Treatment of hidradenitis suppurativa using jak inhibitors

Abstract

To provide methods of treating hidradenitis suppurativa using compounds that modulate activity of Janus kinase 1 and / or 2.SOLUTION: A method of treating hidradenitis suppurativa in a patient in need of treatment comprises, e.g., administering to the patient a therapeutically effective amount of a compound selected from among: ruxolitinib; ruxolitinib where one or more hydrogen atoms are replaced with deuterium atoms; {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; 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; and the like.SELECTED DRAWING: None

Description

[Technical field]

[0001] This application claims the benefit of U.S. Provisional Application No. 62 / 650,600, filed March 30, 2018, which is incorporated by reference in its entirety.

[0002] The present application provides methods of treating hidradenitis suppurativa (HS) using compounds that modulate the activity of Janus kinase (JAK) 1 and / or 2. [Background technology]

[0003] Protein kinases (PKs) regulate diverse biological processes including cell proliferation, survival, differentiation, organogenesis, morphogenesis, neovascularization, tissue repair, and regeneration, among others. Protein kinases also play specialized roles in the host in many human diseases, including cancer. Cytokines, small polypeptides or glycoproteins, regulate many pathways involved in the host inflammatory response to sepsis. Cytokines affect cell differentiation, proliferation, and activation and can regulate both pro- and anti-inflammatory responses, allowing the host to respond appropriately to pathogens. Signal transduction of a wide range of cytokines includes the Janus kinase family of protein tyrosine kinases (JAKs) and signal transducers and activators of transcription (STATs). There are four known mammalian JAKs, namely JAK1 (Janus kinase-1), JAK2, JAK3 (also known as leukocyte Janus kinase, JAKL and L-JAK), and TYK2 (protein tyrosine kinase 2).

[0004] Cytokine-stimulated immune and inflammatory responses contribute to disease pathogenesis as follows: conditions such as severe combined immunodeficiency (SCID) result from suppression of the immune system, while overactive or inappropriate immune / inflammatory responses contribute to the pathology of autoimmune diseases (e.g., asthma, systemic lupus erythematosus, thyroiditis, myocarditis) as well as diseases such as scleroderma and osteoarthritis (Ortmann, RA, T. Cheng, et al. (2000) Arthritis Res 2(1):16-32).

[0005] Loss of JAK expression is associated with many disease states. For example, Jak1- / - mice are stunted at birth, fail to lactate, and die perinatally (Rodig, SJ, MAMeraz, et al. (1998) Cell 93(3):373-83). Jak2- / - mouse embryos are anemic and die approximately 12.5 days post-coitum due to the absence of distinct erythropoiesis.

[0006] The JAK / STAT pathway, particularly all four JAKs, is believed to play a role in the pathogenesis of asthma responses, chronic obstructive pulmonary disease, bronchitis, and other related inflammatory diseases of the lower airways. Several cytokines that signal through JAKs have been linked to inflammatory diseases / pathologies of the upper airways, including those affecting the nose and paranasal sinuses (e.g., rhinitis and sinusitis), whether or not they are classical allergic reactions. The JAK / STAT pathway has also been implicated in inflammatory diseases / pathologies of the eye and chronic allergic reactions.

[0007] Activation of JAK / STAT in cancer can occur by cytokine stimulation (e.g., IL-6 or GM-CSF) or by reduction of endogenous inhibitors of JAK signaling, such as SOCS (suppressor of cytokine signaling) or PIAS (protein inhibitor of activated STAT) (Boudny, V., and Kovarik, J., Neoplasm. 49:349-355, 2002). Activation of STAT signaling, as well as other pathways downstream of JAK (e.g., Akt), correlates with poor prognosis in many cancer types (Bowman, T., et al. Oncogene 19:2474-2488, 2000). Elevated levels of circulating cytokines that signal through JAK / STAT play a causal role in cachexia and / or chronic fatigue. Therefore, JAK inhibition may be beneficial to cancer patients for reasons beyond potential antitumor activity.

[0008] JAK2 tyrosine kinase may be beneficial for patients with myeloproliferative syndromes, such as polycythemia vera (PV), essential thrombocythemia (ET), and myeloid metaplasia with myelofibrosis (MMM) (Levin, et al., Cancer Cell, vol. 7, 2005: 387-397). Inhibition of JAK2V617F kinase reduces hematopoietic cell proliferation, suggesting JAK2 as a possible target for pharmacological inhibition in patients with PV, ET, and MMM.

[0009] Inhibition of JAKs may benefit patients suffering from skin immune disorders and skin sensitization, such as psoriasis. The persistence of psoriasis is thought to depend on several inflammatory cytokines in addition to various chemokines and growth factors (JCI, 113:1664-1675), many of which signal through JAKs (Adv Pharmacol. 2000; 47:113-74).

[0010] Thus, new or improved agents that inhibit kinases such as JAKs are continually needed to develop new, more effective medicines aimed at enhancing or suppressing immune and inflammatory pathways, such as for the treatment of hidradenitis suppurativa. This application is directed to that need and others. Summary of the Invention

[0011] The present application provides a method for treating hidradenitis suppurativa in a patient in need of treatment, comprising administering to the patient a therapeutically effective amount of a compound that inhibits JAK1 and / or JAK2, or a pharma- ceutically acceptable salt thereof.

[0012] In some embodiments, the compound or salt is selective for JAK1 and JAK2, which is more selective than JAK3 and TYK2.

[0013] In some embodiments, the compound or salt is selective for JAK1 over JAK2, JAK3 and TYK2.

[0014] In some embodiments, the compound is ruxolitinib or a pharma- ceutically acceptable salt thereof.

[0015] In some embodiments, the compound is ruxolitinib, or a pharma- ceutically acceptable salt thereof, in which one or more hydrogen atoms have been replaced with deuterium atoms.

[0016] In some embodiments, the salt is ruxolitinib phosphate.

[0017] In some embodiments, the compound 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 pharma- ceutically acceptable salt thereof.

[0018] In some embodiments, the salt 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.

[0019] In some embodiments, the compound 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 pharma- ceutically acceptable salt thereof.

[0020] In some embodiments, the salt 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 phosphate.

[0021] In some embodiments, the compound or salt is administered in dosages of 15, 30, 60, or 90 mg based on the free base.

[0022] In some embodiments, the compound 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 pharma- ceutically acceptable salt thereof.

[0023] In some embodiments, the compound 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.

[0024] In some embodiments, the method further comprises administering an additional therapeutic agent (eg, an antibiotic, a retinoid, a corticosteroid, an anti-TNF-alpha agent, or an immunosuppressant).

[0025] In some embodiments, administration of the compound or salt is topical, hi some embodiments, administration of the compound or salt is oral.

[0026] In some embodiments, the method results in a 10%, 20%, 30%, 40%, or 50% improvement in HiSCR (Hidradenitis Suppurativa Clinical Response).

[0027] The present application also provides a compound that inhibits JAK1 and / or JAK2, or a pharma- ceutically acceptable salt thereof, for use in the treatment of hidradenitis suppurativa.

[0028] The present application also further provides the use of a compound that inhibits JAK1 and / or JAK2, or a pharma- ceutically acceptable salt thereof, for the preparation of a medicament for use in the treatment of hidradenitis suppurativa. [Brief description of the drawings]

