JAK1 pathway inhibitors for treating cytokine-related disorders

Abstract

The present disclosure relates to JAK1 pathway inhibitors and their use in treating cytokine-associated diseases or disorders, such as cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), and CAR-T cell-associated encephalopathy (CRES).

Description

[Technical Field]

[0001] The present disclosure relates to JAK1 pathway inhibitors and their use in treating cytokine-associated diseases or disorders. [Background technology]

[0002] Cytokine-associated diseases or disorders are characterized by excessive immune activation and include cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), and CAR-T cell-associated encephalopathy (CRES).

[0003] Cytokine release syndrome (CRS) is a direct consequence of the overproduction of inflammatory cytokines caused by supraphysiological levels of immune activation and manifests as a series of clinical symptoms including fever, nausea, fatigue, myalgia, malaise, hypotension, hypoxia, and capillary leak, thereby resulting in potential multi-organ toxicity.

[0004] CRS is an unwanted side effect of immunotherapy for serious disease states, such as cancer. Immunotherapies that can cause CRS include the administration of monoclonal antibodies (mAbs) and, more recently, adoptive T-cell therapy for cancer. Lee et al. Blood. 2014, 124(2):188-195. For example, chimeric antigen receptor (CAR) T-cell therapy uses engineered T cells to target cancer and has already been approved by the FDA for use in certain forms of refractory non-Hodgkin's lymphoma and relapsed childhood lymphoblastic leukemia (ALL).

[0005] The cytokine profile involved in CRS includes two major cellular sources: T lymphocyte-derived cytokines, including interferon gamma (IFN)-γ, IL-2, IL-6, soluble IL-6 receptor (IL-6R), and granulocyte-macrophage colony-stimulating factor (GM-CSF), and cytokines secreted primarily by monocytes and / or macrophages, including IL-1β, IL-6, IL-12, IL-18, and tumor necrosis factor (TNF)-α. Xu XJ, Tang YM. Cancer Lett. 2014;343:172-8; Zhang Y., et al. Sci China Life Sci. 2016;59:379-85; Brentjens R., et al. Mol Ther. 2010;18:666-8.

[0006] Modulation of the excessive cytokine response that leads to CRS has the potential to provide important clinical benefits. For example, tocilizumab, an antibody against the IL-6 receptor (IL-6R), reduces the incidence of severe CRS and is FDA-approved for use in CRS. However, tocilizumab's mechanism of action is limited to anti-IL-6R.

[0007] Hemophagocytic lymphohistiocytosis (HLH), another syndrome of excessive or uncontrolled immune activation, primarily affects infants between birth and 18 months of age, but can also affect adults. HLH can be primary (familial) or secondary, meaning it occurs in the context of other infectious, malignant, rheumatic, or metabolic conditions. Symptoms of HLH include cytopenias, hepatosplenomegaly, and fever. Schram, A. and Berliner, N. Blood. 2005. 125(19), 2908-2914.

[0008] Macrophage activation syndrome (MAS) presents clinically in a manner similar to HLH (and is further considered secondary or acquired HLH) and develops due to increased inflammation associated with infection, rheumatic disease, or malignancy. Borgia, RE et al. Arthritis Rheumatol., 2018, doi:10.1002 / art.40417 (pre-publication). MAS was initially described as associated with juvenile idiopathic arthritis, but is increasingly recognized as a complication of other diseases, such as childhood-onset systemic lupus erythematosus (cSLE). Shimizu M., et al. Clin Immunol. 2013 Feb;146(2):73-6. The development of MAS is characterized by a substantial increase in multiple pro-inflammatory cytokines, i.e., a cytokine storm. Borgia, RE et al. Arthritis Rheumatol., 2018, doi:10.1002 / art.40417 (pre-publication). MAS is a serious condition with a high mortality rate, generally 8-22% for pediatric autoimmune diseases and 10-22% for MAS complicated by cSLE. Borgia, RE et al. Arthritis Rheumatol., 2018, doi:10.1002 / art.40417 (pre-publication).

[0009] CAR-T cell-associated encephalopathy (CRES) is the second most common adverse event associated with CAR-T cell therapy, after CRS. CRES is typically characterized by a state of toxic encephalopathy accompanied by symptoms of confusion and delirium, as well as seizures and cerebral edema. CRES symptoms are biphasic and can occur within the first 5 days and / or 3-4 weeks after cellular immunotherapy. The pathophysiological mechanism is thought to involve passive diffusion of cytokines into the brain of patients treated with CAR-T cell therapy. Reducing or eliminating this mechanism may be beneficial for such patients. (Neelapu, et al. Nat Rev Clin Oncol. 2018, 15(1) 47-62).

[0010] Thus, there is a need to develop new therapeutic approaches for treating cytokine-related diseases or disorders. This application addresses this and other needs. [Brief explanation of the drawings]

[0011] [Figure 1] 1 shows dose-dependent inhibition of IL-6 concentrations upon administration of Compound 1 into the blood compartment during anti-CD3 antibody-induced cytokine release syndrome (see Example B). [Figure 2A] 1 shows dose-dependent inhibition of T cell-derived cytokines (i.e., IL-6) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). 1 shows inhibition of IL-6. [Figure 2B] Figure 1 shows dose-dependent inhibition of T cell-derived cytokines (i.e., IFNγ) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). Figure 2 shows inhibition of IFNγ. [Figure 2C] 1 shows dose-dependent inhibition of T cell-derived cytokines (i.e., GM-CSF) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). [Figure 3A] 1 shows dose-dependent inhibition of monocyte- and / or macrophage-derived cytokines (i.e., IL-12) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). 1 shows inhibition of IL-12. [Figure 3B] 1 shows dose-dependent inhibition of monocyte and / or macrophage-derived cytokines (i.e., IL-1β) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). 1 shows inhibition of IL-1β. [Figure 3C] 1 shows dose-dependent inhibition of monocyte- and / or macrophage-derived cytokines (i.e., IL-18) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). 1 shows inhibition of IL-18. [Figure 4]1 shows that the cytokine IL-5 is unaffected by Compound 1 treatment during concanavalin A-induced cytokine release syndrome (see Example C). Summary of the Invention

[0012] Provided herein are methods for treating a cytokine-associated disease or disorder in a subject in need thereof, the methods comprising administering to the patient a therapeutically effective amount of a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof.

[0013] Provided herein is a JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cytokine-associated disease or disorder in a subject in need thereof.

[0014] Provided herein is the use of a JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for use in treating a cytokine-associated disease or disorder in a subject in need thereof.

[0015] The present invention provides, inter alia, a method for treating a cytokine-associated disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof.

