EA042956B1 - JAK1 PATHWAY INHIBITORS FOR THE TREATMENT OF CYTOKINE RELATED DISORDERS - Google Patents

Description

Technical field

The present disclosure relates to inhibitors of the JAK1 pathway and their use in the treatment of cytokine-associated diseases or disorders.

State of the art

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

Cytokine release syndrome (CRS) is a direct result of excessive production of inflammatory cytokines caused by supraphysiological levels of immune activation and manifests as a clinical constellation of symptoms including fever, nausea, fatigue, myalgia, malaise, low blood pressure, hypoxia, hyperpermeability capillaries leading to potential multi-organ toxicity.

CRS is an unwanted side effect of, for example, immunological therapies for severe disease states such as cancer. Immunological therapies that can lead to CRS include the administration of monoclonal antibodies (mAbs) and, more recently, adoptive T-cell-based therapies for cancer treatment. Lee et al. Blood. 2014, 124(2):188-195. For example, chimeric antigen receptor (CAR) T cell therapy uses altered T cells to target neoplasms and has already been approved by the US Food and Drug Administration (FDA) for use in certain forms of refractory non-Hodgkin's lymphoma and recurrent lymph-regional leukemia (ALL) in children.

The cytokine profiles involved in CRS span two major cell sources: T-lymphocyte-derived cytokines including interferon-gamma (IFNy), IL-2, IL-6, soluble IL-6 receptor (IL-6R), and granulocyte-macrophage colony stimulating factor (GM-CSF); and cytokines primarily secreted by monocytes and/or macrophages such as IL-1e, IL-6, IL-12, IL-18, and tumor necrosis factor (TNFa). Xu XJ, Tang Y.M. Cancer Lett. 2014; 343:172-8. Zhang et al. sci. China Life Sci. 2016; 59:379-85. Brentjens R., et al. Mol. Ther. 2010; 18:666-8.

Modulation of an excessive cytokine response leading to CRS has the potential to provide significant clinical benefit. For example, the drug tocilizumab, an antibody against IL-6 receptor (IL-6R), reduces levels of CRS severity and is FDA approved for use in CRS. However, the mechanism of action of tocilizumab is limited only by the anti-IL-6R effect.

Hemophagocytic lymphohistiocytosis (HLH), another syndrome of excessive or uncontrolled immune activation, occurs mainly in infants from birth to 18 months of age, but can also occur in adults. HLH can be primary (familial) or secondary, ie. arising from other infectious, malignant, rheumatological or metabolic conditions. Symptoms of HLH include cytopenias, hepatosplenomegaly, and fevers. Schram, A. and Berliner, N. Blood. 2005. 125(19), 2908-2914.

Macrophage activation syndrome (MAS) is clinically presented in a manner similar to HLH (and even considered secondary or acquired to HLH), and is an episode of increased inflammation associated with infection, rheumatic disease, or malignancy. Borgia, R.E. et al. Arthritis Rheumatol., 2018, doi: 10.1002/art.40417, preliminary publication. MAS was originally described as being associated with juvenile idiopathic arthritis, but is also increasingly recognized as a complication of other diseases such as childhood systemic lupus erythematosus (cSLE). Shimizu M., et al. Clin. Immunol. Feb 2013; 146(2):73-6. The development of MAS is characterized by a significant increase in the amount of pro-inflammatory cytokines, i.e. cytokine storm. Borgia, R.E. et al. Arthritis Rheumatol., 2018, doi: 10.1002/art.40417, preliminary publication. MAS is a life-threatening condition with a high mortality rate: 8-22% in childhood autoimmune diseases in general and 10-22% in MAS complicating cSLE. Borgia, R.E. et al. Arthritis Rheumatol., 2018, doi: 10.1002/art.40417, preliminary publication.

CAR-T cell-associated encephalopathic syndrome (CRES) is the second most common adverse event after CRS associated with CAR-T cell therapy. CRES is typically characterized by a toxic encephalopathic condition with symptoms of confusion and delirium, as well as episodic seizures and cerebral edema. The presentation of CRES may be biphasic with symptoms occurring within the first 5 days and/or 3-4 weeks after cellular immunotherapy. The pathophysiological mechanism is thought to involve passive diffusion of cytokines into the brains of patients treated with CAR-T cell therapy. Reduction or elimination of this mechanism may be beneficial for such patients. Neelapu, et al. Nat. Rev. Clin. oncol. 2018, 15(1):47-62.

Accordingly, there is a need to develop new therapies for the treatment of cytokine-associated diseases or disorders. This invention addresses this and other needs.

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Brief description of graphic materials

In FIG. 1 shows dose-dependent inhibition of IL-6 concentrations following administration of Compound 1 to the blood compartment during anti-CD3 antibody-induced cytokine release syndrome (see Example B).

In FIG. 2A-2C depict dose-dependent inhibition of T cell-derived cytokines (ie, IL-6, IFNy, and GM-CSF) following compound 1 administration during concanavalin A-induced cytokine release syndrome (see Example C).

In FIG. 2A shows IL-6 inhibition.

In FIG. 2B shows IFNy inhibition.

In FIG. 2C shows GM-CSF inhibition.

In FIG. 3A-3C depict dose-dependent inhibition of monocyte- and macrophage-derived cytokines (i.e., IL-12, IL-1e, and IL-18) following administration of compound 1 during concanavalin A-induced cytokine release syndrome (see Example C).

In FIG. 3A shows IL-12 inhibition.

In FIG. 3B shows IL-1β inhibition.

In FIG. 3C shows IL-18 inhibition.

In FIG. 4 shows that the cytokine IL-5 is unaffected by compound 1 during concanavalin A-induced cytokine release syndrome (see Example C).

The essence of the invention

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

Provided herein is a JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, for the treatment of a cytokine-associated disease or disorder in a subject in need thereof.

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

Detailed Description of the Invention

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

The methods described herein use inhibitors of the JAK1 pathway, in particular selective JAK1 inhibitors. A selective JAK1 inhibitor is a compound that preferentially inhibits JAK1 activity over other Janus kinases. JAK1 plays a central role in a number of cytokine and growth factor signaling mechanisms that, when dysregulated, can lead to or contribute to disease states. For example, IL-6 levels are elevated in rheumatoid arthritis, a disease in which they are thought to have deleterious effects (Fonesca, et al., Autoimmunity Reviews, 8:538-42, 2009). Because IL-6 signaling is at least partially mediated through JAK1, IL-6 may indirectly act through inhibition of JAK1, resulting in potential clinical benefit (Guschin, et al. Embo J. 14:1421, 1995; Smolen, et al. al Lancet 371:987, 2008). Moreover, in some cancers, JAK1 is mutated, resulting in constitutive unwanted growth and survival of tumor cells (Mullighan, Proc. Natl. Acad. Sci. USA. 106:9414-8, 2009; Flex, J. Exp. Med. 205: 751-8, 2008). In other autoimmune and cancerous diseases, elevated systemic levels of inflammatory cytokines that activate JAK1 may also contribute to the development of the disease and/or associated symptoms. Therefore, patients with such diseases may benefit from JAK1 inhibition. Selective JAK1 inhibitors may be effective in preventing the unnecessary and potentially unwanted effects of inhibiting other JAK kinases.

JAK1 pathway inhibitor namely Compound 1 (i.e. {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidyl-4-yl}-3-[4-(7H-pyrrolo[2, 3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, see Table 1), provides highly effective dose-dependent modulation of CRS-related inflammatory cytokines (see, for example, Examples B and C and Fig. 1, 2A-2C and 3A-3C). Surprisingly, the therapeutic profile spans multiple pathogenic cytokines and is not limited to the IL-6/IL-6R axis (unlike, for example, tocilizumab). Efficacy is achieved by inhibition of cytokines derived from T cells and monocytes/macrophages, which are of high clinical significance for the pathogenesis of CRS. Additionally, the data presented herein in connection with the JAK1 inhibitory Compound 1 show that treatment benefits are achieved without extensive cytokine immunosuppression (as demonstrated by unchanged IL-5 levels) (FIG. 4).

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

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

In some embodiments, the cytokine-associated disease or disorder is hemophagocytic lymphohistiocytosis (HLH).

