3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-amine (2.5 g, 12.18 mmol) and 4,6-dichloro-N-(methyl-d 3) To a stirred solution of pyridazine-3-carboxamide (3.06 g, 14.62 mmol) in THF (50 mL) was added LiHMDS (48.7 mL, 48.7 mmol, 1 M THF solution). The reaction mixture was stirred at this temperature for 2 h and quenched with aqueous NH 4 Cl solution (50 mL), then extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with saturated brine solution (50 mL), dried over anhydrous Na 2 SO 4 , filtered and concentrated. The crude product was purified by column chromatography on silica gel (230 - 400 mesh), eluting with 90% ethyl acetate / petroleum ether to afford the desired 6-chloro-4-((3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d 3 )pyridazine-3-carboxamide (3.2 g, 7.45 mmol, 61.2% yield) as an off-white solid.

[0249] MS (M+1) m / z: 378.2 (M + H) + . LC retention time 1.39 Min [Method B].

[0250] The following intermediates (3a - 3c) were prepared in a similar manner to Intermediate 3.

[0251]

[0252]

[0253] Intermediate 4

[0254]

[0255] At ambient temperature, 1-chloro-2-isocyanatoethane (1.0 g, 9.48 mmol) was added dropwise to a stirred solution of cyclopropylamine (0.541 g, 9.48 mmol) in THF (20 mL). Then, the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure, the crude product was dissolved in THF (10 mL), and then NaH (0.455 g, 18.95 mmol) was added portionwise at 0 °C. The reaction mixture was stirred at ambient temperature for 3 h. The reaction mixture was quenched with cold water (50 mL). The crude product was extracted with ethyl acetate (2 × 50 mL), washed with brine solution, dried over anhydrous sodium sulfate, and concentrated. The crude product was washed with n-pentane (20 mL) to afford the desired product 1-cyclopropylimidazolidin-2-one (870 mg, 6.83 mmol, 72.0% yield) as an off-white solid.

[0256] GCMS(M) m / z: 127.2 (M) + ; retention time 0.34 min [D].

[0257] Intermediate 5

[0258]

[0259] Step 1:

[0260] At 0 °C, cesium carbonate (11.35 g, 34.8 mmol), p-methoxybenzyl chloride (3.64 g, 23.23 mmol), and potassium iodide (0.771 g, 4.65 mmol) were added to a stirred solution of imidazolidin-2-one (2 g, 23.23 mmol) in DMF (5 mL). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with ice-cold water (50 mL) and extracted with ethyl acetate (2 × 100 mL). The combined organic fractions were dried over anhydrous Na 2 SO 4 and concentrated in vacuo to give the crude product. The crude product was purified by reverse-phase column chromatography using an aqueous solution of 0.1% ammonium formate in ACN as the eluent to give the desired product 1-(4-methoxybenzyl)imidazolidin-2-one (1.5 g, 6.55 mmol, 28.2% yield), as a white crystalline solid.

[0261] MS(M+1) m / z: 207.2 (M+H) + . LC retention time 0.75 [C].[[]END]]

[0262] Step 2:

[0263] At 0 °C, silver trifluoromethanesulfonate (685 mg, 2.67 mmol), (3435 mg, 9.70 mmol), 2-fluoropyridine (0.229 mL, 2.67 mmol), and trimethyl(trifluoromethyl)silane (1724 mg, 12.12 mmol) were added to a stirred solution of 1-(4-methoxybenzyl)imidazolidin-2-one (500 mg, 2.424 mmol) in THF (20 mL) and chlorobenzene (2 mL), and then CsF (1841 mg, 12.112 mmol) was added. Then, it was stirred at ambient temperature for 16 h. The reaction mixture was filtered under reduced pressure and washed with EtOAc (100 mL). The filtrate was concentrated in vacuo. The crude product was purified by reverse-phase column chromatography using an ACN solution of 0.1% ammonium formate to give the desired 1-(4-methoxybenzyl)-3-(trifluoromethyl)imidazolidin-2-one (300 mg, 0.941 mmol, 38.8% yield), as a brown viscous solid.