[0016] The methods described herein utilize JAK1 pathway inhibitors, particularly JAK1-selective inhibitors. JAK1-selective inhibitors are compounds that preferentially inhibit JAK1 activity over other Janus kinases. JAK1 plays a central role in many cytokine and growth factor signaling pathways that, when dysregulated, can lead to or contribute to disease states. For example, IL-6 levels are elevated in rheumatoid arthritis, and it has been suggested that IL-6 has deleterious effects in this disease (Fonesca, et al., Autoimmunity Reviews, 8:538-42, 2009). Because IL-6 signals at least in part through JAK1, IL-6 indirectly presents potential clinical benefits through inhibition of JAK1 (Guschin, et al., Embo J 14:1421, 1995; Smolen, et al., Lancet 371:987, 2008). Furthermore, in some cancers, JAK1 is mutated, leading to constitutive, undesirable tumor cell proliferation and survival (Mullighan, Proc Natl Acad Sci US A. 106:9414-8, 2009; Flex, J Exp Med. 205:751-8, 2008). In other autoimmune diseases and cancers, elevated systemic levels of inflammatory cytokines that activate JAK1 may also contribute to the disease and / or associated symptoms. Therefore, patients with such diseases may benefit from inhibition of JAK1. Selective inhibitors of JAK1 may be beneficial while avoiding the unnecessary and potentially undesirable effects of inhibiting other JAK kinases.

[0017] JAK1 pathway inhibitors, specifically Compound 1 (i.e., {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, see Table 1), achieve highly effective dose-dependent modulation of CRS-associated inflammatory cytokines (see, e.g., Examples B and C and Figures 1, 2A-2C, and 3A-3C). Surprisingly, this therapeutic property encompasses multiple pathogenic cytokines and is not limited to the IL-6 / IL-6R system (e.g., unlike tocilizumab). Efficacy is achieved by inhibiting T cell- and monocyte / macrophage-derived cytokines that are clinically highly relevant to the pathogenesis of CRS. Furthermore, the data presented herein relating to the JAK1 inhibitor Compound 1 demonstrate that the therapeutic benefit is achieved without extensive cytokine immunosuppression (as demonstrated by unchanged IL-5 levels) (Figure 4). DETAILED DESCRIPTION OF THE INVENTION

[0018] In some embodiments, the cytokine-associated disease or disorder is cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell-associated encephalopathy (CRES).

[0019] In some embodiments, the cytokine-associated disease or disorder is cytokine release syndrome (CRS).

[0020] In some embodiments, the cytokine-related disease or disorder is hemophagocytic lymphohistiocytosis (HLH).

[0021] In some embodiments, the cytokine-related disease or disorder is macrophage activation syndrome (MAS). In some embodiments, the macrophage activation syndrome is associated with systemic juvenile idiopathic arthritis. In some embodiments, the macrophage activation syndrome is associated with pediatric systemic lupus erythematosus.

[0022] In some embodiments, the cytokine-associated disease or disorder is CAR-T cell-associated encephalopathy (CRES).

[0023] In some embodiments, the present application provides a method of treating cytokine release syndrome in a subject, the method comprising administering to the subject a CAR-T cell therapy and a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, treating is ameliorating or inhibiting. In some embodiments, treating is preventing.

[0024] In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered simultaneously with CAR-T cell therapy.

[0025] In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered after administration of the CAR-T cell therapy.

[0026] In some embodiments, the CAR-T cell therapy is axicabtagene ciloleucel.

[0027] In some embodiments, the CAR-T cell therapy is tisagenlecleucel.

[0028] In some embodiments, the subject has a B-cell malignancy.

[0029] In some embodiments, the subject has diffuse large B-cell lymphoma (DLBCL), primary mediastinal large B-cell lymphoma, high-grade B-cell lymphoma, transformed follicular lymphoma, or acute lymphoblastic leukemia.

[0030] In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is selective for JAK1 over JAK2, JAK3, and TYK2 (i.e., a JAK1-selective inhibitor). For example, a compound described herein, or a pharmaceutically acceptable salt thereof, preferentially inhibits JAK1 over one or more of JAK2, JAK3, and TYK2. In some embodiments, the compound preferentially inhibits JAK1 over JAK2 (e.g., JAK2 / JAK1 IC 50 In some embodiments, the compound or salt is about 10-fold selective for JAK1 over JAK2. In some embodiments, the compound or salt has an IC50 / IC60 ratio of 1 mM ATP. 50 The antibody is about 3-fold, about 5-fold, about 10-fold, about 15-fold, or about 20-fold selective for JAK1 over JAK2, as calculated by measuring the β-glucan bond (β-glucan bond) (see, eg, Example A).

[0031] In some embodiments, the JAK1 pathway inhibitor is a compound in Table 1 or a pharmaceutically acceptable salt thereof. The compounds in Table 1 are selective JAK1 inhibitors (more selective than JAK2, JAK3, and TYK2). The IC obtained by the method of Example A at 1 mM ATP was 50 The values ​​are shown in Table 1.

[0032] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] + Average ≦10 nM (see Example A for assay conditions) ++Average ≦100 nM (see Example A for assay conditions) +++ Mean ≦300 nM (see Example A for assay conditions) a Enantiomer 1 Data b Enantiomer 2 data

[0033] In some embodiments, the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof.

[0034] In some embodiments, the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate.

[0035] The synthesis and preparation of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile and its adipic acid salt can be found in U.S. Patent Application No. 2011 / 0224190 (filed March 9, 2011), U.S. Patent Application No. 2013 / 0060026 (filed September 6, 2012), and U.S. Patent Application No. 2014 / 0256941 (filed March 5, 2014), each of which is incorporated herein by reference in its entirety.

[0036] In some embodiments, the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof.

[0037] In some embodiments, the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamidophosphate.

[0038] The synthesis and preparation of 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide and its phosphate salts can be found, for example, in U.S. Patent Application No. 2014 / 0343030 (filed May 16, 2014), which is incorporated herein by reference in its entirety.

[0039] In some embodiments, the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile or a pharmaceutically acceptable salt thereof.

[0040] In some embodiments, the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile monohydrate.

[0041] The synthesis of ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile and characterization of its anhydrate and monohydrate forms are described in U.S. Patent Application No. 2014 / 0121198 (filed October 31, 2013) and U.S. Patent Application No. 2015 / 0344497 (filed April 29, 2015), each of which is incorporated herein by reference in its entirety.

