JAK1 pathway inhibitors for treating cytokine-related disorders

Abstract

To provide pharmaceuticals for the treatment of cytokine-related diseases or disorders. [Solution] A pharmaceutical for treating a cytokine-related disease or disorder comprises a JAK1 pathway inhibitor, wherein the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipirazole-1-yl)azetidine-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof, and the cytokine-related disease or disorder is cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell-associated encephalopathy syndrome (CRES).

Description

Technical Field

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

Background Art

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

[0003] Cytokine release syndrome (CRS) is a direct result of the overproduction of inflammatory cytokines caused by supra-physiological levels of immune activation and presents as a series of clinical symptoms such as fever, nausea, fatigue, myalgia, malaise, hypotension, hypoxemia, capillary leakage, etc., thereby resulting in potential multi-organ toxicity.

[0004] CRS is, for example, an unwanted side effect of immunotherapy for severe 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 modified T cells to target cancer and its use in certain forms of refractory non-Hodgkin lymphoma and pediatric relapsed acute lymphoblastic leukemia (ALL) has already been approved by the FDA.

[0005] The cytokine profile involved in CRS includes two main 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 from monocytes and / or macrophages, such as 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] Modifying the excessive cytokine response that leads to CRS has the potential to yield significant clinical benefits. For example, tocilizumab, an antibody against the IL-6 receptor (IL-6R), reduces the incidence of severe CRS, and its use in CRS is approved by the FDA. However, the mechanism of action of tocilizumab is limited to anti-IL-6R only.

[0007] Hemophagocytic lymphohistiocytosis (HLH) is another syndrome of excessive or uncontrolled immune activation, primarily affecting infants from birth to 18 months of age, but can also occur in adults. HLH can be primary (familial) or secondary, meaning it can develop in conjunction with other infectious, malignant, rheumatic, or metabolic conditions. Symptoms of HLH include cytopenia, hepatosplenomegaly, and fever. Schram, A. and Berliner, N.Blood. 2005. 125(19), 2908-2914.

[0008] Macrophage activation syndrome (MAS) presents clinically in a similar manner 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 being 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 numerous 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% in children with autoimmune diseases and 10-22% in MAS complicated with cSLE. Borgia, RE et al. Arthritis Rheumatol., 2018, doi:10.1002 / art.40417 (pre-publication).

[0009] CAR-T cell-associated encephalopathy syndrome (CRES) is the second most common adverse event associated with CAR-T cell therapy, after CRS. CRES is typically characterized by a toxic encephalopathy state with symptoms of confusion and delirium, as well as occasional seizures and cerebral edema. The symptoms of CRES 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 the passive diffusion of cytokines into the brain of patients treated with CAR-T cell therapy. Reduction or elimination of this mechanism may be beneficial for such patients. Neelapu, et al. Nat Rev Clin Oncol. 2018, 15(1)47-62.

[0010] Therefore, there is a need to develop novel therapies for treating cytokine-related diseases or disorders. This application addresses this need and other needs. [Brief explanation of the drawing]

[0011] [Figure 1] During anti-CD3 antibody-induced cytokine release syndrome, administration of compound 1 into the blood compartment showed dose-dependent inhibition of IL-6 concentration (see Example B). [Figure 2A] The drug exhibits 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). It demonstrates inhibition of IL-6. [Figure 2B] The drug exhibits 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). It demonstrates inhibition of IFNγ. [Figure 2C] This 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). It demonstrates inhibition of GM-CSF. [Figure 3A] It exhibits 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). It shows inhibition of IL-12. [Figure 3B] It 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). It shows inhibition of IL-1β. [Figure 3C] It 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). It shows inhibition of IL-18. [Figure 4] This demonstrates that cytokine IL-5 is unaffected by treatment of compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). [Overview of the project]

[0012] This specification provides a method for treating cytokine-related disorders or conditions in subjects requiring treatment, the method comprising administering to the patient a therapeutically effective amount of a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof.

[0013] This specification provides JAK1 pathway inhibitors or pharmaceutically acceptable salts thereof for treating cytokine-related disorders or impairments in subjects requiring treatment.

[0014] This specification provides the use of JAK1 pathway inhibitors or pharmaceutically acceptable salts thereof for the manufacture of drugs used in the treatment of cytokine-related diseases or disorders in subjects requiring treatment. Detailed description of the invention

[0015] In particular, the present invention provides a method for treating cytokine-related disorders or conditions in subjects requiring treatment, 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 the signaling pathways of many cytokines and growth factors that, in dysregulation, can lead to or contribute to disease states. For example, IL-6 levels are elevated in rheumatoid arthritis, and in this disease, IL-6 has been suggested to have adverse effects (Fonesca, et al., Autoimmunity Reviews, 8:538-42, 2009). Since IL-6 signals at least partially via JAK1, IL-6 exists indirectly through JAK1 inhibition, which may provide potential clinical benefits (Guschin, et al., Embo J 14:1421, 1995; Smolen, et al., Lancet 371:987, 2008). Furthermore, in some cancers, JAK1 is mutated, leading to constitutively unwanted 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 JAK1 inhibition. Selective JAK1 inhibitors may be beneficial while avoiding unwanted and potentially undesirable effects caused by inhibiting other JAK kinases.