[0264] MS(M+1) m / z: 275.2 (M+H) + 。LC retention time 1.22 [C].[[]END]]

[0265] Step 3:

[0266] At 0 °C, trifluoroacetic acid (8 mL, 38.3 mmol) was added to 1-(4-methoxybenzyl)-3-(trifluoromethyl)imidazolidin-2-one (350 mg, 1.276 mmol). The reaction mixture was stirred at 50 °C for 4 h. After completion, the reaction mixture was concentrated under reduced pressure and washed with n-pentane (20 mL) to give the desired 1-(trifluoromethyl)imidazolidin-2-one (150 mg, 0.146 mmol, 11.44% yield) as a brown gummy solid. The crude product was used directly without further purification.

[0267] MS(M+1) m / z: 155.2 (M+H) + 。LC retention time 0.56 [C].[[]END]]

[0268] Example 1

[0269]

[0270] To a solution of 6-chloro-4-((3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d 3 )nicotinamide (0.2 g, 0.531 mmol) in 1,4-dioxane (10 mL) in a 40 mL pressure release vial was added 1-isopropylimidazolidin-2-one (0.068 g, 0.531 mmol) and Pd 2 dba 3 (0.024 g, 0.027 mmol), 1,1'-bis(dicyclohexylphosphino)ferrocene (0.015 g, 0.027 mmol), and then Cs 2 CO 3 (0.432 g, 1.327 mmol). The reaction mixture was degassed with N 2 for 5 min. The resulting reaction mixture was heated to 110 °C for 3 h. After completion, the reaction mixture was diluted with ethyl acetate (100 mL), filtered through a pad, and washed thoroughly with ethyl acetate (100 mL). The filtrate was washed with water (50 mL), then with brine (50 mL), and dried over anhydrous Na 2 SO 4Drying. The solvent was evaporated under reduced pressure, and the resulting crude residue was purified by reverse-phase column chromatography (C18 column) using an acetonitrile solution of 50% eluent (aqueous solution of 1% ammonium formate) to obtain the desired product 6-(3-isopropyl-2-oxoimidazolidin-1-yl)-4-((3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d 3 )nicotinamide (80 mg, 0.167 mmol, 31.4% yield), which was a grayish-white solid. MS (M+1) m / z: 469.2 (M+H) + . LC retention time 1.76 [A].[[]]END]]

[0271] 1H-NMR (400 MHz, DMSO-d6): δ 12.33 (s, 1H), 9.96 (s, 1H), 9.29 (s, 1H), 8.31 (s, 1H), 8.16 (d, J = 5.20 Hz, 1H), 7.46 (d, J = 5.20 Hz, 1H), 4.28 (s, 3H), 4.18 - 4.11 (m, 3H), 3.82 (s, 3H), 3.50 (t, J = 7.60 Hz, 2H), 1.17 (d, J = 6.40 Hz, 6H).

[0272] The following Examples (2 - 6) were prepared in a similar manner to the preparation of Example 1.

[0273]

[0274]

[0275]

[0276]

[0277] Example 8

[0278]

[0279] Step 1: 4-Bromo-6-methyl-2-nitropyridin-3-ol

[0280] Concentrated sulfuric acid (1 mL) was added dropwise to solid 4-bromo-6-methylpyridin-3-ol (0.267 g, 1.420 mmol) in a flask at -10 °C [in a salt and ice bath]. Subsequently, fuming nitric acid (0.063 mL, 1.420 mmol) was added dropwise. The reaction mixture was stirred overnight and the reaction slowly warmed to room temperature. The reaction mixture was poured onto approximately 50 g of ice. After the ice melted, the mixture was transferred to a separatory funnel and extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated to give 4-bromo-6-methyl-2-nitropyridin-3-ol as a yellow solid (165 mg, 0.708 mmol, 49.9% yield). 1 1H NMR (400 MHz, chloroform-d) δ 10.63 (s, 1H), 7.75 (s, 1H), 2.57 (s, 3H).

[0281]

[0282] Step 2: 4-Bromo-3-methoxy-6-methyl-2-nitropyridine

[0283] A mixture of 4-bromo-6-methyl-2-nitropyridin-3-ol (160 mg, 0.687 mmol), potassium carbonate (474 mg, 3.43 mmol) and MeI (0.215 mL, 3.43 mmol) in DMF was stirred overnight at room temperature. The reaction mixture was partitioned between EtOAc (30 mL) and water (30 mL). The organic layer was washed with 10% LiCl (2 x 30 mL) and brine (30 mL), dried over anhydrous sodium sulfate and concentrated to give 4-bromo-3-methoxy-6-methyl-2-nitropyridine as a brown solid (133 mg, 0.538 mmol, 78% yield). MS (M+1) m / z: 247.0 (249.0) (M+H) + LC retention time 1.02 [E].