[0042] In some embodiments, the compounds in Table 1 are those disclosed in U.S. Patent Application No. 2011 / 0224190 (filed March 9, 2011), U.S. Patent Application No. 2014 / 0343030 (filed May 16, 2014), U.S. Patent Application No. 2014 / 0121198 (filed October 31, 2013), U.S. Patent Application No. 2010 / 0298334 (filed May 21, 2010), U.S. Patent Application No. 2011 / 0059951 (filed August 31, 2010), U.S. Patent Application No. 2012 / 01 No. 49681 (filed November 18, 2011), U.S. Patent Application No. 2012 / 0149682 (filed November 18, 2011), U.S. Patent Application No. 2013 / 0018034 (filed June 19, 2012), U.S. Patent Application No. 2013 / 0045963 (filed August 17, 2012), and U.S. Patent Application No. 2014 / 0005166 (filed May 17, 2013), each of which is incorporated herein by reference in its entirety.

[0043] In some embodiments, the JAK1 pathway inhibitor is a compound disclosed in U.S. Patent Application No. 2011 / 0224190 (filed March 9, 2011), U.S. Patent Application No. 2014 / 0343030 (filed May 16, 2014), U.S. Patent Application No. 2014 / 0121198 (filed October 31, 2013), U.S. Patent Application No. 2010 / 0298334 (filed May 21, 2010), U.S. Patent Application No. 2011 / 0059951 (filed August 31, 2010), U.S. Patent Application No. 2012 / 0149 No. 681 (filed November 18, 2011), U.S. Patent Application No. 2012 / 0149682 (filed November 18, 2011), U.S. Patent Application No. 2013 / 0018034 (filed June 19, 2012), U.S. Patent Application No. 2013 / 0045963 (filed August 17, 2012), and U.S. Patent Application No. 2014 / 0005166 (filed May 17, 2013), or a pharmaceutically acceptable salt thereof, each of which is incorporated herein by reference in its entirety.

[0044] In some embodiments, the JAK1 pathway inhibitor is a compound of Formula I [ka] or a pharmaceutically acceptable salt thereof, wherein: X is N or CH; L is C(=O) or C(=O)NH; A is phenyl, pyridinyl, or pyrimidinyl, each of which is selected from one or two independently selected R 1 optionally substituted with a group; Each R 1 is independently fluoro or trifluoromethyl.

[0045] In some embodiments, the compound of Formula I is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof.

[0046] In some embodiments, the compound of Formula I is 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-[4-fluoro-2-(trifluoromethyl)phenyl]piperidine-1-carboxamide or a pharmaceutically acceptable salt thereof.

[0047] In some embodiments, the compound of Formula I is [3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(1-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)azetidin-3-yl]acetonitrile or a pharmaceutically acceptable salt thereof.

[0048] In some embodiments, the JAK1 pathway inhibitor is a compound of Formula II [ka] or a pharmaceutically acceptable salt thereof, wherein: R 2 is C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, or C 3-6 Cycloalkyl-C 1-3 alkyl, where C 1-6 Alkyl, C 3-6 Cycloalkyl and C 3-6 Cycloalkyl-C 1-3 the alkyl is optionally substituted with 1, 2, or 3 substituents independently selected from fluoro, —CF3, and methyl; R 3 is H or methyl, R 4 is H, F or Cl, R 5 is H or F, R 6 is H or F, R 7 is H or F, R 8 is H or methyl, R 9 is H or methyl, R 10 is H or methyl, and R 11 is H or methyl.

[0049] In some embodiments, the compound of formula II is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof.

[0050] In some embodiments, the JAK1 pathway inhibitor is a compound of Formula III [ka] or a pharmaceutically acceptable salt thereof, wherein: Cy 4 CN, OH, F, Cl, C 1-3 Alkyl, C 1-3 Haloalkyl, Cyano-C 1-3 Alkyl, HO-C 1-3 Alkyl, Amino, C 1-3 Alkylamino and di(C 1-3 a tetrahydro-2H-pyran ring optionally substituted with one or two groups independently selected from (alkyl)amino, wherein 1-3 Alkyl and di(C 1-3 Alkyl)amino is F, Cl, C 1-3 Alkylaminosulfonyl and C 1-3 optionally substituted with 1, 2, or 3 substituents independently selected from alkylsulfonyl; R 12 is -CH2-OH, -CH(CH3)-OH or -CH2-NHSO2CH3.

[0051] In some embodiments, the compound of formula III is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile or a pharmaceutically acceptable salt thereof.

[0052] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in an amount of about 100 mg to about 600 mg per day, based on the free base. Thus, in some embodiments, the selective JAK1 pathway inhibitor is administered in an amount of about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, or about 600 mg per day, based on the free base.

[0053] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in an amount of about 200 mg per day on a free base basis.

[0054] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in an amount of about 300 mg per day on a free base basis.

[0055] In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered in an amount of about 400 mg per day on a free base basis.

[0056] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in an amount of about 500 mg per day on a free base basis.

[0057] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in an amount of about 600 mg per day on a free base basis.

[0058] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in an amount of about 200 mg once daily on a free base basis.

[0059] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in an amount of about 300 mg once daily on a free base basis.

[0060] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in an amount of about 400 mg once daily on a free base basis.

[0061] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in an amount of about 500 mg once daily on a free base basis.

[0062] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in an amount of about 600 mg once daily on a free base basis.

[0063] In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered in one or more sustained release dosage forms, each comprising the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof.

[0064] Provided herein are methods of treating a cytokine-related disease or disorder in a subject in need thereof, the methods comprising administering to the subject between about 100 mg and 600 mg per day, on a free base basis, of a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof, wherein the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in one or more sustained-release dosage forms comprising the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof.

[0065] The embodiments described herein are intended to be combined in any suitable combination as if the embodiments were multiple dependent claims (e.g., embodiments directed to selective JAK1 pathway inhibitors and dosages thereof, embodiments directed to any salt form of the compounds disclosed herein, embodiments directed to individual types of cytokine-related diseases or disorders, and composition and / or administration embodiments that may be combined in any combination).

[0066] For example, provided herein is a method for treating a cytokine-related disease or disorder in a subject selected from the group consisting of cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell-associated encephalopathy (CRES), comprising administering to the subject {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole

[0023] The present invention relates to a method for treating a rheumatoid arthritis, comprising administering once daily about 200 mg of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof at a dose of about 200 mg on a free base basis, wherein the dose includes one or more sustained release dosage forms each comprising {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof.

[0067] A sustained release dosage form of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof (Table 1, Compound 1) can be found in U.S. Patent No. 2015 / 0065484 (filed August 6, 2014), which is incorporated herein by reference in its entirety.

[0068] All possible combinations are not shown separately herein merely for the sake of brevity.

[0069] The compounds described herein may be asymmetric (e.g., possess one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are intended. Compounds containing asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically inactive starting materials are well known in the art, for example, by resolution of racemic mixtures or stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in 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 separated isomeric forms.

[0070] In some embodiments, the compound has the (R) configuration. In some embodiments, the compound has the (S) configuration.

