EA047032B1 - SUBSTITUTED HETEROCYCLIC COMPOUNDS - Google Patents

Description

Cross references to related applications

This application claims the benefit of U.S. Provisional Application No. 63/188,498, filed May 14, 2021, and U.S. Provisional Application No. 63/318,508, filed March 10, 2022, the disclosures of which are incorporated herein by reference in their entirety.

Technical field of the invention

The present invention provides compounds useful in modulating IL-12, IL-23 and/or IFNa by targeting Tyk-2 to provide inhibition of signal transduction. Provided herein are substituted heterocyclic compounds, compositions containing such compounds, and methods of using them. The invention further relates to pharmaceutical compositions containing at least one compound in accordance with the present invention, which are useful for the treatment of conditions associated with modulation of IL-12, IL-23 and/or IFNa in a mammal. In particular, the present invention relates to compounds useful for the treatment of neurodegenerative diseases.

State of the art

The heterodimeric cytokines interleukin (IL)-12 and IL-23, which are characterized by the presence of a common p40 subunit, are produced by activated antigen-presenting cells and play a critical role in the differentiation and proliferation of Th1 and Th17 cells, two lineages of effector T cells that play key roles in autoimmunity. IL-23 consists of a p40 subunit along with a unique p19 subunit. IL-23, acting through a heterodimeric receptor consisting of IL-23R and IL-12Re1, is required for the survival and expansion of Th17 cells that produce proinflammatory cytokines such as IL-17A, IL-17F, IL-6 and TNF-a (McGeachy, M.J. et al., The link between IL-23 and Th17 cell-mediated pathologies, Semin. Immunol, 19:372-376 (2007)). These cytokines are important links in the pathobiology of a number of autoimmune diseases, including rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, and lupus. IL-12, in addition to the p40 subunit, like IL-23, contains the p35 subunit and acts through a heterodimeric receptor consisting of IL-12Re1 and IL-12Re2. IL-12 is required for the development of Th1 cells and the secretion of IFNy, a cytokine that plays a critical role in immunity by stimulating the expression of the major histocompatibility complex (MHC), class switching of B cells to IgG subclasses and activation of macrophages (Gracie, J.A. et al., Interleukin-12 induces interferon-gamma-dependent switching of IgG alloantibody subclass, Eur. J. Immunol., 26:1217-1221 (1996); and functions, J. Leukoc. Biol, 75(2): 163-189 (2004).

The importance of p40-containing cytokines in the autoimmune response is demonstrated by the finding that mice deficient in either p40, p19, or IL-23R are protected from disease in models of multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease, lupus, and psoriasis, among others (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); 12, independently of IFNgamma, plays a crucial role in the pathogenesis of a murine psoriasis like skin disorder, J. Immunol, 162:74807491 (1999); intestinal inflammation, J. Exp. Med, 203:2473-2483 (2006); Cua, DJ. et al., Interleukin-23 rather than interleukin-12 is the critical cytokine for autoimmune inflammation of the brain, Nature, 421:744. -748 (2003); Murphy, S. A. et al., Divergent pro-and anti-inflammatory roles for IL-23 and IL-12 in joint autoimmune inflammation, J. Exp. Med, 198:19511957(2003)).

In human disease, high expression of p40 and p19 has been measured in psoriatic lesions, and Th17 cells have been identified in active brain lesions in patients with multiple sclerosis (MS) and in the intestinal mucosa of patients with active Crohn's disease (Lee, E. et al ., Increased expression of interleukin 23 p19 and p40 in lesional skin of patients with psoriasis vulgaris, J. Exp. Med., 199:125-130 (2004); infiltrating T cells and glial cells is associated with active disease in multiple sclerosis, Am. J. Pathol., 172:146-155 (2008)). It has also been shown that p19, p40 and p35 mRNA levels are significantly higher in patients with active SLE compared with those in patients with inactive SLE (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).

In addition, genome-wide association studies have identified a number of loci associated with chronic inflammatory and autoimmune diseases that encode factors that function in the IL-23 and IL-12 pathways. Such genes include IL23A, IL2A, IL2B, 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);

- 1 047032

Cho, J.H. et al., Recent insights into the genetics of inflammatory bowel disease, Gastroenterology, 140:17041712 (2011)).

Indeed, anti-p40 treatments that inhibit both IL-12 and IL-23, as well as IL-23-specific anti-P19 treatments, have been shown to effectively target autoimmunity in diseases including psoriasis, Crohn's 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 , placebocontrolled, crossover trial, Lancet, 373:633-640 (2009)). Thus, agents that inhibit the action of IL-12 and IL-23 can be expected to have a therapeutic effect in autoimmune disorders in humans.

Group of type I interferons (IFN), which include representatives of IFNa, as well as IFNe, IFNo:. IFNk and IFN<^ act through the heterodimeric IFNa/β receptor (IFNAR). Type I IFNs have several effects on both the innate and adaptive immune systems, including activation of both cellular and humoral immune responses and increased expression and release of autoantigens (Hall, J.C. et al., Type I interferons: crucial participants in disease amplification in autoimmunity, Nat. Rev. Rheumatol, 6:40-49 (2010).

Patients with systemic lupus erythematosus (SLE), a potentially fatal autoimmune disease, have elevated serum interferon (IFN)a (type I interferon) or increased expression of genes regulated by type I IFN (called the IFNa expression profile) in peripheral blood mononuclear cells and in affected organs were demonstrated in 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 several studies have shown that serum IFNa levels correlate with both activity, and with the severity of the disease (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)). A direct role for IFNa in the pathobiology of lupus is suggested by the observation that administration of IFNa to patients with malignant or viral diseases can induce a lupus-like syndrome. In addition, deletion of IFNAR in lupus-prone mice provides significant 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 lupus-associated loci that encode factors that function 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); implicates IKBKE and IL8 as risk factors for SLE, Eur J. Hum. 19:479–484 (2011). In addition to lupus, there is evidence that abnormal activation of type I interferon-mediated pathways plays an important role in the pathobiology of other autoimmune diseases, such as Sjögren's syndrome and scleroderma (Bave, U. et al., Activation of the type I interferon system in primary Sjogren's 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, agents that inhibit the action of type I interferon responses can be expected to have a therapeutic effect in autoimmune disorders in humans.

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 critical role in regulating the receptor signaling cascade for IL-12, IL-23 and type I interferons 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); al., TYK2 kinase activity is required for functional type I interferon responses in vivo, PLoS One, 7:e39141 (2012)), and in 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, an essential signal that leads to dimerization of STAT proteins and transcription of STAT-dependent proinflammatory genes. Tyk2-deficient mice are resistant to experimental models of colitis, psoriasis, and multiple sclerosis, demonstrating the importance of Tyk2-mediated signaling in autoimmune reactions and related disorders (Ishizaki, M. et al.

- 2 047032 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)).

In humans, subjects expressing the inactive Tyk2 variant are protected from multiple sclerosis and possibly other autoimmune diseases (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 that other Tyk2 variants are 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's Disease and Psoriasis Identifies Seven Shared Susceptibility Loci, Am. J. Hum. Genet., 90:636-647 (2012); 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 for rheumatoid arthritis, Nat. Genet., 44:1336-1340 (2012 )).

TYK2 inhibition may also be used in solid tumors and hematologic malignancies, either as monotherapy or in combination with existing standard treatments, including immunotherapy.

Ex vivo studies in T-cell acute lymphoblastic leukemia (T-ALL) have shown that TYK2 is required for the survival of T-ALL, suggesting a potential direct cancer cell killing mechanism for TYK2 inhibitors in this disease. Sanda, T. et al. TYK2-STAT1-BCL2 Pathway Dependence in T-cell Acute Lymphoblastic Leukemia. Cancer Discov. 3, 564-577 (2013). Numerous TYK2 activating mutations have been discovered and characterized in T-ALL cell lines. An NPM1-TYK2 gene fusion has also been identified in a subset of cutaneous T-cell lymphomas (CTCL), and TYK2 has been shown to be an oncogenic driver of transformation, Kuravi, S. et al. Functional characterization of NPM1TYK2 fusion oncogene. Npj Precis. Oncol. 6, 3 (2022). Loss of signaling through TYK2 may inhibit this transformation potential.

Effective TYK2 inhibitors have been described; however, these compounds tend to be highly polar compounds with high efflux coefficients 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 known that one of the ways drug resistance manifests itself is through 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).

Thus, compounds with lower efflux ratios in in vitro models may have a potentially better chance of effectively treating some oncogenic diseases.

In view of conditions that may benefit from treatment by modulating cytokines and/or interferons, new compounds capable of modulating cytokines and/or interferons, such as IL-12, IL-23 and/or IFNa, and methods of using such compounds may provide significant therapeutic benefits to a wide range of patients in need.

The essence of the invention

The present invention relates to compounds of formula I below, which are useful as modulators of IL-12, IL-23 and/or IFNa by inhibiting Tyk2-mediated signaling.

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

The present invention also provides pharmaceutical compositions comprising a pharmaceutically acceptable carrier and at least one of the compounds of the present invention.

The present invention also provides a method of modulating IL-12, IL-23 and/or IFNa by inhibiting Tyk2-mediated signaling, comprising administering to a host in need of such treatment a therapeutically effective amount of at least one of the compounds of the present invention.

The present invention also provides a method of treating neurodegenerative diseases, comprising administering to a host in need of such treatment a therapeutically effective amount of at least one of the compounds of the present invention.

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

These and other features of the present invention will be set forth in greater detail as we proceed.

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Detailed Description of Embodiments of the Invention

In a first aspect of the present invention there is provided a compound of formula I

or a stereoisomer or pharmaceutically acceptable salt thereof, wherein

X represents -N- or -CH-;

R ¹ is -C(O)R ^1a ;

R ^1a represents C _3-6 cycloalkyl;

R ² represents C _1-6 alkoxy;

R ³ represents C _1-6 alkyl or C _3-6 cycloalkyl.

In a second aspect of the invention, there is provided a compound of formula

or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R ¹ is -C(O)R ^1a ;

R ^1a represents C _3-6 cycloalkyl;

R ² represents C _1-6 alkoxy;

R ³ represents C _1-6 alkyl or _{C 3_6 cycloalkyl} .

In a third aspect of the invention, there is provided a compound of formula

or a stereoisomer or pharmaceutically acceptable salt thereof, wherein

R ¹ is -C(O)R ^1a ;

R ^1a represents C _3-6 cycloalkyl;

R ² represents C _1-6 alkoxy;

R ³ represents C _1-6 alkyl or C _3-6 cycloalkyl.

In another aspect, a compound selected from illustrative examples included within the scope of the first aspect, or a pharmaceutically acceptable salt thereof, is provided.

In another aspect, a compound selected from any subset of compounds included within the scope of any of the aspects described above is provided.

In another aspect, a compound (IUPAC classification) or a pharmaceutically acceptable salt thereof selected from No. (4-((3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridine) is provided -2-yl)amino)-5-propionylpyridin-2yl)cyclopropanecarboxamide, No. (4-((3-methoxy-4-(2-cyclopropyl-2H-1,2,3-triazol-4-yl)pyridin-2 -yl)amino)-5-propionylpyridin-2-yl)cyclopropanecarboxamide, N(4-((2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino )-5-propionylpyridin-2-yl)cyclo

- 4 047032 propanecarboxamide, No. (4-((4-fluoro-2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino)-5-propionylpyridin-2yl )cyclopropanecarboxamide,

H-(4-((3-fluoro-2-methoxy-3-(2-methyl-2H-1.2.3-triazol-4-yl)phenyl)amino)-5-propionylpyridin-2yl)cyclopropanecarboxamide and No(4 -((3-methyl-2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino)-5-propionylpyridin-2yl)cyclopropanecarboxamide.

In another embodiment, the invention provides a pharmaceutical composition comprising one or more compounds of Formula I and a pharmaceutically acceptable carrier or diluent.