In some embodiments, the cytokine-associated disease or disorder is macrophage activation syndrome (MAS). In some embodiments, macrophage activation syndrome is associated with systemic juvenile idiopathic arthritis. In some embodiments, macrophage activation syndrome is associated with systemic lupus erythematosus in childhood.

In some embodiments, the cytokine-associated disease or disorder is CAR-T cell-associated encephalopathic syndrome (CRES).

In some embodiments, the present invention provides a method of treating cytokine release syndrome in a subject, comprising administering to said subject a CAR-T cell therapeutic and a JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof. In some embodiments, the treatment is alleviation or inhibition. In some embodiments, the treatment is prevention.

In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered concurrently with the CAR-T cell therapeutic preparation.

In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered following administration of the CAR-T cell therapeutic.

In some embodiments, the CAR-T cell therapeutic is axicabtagen siloleucel.

In some embodiments, the CAR-T cell therapeutic is tisagenlecleisel.

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

In some embodiments, the subject is ill with diffuse large B cell lymphoma (DLBCL), primary mediastinal large B cell lymphoma, high grade B cell lymphoma, transformed follicular lymphoma, or acute lymphoblastic leukemia.

In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is selective for JAK1 over JAK2, JAK3, and TYK2 (ie, a selective JAK1 inhibitor). For example, the compounds described herein, or pharmaceutically acceptable salts thereof, preferentially inhibit JAK1 over one or more of JAK2, JAK3, and TYK2. In some embodiments, the compounds inhibit JAK1 preferentially over JAK2 (eg, have a JAK2/JAK1 IC ₅₀ ratio >1). In some embodiments, the compounds or salts are about 10-fold more selective for JAK1 than JAK2. In some embodiments, the compounds or salts are about 3-fold, about 5-fold, about 10-fold, about 15-fold, or about 20-fold more selective for JAK1 over JAK2, as calculated by measuring IC ₅₀ at 1 mM. ATP (for example, see example A).

In some embodiments, the JAK1 pathway inhibitor is a compound from Table 1. 1 or a pharmaceutically acceptable salt thereof. Connections in the table. 1 are selective inhibitors of JAK1 (selective over JAK2, JAK3 and TYK2). In table. 1 shows IC50 values obtained by the method of example A at 1 mM ATP.

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Table 1

Comm. No. Prai. Name Structure JAKI IC ₅₀ (nM) JAK2 / JAKI 1 US 2011/0224190 (example 1) {1-{1-[3-Fluoro-2(trifluoromethyl)isonicotinoyl]piperidin4-yl]-3-[4-(7Hpyrrolo[2,3d]pyrimidin-4-yl)1H-pyrazol-1yl]azetidin-3yl ^acetonitrile th 'tn o Τ' + >10 2 US 2011/0224190 (example 154) 4-{3-(Cyanomethyl)-314-(7Н-pyrrolo[2,3d]pyrimidin-4-nl)ΙΗ-pyrazol Γ yl]azetidine-Gyl}-N[4-fluoro-2(trifluoromethyl)phenyl]piperidine -1 carboxamide With. □ ΝΗ Ν X. :<Н Ν-Ν ^Ν Η + >10 3 US 2011/ 0224190 (example 85) L3-L4-(7H- pyrrolo[2,3- d] pyrimidin-4-yl)- 1 H-pyrazol-1 -yl]-1 - (H12- (trifluoromethyl)pyrim idin-4- yl]carbonyl]piperide in-4-yl)azetidine-3- ⁰ /=\ Ν ν Q NN j/ \ oz + >10

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yl]acetonitrile 4 US 2014/03430 30 (example 7) 4-(3-(cyanomethyl)-3(3'.5'-dimethyl-1Н,Н4,4'-bipyrazol-1 yl)azetidin-1-yl]-2,5difluoro-I-[(1S )-2.2 .2trifluoro-1methylethyl]benzamide X ^FF HN N +++ >10 5 US 2014/01211 98 (example 20) ((2R,5S)-5-(2-(( 1R)-1hydroxyethyl]-1H imidazo[4.54]thieno[3,2- b] pyridin-1 yl}tetrahydro-2Hpyran-2yl)acetonitrile H'i Ho Xv >10 6 US 2010/ 0298334 (Example 2)* ¹ 3-(1-(6-chloropyridin2-yl)pyrrolidin-3yl]-3-[4-(7H- pyrrolo[2,3- d] pyrimidin-4-yl)1H-pyrazol-1yl]propanenitrile and % „and + >10 7 US 2010/0298334 (Example 13c) 3-(1-[1,3]oxazolo[5,4b]pyridin-2ylpyrrolidin-3-yl)3-(4-(7H-pyrrolo[2,30]pyrimidin-4-yl)1H-pyrazol-1yl] propanenitrile 5 + >10

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8 US 2011/0059951 (Example 12) 4-[(4-{3-cyano-2-14- (7Н-pyrrolo[2,3- <1]pyrimidin-4-yl)1H-pyrazol-1yl]propyl)piperazin-1-yl)carbonyl]-3fluorobenzonitrile Oh oh + >10 9 US 2011/0059951 (Example 13) 4-[(4-{3-cyano-2-[3- (7Н-pyrrolo[2,3- <1]pyrimidin-4-yl)1H-pyrrol-1yl]propyl| piperazine -1-yl)carbonyl]-3fluorobenzonitrile F O >=> o N— ^/ l Oe ^Ν Η + >10 10 US 2012/ 0149681 (example 7b) [trans-1-[4-(7Hpyrrolo[2,3e]pyrimidin-4-yl)- 1Н-pyrazol-1-yl]-3- (4-C2- (trifluoromethyl)pyrim idine-4- yl]carbonyl’piperase in-1- yl) cyclobutyl] aiethone itrile Ν o 0 ....... NN Ν 1: ^Ν Η + >10

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eleven US 2012/0149681 (example 157) {trans[4-](3hydroxyazetidin-1yl)methyl]-6(trifluoromethyl)pyride in-2- yl]oxy (piperidin-1yl)-1-[4-(7Hpyrrolo[2,3- d] pyrimidi n-4-yl)1H-pyrazol-1yl]cyclobutyl(acetonitrile □ ° ^H N ^J ϋ ^Ν f ^F 0 9./ NN wn NH + >10 12 US 2012/0149681 (example 161) \trans-3-{ ₃ -\ [4{L(2S)-2(hydroxymethyl)pyrr olidin-1 -yl]methyl] 6(trifluoromethyl)pyride in-2-yl]oxy(piperidin-1yl)-1- [4-(7Hpyrrolo[2,30]pyrimidin-4-yl)1H-pyrazol-1-yl]cyclobutyl Jacetonitrile ^H \£^-OH -. ^ro ? A- N ΤΛ LI / n N H >10

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13 US 2012/0149681 (example 162) {wpanc-3-(4-i[4{[(2R)-2- (hydroxymethyl)pyrr olidin-1-yl]methyl}6- (trifluoromethyl) pyride in-2- yl]oxy | piperidin-1 yl)-1-|4-(7Hpyrrolo[2,3c1]pyrimidin-4-yl)1H-pyrazol-1u l]cyclobutyl}acetonitrile N'/ ί ^OH □ '“U with V And 4- >10 14 US 2012/0149682 (example 2O) ^b 4-(4-{3[(dimethylamino)methy l]-5- fluorophenoxy}piperidin-1-yl)-3-[4-(7Hpyrrolo[2,3c!]pyrimidin-4-yl)1H-pyrazol-1yl]butanenitrile Y y _______n + >10 15 US 2013/ 0018034 (example 18) 5-(3-(cyanomethyl)-3[4-(7H-pyrrolo[2.3c!]pyrimidin-4-yl)1H-pyrazol-1nl]azetidine-1-yl'-Nisopropylpyrazine-2carboxamide N——N— O NN' I.W. w n N Η + >10 16 US 2013/ 0018034 (example 28) 4-{3-(cyanomethyl)-3[4-(7H-pyrrolo[2,3c!]pyrimidin-4-yl)1H-pyrazol-1yl]azetidin-1-yl}- ___; o N - y z, / = ^-. // \·' Ή— 4 ,ν--\ Ν—μ Η ί) \ ρ ^; 111·····:. F Y. Υ Ί 7-th + >10