[0284]

[0285] Step 3: 4-Bromo-3-methoxy-6-methylpyridin-2-amine

[0286] At 0 °C, iron powder (210 mg, 3.77 mmol) was added to a stirred solution of 4-bromo-3-methoxy-6-methyl-2-nitropyridine (133 mg, 0.538 mmol) in ethanol (0.6 mL), acetic acid (0.3 mL) and water (0.6 mL). The resulting mixture was warmed to room temperature and stirred for a total of 2 hours. The reaction mixture was passed through Filter and wash the cake with EtOAc and water. Transfer the filtrate to a separatory funnel and add 50 mL of 1.5 M potassium hydrogen phosphate. After shaking, separate the layers and wash the organic layer with brine (50 mL), dry over anhydrous sodium sulfate and concentrate to give 4-bromo-3-methoxy-6-methylpyridin-2-amine as a cream solid (101 mg, 0.465 mmol, 86% yield). MS (M+1) m / z: 217.0 (219.0) (M+H) + LC retention time 0.64 [E].

[0287]

[0288] Step 4: 3-Methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-amine

[0289] A stirred mixture of (2-methyl-2H-1,2,3-triazol-4-yl)boronic acid (Intermediate 19) (89 mg, 0.698 mmol), 4-bromo-3-methoxy-6-methylpyridin-2-amine (101 mg, 0.465 mmol) and PdCl 2 (dppf)-CH 2 Cl 2 adduct (19.00 mg, 0.023 mmol) in dioxane (3.5 mL) was degassed by bubbling nitrogen through the mixture for 5 minutes. 2 M K 3 PO 4 (aq) (0.698 mL, 1.396 mmol) was added rapidly and the reaction mixture was heated at 100 °C for 0.75 h. After cooling to room temperature, the reaction mixture was partitioned between EtOAc (30 mL) and brine (20 mL). After drying over anhydrous sodium sulfate solution, the organic layer was concentrated and the residue was chromatographed on a 12 gm ISCO silica gel column, eluting with a 0-100% EtOAc / Hex gradient. The pure fractions were concentrated to give 3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-amine as a pale yellow solid (75 mg, 0.342 mmol, 73.5% yield). MS (M+1) m / z: 220.2 (M+H) + LC retention time 0.68 [E].

[0290]

[0291] Step 5: 6-Chloro-4-((3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d 3 )pyridazine-3-carboxamide

[0292] At room temperature, within 10 minutes, 1 M KHMDS (1.539 mL, 1.539 mmol) in THF was added dropwise to a solution of 3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-amine (75 mg, 0.342 mmol) and 4,6-dichloro-N-(methyl-d 3 )pyridazine-3-carboxamide (143 mg, 0.684 mmol) in THF. The reaction mixture was stirred at room temperature for 30 minutes. After quenching with 2 mL of saturated ammonium chloride solution, the organics were removed on a rotary evaporator and the residue was diluted with water. Filtration, rinsing the filter cake with ether and drying gave 6-chloro-4-((3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d 3 )pyridazine-3-carboxamide (40 mg, 0.102 mmol, 29.8% yield) as a tan solid. MS (M+1) m / z: 392.2 (M+H) + . LC retention time 1.13 [E].

[0293] Example 8: 6-(3-Isopropyl-2-oxoimidazolidin-1-yl)-4-((3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d 3 )pyridazine-3-carboxamide