[0071] Resolution of a racemic mixture of compounds can be carried out by any of a number of methods known in the art. Exemplary methods include fractional recrystallization using a chiral resolving acid, which is an optically active, salt-forming organic acid. Suitable resolving agents for fractional recrystallization are optically active acids, such as the D- and L-forms of various optically active camphorsulfonic acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, or β-camphorsulfonic acid. Other resolving agents suitable for fractional crystallization include stereoisomerically pure forms (e.g., S- and R-forms or diastereomerically pure forms) of α-methylbenzylamine, 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, and the like.

[0072] Resolution of racemic mixtures can also be carried out by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). Suitable elution solvent compositions can be determined by one skilled in the art.

[0073] The compounds described herein also include tautomeric forms. Tautomeric forms result from the interchange of adjacent double and single bonds with the concomitant migration of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions on a heterocyclic ring system, such as 1H-imidazole, 3H-imidazole, 1H-1,2,4-triazole, 2H-1,2,4-triazole and 4H-1,2,4-triazole, 1H-isoindole and 2H-isoindole, and 1H-pyrazole and 2H-pyrazole. Tautomeric forms may be in equilibrium or sterically locked into one form by appropriate substitution.

[0074] The compounds described herein can also include isotopically labeled compounds of the present disclosure. An "isotopically" or "radiolabeled" compound is a compound of the present disclosure in which one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number that is usually found in nature (i.e., occurring in nature). Suitable radionuclides that can be incorporated into the compounds of the present disclosure include: 2 H (also written with D as deuterium), 3 H (also written as T for tritium), 11 C. 13 C. 14 C. 13 N, 15 N, 15 O. 17 O. 18 O. 18 F, 35 S, 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I, 124 I,125 I and 131 For example, one or more hydrogen atoms in the compounds of the present disclosure can be replaced with a deuterium atom (e.g., C of a compound of formula (I), (II), or (III) or Table 1). 1-6 One or more hydrogen atoms of an alkyl group can be optionally replaced with a deuterium atom, such as -CD3 being substituted for -CH3. As used herein, the term "compound" is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes for the depicted structure, unless the name indicates a specific stereoisomer. Compounds herein identified by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise specified.

[0075] All compounds and pharmaceutically acceptable salts thereof may be found together with other substances such as water and solvents (eg, hydrates and solvates) or may be isolated.

[0076] In some embodiments, the compounds described herein and salts thereof are substantially isolated. By "substantially isolated," it is meant that the compound is at least partially or substantially separated from the context in which it was formed or detected. Partial isolation can include, for example, compositions enriched in the compounds described herein. Substantial isolation can include compositions containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of the compounds described herein or salts thereof. Methods for isolating compounds and salts thereof are routine in the art.

[0077] As used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0078] As used herein, the expressions "ambient temperature" and "room temperature" or "rt" are art-recognized and generally refer to a temperature near the temperature of the room in which the reaction is carried out, e.g., from about 20°C to about 30°C, e.g., the reaction temperature.

[0079] The present invention also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, a "pharmaceutically acceptable salt" refers to a derivative of a disclosed compound, in which the parent compound has been modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, and the like. The pharmaceutically acceptable salts of the present invention include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or a mixture of the two. Generally, non-aqueous solvents such as ether, ethyl acetate, alcohols (e.g., methanol, ethanol, isopropanol, or butanol), or acetonitrile (ACN) are preferred. Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

[0080] As used herein, the terms "subject," "individual," or "patient," used interchangeably, refer to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, or primate, most preferably a human. In some embodiments, the "subject," "individual," or "patient" is in need of the above-described treatment.

[0081] In some embodiments, the inhibitor is administered in a therapeutically effective amount. As used herein, the phrase "therapeutically effective amount" refers to an amount of an active compound or pharmaceutical agent that elicits the biological or pharmacological response sought by a researcher, veterinarian, physician, or other clinician in a tissue, system, animal, individual, or human. In some embodiments, the dosage of the compound or a pharmaceutically acceptable salt thereof administered to a patient or individual is about 1 mg to about 2 g, about 1 mg to about 1000 mg, about 1 mg to about 500 mg, about 1 mg to about 200 mg, about 1 mg to about 100 mg, about 1 mg to 50 mg, or about 50 mg to about 500 mg. In some embodiments, the dosage of the compound or a pharmaceutically acceptable salt thereof is about 200 mg.

[0082] As used herein, the term "treating" or "treatment" refers to one or more of: (1) inhibiting a disease, e.g., inhibiting a disease, symptom, or disorder (i.e., arresting further onset of the pathology and / or symptomology) in an individual experiencing or exhibiting the pathology or symptomology of the disease, condition, or disorder; (2) ameliorating a disease, e.g., ameliorating a disease, symptom, or disorder (i.e., halting the progression of the pathology and / or symptomology) in an individual experiencing or exhibiting the pathology or symptomology of the disease, symptom, or disorder, such as reducing the severity of the disease; or (3) preventing a disease, symptom, or disorder in an individual who is susceptible to the disease, symptom, or disorder but who has not yet experienced or exhibited the pathology or symptomology of the disease. In some embodiments, treatment refers to inhibiting or ameliorating a disease. In some embodiments, treatment is preventing a disease.

[0083] Combination therapy The methods described herein can further include administering one or more additional therapeutic agents. The one or more additional therapeutic agents can be administered to the patient simultaneously or sequentially.

[0084] In some embodiments, the additional therapeutic agent is an IL-6 antagonist or receptor antagonist. In some embodiments, the IL-6 receptor antagonist is tocilizumab.

[0085] In some embodiments, the additional therapeutic agent is an inhibitor of MCP-1. In some embodiments, the additional therapeutic agent is an inhibitor of MIP1B. In some embodiments, the additional therapeutic agent is an inhibitor of IL-2R. In some embodiments, the additional therapeutic agent is an inhibitor of IL-1R. In some embodiments, the additional therapeutic agent is an inhibitor of TNF-α.

[0086] In some embodiments, the additional therapeutic agent is an anti-CD25 antibody. In some embodiments, the anti-CD25 antibody is daclizumab.

[0087] In some embodiments, the additional therapeutic agent is an antagonist of IL-1β.

[0088] In some embodiments, the additional therapeutic agent is an IL1 receptor antagonist (IL1Ra). In some embodiments, the IL1 receptor antagonist (IL1Ra) is anakinra.

[0089] In some embodiments, the additional therapeutic agent is a corticosteroid. In some embodiments, the corticosteroid is prednisone.

[0090] In some embodiments, any of the aforementioned additional therapeutic agents is further used in combination with a corticosteroid (eg, prednisone).

[0091] In some embodiments, the additional therapeutic agents comprise tocilizumab and a corticosteroid, hi some embodiments, the additional therapeutic agents comprise tocilizumab and prednisone.