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2,5-difluoro-M-[(15)- 2,2,2-trifluoro-1 methylethyl]benzamide 17 US 2013/ 0018034 (example 34) 5-{3-(cyanomethyl)-3[4-(1H-pyrrolo[2,3b]pyridin-4-yl)-1 Hpyrazol-1- yl]azetidin-1-yl}-Nisopropylpyrazine-2carboxamide Ν'-. - N' / U. U- 4? ^<: . .. Ν—Ν -Ν r> N and + >10 18 US 2013/ 0045963 (example 45) {1-(^"c-4-{ (6-(2hydroxyethyl)-2(trifluoromethyl)pyrim idine-4- yl]oxy} cyclohexyl) -3-[4-(7Hpyrrolo[2.3c!]pyrimidin-4-yl)1H-pyrazol-1yl]azetidin-3yl}acetonitrile Xr-N'Vn to A y ^h Η >10 19 US 2013/ 0045963 (example 65) {([4- [(ethylamino)methyl]-6(trifluoromethyl)pyride in-2- yl]oxy} cyclohexyl) -3-[4-(7H- pyrrolo[2,3- <1]pyrimidin-4-yl)- 1Н-pyrazole-1- yl]azetidin-3yl{acetonitrile 1 + >10

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20 US 2013/ 0045963 (example 69) {1-(ipi>4-j [4-(1hydroxy-1 methylethyl)-6(trifluoromethyl) pyride in-2- yl]oxy) cyclohexyl) -3-[4-(7Hpyrrolo[2.3c!]pyrimidin-4-yl)1H-pyrazole-1- yl]azet idin-3yl)acetonitrile ηLOH V.O γΛ + >10 21 US 2013/ 0045963 (example 95) [l-(W-4-([4-[[(3R)3- hydroxypyrrolidin1-yl]methyl)-6- (trifluoromethyl) pyride in-2- yl]oxy} cyclohexyl) -3-[4-(7H- pyrrolo[2,3<1]pyrimidin-4-yl)1H-pyrazol-1yl]azet idin-3yl}acetonitrile at I + >10 22 US 2013/ 0045963 (example 95) {1-(grl>4-{ [4-{ [(3S)-3hydroxypyrrolidin1-yl]methyl[-6(trifluoromethyl)pyride in-2- il]oxy! cyclohexyl) -3-[4-(7Hpyrrolo[2,3- s!]pyrimidin-4-yl)- 1Н-pyr azole-1- y-Ό-ΟΗ \ ,__ g nA / \-NX ^F J ^4/ NN |l J ) H + >10

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yl]azetidine-3- yl} acetonitrile yl 23 US 2014/ 0005166 (example 1) {trans-3-(4-([4- ({[(15)-2-hydroxy-1methylethyl]amino}methyl)-6- (trifluoromethyl) pyride in-2- yl]oxy}piperidin-1yl)-1-[4-(7Hpyrrolo[2,3c!]pyrimidin-4-yl)1H-pyrazol-1 and l]cyclobutyl}acetonitrile NH o ^N F ^F 0 I NN ^Ν^Ν Η + >10 24 US 2014/ 0005166 (example 14) {t/ishs-3-(4-{ [4- Gl(2R)-2- hydroxypropyl]amine o 1 methyl)-6(trifluoromethyl)pyride in-2- yl]oxy) piperidine-1 - yl)-1-[4-(7H- pyrrolo[2,3c!]p11rimidin-4-yl)1H-pyrazol-1- and l]cyclobutyl}aceto nitrile Dr. NH fV-F I I NN ϋ n N Η + >10

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25 US 2014/ 0005166 (example 15) (trans-3 -(4-( [4- ({[(2S)-2- hydroxypropyl]amine o(methyl)-6- (trifluoromethyl) pyride in-2- yl]oxy} piperidin-1 yl)-1-[4-(7Hpyrrolo[2,3- 4]pyrimidin-4-yl)III-pyrazol-1- yl]cyclobutyl(acetonitrile OH NH Af ^on f ^F 0 9/ NN Η 4- >10 26 US 2014/ 0005166 (example 20) (trans-3 -(4-([4-<2hydroxyethyl)-6(trifluoromethyl)pyride in-2- yl]oxy}piperidin-1 yl)-1-[4-(7Hpyrrolo[2,3- 4]pyrimidin-4-yl)III-pyrazole-G yl]qi clobutyl]aceto nitrile but o ^s f ^f 0 9./ Η Ν ν ΤΑ Ν Ν Η 4- >10

+ means <10 nM (see Example A for assay conditions).

++ means <100 nM (see Example A for assay conditions).

+++ means <300 nM (see Example A for assay conditions).

^a Data for enantiomer 1.

^b Data for enantiomer 2.

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

In some embodiments, the JAK1 pathway inhibitor is a salt of adipic acid and {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7Hpyrrolo[2,3- d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile.

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

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

In some embodiments, the JAK1 pathway inhibitor is a salt of phosphoric acid and 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1Ή-4,4'-bipyrazol-1-yl) azetidin-1-yl]-2,5difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide.

Synthesis and preparation of 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1Н,1Ή-4,4'-bipyrazol-1-yl)azetidin-1-yl]2,5-difluoro -N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide and its salts of phosphoric acid can be found, for example, in publ. US Patent No. 2014/0343030, filed May 16, 2014, which is incorporated herein by reference in its entirety.

In some embodiments, the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b] pyridin-1-yl}tetrahydro-2H-pyran2-yl)acetonitrile or a pharmaceutically acceptable salt thereof.

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In some embodiments, the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b] pyridin-1-yl}tetrahydro-2H-pyran2-yl)acetonitrile monohydrate.

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 the determination of the properties of its anhydrous and monohydrate forms are described in publ. U.S. Patent No. 2014/0121198, filed October 31, 2013, and publ. US Patent No. 2015/0344497, filed April 29, 2015, each of which is incorporated herein by reference in its entirety.

In some embodiments of the invention, the compounds from the table. 1 obtained by the synthesis procedures described in publ. U.S. Patent No. 2011/0224190, filed March 9, 2011, publ. U.S. Patent No. 2014/0343030, filed May 16, 2014, publ. U.S. Patent No. 2014/0121198, filed October 31, 2013, publ. U.S. Patent No. 2010/0298334, filed May 21, 2010, publ. U.S. Patent No. 2011/0059951, filed August 31, 2010, publ. U.S. Patent No. 2012/0149681, filed November 18, 2011, publ. U.S. Patent No. 2012/0149682, filed November 18, 2011, publ. U.S. Patent 2013/0018034, filed June 19, 2012, publ. U.S. Patent No. 2013/0045963, filed August 17, 2012, publ. US Patent No. 2014/0005166, filed May 17, 2013, each of which is incorporated herein by reference in its entirety.

In some embodiments, the JAK1 pathway inhibitor is selected from the compounds or pharmaceutically acceptable salts thereof from Pub. U.S. Patent No. 2011/0224190, filed March 9, 2011, publ. U.S. Patent No. 2014/0343030, filed May 16, 2014, publ. U.S. Patent No. 2014/0121198, filed October 31, 2013, publ. U.S. Patent No. 2010/0298334, filed May 21, 2010, publ. U.S. Patent No. 2011/0059951, filed August 31, 2010, publ. U.S. Patent No. 2012/0149681, filed November 18, 2011, publ. U.S. Patent No. 2012/0149682, filed November 18, 2011, publ. U.S. Patent 2013/0018034, filed June 19, 2012, publ. U.S. Patent No. 2013/0045963, filed August 17, 2012, publ. US Patent No. 2014/0005166, filed May 17, 2013, each of which is incorporated herein by reference in its entirety.

In some embodiments, the JAK1 pathway inhibitor is a compound of Formula I, or a pharmaceutically acceptable salt thereof, wherein X is N or CH;

L is C(=O) or C(=O)NH;

A is phenyl, pyridinyl or pyrimidinyl, each of which is optionally substituted with 1 or 2 independently selected R ¹ groups; and each R is independently fluoro or trifluoromethyl.

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-4yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof.