The present invention also relates to pharmaceutical compositions for the treatment of diseases associated with the modulation of IL-12, IL-23 and/or IFNa by targeting Tyk-2 to provide inhibition of signal transduction, comprising compounds of formula I or pharmaceutically acceptable salts thereof and pharmaceutically acceptable carriers or thinners.

The invention further relates to methods of treating diseases associated with modulation of IL-12, IL-23 and/or IFNa, comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I.

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

The present invention also provides a method of treating proliferative, metabolic, allergic, autoimmune and inflammatory diseases (or using the compounds of the present invention to make 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 of the compounds of the present invention.

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

The present invention also provides a method of treating a disease (or using the compounds of the present invention for the manufacture of a medicament for the treatment of these diseases), which includes administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I, wherein the disease is 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 scleroderma, ulcerative colitis, Graves' disease, discoid lupus erythematosus, adult-onset Still's disease, juvenile idiopathic arthritis with systemic onset, gout, gouty arthritis, type 1 diabetes mellitus, non-insulin-dependent diabetes mellitus, sepsis, septic shock, shigellosis, pancreatitis (acute or chronic), glomerulonephritis, autoimmune gastritis, diabetes mellitus, 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-Barré syndrome, autoimmune pneumonia, autoimmune thyroiditis, autoimmune inflammatory eye disease and chronic demyelinating polyneuropathy.

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

The present invention also provides a method for treating rheumatoid arthritis (or using the compounds of the present invention for the manufacture 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.

In addition, the present invention also provides a method of treating a condition (or using the compounds of the present invention for the manufacture of a medicament for treating such conditions), which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I, the condition selected from

- 5 047032 acute myeloid leukemia, chronic myeloid leukemia, metastatic melanoma, Kaposi's sarcoma, multiple myeloma, solid tumors, ocular neovascularization and infantile hemangiomas, B-cell lymphoma, systemic lupus erythematosus (SLE), rheumatoid arthritis, psoriatic arthritis, multiple vasculitis, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, allergic rhinitis, multiple sclerosis (MS), transplant rejection, type I diabetes mellitus, membranous nephritis, inflammatory bowel disease, autoimmune hemolytic anemia, autoimmune thyroiditis, cold and heat agglutination diseases, Evans syndrome, hemolytic -uremic syndrome/thrombotic thrombocytopenic purpura (HUS/TTP), sarcoidosis, Sjögren's syndrome, peripheral neuropathies, pemphigus vulgaris and bronchial asthma.

The present invention also provides a method of treating an IL-12, IL-23, and/or IFNa-mediated disease (or using the compounds of the present invention to make a medicament for the treatment of such diseases), which comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I .

The present invention also provides a method of treating an IL-12, IL-23, and/or IFNa-mediated disease (or using the compounds of the present invention for the manufacture of a medicament for treating such 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 IFNa-mediated disease is a disease modulated by IL-12, IL-23 and/or IFNa.

The present invention also provides a method of treating diseases, comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I in combination with other therapeutic agents.

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

In another embodiment, the compounds of Formula I are selected from exemplary compounds or combinations of exemplary compounds or other embodiments described herein.

In another embodiment of the invention, compounds are provided having an IC50 < 1000 nM in at least one of the assays described below.

The present invention may be embodied in other specific forms without departing from the spirit or essential features of the present invention. The present invention covers all combinations of preferred aspects and/or embodiments of the present invention mentioned 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. Moreover, it should be understood that each individual element of the preferred embodiments represents its own independent preferred embodiment. In addition, any element of an embodiment is intended to be combined with any and all other elements from any embodiment to describe a further embodiment.

Detailed Description of the Invention

The following are definitions of terms used in this specification and the accompanying claims. Unless otherwise indicated, the initial definition provided for a group or term herein applies to such group or term throughout the specification and claims, alone or as part of another group.

The compounds of the present invention may have one or more centers of asymmetry. Unless otherwise indicated, all chiral (enantiomeric and diastereomeric) and racemic forms of the compounds of the present invention are included in the present invention. Compounds may also have multiple geometric isomers of olefins, C=N double bonds, and the like, and all such stable isomers are contemplated by the present invention. Cis and trans geometric isomers of the compounds of the present invention are described and can be isolated as a mixture of isomers or as individual isomeric forms. The compounds of the present invention can be isolated in optically active or racemic forms. It is well known in the art how to obtain optically active forms, for example by resolution of racemic forms or by synthesis from optically active starting materials. Unless specific stereochemistry or isomeric form is specifically indicated, the invention provides for all chiral (enantiomeric and diastereoisomeric) and racemic forms and all geometric isomeric forms of the structure.

If any variable (for example, R ³ ) appears more than once in any constituent or formula of a compound, its definition in each occurrence is independent of its definition in each other occurrence. Thus, for example, if a group is shown to be substituted with 0-2 R ³ , then said group may

- 6 047032 be optionally substituted with up to two R ³ groups, and R ³ in each case is independently selected from the definition of R ³ . In addition, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

If a bond with a substituent is shown to cross a bond connecting two atoms in the ring, then that substituent can be bonded to any atom in the ring. If substituents are listed without designating the atom through which such substituent is linked to the residue of a compound of this formula, then such substituent may be linked through any atom in such substituent. Combinations of substituents and/or variables are permitted only if such combinations result in stable compounds.

Where nitrogen atoms (eg, amines) are present in the compounds of the present invention, they can be converted to N-oxides by treatment with an oxidizing agent (eg, MCPBA and/or hydrogen peroxide) to produce other compounds of the present invention. Thus, all nitrogen atoms shown and claimed are considered to include both the nitrogen shown and its Noxide (N^G).

In accordance with the rule used in the art, it is used in the presented structural formulas to represent the bond that is the point of attachment of the moiety or substituent to the core or backbone structure.

The - line, which is not between two letters or symbols, is used to indicate the point of attachment of a substituent. For example, -CONH2 is attached via a carbon atom.

The term optionally substituted with respect to a particular moiety of a compound of formula I (eg, an optionally substituted heteroaryl group) refers to a moiety containing 0, 1, 2 or more substituents. For example, optionally substituted alkyl includes both alkyl and substituted alkyl, as defined below. One skilled in the art should understand with respect to any group containing one or more substituents that such groups are not intended to represent any substitution or substitution pattern that is sterically impracticable, sterically impossible, and/or inherently unstable.

As used herein, the term at least one chemical entity is interchangeable with the term compound.

The term alkyl or alkylene, as used herein, is intended to include saturated aliphatic hydrocarbon groups, both straight and branched chain, containing the number of carbon atoms indicated. For example, Cyoalkyl (or alkylene) is intended to include C1, _C2 , _C3 , _C4 , C5, _C6 , _C7 , C8, C9 and _C10 alkyl groups. In addition, for example, C1- _C6 alkyl means alkyl having from 1 to 6 carbon atoms. Alkyl groups may be unsubstituted or substituted, such that one or more of the hydrogen atoms of such groups is replaced by another chemical group. Example 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, t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl) and the like.

One skilled in the art will understand that when used in this document, the designation CO ₂ is intended to be in the group -c-o-

When the term alkyl is used in conjunction with another group, such as arylalkyl, this configuration more specifically defines at least one of the substituents that will contain the substituted alkyl. For example, arylalkyl refers to a substituted alkyl group, as defined above, in which at least one of the substituents is aryl, such as benzyl. Thus, the term aryl( _C0-4 )alkyl includes substituted lower alkyl containing at least one aryl substituent, and also includes aryl directly linked to another group, i.e. aryl( _C0 )alkyl. The term heteroarylalkyl refers to a substituted alkyl group, as defined above, in which at least one of the substituents is heteroaryl.

The term alkoxy refers to an oxygen atom substituted with alkyl or substituted alkyl as defined herein. For example, the term alkoxy includes an -O-C _1-6 alkyl group such as methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, t-butoxy, pentoxy, 2-pentyloxy, isopentoxy, neopentoxy, hexoxy , 2-hexoxy, 3-hexoxy, 3-methylpentoxy and the like. Lower alkoxy refers to alkoxy groups containing from one to four carbon atoms.

It should be understood that the selection of all groups, including, for example, alkoxy, thioalkyl and aminoalkyl, will be made by one skilled in the art to ensure stable compounds.

As used herein, the term substituted means that any one or more hydrogen atoms on the designated atom or group are replaced by a selection from the designated group, provided that the normal valence of the designated atom is not exceeded. If the substituent is oxo or keto, (i.e. =O), then 2 hydrogen atoms are replaced per atom. Keto substituents are not present on aromatic moieties. Unless otherwise noted, substituents are indicated in the main structure. For example, it should be understood that if (cycloalkyl)alkyl is specified as a possible substituent, then the point of attachment of this substituent to the main structure is at the alkyl moiety. Used in

- 7 047032 herein, double ring bonds are double bonds that are formed between two adjacent ring atoms (eg, C=C, C=N or N=N).

Combinations of substituents and/or variables are permitted only if such combinations result in stable compounds or useful synthesis intermediates. A stable compound or stable structure is meant to mean a compound that is sufficiently stable to persist after being isolated to an acceptable degree of purity from a reaction mixture and subsequently formulated into an effective therapeutic agent. Preferably, the compounds currently listed do not contain an N-halogen, S(O) ₂ H or S(O)H group.

The term cycloalkyl refers to cyclized alkyl groups, including mono-, bi- or polycyclic ring systems. _C3-7 cycloalkyl is intended to include _C3 , _C4 , _C5 , C6 and _C7 cycloalkyl groups. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl, and the like. As used herein, a carbocycle or carbocyclic moiety 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, cycloheptenyl, 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 tetrahydronafil (tetralin). As shown above, bridged rings are also included in the definition of a carbocycle (e.g., [2.2.2]bicyclooctane). Preferred carbocycles, unless otherwise noted, are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and phenyl. When used, the term carbocycle is intended to include aryl. A bridged ring occurs when one or more carbon atoms link two non-adjacent carbon atoms. Preferred bridging bonds are one or two carbon atoms. It is noted that a bridging bond always converts a monocyclic ring into a bicyclic ring. If the ring contains bridging bonds, the substituents listed for the ring may also be present in the bridge.

The term aryl refers to monocyclic or bicyclic aromatic hydrocarbon groups containing from 6 to 12 carbon atoms in the ring portion, such as phenyl and naphthyl groups, each of which may be substituted.

Accordingly, in compounds of formula I, the term cycloalkyl includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclooctyl, etc., as well as the following ring systems:

and the like, which may optionally be substituted at any of the available atoms of the ring(s).

Preferred cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

The term halo or halogen refers to chlorine, bromine, fluorine and iodine.

The term haloalkyl means a substituted alkyl containing one or more halogen substituents. For example, haloalkyl includes mono-, bi- and trifluoromethyl.

The term haloalkoxy means an alkoxy group containing one or more halogen substituents. For example, haloalkoxy includes OCF ₃ .

The terms heterocycle, heterocycloalkyl, heterocyclo, heterocyclic or heterocyclyl can be used interchangeably and refer to substituted and unsubstituted 3-7 membered monocyclic groups, 7-11 membered bicyclic groups and 10-15 membered tricyclic groups in which at least one of the rings contains at least one heteroatom (O, S or N), wherein said heteroatom-containing ring preferably contains 1, 2, or 3 heteroatoms selected from O, S and N. Each heteroatom-containing ring of such a group may contain one or two an oxygen or sulfur atom and/or one to four nitrogen atoms, provided that the total number of heteroatoms in each ring is four or less, and further provided that the ring contains at least one carbon atom. The nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen atoms may optionally be quaternized. The fused rings that terminate bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or completely unsaturated. Heterocyclogroup mo

- 8 047032 can be attached to any available nitrogen or carbon atom. As used herein, the terms heterocycle, heterocycloalkyl, heterocyclo, heterocyclic and heterocyclyl include heteroaryl groups as defined below.