[0017] JAK1 pathway inhibitors, specifically compound 1 (i.e., {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-3-yl}acetonitrile, see Table 1), achieve highly effective dose-dependent modulation of CRS-related inflammatory cytokines (see, e.g., Examples B and C, and Figures 1, 2A-2C, and 3A-3C). Surprisingly, this therapeutic property includes multiple pathogenic cytokines and is not limited to the IL-6 / IL-6R system (e.g., tocilizumab is different). Efficacy is achieved by inhibiting T cell and monocyte / macrophage-derived cytokines that are clinically highly associated with the development of CRS. Furthermore, the data presented herein in relation to JAK1 inhibitor compound 1 demonstrate that therapeutic benefits are achieved without extensive cytokine immunosuppression (as demonstrated by the fact that IL-5 levels remained unchanged) (Figure 4). [Modes for carrying out the invention]

[0018] In some embodiments, cytokine-related disorders or conditions include cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell-associated encephalopathy syndrome (CRES).

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

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

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

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

[0023] In some embodiments, the present application provides a method for treating a cytokine release syndrome in a subject, the method comprising administering CAR-T cell therapy and a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof to the subject. In some embodiments, treatment means remission or inhibition. In some embodiments, treatment means prevention.

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

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

[0026] In some embodiments, CAR-T cell therapy is axicaptagene siloleucel.

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

[0028] In some embodiments, the subjects are suffering from B-cell malignancies.

[0029] In some embodiments, the subjects have 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, JAK1 pathway inhibitors or pharmaceutically acceptable salts thereof are selective to JAK1 over JAK2, JAK3, and TYK2 (i.e., JAK1 selective inhibitors). For example, the compounds described herein, or pharmaceutically acceptable salts thereof, preferentially inhibit JAK1 over one or more JAK2, JAK3, and TYK2. In some embodiments, the compounds preferentially inhibit JAK1 over JAK2 (e.g., JAK2 / JAK1 IC2). 50 Ratio > 1). In some embodiments, the compound or salt is about 10 times more selective to JAK1 than to JAK2. In some embodiments, the compound or salt IC2 at 1 mM ATP. 50 Calculations based on measurements show that JAK2 is approximately 3 times, 5 times, 10 times, 15 times, or 20 times more selective than JAK1 (see, for example, Example A).

[0031] In some embodiments, the JAK1 pathway inhibitor is a compound from Table 1 or a pharmaceutically acceptable salt thereof. The compounds in Table 1 are selective JAK1 inhibitors (more selective than JAK2, JAK3, and TYK2). IC obtained by the method of Example A with 1 mM ATP 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) +++ Average ≤ 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]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-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]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-3-yl}acetonitrile adipinate.

[0035] The synthesis and preparation of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-3-yl}acetonitrile and its adipate are documented 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'-bipirazole-1-yl)azetidine-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'-bipirazole-1-yl)azetidine-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'-bipyrazole-1-yl)azetidine-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide and its phosphate 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]pyridine-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]pyridine-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]pyridine-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile and the characterization of its anhydrous 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 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 The materials are prepared by the synthesis methods described in U.S. Patent Application 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 described 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), and U.S. Patent Application No. 2012 / 0149 Selected from the compounds or pharmaceutically acceptable salts thereof of U.S. Patent Application 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), each of these applications is incorporated herein by reference in its entirety.

[0044] In some embodiments, the JAK1 pathway inhibitor is a compound of formula I. [ka] Alternatively, a pharmaceutically acceptable salt thereof, in the formula, X is N or CH, L is C(=O) or C(=O)NH, A is phenyl, pyridinyl, or pyrimidinyl, each of which is one or two independently selected R 1 It is arbitrarily substituted in the base, and further Each R 1 These are independently fluoromethyl or trifluoromethyl.

[0045] In some embodiments, the compound of formula I is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-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]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-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]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(1-{[2-(trifluoromethyl)pyrimidine-4-yl]carbonyl}piperidine-4-yl)azetidine-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, wherein this C 1-6 alkyl, C 3-6 cycloalkyl and C 3-6 cycloalkyl-C 1-3 alkyl is optionally substituted with one, two, or three 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 further 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

Chemical formula

[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]pyridine-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile or a pharmaceutically acceptable salt thereof.

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

[0053] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered at a dose of approximately 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 at a dose of approximately 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 at a dose of approximately 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 at a dose of approximately 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 at a dose of approximately 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 once daily at a dose of approximately 200 mg on a free base basis.