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-fluoro2-(trifluoromethyl)phenyl]piperidine-1-carboxamide or a pharmaceutically acceptable salt thereof.

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

In some embodiments, the JAK1 pathway inhibitor is a compound of Formula II

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or a pharmaceutically acceptable salt thereof, wherein R ² is C ₁ . ₆ αkyl, C ₁ . ₆ haloalkyl, C ₃ . ₆ cycloalkyl or C ₃ . ₆ cycloalkyl-C ₁ . ₃ αkyl, with said C ₁ . ₆ alkyl, C ₃ . ₆ cycloalkyl and C ₃ . ₆ cycloalkyl-C ₁ . ₃ alkyl are each optionally substituted with 1, 2 or 3 substituents independently selected from fluorine, -CF3 and methyl;

R ³ is H or methyl;

R ⁴ is H, F or Cl;

R ⁵ is H or F;

R ⁶ is H or F;

R ⁷ is H or F;

R ⁸ is H or methyl;

R ⁹ is H or methyl;

R ¹⁰ is H or alkyl;

R ¹¹ is H or methyl.

In some embodiments, the compound of formula II is 4-[3-(cuαnomethyl)-3-(3',5'-gumethyl-1H,1Ή-4,4'-bunyrazol-1-yl)αzetugun-1-ul ]-2,5-gufluoro-N-[(1S)-2,2,2trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof.

In some embodiments, the JAK1 pathway inhibitor is a compound of Formula III

or a pharmaceutically acceptable salt thereof, wherein Cy ⁴ is a tetrahydro-2H-pyranol ring which is optionally substituted with 1 or 2 groups independently selected from CN, OH, F, Cl, C ₁ . ₃ αkyl, C _1-3 haloalkyl, cyano-C _1-3 alkyl, HO-C _1-3 alkyl, amino, C _1-3 alkylamino and di(C _1-3 alkyl)amino, wherein said C ₁ . ₃ αalkyl and di(C _1-3 αalkyl)amino are optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, C ₁ . ₃ alkylaminosulfonyl and C ₁ . ₃ alkylsulfonyl; And

R ¹² is -CH2-OH, -CH(CH3)-OH or -CH2-NHSO2CH3.

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-pyran2-yl)acetonitrile or a pharmaceutically acceptable salt thereof.

In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered in a daily amount of about 100 to about 600 mg as free base. Accordingly, in some embodiments, the selective JAK1 pathway inhibitor is administered in a daily amount of about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg , about 550 mg or about 600 mg as free base.

In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered in a daily amount of about 200 mg as free base.

In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered in a daily amount of about 300 mg as free base.

In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered in a daily amount of about 400 mg as free base.

In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered in a daily amount of about 500 mg as free base.

In some embodiments, the JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, is administered in a daily amount of about 600 mg as free base.

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

- 14 042956

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

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

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

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

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

Provided herein is a method of treating a cytokine-associated disease or disorder in a subject in need thereof, comprising administering to the subject a daily dose of about 100 to 600 mg, as free base, of a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof, wherein the JAK1 pathway inhibitor or its pharmaceutically acceptable salt is administered as one or more sustained release dosage forms containing a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof.

The embodiments of the invention described herein are intended to be combined in any suitable combination as if those embodiments were a plurality of dependent claims (e.g., embodiments relating to a selective JAK1 pathway inhibitor and dosage thereof, embodiments relating to any salt forms of the compounds described herein, embodiments relating to particular types of cytokine-associated diseases or disorders, and embodiments relating to composition and/or administration may be combined in any combination).

For example, provided herein is a method of treating a subject for a cytokine-associated disease or disorder selected from the group consisting of cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or associated with CAR-T cells. encephalopathic syndrome (CRES), the method comprises administering to the subject a single daily dose of about 200 mg, based on the free base, {1-{1-[3-fluoro-2(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[ 4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof, wherein the dose comprises one or more sustained release dosage forms , each contains {1-{1-[3-fluoro-2(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)- 1H-pyrazol1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof.

Sustained release dosage forms {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-3yl}acetonitrile or a pharmaceutically acceptable salt thereof (Table 1, compound 1) can be found in publ. US No. 2015/0065484, filed August 6, 2014, which is hereby incorporated by reference in its entirety.

All possible combinations are not listed individually in this document for the sake of brevity only.

The compounds described herein may be asymmetric (eg, have one or more stereocenters). All stereoisomers are meant, such as enantiomers and diastereomers, unless otherwise indicated. Compounds that contain asymmetrically substituted carbon atoms may be isolated in optically active or racemic form. Methods for preparing optically active forms from optically inactive starting materials are known in the art, for example by separation of racemic mixtures or by stereoselective synthesis. Many geometric olefin isomers, 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. The cis and trans geometric isomers of the compounds of this invention are described and may be isolated as a mixture of isomers or as separate isomeric forms.

In some embodiments, the compound has the (I)-configuration. In some embodiments of the invention, the connection has a ^-configuration.

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

Separation of racemic mixtures can also be carried out by elution on a column filled with an optically active resolving agent (eg dinitrobenzoylphenylglycine). A suitable composition of solvents for elution can be determined by a person skilled in the art.

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

The compounds described herein may also include isotopically labeled compounds as described herein. An isotopically or radioisotopically labeled compound is a compound as defined herein in which one or more atoms are replaced or replaced by an atom whose atomic mass or mass number differs from the atomic mass or mass number normally found in nature (i.e., those occurring in nature). Suitable radionuclides that may be included in the compounds of the present disclosure include, but are not limited to ² H (also written as D for deuterium), ³ H (also written as T for tritium), ^P C, ¹³ C, ¹⁴ C , ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ¹⁸ F ^{, 35} S, ³⁶ Cl, 82 Br, ⁷⁵ Br, ⁷⁶ Br, ⁷⁷ Br, ¹²³ I, ¹²⁴ I, ¹²⁵ I and ¹³¹ I. For example , one or more hydrogen atoms in the compound of the present description may be substituted by a deuterium atom (for example, one or more hydrogen atoms of the C ₁ . ₆ alkyl group of formulas (I), (II) or (III) or the compounds of Table 1 may be optionally substituted deuterium atom, for example -CH ₃ replaced by -CD ₃ ).

The term compound, as used herein, means to include all stereoisomers, geometric isomers, tautomers, and isotopes of the depicted structures, unless the name indicates a specific stereoisomer. Compounds identified herein by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise indicated.

All compounds and their pharmaceutically acceptable salts can be found together with other substances such as water and solvents (eg hydrates and solvates), or they can be isolated.

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

The phrase pharmaceutically acceptable is used herein to refer to those compounds, substances, compositions and/or dosage forms which, within the framework of rational medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reaction or other problem, or complications, commensurate with a reasonable benefit/risk ratio.

The expressions ambient temperature and room temperature, or rt, as used herein, are understood in the art and refer generally to a temperature, such as a reaction temperature, which is approximately the temperature at which a reaction takes place, such as a temperature of about 20 to around 30°C.

The present invention also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, the term pharmaceutically acceptable salts refers to derivatives of the disclosed compounds, wherein the parent compound is modified by converting its acidic or basic moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, salts of mineral or organic acids and basic residues such as amines; alkaline or organic salts of acid residues,

- 16 042956 such as carboxylic acids; and so on. Pharmaceutically acceptable salts of the present invention include conventional non-toxic salts of the parent compound, formed from, for example, non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present invention can be synthesized from a parent compound that contains a basic or acid moiety by conventional chemical methods. In general, such salts may be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or mixtures thereof; generally non-aqueous media such as ether, ethyl acetate, ethanol, alcohols (eg methanol, ethanol, isopropanol or butanol) or acetonitrile (ACN) are preferred.

A list of suitable salts is provided in Remington's Pharmaceutical Sciences, ^17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

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

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

As used herein, the term treat or treatment refers to one or more of (1) suppression of a disease; for example, suppressing a disease, condition, or disorder in a person who is experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder (ie, stopping further development of the pathology and/or symptomatology); (2) alleviate the disease; for example, alleviating a disease, condition, or disorder in an individual who is experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder (ie, reversing the course of the pathology or symptom), such as reducing the severity of the disease; or (3) preventing a disease, condition, or disorder in an individual who may be predisposed to that disease, condition, or disorder but is not yet experiencing or exhibiting the pathology or symptoms of that disease. In some embodiments of the invention, the treatment refers to the suppression or alleviation of the disease. In some embodiments, the treatment is the prevention of a disease.