In addition to the heteroaryl groups described below, exemplary monocyclic heterocyclyl groups include azetidinyl, pyrrolidinyl, oxetanyl, imidazolinyl, oxazolidinyl, isoxazolinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, piperidyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidyl, 2 -oxopyrrolodinyl, 2-oxoazepinyl, azepinyl, 1-pyridonyl, 4-piperidonyl, tetrahydropyranyl, morpholinyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, 1,3-dioxolane and tetrahydro-1,1-dioxothienyl and the like. Exemplary bicyclic heterocyclogroups include quinuclidinyl.

Additional monocyclic heterocyclyl groups include and ^N '

The term heteroaryl refers to substituted and unsubstituted aromatic 5- or 6-membered monocyclic groups, 9- or 10-membered bicyclic groups, and 11- to 14-membered tricyclic groups that contain at least one heteroatom (O, S, or N) of at least at least one of the rings, wherein said heteroatom-containing ring preferably contains 1, 2 or 3 heteroatoms selected from O, S and N. Each ring of a heteroaryl group containing a heteroatom may contain one or two oxygen or sulfur atoms and/or one to four nitrogen atoms, provided that the total number of heteroatoms in each ring is four or less and that each ring contains at least one carbon atom. The fused rings that terminate bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or completely unsaturated. The nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen atoms may optionally be quaternized. Heteroaryl groups that are bicyclic or tricyclic must include at least one fully aromatic ring, and the other fused ring or rings may be aromatic or non-aromatic. A heteroaryl group can be attached at any available nitrogen or carbon atom on any ring. Where valence permits, if said additional ring is cycloalkyl or heterocyclo, it is further optionally substituted with =O (oxo).

Exemplary monocyclic heteroaryl groups include pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, thienyl, oxadiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl and the like.

Exemplary bicyclic heteroaryl groups include indolyl, benzothiazolyl, benzodioxolyl, benzoxazolyl, benzothienyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, ridyl, furopyridyl, dihydroisoindolyl, tetrahydroquinolinyl and the like.

Exemplary tricyclic heteroaryl groups include carbazolyl, benzindolyl, phenanthrolinyl, acridinyl, phenanthridinyl, xanthenyl and the like.

In compounds of formula I, preferred heteroaryl groups include the like, which may be optionally substituted at any available carbon or nitrogen atom.

Unless otherwise indicated, when referring to a specifically named aryl (e.g., phenyl), cycloalkyl (e.g., cyclohexyl), heterocyclo (e.g., pyrrolidinyl, piperidinyl, and morpholinyl), or heteroaryl (e.g., terazolyl, imidazolyl, pyrazolyl, triazolyl, thiazolyl, and furyl ), the reference provides for the inclusion of rings having from 0 to 3, preferably from 0 to 2, substituents selected from those listed above, as appropriate, for aryl, cycloalkyl, heterocyclo and/or heteroaryl groups.

The term carbocyclyl or carbocyclic refers to a saturated or unsaturated monocyclic or bicyclic ring in which all the atoms of all rings are carbon atoms

- 9 047032 leroda. Thus, the term includes cycloalkyl and aryl rings. Monocyclic carbocycles contain from 3 to 6 ring atoms, more typically 5 or 6 ring atoms. Bicyclic carbocycles contain 7 to 12 ring atoms, for example arranged as a bicyclo [4,5], [5,5], [5,6] or [6,6] system, or 9 or 10 ring atoms arranged as a bicyclo [5,6] or [6,6] system. Examples of mono- and bicyclic carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex1-enyl, 1- cyclohex-2-enyl, 1-cyclohex-3-enyl, phenyl and naphthyl. The carbocyclic ring may be substituted, in which case the substituents are selected from those listed above for the cycloalkyl and aryl groups. The term heteroatoms includes oxygen, sulfur and nitrogen.

When the term unsaturated as used herein refers to a ring or group, the ring or group may be fully unsaturated or partially unsaturated.

Throughout the description, groups and their substituents can be selected by one skilled in the art to provide stable moieties and compounds, and compounds useful as pharmaceutically acceptable compounds and/or intermediates useful in the preparation of pharmaceutically acceptable compounds.

The compounds of formula I may exist in free form (without ionization) or may form salts, which are also within the scope of the present invention. Unless otherwise indicated, reference to the compounds of the present invention is intended to include reference to their free form and their salts. The term salt(s) means acid and/or basic salts formed with inorganic and/or organic acids and bases. In addition, the term salt(s) may include zwitterions (internal salts), for example, when a compound of formula I contains both a basic moiety, such as an amine or a pyridine or imidazole ring, and an acidic moiety, such as a carboxylic acid. Pharmaceutically acceptable (ie, non-toxic, physiologically acceptable) salts are preferred, such as, for example, acceptable metal and amine salts in which the cation does not significantly contribute to the toxicity or biological activity of the salt. However, other salts may be used, for example in isolation or purification steps, which may be used during production and are thus provided within the scope of the present invention. Salts of compounds of formula I can be formed, for example, by reacting a compound of formula I with an amount of acid or base, such as an equivalent amount, in a medium, for example a salt-precipitating medium, or an aqueous medium, followed by lyophilization.

Exemplary acid addition salts include acetates (such as those formed with acetic acid or trihaloacetic acid, 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 coated with maleic acid), methanesulfonates (formed with methanesulfonic acid), 2-naphthalene sulfonates, 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.

Examples of basic salts include ammonium salts, alkali metal salts such as sodium, lithium and potassium salts; alkaline earth metal salts such as calcium and magnesium salts; barium, zinc and aluminum salts; salts of organic bases (eg organic amines) such as trialkylamines, eg triethylamine, procaine, dibenzylamine, N-benzyl-β-phenethylamine, 1-ephenamine, N.N'-dibenzylethylenediamine. dehydroabiethylamine, N-ethylpiperidine, benzylamine, dicyclohexylamine or similar pharmaceutically acceptable amines and salts with amino acids such as arginine, lysine and the like. Basic nitrogen-containing groups can be quaternized by means such as lower alkyl halides (for example, methyl, ethyl, propyl and butyl chlorides, bromides and iodides), dialkyl sulfates (for example, dimethyl, diethyl, dibutyl and diamyl sulfates), long-chain halides (for example, decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (for example, benzyl and phenethyl bromides) and others. Preferred salts include monohydrochloride, hydrogen sulfate, methanesulfonate, phosphate or nitrate salts.

As used herein, the expression pharmaceutically acceptable refers to those compounds, substances, compositions and/or dosage forms which, within the bounds of sound medical judgment, are suitable for use in contact with human and animal tissues without causing excessive toxicity, irritation, allergic reaction or other problems or complications in accordance with an acceptable benefit/risk ratio.

As used herein, pharmaceutically acceptable salts refer to derivatives of the disclosed compounds wherein the parent compound is modified to provide its acidic or

- 10 047032 basic salts. Examples of pharmaceutically acceptable salts include, but are not limited to, salts of basic inorganic or organic acid groups such as amines; and alkaline or organic salts of acidic groups such as carboxylic acids. Pharmaceutically acceptable salts include conventional non-toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric and nitric; and salts derived from organic acids, such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic and isetinic, etc.

The pharmaceutically acceptable salts of the present invention can be synthesized from a parent compound that contains a basic or acidic moiety by conventional chemical methods. Typically, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the corresponding base or acid in water or an organic solvent, or a mixture of the two; a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile is generally preferred. A list of suitable salts is provided in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, Easton, PA (1990), the disclosure of which is incorporated herein by reference.

The invention provides for all stereoisomers of the compounds of the present invention, whether in mixture, pure, or substantially pure form. Stereoisomers may include compounds that are optical isomers by virtue of containing one or more chiral atoms, as well as compounds that are optical isomers by virtue of limited rotation about one or more bonds (atropoisomers). The definition of the compounds of the present invention covers all possible stereoisomers and mixtures thereof. In a very specific manner it covers racemic forms and isolated optical isomers having the indicated activity. Racemic forms can be separated by physical means, such as, for example, fractional crystallization, separation or crystallization of diastereomeric derivatives, or separation by chiral column chromatography. Individual optical isomers can be prepared from racemates by traditional methods, such as, for example, the formation of a salt with an optically active acid followed by crystallization.

The present invention is intended to include all isotopes of atoms found in the compounds of the present invention. Isotopes include those atoms that have the same atomic number but different mass numbers. By way of general example and without 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 methods known to one skilled in the art, or by methods similar to those described in the present invention, using the appropriate isotopically labeled reagent instead of those used in others cases of non-isotopically labeled reagent.

The invention also provides prodrugs and solvates of the compounds of the invention. The term prodrug means a compound that, after administration to a subject, undergoes a chemical transformation through metabolic or chemical processes to produce a compound of formula I and/or a salt and/or solvate thereof. Any compound that will be converted in vivo to produce a biologically active agent (ie, a compound of Formula I) is a prodrug within the scope and spirit of the present invention. For example, compounds containing a carboxy group can form physiologically hydrolyzable esters, which serve as prodrugs, hydrolyzing in the body to form compounds of formula I per se. Such prodrugs are preferably administered orally, since hydrolysis in many cases occurs primarily through the action of digestive enzymes. Parenteral administration may be used if the ester is active per se, or in cases where hydrolysis occurs in the blood. Examples of physiologically hydrolyzable esters of compounds of formula I include C _1-6 alkylbenzyl, 4-methoxybenzyl, indanyl, phthalyl, methoxymethyl, C _1-6 alkanoyloxy-C _1-6 alkyl, for example acetoxymethyl, pivaloyloxymethyl or propioyloxymethyl, C _1-6 alkoxycarbonyloxy -C _1-6 alkyl, for example methoxycarbonyloxymethyl or ethoxycarbonyloxymethyl, glycyloxymethyl, phenylglycyloxymethyl, (5methyl-2-oxo-1,3-dioxolen-4-yl)methyl and other well-known physiologically hydrolyzable esters used, for example, in the production of penicillins and cephalosporins. Such esters can be prepared by conventional methods known in the art.

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).

The compounds of formula I and their salts may exist in a tautomeric form, in which hydrogen atoms are transferred to other parts of the molecules, and chemical bonds between the atoms of the molecules are sequential

- 11 047032 regrouped. It should be understood that all tautomeric forms, if possible, are included in the present invention. In addition, the compounds of the present invention may contain trans and cis isomers.

It should also be understood that solvates (eg, hydrates) of the compounds of formula I are also within the scope of the present invention. Solvation methods are generally known in the art.

Practical value

The compounds of the invention modulate IL-23-stimulated and IFNa-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.

Accordingly, the compounds of formula I are of practical value for the treatment of conditions associated with modulation of IL-23 and/or IFNa function, and in particular with selective inhibition of IL-23, IL-12 and/or IFNa, by targeting Tyk2 to mediate signal transmission. Such conditions include IL-23, IL-12, or IFNa-associated diseases in which pathogenic mechanisms are mediated by these cytokines and subsequent activation of the Tyk2 pathway with subsequent proinflammatory responses that may occur in the peripheral and/or central regions.

As used herein, the terms treat or treatment include treating a disease state in a mammal, particularly a human, and includes: (a) preventing or delaying the occurrence of a disease state in the mammal, particularly if said mammal is predisposed to such disease condition, but its presence has not yet been diagnosed; (b) inhibition of the disease state, i.e. stopping its development; and/or (c) achieving complete or partial relief of symptoms or disease state and/or alleviation, improvement, reduction or cure of a disease or disorder and/or its symptoms.

In view of their activity as modulators of IL-23-, IL-12- and/or IFNa-stimulated cellular responses, the compounds of formula I are useful in the treatment of IL-23-, IL-12- and/or IFNa-associated diseases, including restrictions 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, cutaneous lupus, lupus nephritis, discoid lupus erythematosus, psoriasis; autoinflammatory diseases, including CAPS, TRAPS, FMF, adult-onset Still's disease, juvenile idiopathic arthritis with systemic onset, gout, gouty arthritis; metabolic diseases, including type 2 diabetes mellitus, atherosclerosis, myocardial infarction; destructive bone diseases such as bone resorption disease, osteoarthritis, osteoporosis, multiple myeloma-associated bone disorders; proliferative diseases such as acute myeloid leukemia, chronic myeloid leukemia; angiogenic disorders, such as angiogenic disorders including solid tumors, ocular neovascularization and infantile hemangiomas; 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 CIS, optic neuritis, neuromyelitis optica), cerebral ischemia or neurodegenerative diseases caused by traumatic injury, cancer and viral diseases such as metastatic melanoma, Kaposi's sarcoma, multiple myeloma and HIV infection and CMV retinitis, AIDS, respectively.