[0092] Pharmaceutical Formulations and Dosage Forms When used as a pharmaceutical, the JAK1 pathway inhibitor or its pharmaceutically acceptable salt can be administered in the form of a pharmaceutical composition. These compositions can be prepared by methods well known in the pharmaceutical art and can be administered by various routes, depending on whether local or systemic treatment is desired and the area to be treated. Administration can be topical (transdermal, epithelial, ocular, and mucosal, such as intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, such as by nebulizer; intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular, or injection or infusion, or intracranial, such as intrathecal or intraventricular, administration. Parenteral administration can be in the form of a single bolus dose or, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

[0093] The present invention also includes pharmaceutical compositions containing, as an active ingredient, a JAK1 pathway inhibitor described herein or a pharmaceutically acceptable salt thereof in combination with one or more pharmaceutically acceptable carriers (excipients). In some embodiments, the compositions are suitable for topical administration. When preparing these compositions, the active ingredient is typically mixed with an excipient, diluted by the excipient, or enclosed in such a carrier, for example, in the form of a capsule, sachet, paper, or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid material that acts as a solvent, carrier, or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid or liquid vehicles), ointments containing up to 10% by weight of the active compound, soft gelatin capsules, hard gelatin capsules, suppositories, sterile injection solutions, and sterile packaged powders.

[0094] When preparing formulation, active compound can be milled to obtain suitable particle size before being combined with other components.If active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh.If active compound is substantially water-soluble, it can be milled to adjust particle size to, for example, about 40 mesh, to provide substantially uniform distribution in formulation.

[0095] The JAK1 pathway inhibitors can be milled using known milling procedures, such as wet milling, to obtain a particle size appropriate for tablet formation and for other formulation types. Finely divided (nanoparticulate) preparations of JAK1 selective inhibitors can be prepared by methods known in the art, see, for example, International Patent Application No. WO2002 / 000196.

[0096] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose.In addition, the preparation may contain lubricants such as talc, magnesium stearate and mineral oil, wetting agents, emulsifying and suspending agents, preservatives such as methylbenzoate and propylhydroxybenzoate, sweeteners and flavoring agents.The composition of the present invention can be formulated to provide rapid, sustained or delayed release of active ingredients after administration to patients by using methods known in the art.

[0097] The compositions may be formulated in unit dosage form, each dosage containing from about 5 to about 1000 mg (1 g), more usually about 100 to about 500 mg, of the active ingredient. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce a desired therapeutic effect, in association with a suitable pharmaceutical excipient.

[0098] In some embodiments, the compositions of the present invention contain about 5 to about 50 mg of the active ingredient. One of skill in the art will recognize that this embodies compositions containing about 5 to about 10, about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50 mg of the active ingredient.

[0099] In some embodiments, the compositions of the present invention contain about 50 to about 500 mg of the active ingredient. One of skill in the art will recognize that this embodies compositions containing about 50 to about 100, about 100 to about 150, about 150 to about 200, about 200 to about 250, about 250 to about 300, about 350 to about 400, or about 450 to about 500 mg of the active ingredient.

[0100] In some embodiments, the compositions of the present invention contain about 500 to about 1000 mg of the active ingredient. One of skill in the art will recognize that this embodies compositions containing about 500 to about 550, about 550 to about 600, about 600 to about 650, about 650 to about 700, about 700 to about 750, about 750 to about 800, about 800 to about 850, about 850 to about 900, about 900 to about 950, or about 950 to about 1000 mg of the active ingredient.

[0101] Similar dosages of the compounds described herein can be used in the methods and uses of the present invention.

[0102] The active compound can be effective over a wide dosage range and is therefore generally administered in a pharmaceutically effective amount. However, it will be understood that the amount of compound actually administered will usually be determined by a physician according to the relevant circumstances, including the condition being treated, the selected route of administration, the compound actually administered, the age, weight and response of the individual patient, the severity of the patient's condition, etc.

[0103] To prepare solid compositions such as tablets, the primary active ingredient is mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of the compound of the present invention. When these preformulation compositions are referred to as homogeneous, the active ingredient is typically dispersed evenly throughout the composition, so that the composition can be readily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules. This solid preformulation is then subdivided into unit dosage forms of the type described above, containing, for example, from about 0.1 to about 1000 mg of the active ingredient of the present invention.

[0104] The tablets or pills of the present invention can be coated or otherwise compounded to provide a dosage form that offers the advantage of prolonged action. For example, the tablet or pill can include an inner dosage component and an outer dosage component, with the outer component forming an envelope around the inner component. These two components can be separated by an enteric layer. This enteric layer functions to protect the inner component from disintegration in the stomach and further allows the inner component to reach the duodenum intact or to be released in a delayed manner. A variety of materials can be used for such enteric layers or coatings, including many polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0105] Liquid forms that may be incorporated to administer the compounds and compositions of the present invention orally or by injection include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, emulsions flavored with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, elixirs and similar pharmaceutical vehicles.

[0106] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, compositions are administered by the oral or nasal respiratory route for local or systemic effect. Compositions may be nebulized using inert gases. Nebulized solutions may be inhaled directly from the nebulizing device, or the nebulizing device may be attached to a face mask, obturator, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered orally or nasally from devices that deliver the formulation in an appropriate manner.

[0107] Topical formulations may contain one or more conventional carriers. In some embodiments, ointments may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, etc. Cream carrier compositions may be based on a combination of water, glycerol, and one or more other components, such as, for example, glycerol monostearate, PEG-glycerol monostearate, and cetylstearyl alcohol. Gels may be formulated using isopropyl alcohol and water, preferably in combination with other components, such as, for example, glycerol, hydroxyethylcellulose, etc. In some embodiments, topical formulations contain at least about 0.1, at least about 0.25, at least about 0.5, at least about 1, at least about 2, or at least about 5% by weight of a compound described herein. Topical formulations may be suitably packaged in 100g tubes, optionally with instructions for treating a selected indication, such as psoriasis or other skin conditions.

[0108] The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of the administration, such as prophylaxis or treatment, the condition of the patient, the method of administration, etc. In therapeutic applications, compositions may be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose will depend on the symptoms of the disease being treated, as well as the judgment of the attending clinician, based on factors such as the severity of the disease, the age, weight, and general condition of the patient, etc.

[0109] The compositions administered to patients may be in the form of pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or sterile filtered. Aqueous solutions may be packaged or lyophilized for immediate use, with lyophilized preparations being combined with a sterile aqueous carrier prior to administration. The pH of the compound preparations will typically be between 3 and 11, more preferably between 5 and 9, and most preferably between 7 and 8. It will be appreciated that the use of certain of the aforementioned excipients, carriers, or stabilizers may result in the formulation of pharmaceutical salts.