[0029] [Figure 1] Figure 1 shows the individual gene expression values ​​(MFI) of JAK1 for each experimental replicate in keratinocytes simulated with TNFα and IFN-γ in the presence / absence of compounds A-D. Keratinocytes were stimulated with TNFα (25 ng / mL) and IFNγ (25 ng / mL) in the presence / absence of increasing concentrations of JAK inhibitors. Data are presented as JAK1 expression levels for each group. [Diagram 2] Figure 1 shows the individual gene expression values ​​(MFI) of JAK2 for each experimental replicate in keratinocytes simulated with TNFα and IFN-γ in the presence / absence of compounds A-D. Keratinocytes were stimulated with TNFα (25 ng / mL) and IFNγ (25 ng / mL) in the presence / absence of increasing concentrations of JAK inhibitors. Data are presented as JAK2 expression levels for each group. [Diagram 3] Figure 1 shows individual gene expression values ​​(MFI) of IL-1α for each experimental replicate in keratinocytes simulated with TNFα and IFN-γ in the presence / absence of compounds A-D. Keratinocytes were stimulated with TNFα (25 ng / mL) and IFNγ (25 ng / mL) in the presence / absence of increasing concentrations of JAK inhibitors. Data are presented as IL-1α expression levels for each group. [Figure 4] Figure 1 shows individual gene expression values ​​(MFI) of IL-6 for each experimental replicate in keratinocytes simulated with TNFα and IFN-γ in the presence / absence of compounds A-D. Keratinocytes were stimulated with TNFα (25 ng / mL) and IFNγ (25 ng / mL) in the presence / absence of increasing concentrations of JAK inhibitors. Data are presented as IL-6 expression levels for each group. [Diagram 5]Figure 1 shows the individual protein concentrations (pg / mL) of IL-1α for each experimental replicate in keratinocytes simulated with TNFα and IFN-γ in the presence / absence of compounds A-D. Keratinocytes were stimulated with TNFα (25 ng / mL) and IFNγ (25 ng / mL) in the presence / absence of increasing concentrations of JAK inhibitors. Data are presented as IL-1α concentration for each group. [Figure 6] Figure 1 shows the individual protein concentrations (pg / mL) of IL-6 for each experimental replicate in keratinocytes simulated with TNFα and IFN-γ in the presence / absence of compounds A-D. Keratinocytes were stimulated with TNFα (25 ng / mL) and IFNγ (25 ng / mL) in the presence / absence of increasing concentrations of JAK inhibitors. Data are presented as IL-6 concentration for each group. [Figure 7] Figure 1 shows gene expression (MFI) of JAK1, JAK3, and TYK2 in the skin of healthy controls and subjects with hidradenitis suppurativa. Data are presented as JAK1, JAK3, or TYK2 gene expression levels for each healthy control (n=4) and hidradenitis suppurativa (n=41) subject. [Figure 8] 1 shows gene expression (MFI) of STAT1, STAT2, and STAT3 in the skin of healthy controls and subjects with hidradenitis suppurativa. Data are presented as STAT1, STAT2, or STAT3 gene expression levels for each healthy control (n=4) and hidradenitis suppurativa (n=41) subject. [Figure 9] Figure 1 shows gene expression (MFI) of IRAK1, IRAK2, and IRAK4 in the skin of healthy controls and subjects with hidradenitis suppurativa. Data are presented as IRAK1, IRAK2, or IRAK4 gene expression levels for each healthy control (n=4) and hidradenitis suppurativa (n=41) subject. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0030] The present application provides, inter alia, a method for treating hidradenitis suppurativa in a patient in need of treatment, comprising administering a therapeutically effective amount of a compound that inhibits JAK1 and / or JAK2, or a pharma- ceutically acceptable salt thereof.

[0031] The methods described herein utilize compounds or salts that are inhibitors of JAK1 and / or JAK2. In some embodiments, the compound is: Ruxolitinib; Ruxolitinib in which one or more hydrogen atoms have been replaced with deuterium atoms; {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; 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; [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; 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; ((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; 3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; 4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; [trans-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-3-(4-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperazin-1-yl)cyclobutyl]acetonitrile; {trans-3-(4-{[4-[(3-hydroxyazetidin-1-yl)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-{[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-{[(2R)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; 4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile; 5-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide; 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide; 5-{3-(cyanomethyl)-3-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide; {1-(cis-4-{[6-(2-hydroxyethyl)-2-(trifluoromethyl)pyrimidin-4-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-[(ethylamino)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-(1-hydroxy-1-methylethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-{[(3R)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-{[(3S)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {trans-3-(4-{[4-({[(1S)-2-hydroxy-1-methylethyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-({[(2R)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-({[(2S)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-(2-hydroxyethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; or a pharma- ceutically acceptable salt of any of the foregoing.

[0032] In some embodiments, the compound or salt is selective for JAK1 and JAK2 over JAK3 and TYK2. In some embodiments, the compound is 3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile (ruxolitinib), or a pharma- ceutically acceptable salt thereof. Ruxolitinib has an IC of less than 10 nM at 1 mM ATP (assay A) for JAK1 and JAK2. 50 3-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile and ruxolitinib can be prepared by the procedure described in US Pat. No. 7,598,257, filed Dec. 12, 2006 (Example 67), which is incorporated herein by reference in its entirety. In some embodiments, the inhibitor of JAK1 and / or JAK2 is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile phosphate. The phosphate can be prepared as described in US Pat. No. 8,722,693, which is incorporated herein by reference in its entirety.

[0033] In some embodiments, the compound or salt is a JAK1 inhibitor. In some embodiments, the compound or salt is selective for JAK1 over JAK2, JAK3, and TYK2. For example, some of the compounds described herein, or pharma- ceutically acceptable salts thereof, preferentially inhibit JAK1 over one or more of JAK2, JAK3, and TYK2. JAK1 plays a central role in many cytokine and growth factor signaling pathways that, when dysregulated, may result in or contribute to disease states. For example, IL-6 levels are elevated in rheumatoid arthritis, a disease in which IL-6 has been suggested to have deleterious effects (Fonesca, et al., Autoimmunity Reviews, 8:538-42, 2009). IL-6 signals at least in part through JAK1, and thus IL-6 may indirectly provide potential clinical benefit through JAK1 inhibition (Guschin, et al. Embo J 14:1421, 1995; Smolen, et al. Lancet 371:987, 2008). Furthermore, in some cancers, JAK1 is mutated, resulting in constitutive proliferation and survival of unwanted tumor cells (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. Thus, patients with such diseases may benefit from inhibition of JAK1. A selective inhibitor of JAK1 would be beneficial while avoiding the unnecessary and potentially undesirable effects of inhibiting other JAK kinases.

[0034] Hidradenitis suppurativa is characterized by significant skin inflammation, although there are only limited publications outlining the inflammation (Hoffman et al., PLOS One, September 28, 2018, https: / / doi.org / 10.1371 / journal.pone.0203672). Provided herein are examples supporting the hypothesis that inflammation is driven in large part by the JAK / STAT-mediated pathway. Examples C, D, and E show elevated levels of JAK / STAT gene expression in the skin of HS patients compared to healthy skin. Furthermore, Examples C, D, and E show that proinflammatory cytokines (TNF-alpha and IFN-gamma), known to be elevated in HS, induce the JAK / STAT pathway in cultured keratinocytes, and that this induction can be reduced by the addition of a JAK inhibitor. Thus, patients with HS may benefit from inhibition of JAK1. A selective inhibitor of JAK1 would be beneficial while avoiding the unnecessary and potentially undesirable effects of inhibiting other JAK kinases.

[0035] In some embodiments, the compound preferentially inhibits JAK1 over JAK2 (e.g., IC of JAK2 / JAK1). 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 IC 50 The antibodies are 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-terminal sequence of the antibody (see Example A).

[0036] In some embodiments, the JAK1 inhibitor is a compound in Table 1 or a pharma- ceutically 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 at 1 mM ATP 50 The values ​​are shown in Table 1.

[0037] [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

[0038] In some embodiments, the JAK1 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 pharma- ceutically acceptable salt thereof.

[0039] In some embodiments, the JAK1 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.

[0040] 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 adipate salts can be found in U.S. Patent Publication No. 2011 / 0224190, filed March 9, 2011, U.S. Patent Publication No. 2013 / 0060026, filed September 6, 2012, and U.S. Patent Publication No. 2014 / 0256941, filed March 5, 2014, each of which is incorporated herein by reference in its entirety.

[0041] In some embodiments, the JAK1 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 pharma- ceutically acceptable salt thereof.

[0042] In some embodiments, the JAK1 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 phosphate.

[0043] In some embodiments, JAK1 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 hydrochloride.

[0044] In some embodiments, JAK1 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 hydrobromide.

[0045] In some embodiments, JAK1 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 sulfate.

[0046] 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 Publication No. 2014 / 0343030, filed May 16, 2014, each of which is incorporated herein by reference in its entirety.

[0047] In some embodiments, the JAK1 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 pharma- ceutically acceptable salt thereof.

[0048] In some embodiments, the JAK1 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.

[0049] 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 can be found in U.S. Patent Publication No. 2014 / 0121198, filed October 31, 2013, and U.S. Patent Publication No. 2015 / 0344497, filed April 29, 2015, each of which is incorporated herein by reference in its entirety.

[0050] In some embodiments, the compounds in Table 1 are disclosed in U.S. Patent Publication No. 2011 / 0224190, filed March 9, 2011; U.S. Patent Publication No. 2014 / 0343030, filed May 16, 2014; U.S. Patent Publication No. 2014 / 0121198, filed October 31, 2013; U.S. Patent Publication No. 2010 / 0298334, filed May 21, 2010; U.S. Patent Publication No. 2011 / 0059951, filed August 31, 2010; U.S. Patent Publication No. 2011 / 0063636, filed November 28, 2011; No. 2012 / 0149681, filed on May 18, 2011; U.S. Patent Publication No. 2012 / 0149682, filed on November 18, 2011; U.S. Patent Publication No. 2013 / 0018034, filed on June 19, 2012; U.S. Patent Publication No. 2013 / 0045963, filed on August 17, 2012; and U.S. Patent Publication No. 2014 / 0005166, filed on May 17, 2013, each of which is incorporated herein by reference in its entirety.