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

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

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

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

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

[0064] This specification provides a method for treating a cytokine-related disorder or impairment in a subject requiring treatment, comprising administering to the subject approximately 100 mg to 600 mg per day of a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof, based on free base, wherein the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in one or more sustained-release formulations containing the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof.

[0065] The embodiments described herein are intended to be combined in any preferred combination, as if they were multiple dependent claims (for example, embodiments relating to selective JAK1 pathway inhibitors and their doses, embodiments relating to any salt form of the compounds disclosed herein, embodiments relating to individual types of cytokine-related diseases or disorders, and embodiments relating to compositions and / or administrations that can be combined in any combination).

[0066] For example, this specification provides a method for treating a cytokine-related disorder or impairment 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 syndrome (CRES), wherein the method involves adding {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole The treatment involves administering approximately 200 mg of acetonitrile or a pharmaceutically acceptable salt thereof once daily, based on free base, wherein the dose includes one or more sustained-release dosage forms each containing {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]acetonitrile or a pharmaceutically acceptable salt thereof.

[0067] The sustained-release formulation of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-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 in this specification for the sake of brevity.

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

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

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

[0072] The separation of racemic mixtures can also be carried out by eluting them onto a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). A suitable elution solvent composition can be determined by those skilled in the art.

[0073] The compounds described herein also include tautomeristic forms. Tautomeristic forms are obtained by the exchange of adjacent double and single bonds and the simultaneous transfer of protons. Tautomeristic forms include prototropic tautomers, which are protonated states of isomers having the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imoid acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions in the heterocyclic 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. Tautomeristic forms may be in equilibrium or sterically fixed to one form by appropriate substitution.

[0074] The compounds described herein may also include isotope-labeled compounds of this disclosure. An “isotope-labeled” or “radio-labeled” compound is a compound of this disclosure in which one or more atoms are replaced or substituted with atoms having atomic masses or mass numbers different from those normally found in nature (i.e., naturally occurring). Suitable radionuclides that may be incorporated into the compounds of this disclosure include: 2 H (also written as D as Deuterium), 3 H (also written as T as 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 Formula (I), (II), or (III) or C in the compounds of Table 1 may be replaced by a deutherium atom (e.g., C in formula (I), (II), or (III) or the compounds in Table 1). 1-6 One or more hydrogen atoms of the alkyl group may be optionally substituted with a deutherium atom such that -CD3 is substituted for -CH3. As used herein, the term “compound” means all stereoisomers, geometric isomers, tautomers, and isotopes of the given structure unless the name indicates a specific stereoisomer. Compounds herein identified by name or structure as a particular tautomer are intended to include other tautomers unless otherwise specified.

[0075] All compounds and their pharmaceutically acceptable salts can be identified together with other substances such as water and solvents (e.g., hydrates and solvates) or isolated.

[0076] In some embodiments, the compounds and salts thereof described herein are substantially isolated. “Substantially isolated” means that the compound is separated at least partially or substantially from the circumstances in which it was formed or detected. Partial isolation may include, for example, a composition concentrated in the compounds described herein. Substantial isolation may include a composition 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 compound or salt thereof described herein. Methods for isolating compounds and salts thereof are common in the art.

[0077] As used herein, the term "pharmaceutically acceptable" means a compound, material, composition, and / or dosage form that is suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, within reasonable limits of medical judgment, and that has a reasonable benefit-to-risk ratio.

[0078] As used herein, the terms “ambient temperature” and “room temperature” or “rt” are understood in the art and generally refer to a temperature close to the temperature of the room in which the reaction takes place, for example, a temperature between approximately 20°C and approximately 30°C, e.g., the reaction temperature.

[0079] The present invention also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, “pharmaceutically acceptable salt” is a derivative of the disclosed compound, where the parent compound is modified by converting an existing acidic or base moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. The pharmaceutically acceptable salts of the present invention include, for example, conventional non-toxic salts of parent compounds formed from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acidic or base form of these compounds with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture thereof. Generally, non-aqueous solvents such as ether, ethyl acetate, alcohol (e.g., methanol, ethanol, isopropanol, or butanol), or acetonitrile (ACN) are preferred. A list 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” are interchangeable and refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, most preferably humans. In some embodiments, the “subject,” “individual,” or “patient” is in need of the treatment described above.

[0081] In some embodiments, the inhibitor is administered in a therapeutically effective dose. As used herein, the term “therapeutably effective dose” refers to the amount of the active compound or pharmaceutical agent that elicits a desired biological or pharmacological response in a tissue, system, animal, individual, or human, as determined by a researcher, veterinarian, physician, or other clinician. In some embodiments, the dose of the compound or a pharmaceutically acceptable salt 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 about 50 mg, or about 50 mg to about 500 mg. In some embodiments, the dose of the compound or a pharmaceutically acceptable salt is about 200 mg.