Combined types of therapy.

The methods described herein may further include administering one or more additional therapeutic agents. One or more additional therapeutic agents may be administered to the patient simultaneously or sequentially.

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

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

In some embodiments, the additional drug is an anti-CD25 antibody. In some embodiments, the anti-CD25 antibody is daclizumab.

In some embodiments, the additional therapeutic agent is an IL-1β inhibitor.

In some embodiments, the additional therapeutic agent is an IL1 receptor antagonist (ILIRa). In some embodiments, the IL1 receptor antagonist (ILIRa) is anakinra.

- 17 042956

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

In some embodiments of the invention, any of the previous additional therapeutic agents is used in additional combination with a corticosteroid (eg, prednisone).

In some embodiments of the invention, additional therapeutic agents include tocilizumab and a corticosteroid. In some embodiments of the invention, additional therapeutic agents include tocilizumab and prednisone.

Pharmaceutical formulations and dosage forms.

When used as drugs, the JAK1 pathway inhibitors or their pharmaceutically acceptable salts may be administered in the form of pharmaceutical compositions. These compositions may be prepared in a manner well known in the pharmaceutical art and may be administered in a variety of ways depending on whether topical or systemic treatment is desired and also depending on the site to be treated. Administration may be topical (including transdermal, epidermal, ophthalmic, and mucosal, including intranasal, vaginal, and rectal delivery), pulmonary (eg, by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal or intranasal), oral or parenteral. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, intramuscular or injection or infusion administration; or intracranial, eg intrathecal or intraventricular, administration. Parenteral administration may be in the form of a single bolus dose or may be performed, for example, with a continuous perfusion pump. Pharmaceutical compositions and topical formulations may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powdery or oily bases, thickeners, and the like may be necessary or desirable.

This invention also includes pharmaceutical compositions which contain, as an active ingredient, the JAK1 pathway inhibitor described herein, or a pharmaceutically acceptable salt thereof, in combination with one or more pharmaceutically acceptable carriers (auxiliaries). In some embodiments, the composition is suitable for topical administration. In the preparation of compositions, the active ingredient is usually mixed with an excipient, diluted with an excipient, or enclosed in such a carrier in the form of, for example, a capsule, sachet, paper bag, or other container. When the excipient serves as a diluent, it may be a solid, semi-solid or liquid substance which acts as a carrier, vehicle or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, porridges, elixirs, suspensions, emulsions, solutions, syrups, aerosols (in the form of a solid or liquid medium), ointments containing, for example, up to 10% by weight. weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile powders in the package.

When preparing the formulation, the active compound may be milled to provide an appropriate particle size before being combined with other ingredients. If the active compound is substantially insoluble, it can be ground to a particle size of less than 200 mesh. If the active compound is substantially water soluble, then the particle size can be adjusted by milling to provide a substantially uniform distribution in the formulation, eg about 40 mesh.

JAK1 pathway inhibitors can be milled using known milling procedures such as wet milling to obtain a particle size suitable for tableting and other types of formulations. Finely dispersed (composed of nanoparticles) preparations of selective JAK1 inhibitors can be prepared using processes known in the art, for example, see international application No. WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum arabic, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulations may further include lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl and propyl hydroxybenzoates; sweetening agents; and flavoring agents. The compositions of this invention may be formulated to provide rapid, delayed or delayed release of the active ingredient after administration to a patient using procedures known in the art.

The composition may be formulated in unit dosage form, with each dosage containing from about 5 to about 1000 mg (1 g), more typically from about 100 to about 500 mg, of the active ingredient. The term unit dosage forms refers to physically discrete units suitable as unit dosages for humans and other mammals, each unit containing

- 18 042956 a predetermined amount of the active substance, calculated to obtain the desired therapeutic effect, in combination with a suitable pharmaceutical excipient.

In some embodiments, the compositions of this invention contain from about 5 to about 50 mg of the active ingredient. One of ordinary skill in the art will appreciate that compositions comprising about 5 to about 10, about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50 mg of active ingredient.

In some embodiments of the invention, the compositions of this invention contain from about 50 to about 500 mg of the active ingredient. One of ordinary skill in the art will appreciate that compositions comprising about 50 to about 100, about 100 to about 150, about 150 to about 200, about 200 to about 250, about 250 to about 300, about 350 to about 400, or about 450 to about 500 mg of active ingredient.

In some embodiments of the invention, the compositions of this invention contain from about 500 to about 1000 mg of the active ingredient. One of ordinary skill in the art will appreciate that compositions comprising about 500 to about 550, about 550 to about 600, about 600 to about 650, about 650 to about 700, about 700 to about 750, about 750 to about 800, about 800 to about 850, about 850 to about 900, about 900 to about 950, or about 950 to about 1000 mg of active ingredient.

At similar dosages, the compounds described herein can be used in the methods and uses of this invention.

The active compound may be effective over a wide dosage range and is usually administered in a pharmaceutically effective amount. However, it is understood that the amount of compound actually administered will generally be determined by the clinician, taking into account relevant circumstances, including the condition being treated, the chosen route of administration, the particular compound being administered, the age, weight and response of the particular patient, the severity of the patient's symptoms. and so on.

To prepare solid compositions such as tablets, the main active ingredient is mixed with a pharmaceutical excipient to form a solid preform composition containing a homogeneous mixture of the compound of the present invention. By homogenizing these preform compositions, the active ingredient is generally evenly distributed throughout the composition so that the composition can be easily divided into equally effective unit dosage forms such as tablets, pills and capsules. This solid preform mixture is then divided into unit dosage forms of the type described above, containing, for example, from about 0.1 to about 1000 mg of the active ingredient of the present invention.

The tablets or pills of the present invention may be coated or otherwise formulated to provide a dosage form that provides the advantage of sustained release. For example, a tablet or pill may contain an internal dosage component and an external dosage component, the latter being encapsulated on top of the former. The two components can be separated by an enteric layer which provides resistance to disintegration in the stomach and allows the internal component to pass unchanged into the duodenum or be delayed upon release. Various substances can be used for such enteric layers or coatings, such substances include a number of polymeric acids and mixtures of polymeric acids with substances such as shellac, cetyl alcohol and cellulose acetate.

Liquid forms which may include compounds and compositions for oral or injectable administration include aqueous solutions, suitable flavored syrups, aqueous or oily suspensions, and flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil. , as well as elixirs and similar pharmaceutical carriers.

Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures and powders thereof. The liquid or solid composition may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments of the invention, the compositions are administered orally or nasally through the respiratory tract to obtain a local or systemic effect. The compositions can be sprayed using inert gases. Nebulization solutions may be inhaled directly from a nebulization device, or the nebulization device may be attached to a face mask, cape, or intermittent positive pressure breathing apparatus. Solution, suspension, or powder compositions may be administered orally or nasally from formulation delivery devices in an appropriate manner.

Topical formulations may contain one or more conventional carriers. In some embodiments, ointments may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether,

- 19 042956 propylene glycol, white petrolatum, etc. Carrier compositions for creams can be based on water in combination with glycerol and one or more other components, for example glycerol monostearate, PEG-glycerol monostearate and cetyl stearyl alcohol. Gels can be formulated using isopropyl alcohol and water, respectively, in combination with other ingredients such as, for example, glycerol, hydroxyethyl cellulose, and the like. In some embodiments, topical formulations contain at least about 0.1; at least about 0.25; at least about 0.5; at least about 1; at least about 2 or at least about 5 wt. % of the compound described in this document. Formulations for topical application may suitably be packaged in tubes of, for example, 100 g, which may optionally be supplemented with instructions for the treatment of the selected indication, for example psoriasis or other skin disease.

The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of administration such as prophylaxis or therapy, the condition of the patient, the route of administration, and the like. 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 relieve the symptoms of the disease and its complications. Effective doses will depend on the disease state being treated, as well as the judgment of the attending physician, depending on such factors as the severity of the disease, the age, body weight and general condition of the patient, and the like.