More specifically, specific conditions or diseases that may be treated with the compounds of the present invention include, but are not limited to, pancreatitis (acute or chronic), asthma, allergies, 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 mellitus, 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-related arthritis, acute synovitis, P-cell disease pancreas; diseases characterized by massive infiltration of neutrophils; rheumatoid spondylitis, gouty arthritis and other arthritic conditions, cerebral malaria, chronic pulmonary inflammatory disease, silicosis, pulmonary sarcoidosis, bone resorption disease, allograft rejection, fever and myalgias due to infection, cachexia, keloid formations, scar tissue formation, ulcerative colitis , fever, flu, osteoporosis, osteoarthritis, acute myeloblastic leukemia, chronic

- 12 047032 myeloid 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 multiple sclerosis, including CIS, optic neuritis, neuromyelitis optica), cerebral ischemia or traumatic injury-induced neurodegenerative diseases; angiogenic disorders, including solid tumors, ocular neovascularization, and infantile hemangiomas; viral diseases including acute hepatitis infection (including hepatitis A, hepatitis B and hepatitis C), HIV infection and CMV retinitis, AIDS, ARC or malignancy and herpes; stroke, myocardial ischemia, ischemia in stroke-induced acute heart failure, organ hypoxia, vascular hyperplasia, 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 are methods of treatment in which the disease is selected from Alzheimer's disease, Parkinson's disease, ALS, multiple sclerosis (RMS and/or progressive multiple sclerosis, including CIS, optic neuritis, neuromyelitis optica).

When the terms IL-23-, IL-12- and/or IFNa-associated condition or IL-23-, IL-12- and/or IFNa-associated disease or disorder are used herein, each is intended to cover all the conditions defined above, as if repeated in full detail, as well as any other condition influenced by IL-23, IL-12 or IFNa.

Therefore, the present invention provides methods for treating such conditions, comprising administering to a subject in need thereof a therapeutically effective amount of at least one compound of Formula I or a pharmaceutically acceptable salt thereof. A therapeutically effective amount is intended to encompass an amount of a compound of the present invention that is effective when administered alone or in combination to inhibit the function of IL-23, IL-12 or IFNa and/or treat diseases.

Methods of treating IL-23-, IL-12-, or IFNa-associated conditions may include administering compounds of Formula I alone or in combination with each other and/or with other suitable therapeutic agents used in the treatment of such conditions. Accordingly, a therapeutically effective amount also includes an amount of a combination of the claimed compounds that is effective for inhibiting the function of IL-23, IL-12 or IFNa and/or treating diseases associated with IL-23, IL-12 or IFNa.

Examples of such other therapeutic agents include corticosteroids, rolipram, calphostin, cytokine suppressive anti-inflammatory drugs (CSAID), interleukin 10, glucocorticoids, salicylates, nitric oxide and other immunosuppressants; nuclear translocation inhibitors such as deoxyspergualin (DSG); nonsteroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen, celecoxib and rofecoxib; steroids such as prednisolone or dexamethasone; antivirals such as abacavir; antiproliferative agents such as methotrexate, leflunomide, FK506 (tacrolimus, PROGRAF®); antimalarial drugs such as hydroxychloroquine; cytotoxic drugs such as azatiprine and cyclophosphamide; TNF-a inhibitors such as tenidap, anti-TNF antibodies or soluble TNF receptor and rapamycin (sirolimus or RAPAMUNE®) or derivatives thereof.

The above other therapeutic agents, when used in combination with the compounds of the present invention, can be used, for example, in the amounts specified in the Physicians' Desk Reference (PDR) or otherwise determined by one of ordinary skill in the art. In the methods of the present invention, such other therapeutic agent(s) may be administered prior to, concurrently with, or after administration of the compounds of the present invention. The present invention also provides pharmaceutical compositions capable of affecting IL-23-, IL-12-, or IFNa-associated conditions by inhibiting Tyk2-mediated signaling, including IL-23, IL-12, or IFNa-mediated diseases, as described above.

The compositions of the present invention may contain other therapeutic agents described above and may be prepared, for example, using conventional solid or liquid excipients or diluents, as well as pharmaceutical additives of the type appropriate to the type of administration desired (for example, excipients, binders, preservatives , stabilizers, flavorings, etc.) in accordance with techniques that are well known in the art of pharmaceutical preparations.

Accordingly, the present invention further includes compositions containing one or more compounds of Formula I and a pharmaceutically acceptable carrier.

The term pharmaceutically acceptable carrier refers to media accepted in the art for the delivery of biologically active agents to animals, in particular mammals. Pharmaceutically acceptable carriers are formulated according to a number of factors under consideration.

- 13 047032 within the competence of specialists in this field of technology. These include, without limitation, the type and nature of the active agent formulated; the subject to whom the composition containing the agent is to be administered; the intended route of administration of the composition and the target therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous liquid vehicles, as well as a variety of solid and semi-solid dosage forms. Such carriers may include a variety of different ingredients and additives in addition to the active agent, such additional ingredients being included in the dosage form for a variety of reasons, for example, to stabilize the active agent, binders, etc., well known to one skilled in the art. this field of technology. Descriptions of suitable pharmaceutically acceptable carriers and factors involved in their selection can be found in various readily available sources, such as, for example, Remington's Pharmaceutical Sciences, 17th Edition (1985), which is incorporated herein by reference in its entirety.

The compounds of Formula I can be administered by any route suitable for the condition being treated, which may depend on the need for localized treatment or the amount of drug delivered. Topical administration is generally preferred for skin-related diseases, and systemic treatment is preferred for malignant or pre-carcinogenic conditions, although other modes of delivery are contemplated. For example, the compounds can be delivered orally, for example, in the form of tablets, capsules, granules, powders or liquid dosage forms, including syrups; topically, for example, in the form of solutions, suspensions, gels or ointments; sublingual; buccal; parenterally, for example, by subcutaneous, intravenous, intramuscular or intrathoracic injection or infusion routes (for example, in the form of sterile injectable aqueous or non-aqueous solutions or suspensions); intranasally, for example, by means of an inhalation spray; topically, for example, in the form of a cream or ointment; rectally, for example, in the form of suppositories; or by encapsulation in liposomes. Unit dosage forms containing non-toxic, pharmaceutically acceptable carriers or diluents may be administered. The compounds may be administered in a form suitable for immediate release or sustained release. Immediate release or sustained release can be achieved with suitable pharmaceutical compositions or, in particular in the case of sustained release, with devices such as subcutaneous implants or osmotic pumps.

Typical compositions for topical administration include a topical carrier such as PLASTIBASE® (mineral oil thickened with polyethylene).

Exemplary compositions for oral administration include suspensions that may contain, for example, microcrystalline cellulose for bulking, 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. in this field of technology; and immediate release tablets, which may contain, for example, microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and/or lactose and/or other excipients, binders, fillers, disintegrants, diluents and lubricants, such as those known in the art this field of technology. The compounds of the present invention can also be administered orally by sublingual and/or buccal administration, for example, with molded, compressed or freeze-dried tablets. Typical compositions may include instant diluents such as mannitol, lactose, sucrose and/or cyclodextrins. In addition, such formulations may contain high molecular weight excipients such as celluloses (AVICEL®) or polyethylene glycols (PEG); mucosal adhesion auxiliaries such as hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), sodium carboxymethylcellulose (SCMC) and/or maleic anhydride copolymer (eg GANTREZ®); and release control agents such as polyacrylic copolymer (eg CARBOPOL 934®). Lubricants, glidants, flavors, coloring agents and stabilizers may also be added for ease of manufacture and use.

Typical compositions for nasal aerosol or inhalation administration include solutions that may contain, for example, benzyl alcohol or other suitable preservatives, absorption accelerators to enhance absorption and/or bioavailability, and/or other solubilizing or dispersing agents, such as those known in the art.

Typical 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 dispersants or humectants and suspending agents, including synthetic mono- or diglycerides and fatty acids, including oleic acid.

Typical compositions for rectal administration include suppositories, which may

- 14 047032 contain, for example, suitable non-irritating excipients such as cocoa butter, synthetic glycerol esters or polyethylene glycols, which are solid at ordinary temperatures but liquefy and/or dissolve in the rectal cavity to release the drug.

The therapeutically effective amount of a compound of the present invention can be determined by one skilled in the art and includes typical dosages for a mammal ranging from about 0.05 to 1000 mg/kg; from 1 to 1000 mg/kg; from 1 to 50 mg/kg; from 5 to 250 mg/kg; from 250 to 1000 mg/kg body weight of the active compound per day, which can be administered as a single dose or in individual divided doses, for example 1 to 4 times a day. It should be understood that the specific dosage and dosing frequency for any particular subject may vary and will depend on a number of factors, including the potency of the particular compound used, the metabolic stability and duration of action of the compound, species, age, body weight, general health, sex, and the subject's diet, route and time of administration, rate of elimination, drug combinations, and severity of the particular condition. Preferred subjects for treatment include animals, most preferably mammalian species such as humans and domestic animals such as dogs, cats, horses and the like. Therefore, when the term patient is used herein, the term is intended to include all subjects, most preferably mammalian species, that are dependent on modulation of IL-23-, IL12- and/or IFNa-mediated functions.

The compounds of the present invention can be prepared by a number of methods well known to one skilled in the art of organic synthesis. The compounds of the present invention can be synthesized using the procedures described below, in conjunction with synthetic methods known in the field of synthetic organic chemistry, or variations thereof, as will be apparent to those skilled in the art. Preferred methods include, but are not limited to, those described below. All references cited herein are incorporated herein in their entirety by reference.

The compounds of the present invention can be prepared using the reactions and procedures described in this section. Reactions are carried out in solvents appropriate to the reagents and materials used and are suitable for the transformations being carried out. In addition, in describing the synthesis procedures described below, it should be understood that all proposed reaction conditions, including solvent selection, reaction atmosphere, reaction temperature, experimental duration, and work-up procedures, are chosen as standard conditions for a given reaction, which should be readily recognizable specialist in this field of technology. One skilled in the art of organic synthesis will appreciate that the functionality present in the various parts of the molecule must be compatible with the proposed reagents and reactions. Such limitations on substituents that are compatible with the reaction conditions will be apparent to one skilled in the art and alternative techniques must be used. Sometimes this will require a decision to change the order of synthetic steps or to select one particular process design over another in order to obtain the target compound of the invention. It should also be recognized that another important consideration when planning any synthetic route in the art is the rational choice of the protecting group used to protect the reactive functional groups present in the compounds described in the present invention. An authoritative source describing the many alternatives for the qualified practitioner is Greene and Wuts (Protective Groups In Organic Synthesis, Third Edition, Wiley and Sons, 1999).

The key intermediates shown in FIG. 1 can be combined to produce compound 1 by various methods known to those skilled in the art of synthetic organic chemistry.

- 15 047032

Fig.

Scheme 1 shows how intermediate Ia, where X = halogen, such as iodide, in cases where I = simple alkyl (methyl, ethyl, etc.), and intermediate Ib can be combined in the presence of an appropriate base, preferably potassium carbonate, in a suitable solvent, preferably DMF, to obtain the intermediate compounds of formula II. In the case where R=cyclopropyl, Ib can be treated with cyclopropylboronic acid in the presence of copper(II) acetate, 2,2'-bipyridine and sodium carbonate in dichloroethane at elevated temperature. The compound of Formula II can then be monodebrominated in the presence of a strong reducing base, in particular isopropylmagnesium bromide, with a solution of THF in ether at low temperature to give the intermediates of Formula IIa. The compound of formula II can also be used as is to prepare higher-substituted 1,2,3-triazoles. The compound of formula IIa can be used as is or converted to the corresponding boronic acid (IIb) by metal halide exchange followed by quenching with a trialkyl borate, in particular trimethyl borate or triisopropyl borate. The preferred base for metal-halogen exchange may be the isopropyl magnesium chloride-lithium chloride complex in THF at low temperature.