[0082] As used herein, the terms “to treat” or “to cure” mean one or more of the following: (1) inhibiting a disease, for example, inhibiting a disease, symptom or disorder in an individual who is experiencing or exhibiting the pathology or overall symptoms of a disease, symptom or disorder (i.e., preventing further development of the pathology and / or overall symptoms); (2) relieving a disease, for example, reducing the severity of a disease, symptom or disorder in an individual who is experiencing or exhibiting the pathology or overall symptoms of a disease, symptom or disorder (i.e., halting the progression of the pathology and / or overall symptoms); or (3) preventing a disease, symptom or disorder in an individual who is susceptible to the disease, symptom or disorder but has not yet experienced or exhibited the pathology or overall symptoms of that disease. In some embodiments, treatment means inhibiting or relieving a disease. In some embodiments, treatment means preventing a disease.

[0083] Combination therapy The methods described herein may further include administering one or more additional therapeutic agents. These additional therapeutic agents may 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 MCP-1 inhibitor. In some embodiments, the additional therapeutic agent is an MIP1B inhibitor. In some embodiments, the additional therapeutic agent is an IL-2R inhibitor. In some embodiments, the additional therapeutic agent is an IL-1R inhibitor. In some embodiments, the additional therapeutic agent is a TNF-α inhibitor.

[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 IL-1β antagonist.

[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 are used in combination with a corticosteroid (e.g., prednisone).

[0091] In some embodiments, the additional therapeutic agent comprises tocilizumab and a corticosteroid. In some embodiments, the additional therapeutic agent comprises tocilizumab and prednisone.

[0092] Pharmaceutical preparations and dosage forms When used as a pharmaceutical, JAK1 pathway inhibitors or pharmaceutically acceptable salts thereof can be administered in the form of pharmaceutical compositions. These compositions can be prepared by methods well known in the pharmaceutical field and can be administered by various routes depending on whether topical or systemic treatment is preferred and the area being treated. Administration can be topical (transdermal, epithelial, ocular, and mucosal, such as intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or gas injection of powders or aerosols using a sprayer; intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, intramuscular, or injection or infusion, or intracranial administration such as in the subarachnoid space or ventricles. Parenteral administration may be in the form of a single bolus or, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous bases, powder bases, oily bases, or thickeners may be essential or desirable.

[0093] The present invention also includes pharmaceutical compositions containing, as an active ingredient, a JAK1 pathway inhibitor as 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 encapsulated in such carriers, for example, in the form of capsules, pouches, paper or other containers. When the excipient functions as a diluent, it may be a solid, semi-solid or liquid material acting as a solvent, carrier or medium for the active ingredient. Thus, the compositions may be in the form of tablets, pills, powders, lozenges, pouches, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid medium or in a liquid medium), ointments, soft gelatin capsules, hard gelatin capsules, suppositories, sterile injections and sterile packaged powders, for example, containing up to 10% by weight of the active compound.

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

[0095] JAK1 pathway inhibitors can be milled using known milling procedures, such as wet milling, to obtain particle sizes suitable for tablet formation and other formulations. Precisely separated (nanoparticle) preparations of JAK1 selective inhibitors can be prepared by methods well known in the art; see, for example, International Patent Application 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. Furthermore, formulations may contain lubricants, wetting agents, emulsifiers, and suspending agents such as talc, magnesium stearate, and mineral oil, as well as preservatives such as methyl benzoate and propyl hydroxybenzoate, sweeteners, and flavorings. Compositions of the present invention can be formulated using methods known in the art to provide rapid, sustained-release, or delayed release of the active ingredient after administration to a patient.

[0097] The composition may be formulated in unit dosage forms, each dose containing approximately 5 to approximately 1000 mg (1 g), more commonly, approximately 100 to approximately 500 mg of the active ingredient. The term "unit dosage form" refers to a physically distinct unit suitable as a unit dose for human subjects and other mammals, each unit containing a predetermined amount of the active substance calculated to produce the desired therapeutic effect in relation to a suitable pharmaceutically acceptable excipient.

[0098] In some embodiments, the compositions of the present invention contain about 5 to about 50 mg of the active ingredient. Those skilled in the art will understand that this is an embodiment of 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. Those skilled in the art will understand that this is an embodiment of 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. Those skilled in the art will understand that this is an embodiment of 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] In the methods and uses of the present invention, similar dosages of the compounds described herein can be used.

[0102] Since active compounds can be effective across a wide range of doses, they are generally administered in pharmaceutically effective amounts. However, it should be understood that the actual amount of compound administered is usually determined by the physician, taking into account relevant circumstances including the symptoms being treated, the chosen route of administration, the compound actually administered, the individual patient's age, weight and response, and the severity of the patient's symptoms.