[0294] A mixture of 6-chloro-4-((2-methoxy-5-(methoxymethyl)-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino)-N-(methyl-d 3 )pyridazine-3-carboxamide (25 mg, 0.059 mmol), 1-isopropylimidazolidin-2-one (40.9 mg, 0.319 mmol), Pd 2 (dba) 3 chloroform adduct (6.59 mg, 6.38 μmol), xantphos (7.38 mg, 0.013 mmol) and Cs 2 CO 3 (83 mg, 0.255 mmol) in dioxane (0.5 mL) was bubbled with N 2Degas the mixture for 5 minutes. Seal the reaction vessel and heat it to 130 °C for 30 minutes. Dilute the reaction mixture with DMSO and filter. Purify the filtrate by preparative LC / MS under the following conditions: Column: XBridge C18, 200 mm x 19 mm, 5-μm particles; Mobile phase A: 5:95 acetonitrile: water, containing 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water, containing 10 mM ammonium acetate; Gradient: Hold at 28% B for 0 minutes, hold at 28 - 70% B for 20 minutes, then hold at 100% B for 4 minutes; Flow rate: 20 mL / min; Column temperature: 25 °C. Collect fractions triggered by the MS signal. Combine the fractions containing the desired product and dry them by centrifugal evaporation to obtain 6-(3-isopropyl-2-oxoimidazolidin-1-yl)-4-((3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d 3 )pyridazine-3-carboxamide (10.8 mg; 35% yield). MS (M+1) m / z: 484.7 (M+H) + . LC retention time 2.04 [F]. 1 H NMR (500 MHz, DMSO-d 6 ) δ 12.20 (s, 1H), 10.18 (s, 1H), 9.22 (s, 1H), 8.25 (s, 1H), 7.30 (s, 1H), 4.25 (s, 3H), 4.16 - 4.08 (m, 3H), 3.76 (s, 3H), 1.15 (d, J = 6.8 Hz, 6H)) 2 protons are buried under the water peak and the methyl group is buried under the DMSO peak.

[0295] Example 9

[0296]

[0297] Example 9: 6-(3-cyclopropyl-2-oxoimidazolidin-1-yl)-4-((3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d3)pyridazine-3-carboxamide. The compound was prepared using the same procedure as in Example 8. MS (M+1) m / z: 482.1 (M+H) + . LC retention time 1.65 [G]. 1 H NMR (500 MHz, DMSO-d 6)δ12.23(s,1H),10.17(s,1H),9.26(s,1H),8.27(s,1H),7.32(s,1H),4.27(s,3H),4.07(t,J=7.9Hz,2H),3.78(s,3H),3.49(t,J=8.0Hz,2H),2.70-2.64(m,1H),2.53(s,3H),0.72(s,4H).

[0298] Biological assay

[0299] The following assays were used to demonstrate the activity of the compounds of the present invention.

[0300] In vivo brain penetration assay

[0301] Pharmacokinetic studies were performed using C57BL6 wild-type mice (n = 3 per experiment) to determine the brain and plasma exposure of the compounds of the present invention. The compounds were orally administered in the following solution: 5% ethanol; 90% PEG 300; 5% TPGS, 5 mL / kg, final concentration 10 mg / kg. Mice were euthanized 1 hour after dosing, and plasma and brain were collected and frozen for analysis. Brain tissue was homogenized in an equal volume of blank C57BL6 mouse plasma. The concentrations of the compounds in plasma and brain homogenates were determined by LC-MS analysis.

[0302] Bidirectional Permeability Assay of Caco-2 Cells

[0303] Overview

[0304] The described compounds were tested in a Caco-2 bidirectional permeability assay to evaluate their permeability and efflux substrate potential. The compounds (3 μM, in triplicate) were incubated with Caco-2 cells in assay buffer at pH 7.4 (containing 0.5% bovine serum albumin [BSA]) at 37 °C for 2 hours, then extracted for LC-MS analysis to determine their concentration in the reaction mixture and to calculate the permeability coefficient, efflux ratio, and recovery.

[0305] Materials and methods

[0306] Caco-2 (Caucasian colon adenocarcinoma) cells were obtained from the American Type Culture Collection (Manassas, Virginia). Dulbecco's Modified Eagle's Medium (DMEM), N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) buffer, non-essential amino acids, L-glutamine, penicillin-G-streptomycin, and heat-inactivated fetal bovine serum (FBS) were purchased from GIBCO / Invitrogen (Carlsbad, California). 96-well Transwell plates with 0.4-μm pore size polycarbonate membranes (surface area: 0.11 cm 2 ) and low-binding transwell cluster plates were purchased from Sigma Aldrich (St. Louis, Missouri). Low-binding 96-well plates were purchased from Corning (Corning, New York). Modified Hank's Balanced Salt Solution (MHBSS) was prepared by adjusting Hank's Balanced Salt Solution (HBSS) to pH 7.4 with HEPES. HBSS, digoxin, and bovine serum albumin (BSA) were purchased from Sigma (St. Louis, Missouri). Filter blocks (2 mL, 96-well) were purchased from Whatman (Freiburg, Germany). All solvents were of analytical grade.