Scheme 1

Scheme 2 shows how one skilled in the art can combine intermediate IIa or IIb with intermediate Ic, where Y = boronate, in the case of reaction with IIa, or halide, in the case of reaction with IIb, to obtain intermediates of general formula III (intermediates of general formula Ic are commercially available or can be prepared by methods well known to those skilled in the art of organic synthesis). The conversion can be accomplished by those skilled in the art by a transition metal-catalyzed coupling reaction of the appropriate boronate with the appropriate halide. More specifically, this conversion can be accomplished by a Suzuki coupling reaction using PdCl ₂ (dppf)[DCM] as a catalyst and aqueous tribasic potassium phosphate as a base in solvents such as 1,4-dioxane at elevated temperatures. In a similar manner, fully substituted 1,2,3-triazoles (intermediates of general formula IIa) can be prepared with intermediate II. In these cases, it is necessary to take the appropriate bromotriazole and subject it to an additional palladium-catalyzed coupling reaction with alkyl or alkenyl boronates (in this case, followed by reduction of olefins by methods known in the art, i.e., catalytic hydrogenation).

- 16 047032

Scheme 2

Scheme 3 shows how one skilled in the art of organic synthesis can combine intermediates of general formula Id (see WO 2020/086616) with amides/amines to obtain intermediates of general formula IV or IVa. In particular, favorable conditions for this reaction involve the use of a Buchwald-type coupling reaction using Pd ₂ (dba) ₃ as a catalyst, xanphos as a ligand and CS ₂ CO ₃ as a base in 1,4dioxane as a solvent, at elevated temperatures . This catalyst/ligand/base system can be modified in ways known to those skilled in the art.

Scheme 3

Scheme 4 shows how one skilled in the art of organic synthesis can combine compound IV with an appropriate substrate to produce compounds of general formula 1. Compounds of general formula IV are combined with primary amides of general formula Ig or aromatic amines of general formula Ih under transition metal-catalyzed conditions. In particular, favorable conditions for this reaction involve the use of a Buchwald-type coupling reaction using Pd2(dba)3 as catalyst, xantphos as ligand and CS2CO3 as base in 1,4dioxane as solvent, at elevated temperatures. This catalyst/ligand/base system can be modified in ways known to those skilled in the art.

- 17 047032

Scheme 4

Receipt

All reagents purchased from commercial sources were used without further purification unless otherwise stated. All reactions with air- or moisture-sensitive reagents were carried out under an inert atmosphere. Proton and carbon magnetic resonance spectra (1H and ^13C NMR) were recorded on a Bruker Avance 400 or JEOL Eclipse 500 spectrometer and are presented in ppm relative to the reference solvent of the sample in which they were performed. HPLC and LCMS analyzes were performed on a Shimadzu LC-10AS liquid chromatograph with an SPDUV-vis detector at 220 or 254 nm, and MS detection was performed on a Micromass Platform LC spectrometer.

LCMS-Method A:

A linear gradient from 20% to 100% solvent B over 4 min with a 0.6 min hold at 100% B and then a 0.1 min gradient to 20% B and a 0.3 min hold at 20% B.

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

Solvent: B: ACN: buffer (98:02)

Flow rate:1.0ml/min

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

Ultraviolet (UV) imaging at 220 nanometers (nm).

LCMS-Method B:

Linear gradient from 5% to 95% solvent B over 2.5 min with a 1.5 min hold at 95% B and then a 0.5 min gradient to 5% B and a 1.5 min hold at 5% B .

Solvent A: 0.1% TFA in _H2O

Solvent: B: 0.1% TFA in ACN

Flow rate:1.5ml/min

Column: XBridge C8 (50 x 4.6) mm, 3.5 µm

Ultraviolet (UV) imaging at 220 nanometers (nm).

GCMS method:

Chromatographic column: HP-5 (30 m x 320 µm x 0.25 µm)

Column length 30 m, internal diameter 0.32 mm, thickness 0.25 µm

Inlet temperature: 250°C;

Carrier gas: Not;

Detector temperature: 300°C;

column flow 2 ml/min;

air flow 400 ml/min;

H2 flow 40 ml/min;

Heating schedule: 120°C, hold for 3 minutes;

then increase the temperature to 300°C at a rate of 40°C/min and hold for 2 minutes, source temperature: 230°C.

- 18 047032

Abbreviation Meaning ACN Acetonitrile DIPEA Diisopropylamine LiHMDS Lithium bis(trimethylsilyl)amide EtOH Ethanol EtOAc Ethyl acetate THF T ethrahydrofuran DCM Dichloromethane TBAF Tetra-n-butylammonium fluoride DMF Ν,Ν' -Dimethylformamide T.F.A. Trifluoroacetic acid DAST Diethylaminosulfur trifluoride _Tf2O Trifluoromethanesulfonic anhydride dba Xantphos dcpf dppf MeOH DIC HPLC DIAD LC MS rt Pd/C Et Me h °C PBSF HATU DMA MW AcOH DMAP dibenzylideneacetone 4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene 1,1 '-Bis(dicyclohexylphosphino)ferrocene 1,1 '-Bis(diphenylphosphino)ferrocene Methanol Ν, N'-Diisopropylcarbodiimide high performance liquid chromatography Diisopropyl azodicarboxylate liquid chromatography mass spectrometry Room temperature palladium on carbon Ethyl Methyl clock °C Celsius Perfluorobutanesulfonyl fluoride 1-[Bis(dimethylamino)methylene]-7//-1,2,3-triazolo[4,5/>]pyridinium 3-oxide hexafluorophosphate dimethylacetamide microwave Acetic acid 4-dimethyl-aminopyridine Boc Tert-butoxycarbonyl AcCl Acetyl chloride min minutes MHz megahertz m-CPBA meta-chloroperoxybenzoic acid

Intermediate-1:

Stage-1.

4-19 047032 bromopyridin-3-ol (1.70 g, 9.77 mmol) was added to a 250 mL three-neck round bottom flask cooled to -10°C. Concentrated sulfuric acid (5 ml) was added dropwise over 10 min at -10°C with slow stirring under N2 atmosphere. The mixture was continued to stir at the same temperature for 10 min, 4-bromopyridin-3-ol was completely dissolved to form a clear solution. Nitric acid (fuming, 437 μL, 9.77 mmol) was added dropwise over 10 min at −10°C. The resulting mixture was gradually (~1.5 h) brought to room temperature and stirred for 10 h. The reaction mixture was very carefully poured into crushed ice (~150 g). After complete cooling, the mixture was extracted with CH ₂ Cl ₂ (3x50 ml). The resulting organic layer was washed with brine (30 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give 4-bromo-2-nitropyridin-3-ol (1.01 g crude). It was used for the next step without further purification.

GCMS (M) m/z: 218.0 [M] ⁺ . GC retention time 7.36 min.

¹ H-NMR (400 MHz, MeOH-d ₄ ): δ 8.00 (d, J=4.8 Hz, 1H), 7.94 (d, J=4.8 Hz, 1H).

Stage-2.

4-Bromo-2-nitropyridin3-ol (6 g, 27.4 mmol) and DMF (100 mL) were placed in a 250 mL three-neck round bottom flask with a stirrer. The mixture was stirred at room temperature until a clear solution formed (~5 min). K ₂ CO ₃ (7.57 g, 54.8 mmol) was added to the resulting solution in portions and the mixture was stirred at room temperature for 10 minutes. Methyl iodide (3.43 ml, 54.8 mmol) was added dropwise over 5 minutes and the mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with water (60 ml), extracted with EtOAc (3x100 ml). The combined organic layers were washed successively with ice water (2x100 ml) and brine (100 ml). The resulting organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica column chromatography using 0-25% EtOAc in petroleum ether as the mobile phase to give 4-bromo-3-methoxy-2-nitropyridine as a white solid (4.61 g, 71% yield) %).

MS (M+1) m/z. 234.9 [M+H] ⁺ . LC retention time 0.66 min [Method B].

1H-NMR (400 MHz, DMSO-d6): δ 8.25 (d, J= 5.2 Hz, 1H), 8.20 (d, J= 5.2 Hz, 1H), 3.97 (s ,3H).

Stage 3.

4-Bromo-3-methoxy-2nitropyridine (4.70 g, 21.5 mmol), AcOH (20 mL), EtOH (20 mL) and water (10 mL) were placed in a 250 mL three-neck round bottom flask with a stirrer. The mixture was stirred at room temperature until a clear solution formed (~5 min). The mixture was cooled to 0°C. Iron powder (12.0 g, 151 mmol) was added in portions over 10 min at 0°C. The mixture was warmed to room temperature and stirred for 4 hours. The mixture was filtered through a celite pad, washed with EtOAc (2x100 ml). The filtrate was washed successively with saturated aqueous NaHCO ₃ (2x100 ml) and brine (50 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by silica column chromatography (230-400 mesh) using 0 to 60% ESA Ac in petroleum ether to give 4-bromo-3-methoxypyridin-2-amine (3.5 g, 80% yield) in as a white solid.

MS (M+1) m/z: 205.0 [M+H] ⁺ .

LC retention time 0.66 min [Method B].

1H-NMR (400 MHz, DMSO-d ₆ ): δ 7.54 (d, J= 5.2 Hz, 1H), 6.72 (d, J= 5.2 Hz, 1H), 3.69 ( s, 3H). Intermediate 2.

Br ^Meo vS / , / NN / iPrMgCl.LiCI, _K1 ./ _u // j..

NN (MeO) ₃ B, THF У ^H 2 ^NN

-20 °C, 1h. I Pd(PPh ₃ ) ₄ ^Me °yA

Br Stage-l B(OH) ₂ Cs ₂ CO ₃ , 1,4-Dioxane, JL JJ

120 °C, 3h. _H2NN

Stage-2

Stage 1.

To a stirred solution of 4-bromo-2-methyl-2H-1,2,3-triazole (5.0 g, 30.9 mmol) in THF (50 mL), isopropylmagnesium chloride-lithium chloride complex (3.17 g, 30.9 mmol) at 0°C. The reaction mixture was stirred for 2 hours at this temperature and then cooled to -20°C. Trimethyl borate (0.64 mL, 5.7 mmol) was slowly added to this solution. The reaction mixture was stirred at -20 °C for 1 hour, after which the reaction mixture was acidified with an aqueous solution of 1N HO to pH ~5. The resulting mixture was stirred for 10 minutes at 0°C. The reaction mixture was partitioned between EtOAc (50 ml) and water (50 ml). The organic layer was collected, the aqueous layer was again extracted with EtOAc (2x100 ml) and the combined organic layers were washed with brine (50 ml), then dried over anhydrous Na ₂ SO ₄ . The organic solvent was removed under pressure to obtain the crude product. The resulting crude product was washed with 20 ml of n-pentane to obtain the desired (2-methyl-2H-1,2,3-triazol-4-yl)boronic acid (2.6 g, 66.3% yield).

- 20 047032

MS (M+1) m/z: 128.0 [M+H] ⁺ .

LC retention time 0.66 min [Method B].

1H-NMR (400 MHz, DMSO-d6): δ 8.34 (s, 2H), 7.89 (s, 1H), 4.12 (S, 3H).

Stage 2.