[0103] To prepare solid compositions such as tablets, the main active ingredient is mixed with pharmaceutically acceptable excipients to form a solid preliminary formulation composition containing a homogeneous mixture of the compounds of the present invention. When these preliminary formulation compositions are referred to as homogeneous, the active ingredient is typically uniformly dispersed throughout the composition, thereby allowing the composition to be easily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules. Furthermore, this solid preliminary formulation can be subdivided into the aforementioned unit dosage forms, for example, containing 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 provided with a dosage form that offers the advantage of prolonged action by coating or other means of mixing. For example, the tablets or pills may contain an inner and an outer dosing component, with the outer component forming a coating over the inner component. These two components can be separated by an enteric coating. This enteric coating functions to prevent breakdown in the stomach, further allowing the inner component to reach the duodenum intact or enabling delayed release. Various materials can be used for such enteric coatings or coatings, and such materials include many polymer acids, as well as mixtures of polymer acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0105] Liquid forms that may be incorporated for oral or injectable administration of the compounds and compositions of the present invention 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, and elixirs and similar pharmaceutical solvents.

[0106] Compositions for inhalation or gas infusion include solutions and suspensions in pharmaceutically acceptable aqueous solvents or organic solvents or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, compositions are administered orally or via nasal respiration for topical or systemic effects. Compositions may be sprayed using an inert gas. The sprayed solution may be inhaled directly from a spraying device, or the spraying device may be attached to a face mask, embolus, or intermittent positive airway pressure (PAP) respirator. Compositions in solution, suspension, or powder form may be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0107] Topical formulations may comprise one or more conventional carriers. In some embodiments, ointments may comprise water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, etc. Carrier compositions for creams may be based on a combination of water, glycerol, and one or more other components such as glycerin monostearate, PEG-glycerin monostearate, and cetyl stearyl alcohol. Gels may be formulated using isopropyl alcohol and water, preferably in combination with other components such as glycerol, hydroxyethylcellulose, etc. In some embodiments, topical formulations may comprise 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 the compounds described herein. Topical formulations may be suitably packaged in 100g tubes optionally bearing instructions for treating selected indications, 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 administration (such as prevention or treatment), the patient's condition, and the method of administration. In therapeutic applications, a composition may be administered to a patient already suffering from the disease in an amount sufficient to cure or at least partially suppress the symptoms of the disease and its complications. The effective dose depends on the judgment of the attending clinician, based on the symptoms of the disease being treated, as well as factors such as the severity of the disease, the patient's age, weight, and general condition.

[0109] The compositions administered to the patient may be in the form of the pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or aseptically filtered. Aqueous solutions may be packaged or lyophilized for use as is, but lyophilized preparations are combined with a sterile aqueous carrier before administration. The pH of the compound preparation will typically be between 3 and 11, more preferably between 5 and 9, and most preferably between 7 and 8. It will be understood that formulations of pharmaceutical salts can be obtained by using certain excipients, carriers, or stabilizers described above.

[0110] The therapeutic dose of the compounds of the present invention may vary depending, for example, the specific use in which the treatment is performed, the method of administering the compound, the patient's health and symptoms, and the judgment of the prescribing physician. The ratio or concentration of the compounds described herein in a pharmaceutical composition may vary depending on many factors, including the dose, chemical properties (e.g., hydrophobicity), and route of administration. For example, the compounds described herein can be provided for parenteral administration in an aqueous physiological buffer solution containing about 0.1 to about 10% w / v of the compound. Some typical dose ranges are about 1 μg / kg to about 1 g / kg of body weight per day. In some embodiments, the dose range is about 0.01 mg / kg to about 100 mg / kg of body weight per day. The dose may depend on variables such as the type and progression of the disease or disorder, the overall health status of the particular patient, the relative biological efficacy of the selected compound, the formulation of excipients, and the route of administration. The effective dose can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0111] The compositions of the present invention may further include one or more additional pharmaceuticals, such as chemotherapeutic agents, steroids, anti-inflammatory compounds, or immunosuppressants, examples of which are provided herein.

[0112] kit The present invention also includes, for example, a pharmaceutical kit useful in treating and / or preventing cytokine-related diseases or disorders such as CRS, and the kit includes one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of the compounds described herein. Such a kit may further include, as necessary, one or more different conventional pharmaceutical kit components, such as containers containing one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily apparent to those skilled in the art. Instructions indicating the amounts of such components, either as inserts or labels, guidelines for administration and / or guidelines for mixing the components may also be included in the kit. [Examples]

[0113] The present invention will be described in more detail by specific examples. The following examples are provided for illustrative purposes only and are not intended to limit the invention in any way. Those skilled in the art will readily recognize various non-essential parameters that can be changed or modified to produce essentially the same results. The compounds of the examples have been found to be JAK inhibitors by at least one assay described herein.