[0110] Therapeutic amounts of the compounds of the invention can vary according to, for example, the particular therapeutic application, the method of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of a compound described herein in a pharmaceutical composition can vary depending on many factors, including dosage, chemical properties (e.g., hydrophobicity), and route of administration. For example, for parenteral administration, the compounds described herein can be provided in an aqueous physiological buffer solution containing about 0.1 to about 10% w / v of the compound. Some typical dosage ranges are about 1 μg / kg to about 1 g / kg of body weight per day. In some embodiments, the dosage range is about 0.01 mg / kg to about 100 mg / kg of body weight per day. Dosage can depend on variables such as the type and progression of the disease or disorder, the overall health of the particular patient, the relative biological availability of the selected compound, the excipient formulation, and its route of administration. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0111] The compositions of the present invention can further comprise one or more additional pharmaceutical agents, such as chemotherapeutic agents, steroids, anti-inflammatory compounds, or immunosuppressants, examples of which are listed herein.

[0112] kit The present invention also includes pharmaceutical kits useful in treating and / or preventing cytokine-related diseases or disorders, such as CRS, which also include one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound described herein. Such kits can optionally further include one or more of a variety of conventional pharmaceutical kit components, such as, for example, a container containing one or more pharmaceutically acceptable carriers, additional containers, etc., as would be readily apparent to one of skill in the art. Instructions, either as an insert or as a label, indicating the amounts of such components, administration guidelines, and / or guidelines for mixing the components, can also be included in the kit. [Example]

[0113] The present invention will be described in more detail by specific examples.The following examples are provided for illustrative purposes and are not intended to limit the present invention in any way.Those skilled in the art will easily recognize various non-critical parameters that can be changed or modified to achieve essentially the same results.The compound of the examples has been found to be a JAK inhibitor by at least one assay described herein.

[0114] Example A: In vitro JAK kinase assay JAK1 pathway inhibitors, which can be used to treat cytokine-related diseases or disorders, are tested for inhibitory activity against JAK targets according to the following in vitro assay described in Park et al., Analytical Biochemistry 1999, 269, 94-104. The catalytic domains of human JAK1 (aa 837-1142), JAK2 (aa 828-1132), and JAK3 (aa 781-1124) with N-terminal His tags are expressed in insect cells using baculovirus and purified. The catalytic activity of JAK1, JAK2, or JAK3 was assayed by measuring the phosphorylation of a biotinylated peptide. The phosphorylated peptide was detected by homogeneous time-resolved fluorescence (HTRF). The IC of the compound for each kinase was measured in a 40 μL reaction containing enzyme, ATP, and 500 nM peptide in 50 mM Tris (pH 7.8) buffer containing 100 mM NaCl, 5 mM DTT, and 0.1 mg / mL (0.01%) BSA. 50 Measure IC of 1mM 50 For measurements, the ATP concentration in the reaction was 1 mM. The reaction was carried out at room temperature for 1 hour and then stopped with 20 μL of 45 mM EDTA, 300 nM SA-APC, and 6 nM Eu-Py20 in assay buffer (Perkin Elmer, Boston, MA). Binding to the europium-labeled antibody was carried out for 40 minutes, and the HTRF signal was measured using a Fusion plate reader (Perkin Elmer, Boston, MA). The compounds in Table 1 were tested in this assay and showed the IC values ​​seen in Table 1. 50 It was shown that the value of

[0115] Example B: Anti-CD3 antibody-induced cytokine release syndrome in Balb / c mice JAK1 pathway inhibitors can be tested for efficacy against CRS according to the in vivo assay described in Ferran, C. et al., Clin. Exp. Immunol. 1991, 86, 537-543. Specifically, this study tests the ability of compounds to reduce or ameliorate anti-CD3 antibody-induced cytokine release syndrome (CRS) in BALB / c mice. The antibody, clone 145-2C11, is an immunoglobulin G (IgG) hamster MoAb specific for the ε chain of the CD3 mouse molecule (Leo, O. et al., Proc. Natl. Acad. Sci. USA, 1987, 34, 1374). Treatment with 145-2C11 induces high-affinity IL-2 receptors on the surface of splenic T cells, resulting in the release of several cytokines, including tumor necrosis factor (TNF-α), IL-2, IL-3, IL-6, and interferon-gamma (IFN-γ) (Ferran, et al., Eur. J. Immunol. 1990, 20, 509-515, and Algre, M. et al., Eur. J. Immunol., 1990, 707). The release of these cytokines results in behavioral changes in the animals (e.g., inactivity, piloerection, etc.).

[0116] A. Materials and Methods [Table 2] A The synthesis and preparation of compound 1 in Table 1 or {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile and its adipic acid salt can be found in U.S. Patent Application No. 2011 / 0224190 (filed March 9, 2011), U.S. Patent Application No. 2013 / 0060026 (filed September 6, 2012), and U.S. Patent Application No. 2014 / 0256941 (filed March 5, 2014), each of which is incorporated herein by reference in its entirety.

[0117] B. Experimental Design The primary objective of this study was to test the ability of a JAK1 pathway inhibitor (e.g., Compound 1) to reduce or ameliorate anti-CD3 antibody-induced cytokine release syndrome (CRS) in BALB / c mice. A total of 32 BALB / c mice were used in this one-day study. Animals were weighed before administration of the test substance and monitored throughout the experiment. On day 0, 1 hour before anti-CD3 antibody administration, a single dose of vehicle (0.5% methylcellulose) or Compound 1 was administered by oral gavage (PO) to animals in groups 2-4, as detailed in Table 1A. Group 1 served as an untreated control and received no treatment. One hour after pretreatment with vehicle or Compound 1, animals in groups 2-4 received 10 μg of anti-CD3ε antibody (clone 145-2C11) via intravenous injection (IV) to induce CRS. 1.5 hours after anti-CD3 administration, all animals were euthanized by CO2 inhalation. Whole blood was collected by cardiac puncture into K2EDTA tubes and kept on ice until plasma processing. Plasma was collected and stored at -80°C until cytokine multiplex analysis.

[0118] [Table 3]

[0119] C. Experimental Procedure I. Pretreatment with test substance On day 0, animals were administered vehicle, test substance, or Compound 1 as shown in Table 1A. Group 2 received a single dose of vehicle (0.5% methylcellulose) PO at 0.1 mL / 20 g. Group 3 received a single dose of 60 mg / kg Compound 1 PO at 0.1 mL / 20 g. Group 4 received a single dose of 120 mg / kg Compound 1 PO at 0.1 mL / 20 g. Group 1 served as an untreated control and received no treatment.

[0120] II. Administration of anti-CD3ε antibody One hour after administration of the test substance, anti-CD3ε antibody (clone 145-2C11) was administered by intravenous injection to groups 2 to 4. Each animal in groups 2 to 4 received 0.1 mL of 10 μg of anti-CD3ε antibody.