Cesium carbonate (0.963 g, 2.96 mmol), (2-methyl-2H-1,2,3-triazol-4-yl)boronic acid (0.281 g, 2.216 mmol) and purged under N2 gas for 5 min, then tetrakis(triphenylphosphine)palladium( 0) (0.085 g, 0.074 mmol) and heated at 120°C for 3 hours in a sealed tube. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (25 ml), filtered through a celite pad and washed with ethyl acetate (25 ml). The filtrate was washed successively with water (25 ml) and brine (20 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash column chromatography using 0 to 30% EtOAc in petroleum ether to give the desired product 3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin- 2-amine (0.22 g, 72.6% yield) as a yellow solid.

MS (M+1) m/z: 206.2 [M+1] ⁺ . LC retention time 0.36 min [method A].

Intermediate 3.

Η

Ν'\, Br^\

Vg

C^B(OH) ₂

Cu(OAc) ₂ , 2,2-bipy, Na ₂ CO ₃ , 1,2-DCE, 85 °C, 15h.

Stage-1 lk ^N

Br^\ Br 'PrMgCI

THF, 0 °C, 2h.

Stage-2

N'PrMgCI.LiCI

II

Вг (ОМе)зВ, THF, -20 °C, 3h.

Stage-3

N'4

II ,N but- ^B ' ^he

Vg

MeO

_H2N

Pd(PPh ₃ ) ₄ Na ₂ CO ₃ ,1,4-Dioxane, 120 °C, 2h.

Stage-4

_H2N

MeO

Stage 1:

2,2'-Bipyridine (25.2 g, 161 mmol) was added to a stirred suspension of copper (II) acetate (29.3 g, 161 mmol) in 1,2-dichloroethane (500 ml) in a 1000 ml three-neck round bottom flask. and heated the reaction mixture at 80°C for 2 hours. Cyclopropylboronic acid (34.1 g, 397 mmol), 4,5-dibromo-2H-1,2,3-triazole (30 g, 132 mmol) and sodium carbonate (28 .0 g, 264 mmol) in 1,2-dichloroethane (1000 ml) was placed in a 3000 ml three-neck round-bottom flask, the solution of copper (II) complex acetate-2,2'-bipyridine prepared above was added and the reaction mixture was degassed under N2 gas within 5 min. The resulting reaction mixture was purged with _O2 gas for 15 min and then stirred at 85°C for 15 h. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with DCM (1000 ml), filtered through a celite pad and washed thoroughly with DCM (2x500 ml). The resulting filtrate was washed with 1.5 N HCl (2x1000 ml), then brine (1000 ml), dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The resulting crude residue was purified by silica gel column chromatography (230,400 mesh) using 2 to 5% EtOAc in petroleum ether to give 4,5-dibromo-2cyclopropyl-2H-1,2,3-triazole (18 g, 46.4 yield %) in the form of a pale yellow liquid.

GCMS (M) m/z: 266.8 [M] ⁺ . Retention time at GC 3.25 min.

¹ H-NMR (400 MHz, DMSO-d6): δ 4.01-3.98 (m, 1H), 1.38-1.34 (m, 2H), 1.16-1.11 (m, 2H).

Stage 2:

To a stirred solution of 4,5-dibromo-2-cyclopropyl-2H-1,2,3-triazole (18 g, 67.4 mmol) in THF (180 ml) was added isopropyl magnesium chloride (84 ml, 169 mmol) at -20° WITH. The reaction mixture was stirred at this temperature for 30 minutes, then heated to 0°C and stirred for 2 hours. After completion of the reaction, the reaction mixture was quenched by adding saturated aqueous ammonium chloride (50 ml). The reaction mixture was extracted with EtOAc (2x500 ml) and washed with water (500 ml), then brine (500 ml). The collected organic extract was dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography (230-400 mesh) using 5% EtOAc in petroleum ether to give the desired 4-bromo-2-cyclopropyl-2H-1,2,3-triazole (12 g, 90% yield) in the form of a pale yellow liquid.

GCMS (M) m/z: 186.9 [M] ⁺ .

Retention time at GC 2.36 min.

¹ H-NMR (400 MHz, DMSO-d6): δ 7.52 (m, 1H), 4.03-3.97 (m, 1H), 1.45-1.35 (m, 2H), 1 .29-1.14 (m.2H).

Stage 3:

To a stirred solution of 4-bromo-2-cyclopropyl-2H-1,2,3-triazole (12.0 g, 63.8 mmol) in THF

- 21 047032 (100 ml) isopropyl magnesium chloride-lithium chloride complex 1.3 M in THF (58.9 ml, 77 mmol) was slowly added at 10°C. The reaction mixture was stirred for 2 hours at 10°C and then cooled to -20°C. Trimethyl borate (2.487 g, 23.93 mmol) was added to this solution. The resulting reaction mixture was stirred at -20°C for 1 hour. The reaction mixture was acidified with an aqueous solution of 1N HCl until pH ~ 5 was achieved. The resulting mixture was stirred for 10 minutes at 0°C. The reaction mixture was extracted with EtOAc (2x400 ml) and washed with water (200 ml), then brine (200 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure to obtain the crude product. The resulting crude residue was washed with 150 ml of diethyl ether:n-pentane (1:1) to obtain the desired product (2-cyclopropyl-2H-1,2,3-triazol-4yl)boronic acid (6 g, 55.3% yield) as an orange solid.

MS (M+1) m/z: 154.1 [M+1] ⁺ .

LC retention time 1.03 min [Method B].

¹ H-NMR (400 MHz, DMSO-d6): δ 8.34 (s, 2H), 7.89 (s, 1H), 4.14-4.10 (m, 1H), 1.21-1 .19 (m, 2H), 1.151.08 (m, 2H).

Stage 4:

Cesium carbonate (0.353 g, 1.084 mmol) was added to a stirred solution of 4-bromo-3-methoxypyridin-2-amine (0.11 g, 0.542 mmol) in 1,4dioxane (3 mL) and water (0.5 mL). (2-cyclopropyl-2H1,2,3-triazol-4-yl)boronic acid (0.124 g, 0.813 mmol) and purged under N ₂ gas for 5 min, after which tetrakis(triphenylphosphine)palladium(0) (0.031) was added g, 0.027 mmol) and placed in a sealed tube to heat at 120°C for 2 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (25 ml), filtered through a celite pad and washed with ethyl acetate (25 ml). The filtrate was washed successively with water (20 ml) and brine (20 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain the crude product. The crude residue was purified by flash chromatography using 0 to 2% methanol in DCM to give the desired 4-(2-cyclopropyl-2H-1,2,3-triazol-4-yl)-3methoxypyridin-2-amine (103 mg , yield 82%) as a yellow solid.

MS (M+1) m/z: 232.2 [M+1] ⁺ .

LC retention time 1.38 min [method A].

Intermediate 4.

Stage 1:

To a stirred solution of 4,6-dichloronicotinaldehyde (8.5 g, 48.3 mmol) in THF (100 ml) was added ethylmagnesium bromide (48.3 ml, 145 mmol, 3.0 M in diethyl ether) and the solution was stirred at 0°C for 2 hours. The reaction mixture was quenched with saturated aqueous NH ₄ Cl (100 ml) at 0°C and extracted with ethyl acetate (2x200 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain the crude material. The crude compound was purified by silica gel column chromatography (20% EtOAc in hexanes) to give 1-(4,6-dichloropyridin-3-yl)propan-1-one (4.35 g, 38.4% yield) as a pale yellow solid substances.

MS (M+1) m/z: 206.0 [M+H] ⁺ .

LC retention time 1.75 min [Method B].

Stage 2:

Dess-Martin periodinan (17, 91 g, 42.2 mmol) and stirred at rt for 2 hours. The reaction mixture was quenched with 10% Na ₂ CO ₃ solution (50 ml) and extracted with ethyl acetate (2x200 ml). The combined organic layer was dried over Na ₂ SO ₄ and concentrated under reduced pressure to obtain the crude material. The crude compound was purified by silica gel column chromatography (20% EtOAc in hexanes) to give 1-(4,6-dichloropyridin-3-yl)propan-1one (3.7 g, 86% yield) as a pale yellow solid.

MS (m+1) m/z: 204.0 [M+H] ⁺ .

LC retention time 1.44 min [Method B].

Stage 3:

To a stirred solution of 1-(4,6-dichloropyridin-3-yl)propan-1-one (0.2 g, 0.98 mmol) in 1,4dioxane (5 ml) was added cyclopropanecarboxamide (0.1 g, 1. 18 mmol), cesium carbonate (0.96 g, 2.94

- 22 047032 mmol). The reaction mixture was degassed for 5 min under N2 gas, then 1,1'bis(dicyclohexylphosphino)ferrocene (0.68 g, 1.18 mmol), Pd ₂ dba ₃ (0.18 g, 0.196 mmol) was added and further degassed within 5 min. The reaction mixture was sealed and stirred at 80°C for 2 hours. The reaction mixture was filtered through a syringe attachment and washed with ethyl acetate (50 ml). The filtrate was concentrated under reduced pressure to obtain crude material. The crude compound was purified by silica gel column chromatography (10% EtOAc in hexanes) to give N(4-chloro-5-propionylpyridin-2-yl)cyclopropanecarboxamide (0.1 g, 40.4% yield) as a pale yellow solid.

MS (M+1) m/z: 253.0 [M+H] ⁺ .

LC retention time 2.25 min [method B].

Example 1.

Cesium carbonate (516 mg, 1.583 mmol), 3-methoxy-4 -(2-methyl-2H-1,2,3triazol-4-yl)pyridin-2-amine (162 mg, 0.791 mmol) and degassed for 5 min under N ₂ gas, after which 1,1'-bis was added (diphenylphosphino)ferrocene (43.8 mg, 0.079 mmol) and Pd2dba3 (36.2 mg, 0.040 mmol). The reaction mixture was stirred at 130°C for 3 hours in a sealed tube. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (20 ml), filtered through a celite pad and concentrated under reduced pressure. The resulting crude product was purified by flash chromatography using 0 to 2% methanol in DCM and triturated with diethyl ether (20 ml) to give the desired product No. (4-((3-methoxy-4-(2-methyl-2H-1 ,2,3-triazol-4yl)pyridin-2-yl)amino)-5-propionylpyridin-2-yl)cyclopropanecarboxamide (54 mg, 15.9% yield) as a white solid.

MS (M+1) m/z. 422.0 [M+H] ⁺ .

LC retention time 2.29 min [Method B].

¹ H-NMR (400 MHz, DMSO-d6): δ 12.33 (s, 1H), 10.93 (s, 1H), 9.68 (s, 1H), 8.97 (s, 1H), 8.32 (s, 1H), 8.15 (d, J=5.20 Hz, 1H), 7.46 (d, J=5.20 Hz, 1H), 4.28 (s, 3H), 3.82 (s, 3H), 3.18 (q, J=7.20 Hz, 2H), 2.08-2.07 (m, 1H), 1.15 (t, J=7.20 Hz , 3H), 0.87-0.83 (m, 4H).

The following example 2 was prepared in the same way as example 1.

N-l/^Ϋ

O ΗνΑγ ^{0 N} H'S/

Intermediate 5.

- 23 047032

Stage 1:

To a stirred solution of 4,6-dichloronicotinic acid (15.0 g, 78.0 mmol) in DMF (220 ml) was added DIPEA (27.3 ml, 156.0 mmol) and HATU (44.6 g, 117.0 mmol) at 0°C. N,O-dimethylhydroxylamine (5.73 g, 94.0 mmol) was then added portionwise at 0°C. The reaction mixture was stirred at room temperature for 6 hours. Cold water (150 ml) was added to the reaction mixture and extracted with ethyl acetate (3x150 ml). The combined organic extracts were washed with brine (100 ml), dried over Na ₂ SO ₄ and concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography to obtain the desired product 4,6-dichloro-M-methoxy-M-methylnicotinamide (12.5 g, 66.9% yield) as a white solid.

MS (M+1) m/z: 235.4 [M+H], ⁺ LC retention time 1.36 min [Method B].