Compositions administered to a subject may be in the form of the pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization methods or sterile filtered. Aqueous solutions may be packaged for use as is or lyophilized, the lyophilized preparation being combined with a sterile aqueous vehicle prior to administration. Generally, the pH of compound formulations is 3 to 11, more preferably 5 to 9, and most preferably 7 to 8. The use of some of the above excipients, carriers or stabilizers is expected to result in the formation of pharmaceutical salts.

The therapeutic dose of a compound of the present invention may vary according to, for example, the particular application being treated, the route of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of a compound described herein in a pharmaceutical composition may vary depending on a number of factors, including dose, chemical properties (eg, hydrophobicity), and route of administration. For example, the compounds described herein can be provided in an aqueous physiological buffer solution containing from about 0.1 to about 10% wt./about. compounds for parenteral administration. Some usual dosage ranges are 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 to about 100 mg/kg of body weight per day. The dosage is likely to depend on variables such as the type and extent of progression of the disease or disorder, the general health of the individual patient, the relative bioavailability of the compound chosen, the composition of the excipient, and the route of administration. Effective doses can be extrapolated from dose-response curves obtained from in vitro experiments or animal model research systems.

Compositions of this invention may further include one or more additional pharmaceutical agents, such as a chemotherapeutic, steroid, anti-inflammatory compound, or immunosuppressant, examples of which are listed herein.

Sets.

Also included within the present invention are pharmaceutical kits for use in, for example, the treatment and/or prevention of cytokine-associated diseases or disorders such as CRS, which comprise one or more containers containing a pharmaceutical composition containing a therapeutically effective amount of a substance described herein. connections. Such kits may also include, if desired, one or more of the various components of conventional pharmaceutical kits, such as, for example, containers with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be apparent to those skilled in the art. This kit may also include instructions in the form of leaflets, and in the form of labels indicating the amount of input components, recommendations for use and / or recommendations for mixing the components.

Examples

The present invention will be described in more detail using specific examples. The following examples are offered for explanatory purposes and are not intended to limit the present invention in any way. Those skilled in the art will readily recognize a number of non-critical parameters that can be changed or modified to produce substantially identical results. Example compounds that have been identified as JAK inhibitors in accordance with at least one assay described herein.

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EXAMPLE A In vitro JAK kinase activity assay.

JAK1 pathway inhibitors that may be used to treat cytokine related diseases or disorders are tested for inhibitory activity on JAK targets according to the following in vitro assay described in Park et al., Analytical Biochemistry, 1999, 269, 94-104. Express the catalytic domain of human JAK1 (a.k. 837-1142), JAK2 (a.k. 828-1132), and JAK3 (a.k. 781-1124) with the His N-terminal marker using baculovirus in insect cells and purify his. The catalytic activity of JAK1, JAK2 or JAK3 was analyzed by measuring the phosphorylation of the biotinylated peptide. The phosphorylated peptide was detected by homogenous time resolved fluorescence (HTRF). Compound IC ₅₀ concentrations are measured for each kinase in 40 μl reaction mixtures that contain enzyme, ATP and 500 nM peptide in 50 mM Tris (pH 7.8) buffer with 100 mM NaCl, 5 mM DTT and 0.1 mg/ml (0.01%) BSA. With regard to measurements of 1 mm IC ₅₀ , the concentration of ATP in the reaction mixture is 1 mm. Reactions are run at room temperature for 1 hour and then terminated with 20 μl of 45 mM EDTA, 300 nM SA-APC, 6 nM Eu-Py20 in assay buffer (Perkin Elmer, Boston, Massachusetts, USA). Binding to the europium labeled antibody takes 40 minutes and the HTRF signal was measured on a Fusion plate reader (Perkin Elmer, Boston, Massachusetts, USA). Connections in the table. 1 was examined in this assay and shown to have the IC ₅₀ values shown in Table 1. 1.

Example B Anti-CD3 Antibody Induced Cytokine Release Syndrome in BALB/c Mice.

JAK1 pathway inhibitors can be tested for efficacy against CRS according to the in vivo assay described in Ferran, C. et al. Clin. Exp. J. Immunol., 1991, 86, 537-543. In particular, the ability of a compound to reduce or alleviate anti-CD3 antibody-induced cytokine release syndrome (CRS) in BALB/c mice can be tested in this study. The antibody, clone 145-2C11, is a hamster immunoglobulin G (IgG) MoAb that is specific for the ε chain of the mouse CD3 molecule (Leo, O. et al., Proc. Natl. Acad. Sci. USA, 1987, 34, 1374). Treatment with 145-2C11 induces high affinity IL-2 receptors on the surface of spleen T cells and results in the release of several cytokines such as tumor necrosis factor (TNFa), IL-2, IL-3, IL-6, and interferon-gamma (IFNy) (Ferran, et al. Eur. J. Immunol. 1990, 20, 509-515 and Algre, M. et al., Eur. J. Immunol., 1990, 707). The release of these cytokines leads to behavioral changes (eg loss of activity, piloerection, etc.) in animals.

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Mice: male BALB/c

A. Materials and methods.

Kind/line:

Physiological state:

Age/weight range at start

Healthy

6-8 weeks of study:

Animal supplier:

Charles River Laboratories

Number/gender of animals:

Total 32 male mice

Randomization:

Rationale:

Replacement

Mice will be randomized into four (4) groups of eight (S) mice before the start of the study.

As shown in the literature, administration of an anti-SDR antibody (clone 145-2C I I) induces a cytokine release syndrome and serves as a model with which to study the effectiveness of a potential therapy.

Animals will not be replaced during this study.

Anti-POPs

Identification and serial number:

Source:

Storage conditions:

Carrier:

Dose:

Volume / route of administration

Compound:

Storage conditions:

Carrier:

Dose(s):

Volume / route of administration

Frequency and duration of dosing:

Anti-CD3s antibody, clone 145-2C11 BioXCell °C

Sterile saline solution 10 mcg

IV, 100 µl per animal

Compound 1 (Jakl inhibitor) ^A

CT (preparation at CT on a rotator for 0.5% methylcellulose 60 mg/kg and 120 mg/kg

P/O, 0.1 ml/20 g (5 ml/kg)

Once a day per day 0

Synthesis and receipt of compound 1 from the table. 1 or {1-{1-[3-Fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H -pyrazol-1-yl]azetidin-3-yl}acetonitrile and its salts of adipic acid can be found, for example, in publ. U.S. Patent No. 2011/0224190, filed March 9, 2011, publ. U.S. Patent No. 2013/0060026, filed September 6, 2012, and publ. US Patent No. 2014/0256941, filed March 5, 2014, each of which is incorporated herein by reference in its entirety.

B. Scheme of the experiment.

The main aim of this study was to test the ability of a JAK1 pathway inhibitor (eg Compound 1) to reduce or alleviate anti-CD3 antibody-induced cytokine release syndrome (CRS) in BALB/c mice. A total of 32 BALB/c mice were used for this one day study. Animals were weighed prior to dosing with study drug and monitored throughout the course of the experiment. On Day 0, 1 hour prior to administration, anti-CD3 antibodies were given as a single dose of vehicle (0.5% methylcellulose) or Compound 1 by gavage (G/O) to Groups 2-4 animals as shown in Table 1. 1A. Group 1 served as naive controls and received no treatment. After 1 hour pre-treatment with vehicle or compound 1, animals in groups 2-4 were injected with 10 μg of anti-CD3ε antibody (clone 145-2C11) by intravenous injection (IV) to induce CRS. All animals were euthanized by CO2 inhalation 1.5 hours after anti-CD3 antibody administration. Whole blood was collected by cardiac puncture into K2EDTA tubes and stored on ice until plasma isolation was performed. Plasma was collected and stored at -80° C. until multiplex analysis for cytokines was performed.