[0121] III In-life monitoring After administration of anti-CD3 antibodies, animals were closely monitored for signs of distress due to the systemic inflammatory response that developed. Animals that were unable to stand, were cold to the touch, or appeared moribund were euthanized. Moribund animals were euthanized by CO2 inhalation, blood was collected by cardiac puncture, and plasma was stored.

[0122] IV Slaughter One and a half hours after anti-CD3 antibody administration, all animals were euthanized by CO2 inhalation.

[0123] V. Sample Collection Upon sacrifice, whole blood was collected from each animal via cardiac puncture into K2EDTA tubes. The blood was centrifuged, and plasma was collected in cryovials. Plasma was frozen and stored at -80°C for downstream cytokine multiplex assays.

[0124] VI Cytokine Multiplex Analysis Thaw plasma samples on ice and use for cytokine multiplex according to the manufacturer's protocol (ThermoFisher).

[0125] D result Compound 1 dose-dependently inhibited IL-6 levels in the blood compartment (Figure 1). This serves as confirmation of the biological activity observed in the preclinical model of Con A described in Example C below. One-way unpaired analysis of variance (ANOVA) incorporating Sidak's multiple comparison test was performed using GraphPad Prism (version 4.00; GraphPad Software, San Diego, California, USA). A value of p<0.05 was considered significant.

[0126] Example C: Concanavalin A-induced cytokine release syndrome Concanavalin A (Con A) is a selective T lymphocyte mitogen that leads to the release of a wide range of inflammatory cytokines and the proliferation of CD4 and CD8 T cells. Injection of Con-A has been shown to induce cytokine release syndrome, serving as a model for testing the efficacy of treatments for cytokine release syndrome (Gantner, F. at al. Hepatology, 1995, 21, 190-198). The mitogenic response is dependent on the expression of T cell receptors. Animals exhibit behavioral changes, including fever, lethargy, hypotension, hypoxia, capillary leakage, and potential multi-organ toxicity.

[0127] A. Materials and Methods [Table 4]

[0128] B. Experimental Design Specifically, this study examines the ability of a selective JAK1 inhibitor (e.g., Compound 1, Table 1) to reduce or ameliorate Con A-induced cytokine release syndrome (CRS) in BALB / c mice. A total of 40 BALB / c mice were used in this one-day study. Animals were weighed before administration of the test substance and monitored throughout the experiment. On day 0, 60 minutes before Con A administration, a single dose of vehicle (0.5% methylcellulose) or Compound 1 (60 and 120 mg / kg) was administered by oral gavage (PO) to animals in groups 2-4, as detailed in Table 2A. Group 1 served as an untreated control and was therefore untreated. Forty-five minutes after pretreatment with vehicle or Compound 1, animals in groups 2-4 received 20 mg / kg Con A via intravenous injection (IV) to induce CRS. Two hours after Con A administration, all animals were euthanized by CO inhalation. Whole blood was collected by cardiac puncture into K2EDTA tubes and kept on ice until plasma processing. Plasma was collected and stored at -80°C until cytokine multiplex analysis.

[0129] [Table 5]

[0130] C. Experimental Procedure Day 1 The animals were weighed and the dose of Con-A (20 mg / kg) was calculated. The solvent and Compound 1 were prepared at corresponding doses.

[0131] Day 0 I. Pretreatment with test substance On day 0, animals were administered vehicle or Compound 1 as shown in Table 2A. Group 1 served as untreated controls and received no treatment. Group 2 received a single dose of vehicle (0.5% methylcellulose) PO at 0.1 mL / 20 g. Group 3 received a single dose of 60 mg / kg Compound 1 PO at 0.1 mL / 20 g. Group 4 received a single dose of 120 mg / kg Compound 1 PO at 0.1 mL / 20 g.

[0132] II. Administration of Con-A Sixty minutes after administration of the test substance, Con-A was administered by intravenous injection to groups 2 to 4. Each animal in groups 2 to 4 received 0.2 mL of 20 mg / kg of Con-A.

[0133] III In-life monitoring After administration of Con-A, animals were closely monitored for signs of distress due to the systemic inflammatory response that developed.

[0134] IV Slaughter Two hours after Con-A administration, all animals were euthanized by CO2 inhalation.

[0135] V. Sample Collection Upon sacrifice, whole blood was collected from each animal via cardiac puncture into K2EDTA tubes. The blood was centrifuged, and plasma was collected in cryovials. Plasma was frozen and stored at -80°C for downstream cytokine multiplex assays.

[0136] VI Cytokine Multiplex Analysis Thaw plasma samples on ice and use for cytokine multiplex according to the manufacturer's protocol (ThermoFisher).

[0137] D result Compound 1 dose-dependently inhibited IL-6 levels in the blood compartment (Figure 2A). This cytokine is key to the pathophysiology of CRS. T cell-derived IFNγ and GM-CSF cytokines were also significantly inhibited, suggesting that compound 1 has therapeutic potential beyond the mechanism of action limited by tocilizumab (anti-IL-6R only) (Figures 2B and 2C).

[0138] Monocyte- and / or macrophage-derived cytokines were also reduced, with a statistically significant dose-dependent decrease in IL-12 (Figure 3A), as well as trends toward a therapeutic effect for IL-1β (Figure 3B), and IL-18 (Figure 3C), suggesting that JAK1-specific inhibition has therapeutic potential beyond the immune cell types involved in CRS pathology.

[0139] Importantly, the cytokine IL-5 (Figure 4) was unaffected by Compound 1 treatment, is JAK1-independent, and is not involved in CRS pathology. This data suggests that the efficacy of Compound 1 is not mediated by broad, nonspecific immunosuppression.

[0140] One-way unpaired analysis of variance (ANOVA) incorporating Sidak's multiple comparison test was performed using GraphPad Prism (version 4.00, GraphPad Software, San Diego, California, USA). A value of p<0.05 was considered significant.

[0141] Example D: Preparation of sustained release formulation of Compound 1 Extended-release tablets containing Compound 1 were prepared with excipients in the amounts shown in the table below. Protocol A was used for SR1 tablets, Protocol B for SR2 tablets, Protocol C for SR3 tablets and 25 mg SR tablets, and Protocol D for SR4 tablets. These procedures are disclosed in U.S. Patent Application No. 2015 / 0065484, which relates to extended-release dosage forms of Compound 1.

[0142] Protocol A Step 1 Compound 1 adipate salt, microcrystalline cellulose, hypromellose (Methocel K100 LV and Methocel K4M) and lactose monohydrate are sieved separately. Step 2: Transfer the sieved ingredients from step 1 to a suitable blender and mix. Step 3: Transfer the mixture from step 2 into a suitable granulator and mix. Step 4 Add purified water while mixing. Step 5: Transfer the granules from Step 4 into a suitable dryer and dry until the LOD is less than 3%. Step 6 Sift the granules from step 5. Step 7 Blend the granules from Step 6 with the sieved magnesium stearate in a suitable blender. Step 8 Compress the final blend from Step 7 on a suitable rotary tablet press.