Stage 2:

To a stirred solution of 4,6-dichloro-M-methoxy-M-methylnicotinamide (0.8 g, 3.40 mmol) and 2methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl )aniline (0.69 g, 3.40 mmol) in THF (20 ml) was added LiHMDS (10.21 ml, 10.21 mmol, 1 M solution in THF) at 0°C and stirred for 2 hours at rt . The reaction mixture was cooled to 0°C, quenched with a saturated aqueous solution of NH ₄ Cl (30 ml), and extracted with ethyl acetate (2x100 ml). The organic layer was dried over Na ₂ SO ₄ and then concentrated under reduced pressure to obtain a crude residue. The crude compound was purified by silica gel column chromatography (25% EtOAc in petroleum ether) to give 6-chloro-M-methoxy-4-((2methoxy-3-(2-methyl-2H-1,2,3-triazol-4- yl)phenyl)amino)-M-methylnicotinamide (0.85 g, 58.6% yield) as an orange solid.

MS (M+1) m/z: 403.1 [M+H] ⁺ , LC retention time 2.06 min [Method B].

Stage 3:

To a stirred solution of 6-chloro-M-methoxy-4-((2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4yl)phenyl)amino)-Nmethylnicotinamide (0.2 g , 0.5 mmol) in THF (10 ml) was added ethylmagnesium bromide (0.5 ml, 1.5 mmol, 3.0 M solution in diethyl ether) at 0°C and stirred at this temperature for 2 hours. Reaction the mixture was quenched with saturated aqueous NH ₄ Cl (20 ml) at 0°C and extracted with ethyl acetate (50 ml). The organic layer was dried over Na ₂ SO ₄ and concentrated under reduced pressure to obtain the crude material. The crude compound was purified by silica gel column chromatography (20% ethyl acetate in hexanes) to give 1-(6-chloro-4-((2methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl) phenyl)amino)pyridin-3-yl)propan-1-one (0.14 g, 72.5% yield) as a pale yellow solid.

MS (M+1) m/z: 372.1 [M+H] ⁺ , LC retention time 2.17 min [Method B].

Example 3.

NN ^Z NN ^Z

To a stirred solution of 1-(6-chloro-4-((2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino)pyridin-3-yl)propane -1-one (150 mg, 0.403 mmol) in 1,4-dioxane (3 ml) was added with cesium carbonate (329 mg, 1.009 mmol) and cyclopropanecarboxamide (68.7 mg, 0.807 mmol) at ambient temperature. The reaction mixture was degassed under _N2 for 5 minutes. Pd ₂ dba ₃ (73.9 mg, 0.081 mmol) and Xantphos (46.7 mg, 0.081 mmol) were added to the reaction mixture and degassed for 5 min. The resulting reaction mixture was stirred at MW at 130°C for 2 hours. The reaction mixture was filtered through a pad of celite, washed with EtOAc (50 ml) and concentrated under reduced pressure to obtain the crude product. The crude residue was purified by preparative reverse phase HPLC to give N(4-((2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino)-5propionylpyridin-2- yl)cyclopropanecarboxamide (40 mg, yield 23.16%) as a white solid.

MS (M+1) m/z: 421.0 [M+H] ⁺ , LC retention time 1.18 min [Method A].

1H-NMR (400 MHz, DMSO-d6): δ 11.06 (s, 1H), 10.93 (s, 1H), 8.90 (s, 1H), 8.13 (s, 1H), 8 .03 (s, 1H), 7.70 (d, J=7.6 Hz, 1H), 7.48 (d, J=7.6 Hz, 1H), 7.27-7.31 (m, 1H), 4.24 (s, 3H), 3.64 (s, 3H), 3.14 (q, J=7.2 Hz, 2H), 2.00-2.03 (m, 1H), 1.13 (t, J=7.2 Hz, 3H), 0.78-0.79 (m, 4H).

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Intermediate 6.

Stage 1:

To a stirred solution of 3-bromo-4-fluoro-2-methoxyaniline (800 mg, 3.64 mmol) and (2-methyl2H-1,2,3-triazol-4-yl)boronic acid (554 mg, 4.36 mmol) in 1,4-dioxane (10 ml) was added an aqueous 2N K ₃ PO ₄ solution (3.6 ml, 7.28 mmol) and degassed under N ₂ for 5 min. PdCl ₂ (dppf)-DCM adduct (297 mg, 0.364 mmol) was then added to the reaction mixture and stirred at 90°C for 6 hours in a sealed tube. The reaction mixture was diluted with ethyl acetate (30 ml) and filtered through a celite pad, the celite pad was washed with ethyl acetate (30 ml). The filtrate was washed successively with water (40 ml) and brine (40 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain the crude product. The crude residue was purified by silica gel column chromatography (100-200 mesh) using 30-35% EtOAc in petroleum ether as eluent to give the desired product 4-fluoro-2-methoxy-3(2-methyl-2H-1,2, 3-triazol-4-yl)aniline (520 mg, 64.4% yield) as a brown solid.

MS (M+1) m/z: 223.2 [M+H] ⁺ , LC retention time 1.16 min [Method A].

Intermediate 7.

Stage 1:

To a stirred solution of 3-bromo-5-fluoro-2-methoxyaniline (2 g, 9.09 mmol) in 1,4-dioxane (20 ml) in a sealed tube at ambient temperature was added bispin (2.308 g, 9.09 mmol ) and KOAc (0.892 g, 9.09 mmol). The reaction mixture was purged with N2 gas for 5 min, after which PdCl2(dppf)-DCM adduct (0.742 g, 0.91 mmol) was added. The reaction mixture was stirred at 90°C for 5 hours and cooled to room temperature. The reaction mixture was diluted with EtOAc (100 ml) and filtered through a celite pad. The filtrate was washed with water (50 ml) and brine (50 ml). The collected organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure to obtain the crude product. The crude residue was purified by silica gel column chromatography (100-200 mesh) using 20-25% EtOAc in petroleum ether to give the desired product 5-fluoro-2-methoxy-3-(4,4,5,5-tetramethyl-1,3 ,2-dioxaborolan-2-yl)aniline (2 g, 82% yield) as a white solid.

MS (M+1) m/z: 267.8 (M+H) ⁺ , LC retention time 2.43 min [Method A].

Stage 2:

To a stirred solution of 5-fluoro-2-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2yl)aniline (989 mg, 3.70 mmol) and 4-bromo-2 -methyl-2H-1,2,3-triazole (500 mg, 3.09 mmol) in 1,4-dioxane (10 ml), aqueous 2N K3PO4 solution (4.63 ml, 9.26 mmol) was added and purged with N2 gas within 5 min. PdCl ₂ (dppf)-DCM adduct (252 mg, 0.309 mmol) was added to the reaction mixture and stirred at 90°C for 6 hours. The reaction mixture was diluted with ethyl acetate (50 ml) and washed with brine (25 ml). The orange layer was collected, dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography (100-200 mesh) using 35% EtOAc in petroleum ether to give the desired product 5-fluoro-2-methoxy-3-(2-methyl-2H-1,2,3-triazol- 4-yl)aniline (0.6 g, 38% yield) as a brown solid.

MS (M+1) m/z: 223.0 (M+H) ⁺ , LC retention time 1.95 min [Method A].

The following intermediate 8 was prepared from 3-bromo-2-methoxy-5-methylaniline in a similar manner to the preparation of intermediate 7.

- 25 047032

Intermediate connection No. MW m/z [M+H] ⁺ Rt (min) [Method] 8 218.26 219.2 1.04 [A]

Example 4.

A mixture of 4-fluoro-2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)aniline (376 mg, 1.691 mmol), N-(4chloro-5-propionylpyridin-2- yl)cyclopropanecarboxamide (200 mg, 0.791 mmol), cesium carbonate (774 mg, 2.374 mmol) in 1,4-dioxane (5 ml) was degassed by bubbling N ₂ gas for 5 min. 1,1'-Bis(dicyclohexylphosphino)ferrocene (45.8 mg, 0.079 mmol), Pd ₂ dba ₃ (36.2 mg, 0.040 mmol) was then added and the reaction mixture was degassed by bubbling N2 gas for 5 min. The reaction vessel was sealed and heated to 110°C for 16 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (20 ml), filtered through a 0.45 μm nylon filter and concentrated. The crude residue was purified by reverse phase preparative HPLC to give N(4-((4-fluoro-2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino) -5-propionylpyridin-2-yl)cyclopropanecarboxamide (130 mg, yield 36.8%) as a white solid.

MS (M+1) m/z: 439.2 [M+H] ⁺ , LC retention time 2.517 min [Method B].

¹ H-NMR (400 MHz, DMSO-d6): δ 10.92 (s, 1H), 10.85 (s, 1H), 8.88 (s, 1H), 8.02 (s, 1H), 7.84 (s, 1H), 7.53-7.50 (m, 1H), 7.24 (t, J=9.60 Hz, 1H), 4.25 (s, 3H), 3.55 (s , 3H), 3.13 (q, J=7.20 Hz, 2H), 2.02-1.99 (m, 1H), 1.11 (t, J=7.20 Hz, 3H), 0 .80-0.78 (m, 4H).

The following examples (5-6) were prepared similarly to example 4.

Example No. ^a X MW m/z [M+H] ⁺ Rt (min) [Method] 5 F 438.46 439.0 3.11 [A] 6 СНз 434.5 435.2 2.91 [A]

^a = Rac-BINAP was used as a ligand in the preparation, instead of dcpf.

Example No. 'NMR 2 'H-NMR (400 MHz, DMSO- _d6 ): δ 12.34 (s, 1H), 10.94 (s, 1H), 9.70 (s, 1H), 8.97 (s, 1H) , 8.32 (s, 1H), 8.15 (d, J = 5.60 Hz, 1H), 7.47 (d, J = 5.20 Hz, 1H), 4.30-4.24 ( m, 1H), 3.81 (s, 3H), 3.18 (q, J = 7.20 Hz, 2H), 2.09-2.06 (m, 1H), 1.32-1.29 (m, 2H), 1.18-1.08 (m, 5H), 0.89-0.87 (m, 4H). 5 Ή-NMR (400 MHz, DMSO-d ₆ ): S 11.21 (s, 1H), 11.01 (s, 1H), 8.93 (s, 1H), 8.19 (s, 1H), 8.13 (s, 1H), 7.42-7.36 (m, 2H), 4.25 (s, 3H), 3.65 (s, 3H), 3.15 (q, J = 7, 20 Hz, 2H), 2.08-2.01 (m, 1H), 1.13 (t, J = 7.20 Hz, 3H), 0.86-0.80 (m, 4H). 6 'H-NMR (400 MHz, DMSO- _d6 ): δ 11.15 (s, 1H), 11.05 (s, 1H), 8.87 (s, 1H), 8.11 (s, 1H) , 7.65 (s, 1H), 7.60 (s, 1H), 7.30 (s, 1H), 4.24 (s, 3H), 3.61 (s, 3H), 3.15 (q, J = 7.20 Hz, 2H), 2.35 (s, 3H), 1.97 (m, 1H), 1.13 (t, J = 7.20 Hz, 3H), 0.86-0, 83 (m, 4H).

Biological tests

The following assays are used to demonstrate the activity of the compounds of the invention.

In vivo brain penetration assay

A pharmacokinetic study was conducted in wild-type C57BL6 mice (n=3 per experiment) to determine the brain and plasma effects of the compounds of the invention. The compound was administered orally in a solution of 5% ETHANOL; 90% PEG 300; 5% TPGS at 5 ml/kg to achieve a final concentration of 10 mg/kg. Mice were sacrificed 1 h after drug administration, and plasma and brain were collected and frozen for analysis. Brain tissues were homogenized in a 1:1 volume with pure plasma from C57BL6 mice. The concentration of the compound in plasma and brain homogenate was determined by the method

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LC-MS.

Analysis of bidirectional permeability in Caco-2 cells

Short description

The described compounds were tested using the Caco-2 bidirectional permeability assay to evaluate their permeability and substrate efflux potential. 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]) for 2 h at 37°C and then isolated for LC analysis. MS analysis to determine their concentration in reaction mixtures and calculate the permeability coefficient, efflux coefficient and recovery rate.