[0114] Example A: In vitro JAK kinase assay JAK1 pathway inhibitors that may be used to treat cytokine-related diseases or disorders will be tested for their 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 (aa837-1142), JAK2 (aa828-1132), and JAK3 (aa781-1124) with N-terminal His tags were expressed in insect cells using baculovirus and purified. The catalytic activity of JAK1, JAK2, or JAK3 was assayed by measuring the phosphorylation of biotinylated peptides. Phosphorylated peptides were detected by homogeneous time-resolved fluorescence (HTRF). In a 40 μL reaction mixture containing enzymes, ATP, and 500 nM peptides in 50 mM Tris (pH 7.8) buffer containing 100 mM NaCl, 5 mM DTT, and 0.1 mg / mL (0.01%) BSA, the IC50 of each compound was measured for each kinase. 50 Measure the 1mM IC. 50 For the measurement, the ATP concentration during the reaction was 1 mM. The reaction was carried out at room temperature for 1 hour, and then stopped with an assay buffer solution (Perkin Elmer, Boston, MA) containing 20 μL of 45 mM EDTA, 300 nM SA-APC, and 6 nM Eu-Py20. Binding to the europium-labeled antibody was allowed for 40 minutes, and the HTRF signal was measured using a Fusion plate reader (Perkin Elmer, Boston, MA). When the compounds listed in Table 1 were tested with this assay, the IC2011 results were as shown in Table 1. 50 It was shown that it has the value of [value].

[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 can test the ability of compounds to alleviate or remit anti-CD3 antibody-induced cytokine release syndrome (CRS) in BALB / c mice. The antibody, clone 145-2C11, is an immunoglobin G (IgG) hamster MoAb specific to 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, leading to 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 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]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-3-yl}acetonitrile and its adipate 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 mitigate or alleviate 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. The animals were weighed before administration of the test substance and monitored throughout the experiment. On day 0, one hour before administration of anti-CD3 antibody, a single dose of solvent (0.5% methylcellulose) or compound 1 was administered to groups 2-4 by forced oral administration (PO), as detailed in Table 1A. Group 1 served as an untreated control and received no treatment. One hour after pretreatment with solvent or compound 1, groups 2-4 were administered 10 μg of anti-CD3ε antibody (clone 145-2C11) by intravenous injection (IV) to induce CRS. 1.5 hours after anti-CD3 administration, all animals were euthanized by CO2 inhalation. Whole blood was collected in a K2EDTA blood collection tube via cardiac puncture and stored on ice until plasma processing was performed. The collected plasma was stored at -80°C until cytokine multiplexing was performed.

[0118] [Table 3]

[0119] C. Experimental Procedure I. Pretreatment of Test Substances On day 0, animals were administered either the solvent, the test substance, or compound 1, as shown in Table 1A. Group 2 received a single dose of the solvent (0.5% methylcellulose) at 0.1 mL / 20 g via PO. Group 3 received a single dose of compound 1 at 60 mg / kg via PO at 0.1 mL / 20 g. Group 4 received a single dose of compound 1 at 120 mg / kg via PO at 0.1 mL / 20 g. Group 1 served as an untreated control and therefore 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 intravenously to groups 2-4. Each animal in groups 2-4 received 0.1 mL of anti-CD3ε antibody (10 μg).

[0121] III. Monitoring during survival After administering anti-CD3 antibodies, the animals were carefully monitored for signs of distress due to the systemic inflammatory response that developed. Animals that were unable to get up, were cold to the touch, or were near death were euthanized. Near death animals were euthanized by CO2 inhalation, blood was collected by cardiac puncture, and plasma was preserved.

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

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

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

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

[0126] Example C: Concanavalin A-induced cytokine release syndrome Concanavalin A (Con A) is a selective T lymphocyte mitogen that induces widespread inflammatory cytokine release and proliferation of CD4 and CD8 T cells. Injection of Con-A has been shown in the literature to induce cytokine release syndrome, making it a useful model for testing the efficacy of treatments for cytokine release syndrome (Gantner, F. et al. Hepatology, 1995, 21, 190-198). The mitogen response is dependent on T cell receptor expression. Animals exhibit behavioral changes such as fever, malaise, hypotension, hypoxia, capillary leakage, and potential multi-organ toxicity.

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

[0128] B. Experimental Design In particular, this study tests the ability of selective JAK1 inhibitors (e.g., Compound 1, Table 1) to mitigate or alleviate 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. The 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 solvent (0.5% methylcellulose) or Compound 1 (60 and 120 mg / kg) was administered to groups 2-4 by forced oral administration (PO), as detailed in Table 2A. Group 1 served as an untreated control and received no treatment. Forty-five minutes after pretreatment with solvent or Compound 1, groups 2-4 were administered 20 mg / kg of Con A by intravenous injection (IV) to induce CRS. Two hours after Con A administration, all animals were euthanized by CO2 inhalation. Whole blood was collected in a K2EDTA blood collection tube via cardiac puncture and stored on ice until plasma processing was performed. The collected plasma was stored at -80°C until cytokine multiplexing was performed.