[0051] In some embodiments, the JAK1 inhibitor is selected from the group consisting of those disclosed in U.S. Patent Publication No. 2011 / 0224190, filed March 9, 2011, U.S. Patent Publication No. 2014 / 0343030, filed May 16, 2014, U.S. Patent Publication No. 2014 / 0121198, filed October 31, 2013, U.S. Patent Publication No. 2010 / 0298334, filed May 21, 2010, U.S. Patent Publication No. 2011 / 0059951, filed August 31, 2010, U.S. Patent Publication No. 2011 / 0059951, filed November 18, 2011, and U.S. Patent Publication No. 2011 / 0063664, filed May 18, 2011. No. 2012 / 0149681, U.S. Patent Publication No. 2012 / 0149682, filed November 18, 2011, U.S. Patent Publication No. 2013 / 0018034, filed June 19, 2012, U.S. Patent Publication No. 2013 / 0045963, filed August 17, 2012, and U.S. Patent Publication No. 2014 / 0005166, filed May 17, 2013, or a pharma- ceutically acceptable salt thereof, each of which is incorporated herein by reference in its entirety.

[0052] In some embodiments, the JAK1 inhibitor is a compound of formula I [ka] or a pharma- ceutically 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.

[0053] 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 pharma- ceutically acceptable salt thereof.

[0054] 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 pharma- ceutically acceptable salt thereof.

[0055] 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 pharma- ceutically acceptable salt thereof.

[0056] In some embodiments, the JAK1 inhibitor is a compound of formula II [ka] or a pharma- ceutically 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 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 7is H or F; R 8 is H or methyl; R 9 is H or methyl; R 10 is H or methyl; R 11 is H or methyl.

[0057] 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 pharma- ceutically acceptable salt thereof.

[0058] In some embodiments, the JAK1 inhibitor is a compound of formula III [ka] or a pharma- ceutically acceptable salt thereof, wherein: Cy 4 CN, OH, F, Cl, C 1-3 Alkyl, C 1-3 Haloalkyl, CN-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 the group consisting of aryl, ... 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.

[0059] 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 pharma- ceutically acceptable salt thereof.

[0060] In some embodiments, the JAK1 and / or JAK2 inhibitor is valcitinib, tofacitinib, oclacitinib, filgotinib, gandotinib, lestaurtinib, momelotinib, bacritinib, PF-04965842, upadacitinib, peficitinib, cucurbitacin I, ATI-501 (Aclaris), ATI-502 (Aclaris), JTE052 (Leo Pharma and Japan Tobacco), or CHZ868.

[0061] In some embodiments, the inhibitor of JAK1 and / or JAK2 may be an isotopically labeled compound, or a pharma- ceutically acceptable salt thereof. An "isotopically" or "radiolabelled" compound is a compound of the present disclosure in which one or more atoms have been replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature (i.e., naturally occurring). Suitable radionuclides that may be incorporated into the compounds of the present disclosure include: 2 H (also written as D for 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 131These include, but are not limited to, I. For example, one or more hydrogen atoms in the compounds of the present disclosure can be replaced with a deuterium atom, such that -CH3 is replaced with -CD3.

[0062] One or more constituent atoms of the compounds described herein may be replaced or substituted with natural or non-natural abundance isotopes of the atoms. In some embodiments, the compounds contain at least one deuterium atom. In some embodiments, the compounds contain two or more deuterium atoms. In some embodiments, the compounds contain 1-2, 1-3, 1-4, 1-5, or 1-6 deuterium atoms. In some embodiments, all hydrogen atoms in the compounds may be replaced or substituted with deuterium atoms.

[0063] Synthetic methods for incorporating isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas, New York, NY, Appleton-Century-Crofts, 1971; The Renaissance of H / D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey, Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling by James R. Hanson, Royal Society of Chemistry, 2011). Isotopically labeled compounds can be used in a variety of studies, such as NMR spectroscopy, metabolic experiments, and / or assays.

[0064] Substitution with heavier isotopes, such as deuterium, can provide certain therapeutic advantages, such as increased in vivo half-life or reduced dosage requirements, due to greater metabolic stability, and therefore may be preferred in some cases (see, e.g., A. Kerekes et.al. J. Med. Chem. 2011, 54, 201-210; R. Xu et.al. J. Label Compd. Radiopharm. 2015, 58, 308-312). In particular, substitution at one or more metabolic sites may provide one or more therapeutic advantages.

[0065] Thus, in some embodiments, an inhibitor of JAK1 and / or JAK2 is a compound, or a pharma- ceutically acceptable salt thereof, in which one or more hydrogen atoms in the compound are replaced by deuterium atoms.

[0066] In some embodiments, the inhibitor of JAK1 and / or JAK2 is ruxolitinib, in which one or more hydrogen atoms are replaced by deuterium atoms, or a pharma- ceutically acceptable salt thereof. In some embodiments, the inhibitor of JAK1 and / or JAK2 is any of the compounds described in U.S. Patent No. 9,249,149, which is incorporated herein by reference in its entirety, or a pharma- ceutically acceptable salt thereof. In some embodiments, the inhibitor of JAK1 and / or JAK2 is CTP-543, or a pharma- ceutically acceptable salt thereof.

[0067] In some embodiments, the compound is a compound of formula I [ka] or a pharma- ceutically acceptable salt thereof, wherein: R 1 is selected from H and D; Each R 2 are independently selected from H and D, provided that each R bonded to a common carbon 2 is the same; Each R 3are independently selected from H and D, provided that each R bonded to a common carbon 3 is the same; R 4 is selected from H and D; Each R 5 are the same and selected from H and D; R 6 , R 7 , and R 8 are each independently selected from H and D; 1 is H, and each R 2 and each R 3 is H and R 4 is H and R 6 , R 7 , and R 8 When each is H, each R 5 is D.

[0068] In some embodiments, the inhibitor of JAK1 and / or JAK2 is a compound of formula I selected from the following compounds 100-130 in the table below, where R 6 , R 7 , and R 8 In some embodiments, the inhibitor of JAK1 and / or JAK2 is a compound of formula I selected from the following compounds 200-231 in the table below, where R 6 , R 7 , and R 8 each is D), or a pharma- ceutically acceptable salt thereof. TIFF2024026074000015.tif246152 TIFF2024026074000016.tif204152

[0069] In some embodiments, the inhibitor of JAK1 and / or JAK2 is baricitinib, or a pharma- ceutically acceptable salt thereof, in which one or more hydrogen atoms are replaced by deuterium atoms. In some embodiments, the inhibitor of JAK1 and / or JAK2 is any of the compounds described in U.S. Patent No. 9,540,367, which is incorporated herein by reference in its entirety, or a pharma- ceutically acceptable salt thereof.

[0070] As used herein, the phrase "optionally substituted" means unsubstituted or substituted. As used herein, the term "substituted" means that a hydrogen atom has been removed and replaced with a substituent. It is understood that substitution at a given atom is limited by the valence of the atom.

[0071] As used herein, "C" when used alone or in combination with other terms n-m The term "alkyl" refers to a saturated hydrocarbon group that may be straight or branched chain having n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6 or 1 to 3 carbon atoms. Examples of alkyl moieties include, but are not limited to, chemical groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methyl-1-butyl, 3-pentyl, n-hexyl, 1,2,2-trimethylpropyl, and the like.

[0072] As used herein, the term "alkylene," used alone or in combination with other terms, refers to a divalent alkyl linking group, which may be branched or straight chain, in which two substituents may be attached at any position of the alkylene linking group. Examples of alkylene groups include, but are not limited to, ethane-1,2-diyl, propane-1,3-diyl, propane-1,2-diyl, and the like.

[0073] As used herein, "HO-C 1-3The term "-alkyl" refers to a group of formula -alkylene-OH, said alkylene group having 1 to 3 carbon atoms.

[0074] As used herein, "CN-C 1-3 The term "alkyl" refers to a C 1-3 Refers to alkyl.

[0075] As used herein, the term "amino" refers to a group of formula -NH2.

[0076] As used herein, "di(C 1-3 The term "-N(alkyl)amino" refers to a group of the formula -N(alkyl)2, where each of the two alkyl groups independently has 1 to 3 carbon atoms.

[0077] As used herein, "C 1-3 The term "alkylamino" refers to a group of the formula -NH(alkyl), where the alkyl group has 1 to 3 carbon atoms.

[0078] As used herein, "di(C 1-3 The term "alkyl)aminosulfonyl" refers to a group of formula -S(O)2N(alkyl)2, where each alkyl group independently has 1 to 3 carbon atoms.

[0079] As used herein, "C 1-3 The term "alkylsulfonyl" refers to a radical of the formula -S(O)2-alkyl, where the alkyl radical has 1 to 3 carbon atoms.