[0143] Protocol B Step 1 Compound 1 adipate, microcrystalline cellulose, hypromellose and pregelatinized starch are separately sieved. Step 2: Transfer the sieved ingredients from step 1 to a suitable blender and mix. Step 3: Transfer the mixture from step 2 into a suitable granulator and mix. Step 4 Add purified water while mixing. Step 5: Transfer the granules from Step 4 into a suitable dryer and dry until the LOD is less than 3%. Step 6 Sift the granules from step 5. Step 7 Sift the Polyox, butylated hydroxytoluene and colloidal silicon dioxide separately. Step 8: Transfer the granules from Step 6 and the ingredients from Step 7 to a suitable blender and mix. Step 9 Add the sifted magnesium stearate to the ingredients from Step 8 and continue blending. Step 10 Compress the final blend from Step 9 on a suitable rotary tablet press.

[0144] Protocol C Step 1 Sift lactose monohydrate, Compound 1 adipate salt, microcrystalline cellulose and hypromellose separately through suitable sieves. Step 2: Transfer the sieved ingredients from step 1 to a suitable blender and mix. Step 3: Transfer the mixture from step 2 into a suitable granulator and mix. Step 4 Add purified water while mixing. Step 5 Sift the wet granules through a suitable sieve. Step 6: Transfer the granules from Step 5 into a suitable dryer and dry until the LOD is less than 3%. Step 7: Mill the granules from Step 6. Step 8 Blend the granules from Step 7 with the sieved magnesium stearate in a suitable blender. Step 9 Compress the final blend from Step 8 on a suitable rotary tablet press.

[0145] Protocol D Step 1: Pregelatinized starch, Compound 1 adipate, hypromellose and a portion of the required microcrystalline cellulose are individually sieved through suitable sieves. Step 2: Transfer the sieved ingredients from step 1 to a suitable blender and mix. Step 3: Transfer the mixture from step 2 into a suitable granulator and mix. Step 4 Add purified water while mixing. Step 5 Sift the wet granules through a suitable sieve. Step 6: Transfer the granules from Step 5 into a suitable dryer and dry until the LOD is less than 3%. Step 7: Mill the granules from Step 6. Step 8 Sift the remaining part of the microcrystalline cellulose and half of the sodium bicarbonate. Step 9: Transfer the milled granules from Step 7 and the ingredients from Step 8 to a suitable blender and mix. Step 10: Sift the remaining portion of the sodium bicarbonate and mix with the mixture from Step 9. Step 11 Sift the magnesium stearate and mix with the mixture from Step 10. Step 12 Compress the final blend of Step 11 on a suitable rotary tablet press.

[0146] SR1: Composition of 100 mg sustained-release tablets [Table 6] a The conversion factor for the adipate salt to the free base is 0.7911. b It was added after granulation. c removed during processing.

[0147] SR2: Composition of 100 mg sustained-release tablets [Table 7] a The conversion factor for the adipate salt to the free base is 0.7911. b It was added after granulation. c removed during processing.

[0148] SR3 (100 mg): Composition of 100 mg sustained-release tablets [Table 8] a The conversion factor for the adipate salt to the free base is 0.7911. b It was added after granulation. c removed during processing.

[0149] SR4: Composition of 100 mg sustained-release tablets [Table 9] a The conversion factor for the adipate salt to the free base is 0.7911. b It was added after granulation. c removed during processing. d A portion was added before granulation and a portion was added after granulation.

[0150] 25mg SR: Composition of 25mg sustained-release tablets [Table 10] a The conversion factor for the adipate salt to the free base is 0.7911. b It was added after granulation. c removed during processing.

[0151] Various modifications of the present invention, in addition to those described herein, will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this application, including all patents, patent applications, and publications, is hereby incorporated by reference in its entirety. The present application also includes the following aspects. [Aspect 1] A method for treating a cytokine-related disease or disorder in a subject, comprising administering to the subject a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof. [Aspect 2] The method of aspect 1, wherein the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is selective for JAK1 over JAK2, JAK3, and Tyk2. [Aspect 3] 3. The method of aspect 1 or 2, wherein said cytokine-associated disease or disorder is cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell-associated encephalopathy (CRES). [Aspect 4] The method of embodiment 3, wherein said cytokine-associated disease or disorder is cytokine release syndrome (CRS). [Aspect 5] The method of embodiment 3, wherein said cytokine-associated disease or disorder is hemophagocytic lymphohistiocytosis (HLH). [Aspect 6] The method of embodiment 3, wherein said cytokine-associated disease or disorder is macrophage activation syndrome (MAS). [Aspect 7] The method of embodiment 6, wherein said macrophage activation syndrome (MAS) is associated with systemic juvenile idiopathic arthritis. [Aspect 8] The method of embodiment 3, wherein said cytokine-associated disease or disorder is CAR-T cell-associated encephalopathy (CRES). [Aspect 9] Aspect 9. The method of any one of aspects 1-8, wherein the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof. [Aspect 10] The method of any one of aspects 1 to 8, wherein the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate. [Aspect 11] The method of any one of aspects 1 to 8, wherein the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof. [Aspect 12] The method of any one of aspects 1 to 8, wherein the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamidophosphate. [Aspect 13] Aspect 9. The method of any one of aspects 1-8, wherein the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile or a pharmaceutically acceptable salt thereof. [Aspect 14] Aspect 9. The method of any one of aspects 1-8, wherein the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile monohydrate. [Aspect 15] 15. The method of any one of aspects 1-14, further comprising administering to said subject tocilizumab. [Aspect 16] 15. The method of any one of aspects 1-14, further comprising administering a corticosteroid to said subject. [Aspect 17] Aspect 15. The method of any one of aspects 1-14, further comprising administering prednisone to said subject. [Aspect 18] 15. The method of any one of aspects 1-14, further comprising administering to said subject tocilizumab and a corticosteroid.

Claims

1. A pharmaceutical for treating cytokine release syndrome, comprising a JAK1 pathway inhibitor, wherein the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof.

2. The pharmaceutical composition of claim 1, wherein the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate.

3. The medicament according to claim 1 or 2, wherein the medicament is administered in combination with tocilizumab.

4. 3. The method of claim 1, wherein the medicament is administered in combination with a corticosteroid.

5. 3. The method of claim 1, wherein the medicament is administered in combination with prednisone.

6. The medicament according to claim 1 or 2, wherein the medicament is administered in combination with tocilizumab and a corticosteroid.

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