[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 in the corresponding doses.

[0131] Day 0 I. Pretreatment of Test Substances On day 0, animals were administered either the solvent or compound 1, as shown in Table 2A. Group 1 served as an untreated control and received no treatment. Group 2 received a single dose of the solvent (0.5% methylcellulose) at 0.1 mL / 20 g via PO. Group 3 received a single dose of compound 1 at 60 mg / kg via PO at 0.1 mL / 20 g. Group 4 received a single dose of compound 1 at 120 mg / kg via PO at 0.1 mL / 20 g.

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

[0133] III. Monitoring during survival After administering Con-A, the animals were carefully monitored for signs of distress due to the systemic inflammatory response that developed.

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

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

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

[0137] D result Compound 1 dose-dependently inhibited IL-6 concentration in blood compartments (Figure 2A). This cytokine is a key component in 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] Cytokines derived from monocytes and / or macrophages were also reduced. A statistically significant dose-dependent decrease in IL-12 (Figure 3A), as well as trends in therapeutic effects with IL-1β (Figure 3B) and IL-18 (Figure 3C), were observed, suggesting that JAK1-specific inhibition has therapeutic potential beyond the immune cell types involved in the pathology of CRS.

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

[0140] A 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 p-value of < 0.05 was considered statistically significant.

[0141] Example D: Preparation of a sustained-release formulation of compound 1 Sustained-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 sustained-release formulations of compound 1.

[0142] Protocol A Step 1: The adipine salt of compound 1, microcrystalline cellulose, hypromellose (methocel K100 LV and methocel K4M), and lactose monohydrate are individually sieved. Step 2: Transfer the sifted ingredients from Step 1 to a suitable blender and mix. Step 3: Transfer the mixture from Step 2 to a suitable granulator and mix. Step 4: Add purified water while mixing. Step 5: Transfer the granules from Step 4 to a suitable dryer and dry until the LOD is less than 3%. Step 6: Sift the granules from Step 5. Step 7: Mix the granules from Step 6 with the sieved magnesium stearate in a suitable blender. Step 8: The final mixture from Step 7 is compressed in a suitable rotary tableting machine.

[0143] Protocol B Step 1: The adipine salt of compound 1, microcrystalline cellulose, hypromellose, and pregelatinized starch are sieved individually. Step 2: Transfer the sifted ingredients from Step 1 to a suitable blender and mix. Step 3: Transfer the mixture from Step 2 to a suitable granulator and mix. Step 4: Add purified water while mixing. Step 5: Transfer the granules from Step 4 to 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 individually. 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 The final mixture from Step 9 is compressed in a suitable rotary tableting machine.

[0144] Protocol C Step 1: Lactose monohydrate, adipine of compound 1, microcrystalline cellulose, and hypromellose are individually sieved using a suitable sieve. Step 2: Transfer the sifted ingredients from Step 1 to a suitable blender and mix. Step 3: Transfer the mixture from Step 2 to a suitable granulator and mix. Step 4: Add purified water while mixing. Step 5 Sift the moist granules through a suitable sieve. Step 6: Transfer the granules from Step 5 to a suitable dryer and dry until the LOD is less than 3%. Step 7: Grind the granules from Step 6 into a powder. Step 8: Mix the granules from Step 7 and the sieved magnesium stearate in a suitable blender. Step 9: The final mixture from Step 8 is compressed using a suitable rotary tableting machine.

[0145] Protocol D Step 1: The pregelatinized starch, the adipate of compound 1, hypromellose, and a portion of the required microcrystalline cellulose are individually sieved using a suitable sieve. Step 2: Transfer the sifted ingredients from Step 1 to a suitable blender and mix. Step 3: Transfer the mixture from Step 2 to a suitable granulator and mix. Step 4: Add purified water while mixing. Step 5 Sift the moist granules through a suitable sieve. Step 6: Transfer the granules from Step 5 to a suitable dryer and dry until the LOD is less than 3%. Step 7: Grind the granules from Step 6 into a powder. Step 8: Sift the remaining 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 sodium bicarbonate and mix it with the mixture from Step 9. Step 11 Sift the magnesium stearate and mix it with the mixture from Step 10. Step 12 The final mixture from Step 11 is compressed in a suitable rotary tableting machine.

[0146] Composition of SR1:100mg sustained-release tablets [Table 6] a The conversion factor for adipine salts to free bases is 0.7911. b It was added after granulation. c Removed during processing.

[0147] Composition of SR2:100mg sustained-release tablets [Table 7] a The conversion factor for adipine salts to free bases 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 adipine salts to free bases is 0.7911. b It was added after granulation. c Removed during processing.