[0080] As used herein, "halo" or "halogen," used alone or in combination with other terms, includes fluoro, chloro, bromo, and iodo. In some embodiments, a halo group is fluoro or chloro.

[0081] As used herein, "C" when used alone or in combination with other terms means n-mThe term "haloalkyl" refers to a C alkyl group having up to {2(n-m)+1} halogen atoms which may be the same or different. n-m In some embodiments, the halogen atom is a fluoro atom. In some embodiments, the alkyl group has 1-6 or 1-3 carbon atoms. Examples of haloalkyl groups include CF3, C2F5, CHF2, CCl3, CHCl2, C2Cl5, and the like. In some embodiments, the haloalkyl group is a fluoroalkyl group.

[0082] As used herein, "C 1~3 The term "fluoroalkyl" refers to a C alkyl group that may be partially or fully substituted with fluoro atoms. 1~3 Refers to an alkyl group.

[0083] As used herein, "C" when used alone or in combination with other terms means 3-6 The term "cycloalkyl" refers to a non-aromatic monocyclic hydrocarbon moiety having 3 to 6 carbon atoms that may optionally contain one or more alkenylene groups as part of the ring structure. One or more ring-forming carbon atoms of a cycloalkyl group may be oxidized to form a carbonyl bond. Exemplary C 3-6 Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, etc. In some embodiments, the cyclopropyl group is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0084] As used herein, "C 3-6 Cycloalkyl-C 1-3 The term "alkyl" refers to a group of the formula -C 1-3 Alkylene-C 3-6 Refers to a cycloalkyl group.

[0085] The compounds described herein may be asymmetric (e.g., have 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 inactive starting materials are 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 may 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 may be isolated as a mixture of isomers or as separated isomeric forms. In some embodiments, the compounds have the (R) configuration. In some embodiments, the compounds have the (S) configuration.

[0086] Resolution of racemic mixtures of compounds can be carried out by any of a number of methods known in the art. Exemplary methods include fractional recrystallization using chiral resolving acids that are optically active salt-forming organic acids. Resolving agents suitable for fractional recrystallization are optically active acids such as, for example, 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 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.

[0087] Resolution of racemic mixtures can also be carried out by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). Appropriate elution solvent compositions can be determined by one skilled in the art.

[0088] The compounds described herein also include tautomeric forms. Tautomeric forms are obtained by the exchange of a single bond with an adjacent double bond and the concomitant transfer 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 more than one position of a heterocyclic ring system, such as 1H-imidazole and 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 can be in equilibrium or sterically fixed in one form by appropriate substitution. For example, it will be recognized that the following pyrazole ring can form two tautomers: [ka] The claims are intended to cover both tautomers.

[0089] All compounds and their pharma- ceutically acceptable salts may be found together with other substances such as water and solvents (eg, hydrates and solvates) or may be isolated.

[0090] In some embodiments, the compounds described herein or salts thereof are substantially isolated. By "substantially isolated" it is meant that the compound is at least partially or substantially separated from the environment in which it was formed or detected. Partial separation can include, for example, compositions enriched for the compounds described herein. Substantial separation can include compositions containing at least about 50% by weight, at least about 60% by weight, at least about 70% by weight, at least about 80% by weight, at least about 90% by weight, at least about 95% by weight, at least about 97% by weight, or at least about 99% by weight of the compounds described herein or salts thereof. Methods for isolating compounds and their salts are routine in the art.

[0091] The phrase "pharmacologically acceptable" is used herein to refer to compounds, substances, compositions, and / or dosage forms that are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, within the scope of sound medical judgment, commensurate with a reasonable benefit / risk ratio.

[0092] As used herein, the expressions "ambient temperature" and "room temperature" or "rt" are art-recognized and generally refer to a temperature, e.g., a reaction temperature approaching the temperature of the room in which the reaction is carried out, e.g., a temperature of about 20° C. to about 30° C.

[0093] The present invention also includes pharma- ceutically acceptable salts of the compounds described herein. As used herein, "pharma- ceutically acceptable salts" refers to derivatives of the disclosed compounds, in which the parent compound is modified by converting an acidic or basic moiety present therein into its salt form. Examples of pharma- ceutically 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 pharma- ceutically acceptable salts of the present application include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharma- ceutically acceptable salts of the present application can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. In general, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or in a mixture of the two, generally in non-aqueous media such as ether, ethyl acetate, alcohol (e.g., methanol, ethanol, isopropanol, or butanol) or acetonitrile (ACN). 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.

[0094] As used herein, the term "contacting" refers to bringing the indicated moieties together in an in vitro system or in vivo system. For example, "contacting" a JAK with a compound of the invention includes administering a compound of the application to an individual or patient, such as a human, that has a JAK, as well as introducing a compound of the invention to a sample, including, for example, a cell preparation or purified preparation that contains a JAK.

[0095] As used herein, the terms "subject", "individual", or "patient" are used interchangeably and refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, primates, and most preferably humans. In some embodiments, a "subject", "individual", or "patient" is in need of said treatment.

[0096] 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 pharmaceutical response desired by a researcher, veterinarian, physician, or other clinician in a tissue, system, animal, individual, or human.

[0097] As used herein, the term "treating" or "treatment" refers to one or more of: (1) inhibiting a disease, e.g., inhibiting a disease, condition, or disorder in an individual experiencing or exhibiting a pathology or symptom of the disease, condition, or disorder (i.e., preventing further progression of the pathology and / or symptoms); (2) ameliorating a disease, e.g., ameliorating a disease, condition, or disorder in an individual experiencing or exhibiting a pathology or symptom of the disease, condition, or disorder (i.e., reversing the pathology and / or symptoms), e.g., reducing the severity of the disease; or (3) preventing a disease, condition, or disorder in an individual who may be predisposed to the disease, condition, or disorder but has not yet experienced or exhibited a pathology or symptom of the disease. In some embodiments, treating refers to inhibiting or ameliorating a disease. In some embodiments, treating is preventing a disease.

[0098] Combination therapy The methods described herein may further include administering one or more additional therapeutic agents. The one or more additional therapeutic agents may be administered to the patient simultaneously or sequentially.

[0099] In some embodiments, the additional therapeutic agent is an antibiotic. In some embodiments, the antibiotic is clindamycin, doxycycline, minocycline, trimethoprim-sulfamethoxazole, erythromycin, metronidazole, rifampin, moxifloxacin, dapsone, or a combination thereof. In some embodiments, the antibiotic is clindamycin, doxycycline, minocycline, trimethoprim-sulfamethoxazole, or erythromycin in combination with metronidazole. In some embodiments, the antibiotic is a combination of rifampin, moxifloxacin, and metronidazole. In some embodiments, the antibiotic is a combination of moxifloxacin and rifampin.

[0100] In some embodiments, the additional therapeutic agent is a retinoid, hi some embodiments, the retinoid is etretinate, acitretin, or isotretinoin.

[0101] In some embodiments, the additional therapeutic agent is a steroid. In some embodiments, the additional therapeutic agent is a corticosteroid. In some embodiments, the steroid is triamcinolone, dexamethasone, fluocinolone, cortisone, prednisone, prednisolone, or flumetholone, or the like.

[0102] In some embodiments, the additional therapeutic agent is an anti-TNF-alpha agent. In some embodiments, the anti-TNF-alpha agent is an anti-TNF-alpha antibody. In some embodiments, the anti-TNF-alpha agent is infliximab or etanercept, or adalimumab.

[0103] In some embodiments, the additional therapeutic agent is an immunosuppressant. In some embodiments, the immunosuppressant is methotrexate or cyclosporine A. In some embodiments, the immunosuppressant is mycophenolate mofetil or mycophenolate sodium.

[0104] In some embodiments, the additional therapeutic agent is finasteride, metformin, adapalene, or azelaic acid.

[0105] In some embodiments, the method further comprises administering an additional therapeutic agent selected from an IMiD, an anti-IL-6 agent, a hypomethylating agent, and a biological response modifier (BRM).

[0106] Generally, BRMs are substances produced by living organisms to treat disease and may occur naturally in the body or may be produced in a laboratory. Examples of BRMs include IL-2, interferons, various types of colony stimulating factors (CSF, GM-CSF, G-CSF), monoclonal antibodies such as abciximab, etanercept, infliximab, rituximab, trastuzumab, and high-dose ascorbate.

[0107] In some embodiments, the hypomethylating agent is a DNA methyltransferase inhibitor. In some embodiments, the DNA methyltransferase inhibitor is selected from 5-azacytidine and decitabine.

[0108] Generally, the IMiD is an immunomodulatory agent. In some embodiments, the IMiD is selected from thalidomide, lenalidomide, pomalidomide, CC-11006, and CC-10015.