Materials and methods

Caco-2 cells (human colon adenocarcinoma) were obtained from the American Type Culture Collection (Manassas, Virginia). Dulbecco's modified medium (DMEM), N-2hydroxyethylpiperazine-No.2-ethanesulfonic acid (HEPES) buffer, nonessential amino acids, L-glutamine, penicillin-O-streptomycin, and heat-inactivated fetal bovine serum (FBS) were purchased from GIBCO/Invitrogen (Carlsbad, California). Transwell 96-well plates (surface area: 0.11 cm ² ) with 0.4 μm pore size polycarbonate membrane and low binding transwell cluster plates were purchased from Sigma Aldrich (Saint 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) with HEPES to pH 7.4. HBSS, digoxin, and bovine serum albumin (BSA) were purchased from Sigma (Saint Louis, Missouri). Filtration blocks (2 mL, 96 wells) were purchased from Whatman (Freiburg, Germany). All solvents were of analytical grade.

Cell preparation

14-28 days before analysis, Caco-2 cells were seeded on polycarbonate filter membranes in 96-well plates at a density of 1.8 x ^{10 5} cells/cm ² , approximately 2.0 x 10 ⁴ cells per well. Cells were grown in DMEM culture medium supplemented with 10% fetal bovine serum, 10 mM HEPES, 1% nonessential amino acids, 2 mM L-glutamine, 100 U/ml penicillin-G, and 100 μg/ml streptomycin. The culture medium was replaced every 3 days and the cells were maintained at 37°C in an atmosphere of 95% relative humidity and 5% CO2. Immediately before analysis, cells were assessed for the formation of dense adhesions (see Quality Control section below).

Preparation of compounds

Compounds were solubilized to 10 mM in 100% DMSO. After visual confirmation of complete solubilization, 10 mM compound stock solutions were added to a 96-well plate and further serially diluted in 100% DMSO to create a 100x concentration of 0.3 mM. Four (4) control compounds were tested along with the described compounds, they were added at 100x concentration of 0.3 mM in quadruplicate.

Permeability assessment

The described compounds were tested in triplicate in one experiment at a final concentration of 3 μM. Cell passages used in the assay passed quality control criteria (see Quality Control below). The study was carried out on monolayers of Caco-2 cells cultured for 14-28 days, with a number of cell passages from 20 to 80.

The assay (vehicle) buffer consisted of MHBSS adjusted to pH 7.4 and 0.5% BSA. From a 100x plate of compounds, 8 µl of a 100% stock solution of compounds in DMSO was added to 800 µl of assay buffer, mixed well and filtered to remove precipitate as a final preparation step before incubation. The target final test concentration of the described compounds and control compounds was 3 μM. The filtrate was a stock solution of the starting compounds, which was used as a donor solution for analysis (in both directions). The receiving solution was only the analytical buffer.

Immediately before analysis, each cell monolayer was washed 3 times with assay buffer to remove all traces of culture medium.

Permeability studies began by adding 100 μL of assay buffer plus/minus compound to the apical compartment of the transwell and 200 μL of assay buffer plus/minus compound to the basolateral compartment of the 96-well cluster low-binding transwell plate. To determine apical-basolateral (A^B) permeability (direction of uptake), buffer containing compounds or control compounds (1x donor solution) was placed in the apical compartments (donor wells), and only buffer was placed in the corresponding basolateral compartments (receiver wells). To determine basolateral-apical (B^A) permeability (secretory direction), buffer with compounds or control compounds (1x donor solution) were placed in the basolateral compartments (donor wells), and only buffer was placed in the corresponding apical compartments (receiver wells). The transwell plates were then incubated for 2 hours at 37°C in an atmosphere of 95%

- 27 047032 relative humidity and 5% CO2. After incubation, 75 μl was removed from each apical and basolateral compartment and transferred to 96-well low-binding plates preloaded with 75 μl/well acetonitrile containing 250 nM propranolol, 250 nM diclofenac, and 500 nM tolbutamide as internal standards. The samples were then analyzed by LCMS/MS to determine the concentrations of the reported compounds and control compounds.

Analysis of analyzed samples

The concentrations of the described compounds and control compounds in the analyzed samples were determined by LC-MS/MS. Multiplex AB Sciex 4500/5500/6500 systems consisted of 2 sets of 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 electrospray ionization (ESI) mode. . To obtain optimal SRM conditions for sample analysis, MS/MS optimization was performed for each compound using Discovery Quant™ software (AB Sciex) with saturation control using 5 μM standard solutions in methanol:water (1:1, v/v ), prepared from initial solutions of compounds. Optimization was carried out by flow injection with an injection volume of 40 μl with isocratic elution of 75% mobile phase B (0.2% formic acid in acetonitrile) and 25% mobile phase A (0.2% formic acid in water).

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

Table A: Bidirectional permeability in the Caco-2 cell assay - Mobile phase gradient for sample analysis Time (s) Length(s) Flow (ml/min) Gradient %A %IN 0 5 0.7 Stage 98 2 5 25 0.7 Ramp 2 98 thirty 20 0.7 Stage 2 98 50 thirty 0.7 Stage 98 2

A=0.2% formic acid in water;

B=0.2% formic acid in acetonitrile

Discovery Quant™ automatically determined the optimal ionization polarity (positive or negative), precursor and product ions, declustering potential, and collision energy for subject and reference compounds. Optimized SRM MS/MS conditions were used for sample analysis. For quantitation, the ratios of the peak areas of the described compounds or a control compound to the internal standard were used. The peak area ratio of the compounds in the dosage solution was used to determine the concentration of the compound in the sample.

Data analysis

The following results were obtained for the compounds described: permeability coefficient (Pc [nanometers per second]), efflux coefficient and percent recovery.

The Pc value was calculated using the following equation:

_n ^At ^x

Рс = t.—l---S x C _DQ x t where

C _At is the concentration of the test compound in the acceptor well after time t,

V _A is the volume in the acceptor well,

S - membrane surface area (0.11 cm ² ),

C _D0 is the initial concentration of the test compound in the donor well, t is the incubation time.

The efflux coefficient was calculated as

R C(c—»A}

Efflux coefficient = —---PC(A^B)

The recovery rate (%) was calculated by expressing the total amount (nmol) of test compound present in the donor and receiver assay compartments at the end of the incubation time (together) as a fraction (percentage) of the total amount (nmol) of test compound added to the donor compartment before incubation. The recovery rate (%) was calculated using the following equation:

- 28 047032

Cdc X Vd + CD^ X Sp % Recovery = °---- X 100

C _DQ x V _D where

C _D0 is the initial concentration of the test compound in the donor well,

VD is the volume in the donor well,

C _Dt - concentration in the donor well after time t,

C _At is the concentration in the acceptor well after time t,

V _A is the volume in the acceptor well.

Quality control

Caco-2 cells in one of the transwell plates used on the day of analysis were assessed for the formation of dense adhesions by measuring transepithelial electrical resistance (TEER). TEER was assessed using an EVOM resistance meter (World Precision Instruments, Sarasota, Florida). In each well of the transwell plate, the TEER value was > 600 Ω·οιη2, and the cell passage and all plates of this batch were accepted for analysis.

Four (4) control compounds with Pc values spanning the permeability range were tested along with the compounds described in each experiment. The acceptance criteria for this assay require that results for the 3 µM control compounds be within acceptable previously observed ranges. Acceptable ranges of Pc values and efflux coefficients previously observed for these 4 control compounds are given in Table. IN.

In these studies, results for all control compounds were within the respective previously observed ranges. Thus, the analysis data were adopted for data analysis and evaluation of compounds describing bidirectional permeability in Caco-2 cells.

Table B: Bidirectional Permeability Assay in Caco-2 Cells Previously Observed Results for Control Compounds Compound Рс (А->В) (nm/s) Рс (В->А) (nm/s) Efflux coefficient Digoxin 18 ± 7 265 ± 74 14.7 Nadolol 20 ±9 25 ±11 1.3 Atenolol 19 ± 8 27 ±10 1.4 Verapamil 120 ±20 160 ±40 1.3

Values represent mean ± standard deviation.

Рс=permeability coefficient.

A^B=apical-basolateral direction.

B^A=basolateral-apical direction.

IFNa-induced STAT phosphorylation in human whole blood

After an hour's incubation with the compound, human whole blood (anticoagulated with ACD-A) was stimulated with 1000 U/ml recombinant human LFNa A/D (R&D Systems 11200-2) for 15 min. Stimulation was stopped by adding Fix/Lyse buffer (BD 558049). Cells were stained with CD3 FITC antibody (BD 555916), washed, and permeabilized on ice with Perm III buffer (BD 558050). Cells were then stained with Alexa-Fluor 647 pSTAT5 (pY694) antibody (BD 612599) for 60 min before analysis on the iQue Plus instrument. The magnitude of pSTAT5 expression was assessed by the median fluorescence intensity after gating of the CD3-positive population.

Table 1. Efficacy of Example Compounds in Human Whole Blood Analysis

Example, no. IFNa pSTAT5 in human whole blood IC50 (µM) Saso-2 AB (nm/s) Efflux coefficient for Caco-2 1 0.037 663 OD 2 0.40 180 0.2 3 0.46 183 OD 4 0.28 545 0.4 5 0.71 258 o, s 6 0.77 152 0.3

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Table 2. Comparison of the CNS penetration profiles of compounds of examples 1 and 3 with compounds A and B

2020/086616 A) (Example 163 from /VO 2020/086616 A)

Surprisingly, it has been found that the 1,2,3-substituted triazole compounds of this invention have a significantly higher brain/plasma ratio than the structurally similar 1,2,4-substituted triazoles. Thus, the compounds of the invention are able to penetrate the blood-brain barrier and may be useful for the treatment of certain neurological disorders.

Claims (7)

1. Compound of formula I or a stereoisomer or pharmaceutically acceptable salt thereof, wherein X represents -N- or -CH-; R ¹ is -C(O)R ^1a ; R ^1a represents C _3-6 cycloalkyl; R ² represents C _1-6 alkoxy; R ³ represents C _1-6 alkyl or C _3-6 cycloalkyl. 2. Compound according to claim 1 of the formula or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R ¹ is -C(O)R ^1a ; R ^1a represents C _3-6 cycloalkyl; R ² represents C _1-6 alkoxy; R ³ represents C _1-6 alkyl or C _3-6 cycloalkyl. 3. Compound according to claim 1 of the formula - 30 047032 or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R ¹ is -C(O)R ^1a ; R ^1a represents C _3-6 cycloalkyl; R ² represents C _1-6 alkoxy; R ³ represents C _1-6 alkyl or C _3-6 cycloalkyl. 4. A compound or a pharmaceutically acceptable salt thereof selected from K-(4-((3-methoxy-4-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-5-propionylpyridin-2yl)cyclopropanecarboxamide, K-(4-((3-methoxy-4-(2-cyclopropyl-2H-1,2,3-triazol-4-yl)pyridin-2-yl)amino)-5-propionylpyridin-2-yl)cyclopropanecarboxamide , K-(4-((2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino)-5-propionylpyridin-2-yl)cyclopropanecarboxamide, K-(4-((4-fluoro-2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino)-5-propionylpyridin-2yl)cyclopropanecarboxamide, K-(4-((3-fluoro-2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino)-5-propionylpyridin-2yl)cyclopropanecarboxamide and K-(4-((3-methyl-2-methoxy-3-(2-methyl-2H-1,2,3-triazol-4-yl)phenyl)amino)-5-propionylpyridin-2yl)cyclopropanecarboxamide. 5. A pharmaceutical composition comprising one or more compounds according to any one of claims 14 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent. 6. Use of a compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof for the treatment of a neurodegenerative disease. 7. Use according to claim 6, wherein the neurodegenerative disease is Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis (RMS and/or progressive MS, including CIS, optic neuritis, neuromyelitis optica).