[0149] Composition of SR4:100mg sustained-release tablets [Table 9] a The conversion factor for adipine salts to free bases is 0.7911. b It was added after granulation. c Removed during processing. d Some was added before granulation, and some was added after granulation.

[0150] 25mg SR: Composition of 25mg sustained-release tablets [Table 10] a The conversion factor for adipine salts to free bases is 0.7911. b It was added after granulation. c Removed during processing.

[0151] In addition to those described herein, various modifications of the present invention will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to extend to the claims attached herein. All references, including all patents, patent applications and publications, cited herein are incorporated herein by reference in their entirety. Furthermore, this application includes the following aspects. [Aspect 1] A method for treating a target cytokine-related disorder or impairment, comprising administering a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof to the target. [Aspect 2] The method according to embodiment 1, wherein the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is more selective for JAK1 than for JAK2, JAK3, and Tyk2. [Aspect 3] The method according to embodiment 1 or 2, wherein the cytokine-related disease or disorder is cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell-associated encephalopathy syndrome (CRES). [Aspect 4] The method according to embodiment 3, wherein the cytokine-related disease or disorder is cytokine release syndrome (CRS). [Aspect 5] The method according to embodiment 3, wherein the cytokine-related disease or disorder is hemophagocytic lymphohistiocytosis (HLH). [Aspect 6] The method according to embodiment 3, wherein the cytokine-related disease or disorder is macrophage activation syndrome (MAS). [Aspect 7] The method according to embodiment 6, wherein the macrophage activation syndrome (MAS) is associated with systemic juvenile idiopathic arthritis. [Aspect 8] The method according to embodiment 3, wherein the cytokine-related disease or disorder is CAR-T cell-associated encephalopathy syndrome (CRES). [Aspect 9] The method according to any one of embodiments 1 to 8, wherein the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof. [Aspect 10] The method according to any one of embodiments 1 to 8, wherein the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-3-yl}acetonitrile adipate. [Aspect 11] The method according to any one of embodiments 1 to 8, wherein the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipirazole-1-yl)azetidine-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof. [Aspect 12] The method according to any one of embodiments 1 to 8, wherein the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipirazole-1-yl)azetidine-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide phosphate. [Aspect 13] The method according to any one of embodiments 1 to 8, wherein the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile or a pharmaceutically acceptable salt thereof. [Aspect 14] The method according to any one of embodiments 1 to 8, wherein the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile monohydrate. [Aspect 15] The method according to any one of embodiments 1 to 14, further comprising administering tocilizumab to the subject. [Aspect 16] The method according to any one of embodiments 1 to 14, further comprising administering a corticosteroid to the subject. [Aspect 17] The method according to any one of embodiments 1 to 14, further comprising administering prednisone to the subject. [Aspect 18] The method according to any one of embodiments 1 to 14, further comprising administering tocilizumab and a corticosteroid to the subject.

Claims

1. A pharmaceutical product comprising a JAK1 pathway inhibitor for the treatment of a cytokine-related disorder or impairment, wherein the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipirazole-1-yl)azetidine-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof, and the cytokine-related disorder or impairment is cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell-associated encephalopathy syndrome (CRES).

2. The pharmaceutical product according to claim 1, wherein the cytokine-related disease or disorder is cytokine release syndrome (CRS).

3. The pharmaceutical product according to claim 1, wherein the cytokine-related disease or disorder is hemophagocytic lymphohistiocytosis (HLH).

4. The pharmaceutical product according to claim 1, wherein the cytokine-related disease or disorder is macrophage activation syndrome (MAS).

5. The pharmaceutical product according to claim 4, wherein the macrophage activation syndrome (MAS) is related to systemic juvenile idiopathic arthritis.

6. The pharmaceutical product according to claim 1, wherein the cytokine-related disease or disorder is CAR-T cell-associated encephalopathy syndrome (CRES).

7. The pharmaceutical product according to any one of claims 1 to 6, wherein the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipirazole-1-yl)azetidine-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamidophosphate.

8. The pharmaceutical product according to any one of claims 1 to 7, wherein the pharmaceutical product is administered in combination with tocilizumab.

9. The pharmaceutical product according to any one of claims 1 to 7, wherein the pharmaceutical product is administered in combination with a corticosteroid.

10. The pharmaceutical product according to any one of claims 1 to 7, wherein the pharmaceutical product is administered in combination with prednisone.

11. The pharmaceutical product according to any one of claims 1 to 7, wherein the pharmaceutical product is administered in combination with tocilizumab and a corticosteroid.

12. A pharmaceutical product according to any one of claims 1 to 7, for use as monotherapy in the treatment of the cytokine-related disease or disorder.

13. The pharmaceutical agent according to any one of claims 1 to 12, wherein the treatment remits or inhibits the cytokine-related disease or disorder.

14. The pharmaceutical product according to any one of claims 1 to 12, wherein the treatment includes preventing the cytokine-related disease or disorder.

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