[0109] In some embodiments, the method further comprises administering an additional therapeutic agent selected from antithymocyte globulin, recombinant human granulocyte colony stimulating factor (G CSF), granulocyte-monocyte CSF (GM-CSF), erythropoietin stimulating agents (ESAs), and cyclosporine.

[0110] In some embodiments, the method further comprises administering to the patient an additional JAK inhibitor, in some embodiments, the additional JAK inhibitor is valcitinib, tofacitinib, oclacitinib, filgotinib, gandotinib, lestaltinib, momelotinib, bacritinib, PF-04965842, upadacitinib, peficitinib, fedratinib, cucurbitacin I, or CHZ868.

[0111] One or more additional pharmaceutical agents, such as anti-inflammatory agents, immunosuppressants, and PI3Kδ, mTor, Bcr-Abl, Flt-3, RAF, and FAK kinase inhibitors, such as those described in WO2006 / 056399, which is incorporated herein by reference in its entirety, or other agents, can be used in combination with the compounds described herein for the treatment of a JAK-associated disease, disorder, or condition. The one or more additional pharmaceutical agents can be administered to the patient simultaneously or sequentially.

[0112] Examples of Bcr-Abl inhibitors include the genera and species of compounds disclosed in U.S. Pat. No. 5,521,184, WO 04 / 005281, and U.S. Patent Application No. 60 / 578,491, all of which are incorporated herein by reference in their entireties, and pharma- ceutically acceptable salts thereof.

[0113] Examples of suitable Flt-3 inhibitors include compounds and pharma- ceutically acceptable salts thereof, such as those disclosed in WO03 / 037347, WO03 / 099771, and WO04 / 046120, all of which are incorporated herein by reference in their entirety.

[0114] Examples of suitable RAF inhibitors include compounds and pharma- ceutically acceptable salts thereof, such as those disclosed in WO00 / 09495, and WO05 / 028444, both of which are incorporated herein by reference in their entireties.

[0115] Examples of suitable FAK inhibitors include compounds and pharma- ceutically acceptable salts thereof, such as those disclosed in WO04 / 080980, WO04 / 056786, WO03 / 024967, WO01 / 064655, WO00 / 053595, and WO01 / 014402, all of which are incorporated herein by reference in their entirety.

[0116] In some embodiments, one or more compounds of the present invention can be used in combination with one or more other kinase inhibitors, including imatinib, particularly to treat patients who are resistant to imatinib or other kinase inhibitors.

[0117] In some embodiments, the additional therapeutic agent is fluocinolone acetonide (Retisert®), or rimexolone (AL-2178, Vexol, Alcon).

[0118] In some embodiments, the additional therapeutic agent is cyclosporine (Restasis®).

[0119] In some embodiments, the additional therapeutic agent is Dehydrex™ (Holles Labs), Civamide (Opko), sodium hyaluronate (Vismed, Lantibio / TRB Chemedia), cyclosporine (ST-603, Sirion Therapeutics), ARG101(T) (testosterone, Argentis), AGR1012(P) (Argentis), ecabet sodium (Senju-Ista), gefarnate (Santen), 15-(s)-hydroxyeicosatetraenoic acid (15(S)-HETE), seviremine, doxycycline (ALTY-0501, Alacrity), minocycline, iDestrin™ (NP50301, Nascent Pharmaceuticals), cyclosporine A (Nova22007, Novagali), oxytetracycline (Duramycin, MOLI1901, Lantibio), CF101(2S,3S,4R,5R)-3,4-Dihydroxy-5-[6-[(3-iodophenyl)methylamino]purin-9-yl]-N-methyl-oxolane-2-carbamyl, Can-Fite Biopharma), voclosporin (LX212 or LX214, Lux Biosciences), ARG103 (Agentis), RX-10045 (synthetic resolvin analog, Resolvyx), DYN15 (Dyanmis Therapeutics), rivoglitazone (DE011, Daiichi Sanko), TB4 (RegeneRx), OPH-01 (Ophtalmis Monaco), PCS101 (Pericor Science), REV1-31 (Evolutec), Lacritin (Senju), rebamipide (Otsuka-Novartis), OT-551 (Othera), PAI-2 (University of Pennsylvania and Temple University) The therapeutic agent is selected from the group consisting of rifabutin, tacrolimus, pimecrolimus (AMS981, Novartis), loteprednol etabonate, rituximab, diquafosol tetrasodium (INS365, Inspire), KLS-0611 (Kissei Pharmaceuticals), dehydroepiandrosterone, anakinra, efalizumab, mycophenolate sodium, etanercept (Embrel®), hydroxychloroquine, NGX267 (TorreyPines Therapeutics), Actemra, gemcitabine, oxaliplatin, L-asparaginase, and thalidomide.

[0120] In some embodiments, the additional therapeutic agent is an antiangiogenic agent, a cholinergic agonist, a TRP-1 receptor modulator, a calcium channel blocker, a mucin secretagogue, a MUC1 stimulator, a calcineurin inhibitor, a corticosteroid, a P2Y2 receptor agonist, a muscarinic receptor agonist, an mTOR inhibitor, another JAK inhibitor, a Bcr-Abl kinase inhibitor, an Flt-3 kinase inhibitor, a RAF kinase inhibitor, and a FAK kinase inhibitor, such as those described in WO2006 / 056399, the entirety of which is incorporated herein by reference. In some embodiments, the additional therapeutic agent is a tetracycline derivative (e.g., minocycline or doxycycline). In some embodiments, the additional therapeutic agent binds to FKBP12.

[0121] In some embodiments, the additional therapeutic agent is an alkylating or DNA crosslinking agent; antimetabolite / demethylating agent (e.g., 5-fluorouracil, capecitabine, or azacitidine); anti-hormonal therapy (e.g., hormone receptor antagonists, SERMs, aromatase inhibitors); mitotic inhibitors (e.g., vincristine or paclitaxel); topoisomerase (I or II) inhibitors (e.g., mitoxantrone and irinotecan); apoptosis inducers (e.g., ABT-737); nucleic acid therapy. nuclear receptor ligands (e.g., agonists and / or antagonists: all-trans retinoic acid or bexarotene); epigenetic targeting agents such as histone deacetylase inhibitors (e.g., vorinostat), hypomethylating agents (e.g., decitabine); modulators of protein stability such as Hsp90 inhibitors, ubiquitin and / or ubiquitin-like binding or debinding molecules; or EGFR inhibitors (erlotinib).

[0122] In some embodiments, the additional therapeutic agent includes antibiotics, antivirals, antifungals, anesthetics, anti-inflammatory agents including steroidal and non-steroidal anti-inflammatory drugs, and anti-allergy agents. Examples of suitable pharmaceutical agents include aminoglycosides such as amikacin, gentamicin, tobramycin, streptomycin, netilmicin, and kanamycin; fluoroquinolones such as ciprofloxacin, norfloxacin, ofloxacin, trovafloxacin, lomefloxacin, levofloxacin, and enoxacin; naphthyridines; sulfonamides; polymyxins; chloramphenicol; neomycin; paramomycin; colistimethate; bacitracin ... These include: cyclosporine; tetracyclines; rifampin and its derivatives ("rifampin"); cycloserine; beta-lactams; cephalosporins; amphotericin; fluconazole; flucytosine; natamycin; miconazole; ketoconazole; corticosteroids; diclofenac; flurbiprofen; ketorolac; suprofen; cromolyn; lodoxamide; levocabastine; naphazoline; antazoline; pheniramine; or azalide antibiotics.

[0123] Pharmaceutical Preparations and Dosage Forms When used as a pharmaceutical, the compounds of the present invention can be administered in the form of pharmaceutical compositions. 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 (including transdermal, epidermal, ocular, and mucosal delivery, including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion, or intracranial, e.g., 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.

[0124] In some embodiments, administration is topical, hi some embodiments, administration is topical to the skin.

[0125] In some embodiments, administration is oral.

[0126] The present invention also includes pharmaceutical compositions that contain the compound of the present invention or a pharma- ceutically acceptable salt thereof as an active ingredient in combination with one or more pharma- ceutically acceptable carriers (excipients). In some embodiments, the compositions are suitable for topical administration. In preparing the compositions of the present invention, the active ingredient is typically mixed with an excipient, diluted by an excipient, or enclosed in such a carrier, for example, in the form of a capsule, sachet, paper, or other container. When an excipient functions as a diluent, it can be a solid, semi-solid, or liquid substance that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the composition can be in the form of a tablet, pill, powder, lozenge, sachet, cachet, elixir, suspension, emulsion, solution, syrup, aerosol (as a solid or liquid medium), for example, ointment containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

[0127] When preparing formulation, active compound can be pulverized to obtain suitable particle size before being combined with other components.If active compound is substantially insoluble, it can be pulverized to a particle size of less than 200 mesh.If active compound is substantially water-soluble, particle size can be adjusted by pulverization to obtain substantially uniform distribution in formulation, for example, about 40 mesh.