[0307] Cell Preparation

[0308] Fourteen (14) to twenty-eight (28) days prior to the assay, Caco-2 cells were seeded at a density of 1.8x10 5 cells / cm 2 onto the polycarbonate filter membranes in 96-well transwell plates, approximately 2.0x10 4 cells per well. The cells were grown in a medium consisting of DMEM supplemented with 10% fetal bovine serum, 10 mM HEPES, 1% non-essential amino acids, 2 mM L-glutamine, 100 U / mL penicillin-G, and 100 μg / mL streptomycin. The medium was changed every 3 days, and the cells were maintained at 37 °C, 95% relative humidity, and 5% CO 2 atmosphere. Prior to analysis, the formation of tight junctions in the cells was evaluated (see the Quality Control section below).

[0309] Compound Preparation

[0310] The compound was dissolved in 100% DMSO to 10 mM. After visual confirmation of complete dissolution, the 10 mM compound stock solution was plated into 96-well plates and further serially diluted in 100% DMSO to produce 100x stock solutions at a concentration of 0.3 mM. Four (4) control compounds were tested along with the compound, and they were plated at 100-fold concentration of 0.3 mM, in quadruplicate.

[0311] Permeability assessment

[0312] The compound was tested in triplicate in a single experiment at a final concentration of 3 μM. The cell passages used in the experiment have passed the QC standard (see the Quality Control section below). This study was conducted using a monolayer of Caco-2 cells cultured for 14 to 28 days, with the cell passage number between 20 and 80.

[0313] The assay (transport) buffer consisted of MHBSS adjusted to pH 7.4 and 0.5% BSA. From a 100x compound plate, 8 μL of a 100% DMSO stock solution of the compound was added to 800 μL of assay buffer, mixed well, and filtered to remove any precipitate as the final preparation step before the assay incubation. The target final assay concentration of the described compound and the control compound was 3 μM. The filtrate represented the initial stock compound solution (bidirectional) used as the assay donor solution. The receiving solution was only the assay buffer.

[0314] Just before the assay was performed, each cell monolayer was washed 3 times with assay buffer to remove all traces of the medium. The permeability study was initiated by adding 100 μL of assay buffer with / without compound to the apical transwell compartment of a 96-well transwell low-binding cluster plate and adding 200 μL of assay buffer with / without compound to the basolateral compartment. For apical-to-basolateral (A→B) permeability (absorptive direction), the buffer (1x donor solution) containing the compound or the control compound was placed in the apical compartment (donor well), while only the buffer was placed in the corresponding basolateral compartment (receptor well). For basolateral-to-apical (B→A) permeability (secretory direction), the buffer (1x donor solution) containing the compound or the control compound was placed in the basolateral compartment (donor well), while only the buffer was placed in the corresponding apical compartment (receptor well). Then the Transwell was incubated at 37 °C, 95% relative humidity, and 5% CO 2 atmosphere for 2 hours. After incubation, 75 μL of liquid was taken from each apical and basolateral compartment and transferred to a 96-well low-binding plate that had been pre-added with 75 μL of an acetonitrile solution containing 250 nM propranolol, 250 nM diclofenac, and 500 nM tolbutamide as internal standards per well. Subsequently, the samples were analyzed by LC-MS / MS to determine the described compound and the control compound.

[0315] Analysis of assay samples

[0316] The concentrations of the compound and the reference compound in the sample were determined by LC-MS / MS analysis. The AB Sciex 4500 / 5500 / 6500 multiplex system consists of two Shimadzu 20ADvp binary pumps with SCL-20Avp controllers (for gradient elution), an LS1 autosampler, and an AB Sciex 4500 / 5500 / 6500 triple quadrupole mass spectrometer operating in the electrospray ionization (ESI) mode. To obtain the optimal SRM conditions for sample analysis, DiscoveryQuant TM (AB Sciex) was used to optimize MS / MS for each compound, characterized by saturation control with a 5 μM standard solution in a methanol and water (1:1, v / v) mixture prepared from the compound stock solution. Optimization was performed using flow injection analysis with an injection volume of 40 μL, 75% mobile phase B (0.2% formic acid in acetonitrile), and 25% mobile phase A (aqueous 0.2% formic acid), isocratic elution.