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Table 1A

Study design

Group Quantity animals Preliminary PI treatment (P/O) Introduction Schedule doses Anti-CD3 (10 mcg, (V/V) Schedule for killing/collecting plasma Endpoints 1 8/males "Naive" 60 minutes before anti-POPs - 1.5 h after anti-POP administration Whole blood via cardiac puncture ( _K2EDTD tubes) Soor plasma for multiplex analysis of cytokines 2 8/males Carrier + 3 8/males Compound 1 (60 mg/kg) + 4 8/males Compound 1 (120 mg/kg) +

C. Experimental procedures.

I. Pretreatment with an investigational drug (PI).

On Day 0, animals were dosed with vehicle or study drugs or Compound 1 as shown in Table 1. 1A. Group 2 received one dose of vehicle (0.5% methylcellulose) via 0.1 mg/20 g po. Group 3 received one 60 mg/mL dose of Compound 1 via 0.1 mg/20 g po. Group 4 received a single 120 mg/mL dose of Compound 1 via PO at 0.1 mg/20 g. Group 1 served as naive controls and received no treatment.

II. Introduction of anti-CD3ε antibodies.

One hour after the administration of the study drug, groups 2-4 were injected with anti-CD3ε antibody (clone 145-2C11) by IV injection. Each animal in groups 2-4 received 10 μg of anti-CD3ε antibody in 0.1 ml.

III. Live monitoring.

After the administration of anti-CD3 antibodies, the animals were closely monitored for signs of malaise caused by the resulting systemic inflammatory response. They euthanized animals that could not exist on their own, cold to the touch or dying. Dying animals were euthanized by inhalation of CO ₂ , blood was collected by cardiac puncture, and plasma was separated.

IV. Killing.

1.5 hours after administration of anti-CD3 antibodies, all animals were euthanized by inhalation of CO ₂ .

V. Sampling.

After sacrifice, whole blood was collected from each animal by cardiac puncture into K2EDTA tubes. Blood was centrifuged and plasma was collected in cryo vials. Plasma was frozen and stored at -80°C for subsequent multiplex analysis of cytokines.

VI. Multiplex analysis of cytokines.

Plasma samples are thawed on ice and used for multiplex cytokine analysis according to the manufacturer's protocol (ThermoFisher).

D. Results.

Compound 1 dose-dependently inhibited IL-6 concentrations in the blood compartment (FIG. 1). This confirms the biological activity seen in the preclinical Con A model described in Example C below. Unpaired one-way analysis of variance (ANOVA) including Sidak's multiple comparison test was performed using GraphPad Prism (version 4.00; GraphPad Software, San Diego). , California, USA). A p value <0.05 was considered statistically significant.

Example C Concanavalin A Induced Cytokine Release Syndrome.

Concanavalin A (Con A) is a selective T-lymphocyte mitogen resulting in extensive release of inflammatory cytokines and proliferation of CD4 and CD8 T cells. As shown in the literature, injection of Con-A induces cytokine release syndrome and serves as a model with which to investigate the effectiveness of therapy for cytokine release syndrome.

- 23 042956 (Gantner, F. at al. Hepatology, 1995, 21, 190-198). The response to the mitogen depends on the expression of the T-cell receptor. Animals exhibit behavioral changes such as fever, malaise, hypotension, hypoxia, increased capillary permeability, and potential multiple organ toxicity.

A. Materials and methods.

Species/line: Mice: BALB/c females

Physiological condition: Healthy

Age/weight range at start of 6-8 weeks of study:

Animal supplier: Taconic Number/gender of animals: Total 40 mice Randomization: Mice were randomized into five (5) groups of eight (8) mice prior to study entry. Rationale: As shown in the literature, the administration of Con-A induces a cytokine release syndrome and serves as a model with which to study the efficacy of a potential therapy.

B. Scheme of the experiment.

Specifically, this study examines the ability of a selective JAK1 inhibitor (e.g., Compound 1, Table 1) to reduce or alleviate Con A-induced Cytokine Release Syndrome (CRS) in BALB/c mice. A total of 40 BALB/c mice were used for this one-day study. . Animals were weighed prior to dosing with study drug and monitored throughout the course of the experiment. On day 0, 60 min prior to Con A administration, vehicle (0.5% methylcellulose) or Compound 1 (60 and 120 mg/kg) was given as a single dose by gavage (PO) to Groups 2-4 animals as shown in table. 2A. Group 1 served as naive controls and received no treatment. After 45 min pre-treatment with vehicle or Compound 1, animals in groups 2-4 were treated with 20 mg/kg Con A by intravenous (IV) injection to induce CRS. All animals were euthanized by CO ₂ inhalation 2 hours after Con A administration. Whole blood was collected by cardiac puncture into K2EDTA tubes and stored on ice until plasma isolation was performed. Plasma was collected and stored at -80° C. until multiplex analysis for cytokines was performed.

Table 2A

Study design

Group Quantity animals Preliminary treatment (P/O) Introduction Schedule doses Sop- A (V/V) Schedule for killing/collecting plasma Endpoints

1 10 "Naive" over 60 minutes before Con A - 2 hours after Con A injection Whole blood via cardiac puncture Plasma collection FOR multiplex cytokine analysis 2 10 Carrier + 3 10 Compound 1 (60 mg/kg) + 4 10 Compound 1 (120 mg/kg) +

C. Experimental Procedures Day -1.

The animals were weighed and the dose of Con-A (20 mg/kg) was calculated.

The carrier and compound 1 were prepared at appropriate doses.

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Day 0

I. Pretreatment with an investigational drug (PI).

On day 0, animals were dosed with vehicle or Compound 1 as shown in Table 1. 2A. Group 1 served as naive controls and received no treatment. Group 2 received one dose of vehicle (0.5% methylcellulose) via 0.1 mg/20 g po. Group 3 received one 60 mg/mL dose of Compound 1 via 0.1 mg/20 g po. Group 4 received a single 120 mg/mL dose of Compound 1 via PO at 0.1 mg/20 g.

II. Introduction Con-A.

60 minutes after administration of the study drug, Con-A was administered via IV injection to groups 2-4. Each animal in groups 2-4 received 20 mg/kg Con-A in 0.2 ml.

III. Live monitoring.

After administration of Con-A, the animals were closely monitored for signs of malaise caused by the resulting systemic inflammatory response.

IV. Killing.

2 hours after administration of Con-A, all animals were euthanized by inhalation of CO ₂ .

V. Sampling.

After sacrifice, whole blood was collected from each animal by cardiac puncture into K2EDTA tubes. Blood was centrifuged and plasma was collected in cryo vials. Plasma was frozen and stored at -80°C for subsequent multiplex analysis of cytokines.

VI. Multiplex analysis of cytokines.

Plasma samples are thawed on ice and used for multiplex cytokine analysis according to the manufacturer's protocol (ThermoFisher).

D. Results.

Compound 1 dose-dependently inhibited IL-6 concentrations in the blood compartment (FIG. 2A). This cytokine is a key mediator of the pathophysiology of CRS. The T cell-derived cytokines IFNy and GM-CSF were also significantly inhibited, suggesting that Compound 1 has therapeutic potential beyond the limited mechanism of action of tocilizumab (anti-IL-6R only) (Fig. 2B). and 2C).

The levels of monocyte- and/or macrophage-derived cytokines also decreased. A statistically significant dose-dependent decrease in IL-12 (Fig. 3A) was observed, as well as trends in treatment effect with IL-1e (Fig. 3B) and IL-18 (Fig. 3C), suggesting that JAK1 specific inhibition has a therapeutic effect. potential among immune cell types involved in CRS pathology.

Importantly, the cytokine IL-5 (FIG. 4) was not affected by Compound 1 treatment, is independent of JAK1, and is not involved in the pathology of CRS. These data indicate that Compound 1-based efficacy is not mediated by extensive non-specific immune suppression.

An unpaired one-way analysis of variance (ANOVA) including Sidak's multiple comparison test was performed using GraphPad Prism (version 4.00; GraphPad Software, San Diego, CA, USA). A p value <0.05 was considered statistically significant.

Example D Preparation of Sustained Release Formulations of Compound 1.

Sustained release (SR) tablets containing compound 1 were prepared with excipients in the amounts indicated in the following tables. Protocol A was applied to SR1 tablets, Protocol B was applied to SR2 tablets, Protocol C was applied to SR3 tablets and SR 25 mg tablets, and Protocol D was applied to SR4 tablets. These procedures are described in pub. US patent No. 2015/0065484, which relates to dosage forms of compound 1 with delayed release.