[0128] The compounds of the present invention can be milled using known milling procedures, such as wet milling, to obtain a particle size suitable for tablet formation and other formulation types. Finely divided (nanoparticulate) preparations of the compounds of the present invention can be prepared by processes known in the art, see, for example, the disclosures in International Application No. WO2002 / 000196.

[0129] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose.The formulation may further include 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 quick, sustained, or delayed release of active ingredient after administration to a patient by using methods known in the art.

[0130] In some embodiments, the pharmaceutical composition comprises silicified microcrystalline cellulose (SMCC) and at least one compound described herein, or a pharma- ceutically acceptable salt thereof. In some embodiments, the silicified microcrystalline cellulose comprises about 98% microcrystalline cellulose and about 2% silicon dioxide w / w.

[0131] In some embodiments, the composition is a sustained release composition comprising at least one compound described herein or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier. In some embodiments, the composition comprises at least one compound described herein or a pharma- ceutically acceptable salt thereof, and at least one component selected from microcrystalline cellulose, lactose monohydrate, hydroxypropyl methylcellulose, and polyethylene oxide. In some embodiments, the composition comprises at least one compound described herein or a pharma- ceutically acceptable salt thereof, and microcrystalline cellulose, lactose monohydrate, and hydroxypropyl methylcellulose. In some embodiments, the composition comprises at least one compound described herein or a pharma- ceutically acceptable salt thereof, and microcrystalline cellulose, lactose monohydrate, and polyethylene oxide. In some embodiments, the composition further comprises magnesium stearate or silicon dioxide. In some embodiments, the microcrystalline cellulose is AvicelPH102™. In some embodiments, the lactose monohydrate is Fast-flo 316™. In some embodiments, the hydroxypropyl methylcellulose is hydroxypropyl methylcellulose 2208 K4M (e.g., Methocel K4 M Premier™) and / or hydroxypropyl methylcellulose 2208 K100LV (e.g., Methocel K00LV™). In some embodiments, the polyethylene oxide is polyethylene oxide WSR 1105 (e.g., Polyox WSR 1105™).

[0132] In some embodiments, the compositions are manufactured using a wet granulation process. In some embodiments, the compositions are manufactured using a dry granulation process.

[0133] The compositions can be formulated in unit dosage form, with each dosage containing about 1 to about 1,000 mg, about 1 mg to about 100 mg, about 1 mg to about 50 mg, and about 1 mg to about 10 mg of active ingredient. Preferably, the dosage is about 1 mg to about 50 mg or about 1 mg to about 10 mg of active ingredient. In some embodiments, each dosage contains about 10 mg of active ingredient. In some embodiments, each dosage contains about 50 mg of active ingredient. In some embodiments, each dosage contains about 25 mg of 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 agent calculated to produce a desired therapeutic effect in association with a suitable pharmaceutical excipient.

[0134] In some embodiments, the composition comprises about 1 to about 1,000 mg, about 1 mg to about 100 mg, about 1 mg to about 50 mg, and about 1 mg to about 10 mg of active ingredient. Preferably, the composition comprises about 1 mg to about 50 mg or about 1 mg to about 10 mg of active ingredient. One of skill in the art will appreciate that this embodies compounds or compositions that contain about 1 mg to about 10 mg, about 1 mg to about 20 mg, about 1 mg to about 25 mg, or about 1 mg to about 50 mg of active ingredient.

[0135] In some embodiments, the dosage of the compound or a pharma- ceutically acceptable salt thereof is 15, 30, 60, or 90 mg based on the free base. In some embodiments, the dosage of compound 4 or a pharma- ceutically acceptable salt thereof is 15, 30, 60, or 90 mg based on the free base. In some embodiments, the dosage of the compound or a pharma- ceutically acceptable salt thereof is 15 mg based on the free base. In some embodiments, the dosage of the compound or a pharma- ceutically acceptable salt thereof is 30 mg based on the free base. In some embodiments, the dosage of the compound or a pharma- ceutically acceptable salt thereof is 60 mg based on the free base. In some embodiments, the dosage of the compound or a pharma- ceutically acceptable salt thereof is 90 mg based on the free base.

[0136] The active compound can be effective over a wide dosage range and is generally administered in a pharma- ceutical effective amount. However, it will be understood that the amount of compound actually administered will usually be determined by the 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.

[0137] To prepare solid compositions such as tablets, the primary active ingredient is mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogenous mixture of the compounds of the present application. When these preformulation compositions are referred to as homogenous, the active ingredient is typically evenly dispersed throughout the composition, and the composition can be readily divided into equally effective unit dosage forms such as tablets, pills, and capsules. This solid preformulation composition is then further divided into unit dosage forms of the type described above, for example, containing from about 0.1 to about 1000 mg of the active ingredient of the present application.

[0138] The tablets or pills of the present application can be coated or otherwise compounded to provide a dosage form that provides the advantage of prolonged action. For example, the tablet or pill can include an inner dose and an outer dose component, the latter being in the form of an envelope on a forming agent. The two components can be separated by an enteric layer that resists disintegration in the stomach and allows the inner component to pass intact into the duodenum or be delayed in release. A variety of materials can be utilized as such enteric layers or coatings, including materials including a number of polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0139] Liquid forms for oral or injectable administration into which the compounds and compositions of the present application may be incorporated include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions containing edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0140] Compositions for inhalation or insufflation include solutions and suspensions in pharma- ceutically acceptable aqueous or organic solvents, or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharma- ceutically acceptable excipients as described above. In some embodiments, the compositions are administered by oral or nasal respiratory route for local or systemic effect. Compositions may be nebulized by using inert gases. Nebulized solutions may be breathed directly from the nebulizing device, or the nebulizing device may be attached to a face mask tent or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0141] Topical formulations may include one or more conventional carriers. In some embodiments, ointments may include water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, and the like. The carrier composition of creams may be based on glycerol and water in combination with one or more other ingredients, for example, glycerol monostearate, PEG-glycerol monostearate, and cetylstearyl alcohol. Gels may be formulated using isopropyl alcohol and water, preferably in combination with other ingredients, for example, glycerol, hydroxyethylcellulose, and the like. In some embodiments, topical formulations include 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 of the present invention. Topical formulations may be suitably packaged in 100 g tubes, optionally accompanied by instructions for treating a selected indication, for example, psoriasis or other skin conditions.

[0142] 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, the 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 condition 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 health of the patient.

[0143] The compositions administered to a patient may be in the form of pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or may be sterile filtered. Aqueous solutions may be packaged or lyophilized for use as is, although lyophilized preparations are combined with a sterile aqueous carrier prior to administration. The pH of the compound preparations will typically be 3-11, more preferably 5-9, and most preferably 7-8. It will be appreciated that the use of certain of the aforementioned excipients, carriers, or stabilizers will result in the formulation of pharmaceutical salts.

[0144] The therapeutic dosage of the compounds of the invention may vary according to, for example, the particular application for which the treatment is to be performed, 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 of the invention in a pharmaceutical composition may vary depending on a number of factors, including the dosage amount, chemical characteristics (e.g., hydrophobicity), and the route of administration. For example, the compounds of the invention may be provided in an aqueous physiological buffer solution containing about 0.1 to about 10% w / v of the compound for parenteral administration. Some typical dosages range from about 1 μg / kg to about 1 g / kg of body weight per day. In some embodiments, the dosage range is from about 0.01 mg / kg to about 100 mg / kg of body weight per day. The dosage may depend on such variables as the type and extent of 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 may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0145] The compositions of the invention may further comprise one or more additional pharmaceutical agents, examples of which are listed herein.

[0146] kit The present application also includes pharmaceutical kits useful in treating and / or preventing a cytokine-related disease or disorder, such as, for example, CRS, which include one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound described herein. As will be apparent to one of skill in the art, such kits can optionally further include one or more of a variety of conventional pharmaceutical kit components, such as, for example, a container with one or more pharma- ceutically acceptable carriers, additional containers, etc. Instructions, either as an insert or label, indicating the amounts of components to be administered, administration guidelines, and / or guidelines for mixing the components, can also be included in the kit. EXAMPLES

[0147] 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 manner. Those skilled in the art will easily recognize various non-critical parameters that can be changed or modified to produce essentially the same results.