[0317] A 5 μL aliquot of the sample was injected and then gradient elution separation was performed on a Kinetex XB-C18, 2.6 μm, 2.1 x 30 mm column using a mobile phase consisting of A (aqueous 0.2% formic acid) and B (0.2% formic acid in acetonitrile).

[0318]

[0319] A = aqueous 0.2% formic acid; B = acetonitrile solution of 0.2% formic acid

[0320] DiscoveryQuant TM automatically determines the optimal ionization polarity (positive or negative), precursor and product ions, declustering potential, and collision energy for the compound and the reference compound. The optimized SRM MS / MS conditions were used for sample analysis. The compound or reference compound described by the peak area ratio was quantified against an internal standard. The peak area ratio of the compound in the dosing solution was used to determine the concentration of the compound in the sample.

[0321] Data analysis

[0322] The following results for the described compound were reported: permeability coefficient (Pc [nanometers per second]), efflux ratio, and percentage recovery.

[0323] The Pc value was calculated using the following equation:

[0324]

[0325] Where:

[0326] C At = concentration of the test compound in the receptor well at time t,

[0327] V A = volume in the receptor well,

[0328] S = surface area of the membrane (0.11 cm 2 ),

[0329] C D0 = initial concentration of the test compound in the donor well,

[0330] t = incubation time.

[0331] The efflux ratio is calculated as:

[0332]

[0333] The recovery (%) is calculated by expressing the total amount (nmol) of the test compound present in the donor and receptor assay compartments at the end of the incubation time as a fraction (percentage) of the total amount (nmol) of the test compound added to the donor compartment before the assay incubation. It is calculated using the following equation:

[0334]

[0335] where:

[0336] C D0 = initial concentration of the test compound in the donor well,

[0337] V D = volume in the donor well,

[0338] C Dt = concentration in the donor well after time t,

[0339] C At = concentration in the receptor well after time t,

[0340] V A = volume in the receptor well.

[0341] Quality control

[0342] The formation of tight junctions in Caco-2 cells in the transwell plates used on the day of the assay was evaluated using transepithelial electrical resistance (TEER) measurements. TEER evaluations were performed using an EVOM resistance meter (World Precision Instruments, Sarasota, Florida). Each well of the transwell plate showed a TEER value > 600 Ω·cm 2 , and all plates of this cell passage and seeding batch were suitable for the assay.

[0343] Four (4) control compounds were tested with the compounds described in each experiment, and their Pc values covered a range of permeabilities. The acceptance criteria for this test required that the control compounds be within an acceptable historical range at 3 μM. The Pc values of these 4 controls observed historically and the acceptable ranges for external ratios are shown in Table B.

[0344] In these studies, the results for all control compounds were within their respective historical ranges. Therefore, the acceptance assay data were used for data analysis and evaluation of the bidirectional permeability of the compounds described in Caco-2 cells.

[0345]

[0346] Values are mean ± standard deviation.

[0347] Pc = permeability coefficient. A→B = apical to basolateral. B→A = basolateral to apical.

[0348] IFNα-induced STAT phosphorylation in human whole blood

[0349] After incubation with the compound for one hour, human whole blood (drawn with ACD-A as anticoagulant) was stimulated with 1000 U / mL recombinant human IFNαA / D (R&D Systems 11200-2) for 15 minutes. Stimulation was terminated by adding Fix / Lyse buffer (BD 558049). The cells were stained with CD3 FITC antibody (BD 555916), washed, and permeabilized using Perm III buffer (BD 558050) on ice. The cells were then stained with Alexa-Fluor 647 pSTAT5 (pY694) antibody (BD 612599) for 60 minutes and then analyzed on an iQue Plus. After gating on CD3-positive populations, pSTAT5 expression was quantified by median fluorescence intensity.