[0148] Example A: In vitro JAK kinase assay JAK1 inhibitors that may be used to treat cytokine-related diseases or disorders are tested for inhibitory activity of 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 (amino acids 837-1142), JAK2 (amino acids 828-1132), and JAK3 (amino acids 781-1124) with N-terminal His tags are expressed using baculovirus in insect cells 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 1 mM 50 For the 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, 6 nM Eu-Py20 assay buffer (Perkin Elmer, Boston, MA). Binding to europium-labeled antibodies was carried out for 40 minutes and the HTRF signal was measured on a Fusion plate reader (Perkin Elmer, Boston, MA). The compounds in Table 1 were tested in this assay and showed the IC 50 It was shown that the value of

[0149] Example B: Safety and Efficacy Study of JAK1 and / or JAK2 Inhibitors in Subjects with Moderate to Severe Hidradenitis Suppurativa A randomized, double-blind, placebo-controlled, multicenter study will be conducted in men and women aged 18-75 years with at least 6 months of moderate (Hurley stage II) to severe (Hurley stage III) hidradenitis suppurativa. Hurley stage I is associated with sinus tracts and abscess formation (single or multiple) without scarring. Hurley stage II is associated with recurrent abscesses with duct formation and scarring, single or multiple widely separated lesions. Hurley stage III is associated with diffuse or near-diffuse involvement over the entire area, or multiple interrelated ducts and abscesses. Study participants will be randomized into 5 groups (approximately 50 participants per group) and treated with 15, 30, 60, or 90 mg of an inhibitor of JAK1 and / or JAK2 (e.g., ruxolitinib, Compound 4, or Compound 5, or a pharmacologic acceptable salt thereof), or placebo. At week 16 (primary endpoint), participants in the placebo group will be re-randomized to the active treatment group for 8 weeks. Blinding will be maintained. The primary endpoint is the proportion of subjects achieving hidradenitis suppurativa clinical response (HiSCR) at week 16.

[0150] Secondary endpoints included: (1) proportion of subjects with HiSCR above baseline at each visit; (2) proportion of subjects achieving abscess and inflammatory nodule (AN) count between 0 and 2 at each visit; (3) mean change from baseline in HS Numeric Pain Rating Scale1) at each visit; (4) change in Modified Sartorius Scale at weeks 16 and 24; (5) change in draining fistula count at each visit; (6) proportion of subjects requiring lesion-salvage therapy through week 24; (7) number of episodes of lesion-salvage therapy through week 24; (8) population PK (e.g., apparent clearance, apparent sigma, ... (9) safety and tolerability assessed by monitoring the frequency, duration, and severity of AEs, physical examination, vital signs, and laboratory data of hematology, serum chemistry, and urinalysis; (10) change in Dermatology Life Quality Index (DLQI) assessment; (11) change in disease severity from baseline assessed by IHS43 score at each visit; (12) change in Hidradenitis Suppurativa Quality of Life (HiSQOL) assessment at each visit over baseline; and (13) assessment of dose / exposure response for percent change from baseline in terms of efficacy and safety endpoints during the treatment period.

[0151] HiSCR is defined as at least a 50% reduction in the number of abscesses and inflammatory nodules (AN) without an increase in the number of abscesses and draining fistulas at week 16 compared to baseline. A pain numerical rating scale is used to assess worst and average skin pain due to HS. The two-item rating ranges from 0 (no skin pain) to 10 (worst skin pain imaginable). Ratings are recorded in a daily diary before participants go to bed, based on a "past 24 hours" recall period. A modified Sartorius scale is used to quantify the severity of HS. Points are awarded for 12 body regions (left and right axillae, left and right submammary / inframammary regions, intermammary regions, left and right buttocks, left and right inguinal folds, perianal regions, perineal regions, etc.): nodules (2 points each); abscesses (4 points); fistulas (4 points); scars (1 point); longest distance between two lesions (2-6 points, 0 if no lesions); and, if lesions are separated, points are awarded for acquiring normal skin (0 points yes, 6 points no). The sum of the scores of the 12 regions is the sum of the scores of the 12 regions. Lesion rescue treatment: If an acutely painful lesion requires immediate intervention, the physician has the option to perform rescue intervention. There are only two types of interventions allowed: (1) injection of intralesional triamcinolone acetonide suspension (up to 30 mg in total during the same visit) and / or (2) incision and drainage. Interventions may occur on up to two different lesions at the same visit or on the same lesion at two different study visits. The same lesion cannot be treated twice at the same visit. If subjects require more than two interventions by week 16, they will be discontinued from the study. International Hidradenitis Suppurativa Severity Scoring System (IHS4): IH4 ​​(points) = (number of nodules x 1) + (number of abscesses x 2) + (number of drainage tunnels [fistulas / sinuses] x 4). Mild HS: ≤ 3 points; moderate HS: 4-10 points; severe HS: ≥ 11 points.

[0152] Study Treatment 1 (active) includes oral tablets containing 15 mg 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. Dosage levels include 15 mg (1 tablet), 30 mg (2 tablets), 60 mg (4 tablets), and 90 mg (6 tablets). Study Treatment 2 (placebo) includes oral tablet placebo.

[0153] Blood samples for measurement of plasma concentrations of JAK1 and / or JAK2 inhibitors will be taken before and after study drug administration at pre-dose, 1 hour post-dose, and 2-5 hours post-dose, and at least at weeks 2, 12, 16, 20, and 24. If a subject discontinues prior to week 8, a trough PK sample will be collected, if possible, at the premature discontinuation visit. The date / time of the last prior dose administration will also be recorded.

[0154] Superiority trials of 90, 60, 30, and 15 mg of JAK1 and / or JAK2 inhibitors compared to placebo will be performed using the Hochberg procedure at an overall two-sided α=0.05 level. Comparisons of each active group to placebo at week 16 will be performed using logistic regression. At all dose levels, superiority trials will be significant (e.g., 10%, 20%, 30%, 40%, or 50% improvement in HiSCR (Hidradenitis Suppurativa Clinical Response)), demonstrating efficacy of JAK1 and / or JAK2 inhibitors to treat HS. Trials have shown reduction in nodules and non-inferiority / superiority compared to placebo.

[0155] All secondary and exploratory efficacy measures will be evaluated using descriptive statistics. Clinical safety data (vital signs, routine laboratory tests, and AEs) will be analyzed using descriptive statistics. Exposure-response (ER) relationships between plasma JAK1 and / or JAK2 inhibitor PK exposure and efficacy / safety data will be determined. Interim analyses will be conducted when at least half of randomized subjects reach 16 weeks to estimate treatment response and facilitate planning of future studies.

[0156] Example C. Interferon-gamma and tumor necrosis-alpha induced Janus kinase expression in keratinocytes and subsequent production of inflammatory mediators Transformed human keratinocyte (HaCaT) cells were purchased from AddexBio (catalog number T0020001) and cultured in optimized Dulbecco's modified Eagle's medium (AddexBio, catalog number C0003-02) supplemented with 10% fetal bovine serum (Hyclone, catalog number 16140-071) and 1x penicillin / streptomycin (Gibco, catalog number 15140-122). When the cells reached 80-90% confluence, they were washed with 1x DPBS and then detached from the cell culture flask by incubating with 0.25% trypsin (Gibco, catalog number 25200-056) for 3-5 min at 37 °C / 5% CO2. Cell culture medium was added to the trypsinized cells, and then the cell suspension was transferred to a sterile 15 mL centrifuge tube and spun down at 1300 rpm for 10 min. The trypsin-containing medium was aspirated from the cell pellet, and the pellet was then resuspended in 10 mL of cell medium. Cells were counted using a Countess II automated cell counter and plated at 4 × 10 4 HaCaT cells were seeded at a concentration of 1000 cells / mL and incubated for 48 hours at 37°C / 5% CO2. After 48 hours, the medium was removed and replaced with 500uL of cell medium or a combination stimulus of recombinant human interferon gamma (R&D Systems, Catalog No. 285-IF-100) and recombinant human tumor necrosis factor alpha (R&D Systems, Catalog No. 210-TA-020). HaCaT cells treated with a combination cytokine stimulus were treated with each cytokine at a final concentration of 10ng / mL, 25ng / mL, 50ng / mL, or 100ng / mL. Treated plates were mixed by gentle agitation for 30 seconds and then incubated for 24 hours at 37°C / 5% CO2. At the end of the 24 hour incubation, the medium was immediately removed from each plate.

[0157] RNA was isolated from HaCaT cells using QuantiGene Plex assay reagents and protocols (Affymetrix, Catalog No. QGP-232-M18042302). Cells were washed with 1x DPBS and then lysed by incubation with the provided QuantiGene lysis buffer for 30 min at 50-55°C. Cell lysates were incubated at 55°C for 18-24 h with capture beads and a probe set designed to specifically hybridize to mRNA from the targets of interest. The panel of 32 targets of interest included housekeeping genes that were used to normalize results. After 18-24 h of incubation, signals were amplified utilizing branched DNA methodology according to the manufacturer's procedure (Affymetrix, Catalog No. QGP-232-M18042302). After hybridization and washing steps, assay plates were read on a Luminex 200 and data were expressed as median net fluorescence intensity. Data were then normalized to the median net fluorescence intensity of the housekeeping gene HPRT1 (Table 2).

[0158] [Table 2-1] [Table 2-2]

[0159] Target proteins of interest in the medium were detected and quantified using ProCarta Multiplex Immunoassay reagents and protocols (Invitrogen, Cat. No. EPX450-12171-901). The medium was incubated with antibody-conjugated beads designed to bind to specific target protein epitopes and identify the bound protein via the characteristic spectral pattern of the beads. Biotinylated detection antibodies designed to bind to different epitopes of the same target protein, and streptavidin-PE, were added to the assay plate to quantify the amount of target protein. The assay plate was read on a Luminex 200 and data were expressed as median net fluorescence intensity. The median net fluorescence intensity of the antigen standard curve, generated according to the manufacturer's procedure (Invitrogen, Cat. No. EPX450-12171-901), was plotted against the expected concentration of each standard. The concentration of each protein was estimated from the antigen standard curve and the concentrations were expressed as pg / mL (Table 3).

[0160] [Table 3]

[0161] Example D. Janus kinase inhibitors interfere with interferon-gamma and tumor necrosis-alpha mediated inflammation in keratinocytes Transformed human keratinocyte (HaCaT) cells were purchased from AddexBio (catalog number T0020001) and cultured as outlined in Example C. Four compounds A-D (A: ruxolitinib, B: itacitinib ({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), C: 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]acetonitrile) were cultured as outlined in Example C. ]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide, D:((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) were reconstituted in DMSO and then each compound was serially diluted in cell culture medium to concentrations of 400 nM, 200 nM, 100 nM, and 50 nM. After 48 hours, cell culture medium was removed from the 24-well plate and replaced with 250 uL of medium containing the serially diluted drug, then incubated for 15 minutes at 37°C / 5% CO2. After drug incubation, 250 uL of combination stimulus containing recombinant human interferon gamma (R&D Systems, Catalog No. 285-IF-100) and recombinant human tumor necrosis factor alpha (R&D Systems, Catalog No. 210-TA-020) was added to the plates. Final concentrations of recombinant human interferon gamma and recombinant human tumor necrosis factor alpha were 25 ng / mL of each cytokine. Cytokine stimuli added to drug-containing wells were at final concentrations of 25 nM, 50 nM, 100 nM, and 200 nM for each drug treatment. Treated plates were mixed by gentle swirling for 30 seconds and then incubated for 24 hours at 37° C. / 5% CO2. At the end of the 24 hour incubation, media was immediately removed from each plate.

[0162] RNA was isolated from HaCaT cells using QuantiGene Plex assay reagents and protocols (Affymetrix, Catalog No. QGP-232-M18042302) according to the manufacturer's guidelines. Cells were washed with 1x DPBS and then lysed by incubation with the provided QuantiGene lysis buffer for 30 min at 50-55°C. Cell lysates were incubated for 18-24 h at 55°C with capture beads and a probe set designed to specifically hybridize to mRNA from the target of interest. Genes included housekeeping genes (e.g., HPRT1, GAPDH) that were used to normalize results. After 18-24 h of incubation, signals were amplified utilizing branched DNA methodology according to the manufacturer's procedures (Affymetrix, Catalog No. QGP-232-M18042302). After hybridization and washing steps, the assay plates were read on a Luminex 200 and data was expressed as median net fluorescence intensity. Data was then normalized to the median net fluorescence intensity of the housekeeping gene HPRT1 (Table 4).

[0163] [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4]

[0164] Figures 1-4 show the individual gene expression values ​​(MFI) for JAK1, JAK2, IL-1α, and IL-6 for each experimental replicate in keratinocytes simulated with TNFα and IFN-γ in the presence / absence of JAK inhibitors.

[0165] Target proteins of interest in the medium were detected and quantified using ProCarta Multiplex Immunoassay reagents and protocols (Invitrogen, Cat. No. EPX450-12171-901). The medium was incubated with antibody-conjugated beads designed to bind to specific target protein epitopes and identify the bound protein via the characteristic spectral pattern of the beads. Biotinylated detection antibodies designed to bind to different epitopes of the same target protein, and streptavidin-PE, were added to the assay plate to quantify the amount of target protein. The assay plate was read on a Luminex 200 and data were expressed as median net fluorescence intensity. The median net fluorescence of the antigen standard curve, generated according to the manufacturer's procedure (Invitrogen, Cat. No. EPX450-12171-901), was plotted against the expected concentration of each standard. The concentration of each protein was estimated from the antigen standard curve and the concentrations were expressed as pg / mL (Table 5).

[0166] [Table 5-1] [Table 5-2]

[0167] Figures 5 and 6 show the individual protein concentrations (pg / mL) of IL-1α and IL-6, respectively, for each experimental replicate in keratinocytes simulated with TNFα and IFN-γ in the presence / absence of JAK inhibitors.

[0168] Example E: Hidradenitis Suppurativa Skin Biopsies are Characterized by Increased Janus Kinase Expression Total RNA of healthy control skin from three single donors was purchased from Amsbio (catalog numbers HR101 and R1234218-50). Total RNA of healthy control skin from a pool of donors was purchased from Life Technologies Corporation (catalog number QS0639). Hidradenitis suppurativa skin biopsies (41 donors) were purchased as formalin-fixed paraffin-embedded (FFPE) blocks from which total RNA was purified from Discovery Life Sciences.

[0169] Gene expression from healthy control (n=4) and hidradenitis suppurativa (n=41) skin total RNA samples was measured for the genes outlined in Table 6 using QuantiGene Plex assay reagents and protocols (Life Technologies Corporation, catalog number QGP-277-M19012402). Purified RNA was used in the recommended assay range of 50ng to 500ng and incubated overnight with capture beads designed to specifically hybridize to the mRNA of the selected genes (Table 6). This target panel included several housekeeping genes that were used to normalize the results. After overnight incubation, the signal was amplified using branched DNA methodology according to the manufacturer's procedure (Life Technologies Corporation). The assay plate was read on a Luminex 200 and data was expressed as median net fluorescence intensity (net MFI). Data was normalized to the geometric mean of the net MFI of the housekeeping genes ACTB and GAPDH. Figures 7-9 show gene expression of JAK1, JAK3, TYK2, STAT1, STAT2, STAT3, IRAK1, IRAK2, and IRAK4 in the skin of healthy controls and subjects with hidradenitis suppurativa.

[0170] [Table 6]

[0171] In addition to those described herein, various modifications of the present invention 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 patent publications, is hereby incorporated by reference in its entirety.

Claims

1. 1. A pharmaceutical composition for treating abscesses or inflammatory nodules, or both, resulting from hidradenitis suppurativa, comprising: ruxolitinib; and Ruxolitinib in which one or more hydrogen atoms have been replaced with deuterium atoms or a pharmaceutically acceptable salt thereof.

2. 2. The pharmaceutical composition of claim 1, wherein the compound or a pharmaceutically acceptable salt thereof is ruxolitinib or a pharmaceutically acceptable salt thereof.

3. 3. The pharmaceutical composition of claim 2, wherein the pharmaceutically acceptable salt of the compound is a pharmaceutically acceptable salt of ruxolitinib.

4. The pharmaceutical composition of claim 3, wherein the pharmaceutically acceptable salt of ruxolitinib is ruxolitinib phosphate.

5. 3. The pharmaceutical composition of claim 2, wherein the compound is ruxolitinib free base.

6. 2. The pharmaceutical composition of claim 1, wherein the compound or a pharmaceutically acceptable salt thereof is ruxolitinib, or a pharmaceutically acceptable salt thereof, in which one or more hydrogen atoms have been replaced with deuterium atoms.

7. The pharmaceutical composition according to any one of claims 1 to 6, which is administered together with an additional therapeutic agent.

8. 8. The pharmaceutical composition of claim 7, wherein the additional therapeutic agent is a corticosteroid.

9. 9. The pharmaceutical composition of claim 8, wherein the corticosteroid is triamcinolone, dexamethasone, fluocinolone, cortisone, prednisone, prednisolone, or flumetholone.

10. The pharmaceutical composition according to any one of claims 1 to 9, which is administered topically to the skin.

11. The pharmaceutical composition according to any one of claims 1 to 9, which is administered orally.

12. 12. The pharmaceutical composition of any one of claims 1 to 11, which results in a 10%, 20%, 30%, 40%, or 50% improvement in HiSCR (Hidradenitis Suppurativa Clinical Response).

13. The pharmaceutical composition of any one of claims 1 to 12, wherein said treatment reduces abscesses caused by hidradenitis suppurativa.

14. The pharmaceutical composition of any one of claims 1 to 12, wherein said treatment reduces inflammatory nodules caused by hidradenitis suppurativa.

15. The pharmaceutical composition of any one of claims 1 to 12, wherein said treatment reduces abscesses and inflammatory nodules resulting from hidradenitis suppurativa.

Images