[0350] Table 1: Potency of exemplary compounds in human whole blood assays

[0351] Ex.# IFNa pSTAT5 IC50 in Human Whole Blood (μM) Caco-2 AB (nm / s) Caco-2 Efflux Ratio 1 0.066 494 0.4 2 0.026 233 1.4 3 0.077 287 0.7 4 0.026 576 0.4 5 0.095 361 0.4 6 0.80 118 0.6 7 0.047 139 0.3 8 0.383 154 0.3 9 0.141 137 0.2

Claims

1. A compound of formula I or a stereoisomer or pharmaceutically acceptable salt thereof, wherein X is -N- or -CH-; Y is -N- or -CH-; R is R 1 is CF 3 , C 1-6 alkyl or C 3-6 cycloalkyl; R 2 is H or C 1-6 alkyl; Z is CHR 1 、CH 2 、CR 1 2 、O、NR 1 or C(O); and n is 0, 1, 2 or 3.

2. The compound according to claim 1, which has the following formula or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R is R 1 is CF 3 , C 1-6 alkyl or C 3-6 cycloalkyl; R 2 is H or C 1-6 alkyl; Z is CHR 1 、CH 2 、CR 1 2 、O, NR 1 or C(O); and n is 0, 1, 2 or 3.

3. The compound according to claim 1, which has the following formula or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R is R 1 is CF 3 , C 1-6 alkyl or C 3-6 cycloalkyl; Z is CHR 1 , CH 2 , CR 1 2 , O, NR 1 or C(O); and n is 0, 1, 2 or 3.

4. The compound according to claim 1, which has the following formula or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R is R 1 is CF 3 , C 1-6 alkyl or C 3-6 cycloalkyl; Z is CHR 1 , CH 2 , CR 1 2 , O, NR 1 or C(O); and n is 0, 1, 2 or 3.

5. The compound according to claim 1, which has the following formula or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R is R 1 is CF 3 , C 1-6 alkyl or C 3-6 cycloalkyl; Z is CHR 1 , CH 2 , CR 1 2 , O, NR 1 or C(O); and n is 0, 1, 2 or 3.

6. A compound or a pharmaceutically acceptable salt thereof, selected from 4-{[3-Methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl]amino}-N-(2H3)methyl-6-[2-oxo-3-(propan-2-yl)imidazolidin-1-yl]pyridazine-3-carboxamide; 4-{[3-Methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl]amino}-N-(2H3)methyl-6-[2-oxo-3-(propan-2-yl)imidazolidin-1-yl]pyridine-3-carboxamide; 4-{[2-Methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl]amino}-N-(2H3)methyl-6-[2-oxo-3-(propan-2-yl)imidazolidin-1-yl]pyridine-3-carboxamide; 6-(3-Cyclopropyl-2-oxoimidazolidin-1-yl)-4-{[3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl]amino}-N-(2H3)methylpyridazine-3-carboxamide; 6-(3-Cyclopropyl-2-oxoimidazolidin-1-yl)-4-{[2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl]amino}-N-(2H3)methylpyridazine-3-carboxamide; 6-(3-Isopropyl-2-oxoimidazolidin-1-yl)-4-((3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d 3 )pyridazine-3-carboxamide; 6-(3-Cyclopropyl-2-oxoimidazolidin-1-yl)-4-((3-methoxy-6-methyl-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-N-(methyl-d3)pyridazine-3-carboxamide; 6-(3-Cyclobutyl-2-oxoimidazolidin-1-yl)-4-{[3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl]amino}-N-(2H3)methylpyridazine-3-carboxamide; and 4-{[2-Methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl]amino}-N-(2H3)methyl-6-[2-oxo-3-(trifluoromethyl)imidazolidin-1-yl]pyridine-3-carboxamide.

7. A pharmaceutical composition comprising one or more of the compounds according to claim 1 and a pharmaceutically acceptable carrier or diluent.

8. A pharmaceutical composition comprising one or more of the compounds according to claim 6 and a pharmaceutically acceptable carrier or diluent.

9. A method of treating a disease, comprising administering a therapeutically effective amount of the compound according to claim 1 to a patient in need of such treatment, wherein the disease is a neurodegenerative disease.

10. The method according to claim 9, wherein the neurodegenerative disease is Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, multiple sclerosis, optic neuritis or neuromyelitis optica.

11. The method according to claim 10, wherein MS includes relapsing MS and / or progressive MS, including clinically isolated syndrome (CIS).

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