Protocol A:

Step 1. The adipic acid salt of Compound 1, microcrystalline cellulose, hypromellose (Methocel K100 LV and Methocel K4M) and lactose monohydrate are sieved separately.

Stage 2. Transfer the sieved materials from stage 1 to a suitable mixer and mix.

Stage 3. Transfer the mixture from stage 2 to a suitable granulator and mix.

Step 4. During mixing, purified water is added.

Step 5: Transfer the granules from step 4 to a suitable dryer and dry until the loss on drying (LOD) is less than 3%.

Step 6: Screen the granules from step 5.

Step 7. Mix the screened magnesium stearate with the granules from step 6 in a suitable mixer.

Step 8. Compress the finished mixture from step 7 on a suitable rotary tablet press.

Protocol B:

Step 1. The adipic acid salt and Compound 1, microcrystalline cellulose, hypromellose and pregelatinized starch are sieved separately.

- 25 042956

Stage 2. Transfer the sieved materials from stage 1 to a suitable mixer and mix.

Stage 3. Transfer the mixture from stage 2 to a suitable granulator and mix.

Step 4. During mixing, purified water is added.

Step 5. Transfer the granules from step 4 to a suitable dryer and dry until the loss on drying (LOD) is less than 3%.

Step 6: Screen the granules from step 5.

Step 7. Separately sieved Polyox, butylated hydroxytoluene and colloidal silicon dioxide.

Step 8. Transfer the granules from step 6 and substances from step 7 into a suitable mixer and mix.

Step 9 Add magnesium stearate to the materials from step 8 and continue mixing.

Step 10 Compress the finished mixture from step 9 on a suitable rotary tablet press.

Protocol C:

Step 1. Separately, lactose monohydrate, adipic acid salt of Compound 1, microcrystalline cellulose and hypromelloses are sifted through a suitable sieve.

Stage 2. Transfer the sieved materials from stage 1 to a suitable mixer and mix.

Stage 3. Transfer the mixture from stage 2 to a suitable granulator and mix.

Step 4. During mixing, purified water is added.

Step 5. Wet granules are sifted through a suitable sieve.

Step 6. Transfer the granules from step 5 to a suitable dryer and dry until the loss on drying (LOD) is less than 3%.

Step 7. Crush the granules from step 6.

Step 8. Mix the screened magnesium stearate with the granules from step 7 in a suitable mixer.

Step 9. Compress the finished mixture from step 8 on a suitable rotary tablet press.

Protocol D:

Step 1. Separately, the pregelatinized starch, the adipic acid salt of Compound 1, the hypromellose and a portion of the microcrystalline cellulose are sifted through a suitable sieve.

Stage 2. Transfer the sieved materials from stage 1 to a suitable mixer and mix.

Stage 3. Transfer the mixture from stage 2 to a suitable granulator and mix.

Step 4. During mixing, purified water is added.

Step 5. Wet granules are sifted through a suitable sieve.

Step 6. Transfer the granules from step 5 to a suitable dryer and dry until the loss on drying (LOD) is less than 3%.

Step 7. Crush the granules from step 6.

Step 8. Screen the rest of the microcrystalline cellulose and half of the sodium bicarbonate.

Step 9. Transfer the granules from step 7 and the screened materials from step 8 to a suitable mixer and mix.

Step 10. Sift the rest of the sodium bicarbonate and mix with the mixture from step 9.

Step 11. Sift the magnesium stearate and mix with the mixture from step 10.

Step 12 Compress the finished mixture from step 11 on a suitable rotary tablet press.

SR1. Composition of 100 mg sustained release tablets.

Component Function Weight (mg/tablet) Compound (wt%) Salt of adipic acid and compounds 1 ^L Active ingredient 126.42 21.1 microcrystalline cellulose Filler 60.0 10.0 Hypromellose (Methocel K100LV) Release control 60.0 10.0 Hypromellose (Methocel K4M) Release control 60.0 10.0 Lactose monohydrate Filler 290.58 48.4 Magnesium stearate ^b Lubricant 3.0 0.5

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Component Function Weight (mg/tablet) Compound (wt%) Purified water ^L Fluid for granulation enough. number - Total 600.0 100

^a The conversion factor for adipic salt to free base is 0.7911. ^b Added after granulation.

^c Removed during production.

SR2. Composition of 100 mg sustained release tablets.

Component Function Weight (mg/tablet) Compound (wt%) Salt of adipic acid and compound 1' ^L Active ingredient 126.4 ^a 21.1 Microcrystal and weighty cellulose Filler 180.0 30.0 Hypromellose (Methocel K100LV) Binder 6.0 1.0 Polyethylene oxide (Polyox WRS 1105) ^b Release control 180.0 30.0 pregelatinized starch Filler 101.6 16.9 Colloidal silicon dioxide ^b glidant 3.0 0.5 Butylated ^{hydroxytoluene} Antioxidant 0.012 0.002 Magnesium ^stearate Lubricating substance 3.0 0.5 Purified water ^L Fluid for granulation enough. quantity -- Total 600.0 100.0

^a The conversion factor for adipic salt to free base is 0.7911.

^b Added after granulation.

^c Removed during production.

SR3 (100 mg). Composition of 100 mg sustained release tablets.

Component Function Weight (mg/tablet) Compound (wt%) Salt of adipic acid and compounds 1 ^a Active ingredient 126.4 ^a 21.1 microcrystalline cellulose Filler 108.0 18.0 Hypromellose (Methocel K100LV) Release control 42.0 7.0 Hypromellose (Methocel K4M) Release control 30.0 5.0 Lactose monohydrate Filler 290.6 48.4 Magnesium stearate ’ Lubricant 3.0 0.5 Purified water ^L Fluid for granulation enough. quantity -- Total 600.0 100.0

^a The conversion factor for adipic salt to free base is 0.7911.

^b Added after granulation.

^c Removed during production.

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SR4. Composition of 100 mg sustained release tablets.

Excipient Function Weight (mg/tablet) Compound (wt%) Salt of adipic acid and compound 1 ^:ι Active ingredient ^126.4th 21.1 Microcrystalline cellulose ^d Filler 104.6 17.4 Hypromellose (Methocel K100LV) Release control 210.0 35.0 Pregelatin from and millet starch Filler 60.0 10.0 Sodium ^bicarbonate Substance that promotes passage through the stomach 96.0 16.0 Magnesium ^stearate Lubricating substance 3.0 0.5 Water purified ^from Liquid for granulation enough. quantity -- Total 600.0 100.0

^a The conversion factor for adipic salt to free base is 0.7911.

^b Added after granulation.

^c Removed during production.

^d Part added before and part added after granulation.

SR at 25 mg. Composition of 25 mg sustained release tablets.

Component Function Weight (mg/tablet) Compound (wt%) Salt of adipic acid and compound D ¹ Active ingredient 31.6 ^a 12.6 microcrystalline cellulose Filler 105.0 42.0 Hypromellose, (Methocel K100LV) Release control 25.0 10.0 Hypromellose, (Methocel K4M) Release control 25.0 10.0 Lactose monohydrate Filler 62.15 24.9 Magnesium ^stearate Lubricating 1.25 0.5 substance Water purified ^from Fluid for granulation enough. quantity - Total 250 100.0

^a The conversion factor for adipic salt to free base is 0.7911.

^b Added after granulation.

^c Removed during production.

Various modifications of the invention, in addition to those shown and described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to be within the scope of the appended claims. Each reference, including all patents, patent applications and publications cited herein, is incorporated herein by reference in its entirety.

Claims (8)

1. A method of treating cytokine release syndrome in a subject, said method comprising administering to the subject a selective inhibitor of the JAK1 pathway, which is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}- 3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof. 2. The method of claim 1 wherein the selective JAK1 pathway inhibitor is the adipic acid salt {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-( 7H-pyrrolo[2,3d]pyrimidin-4-ul)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile. 3. The method of claim 1 or 2, wherein the method further comprises administering tocilizumab to said subject. 4. The method of claim 1 or 2, wherein the method further comprises administering a corticosteroid to said patient. 5. The method of claim 1 or 2, wherein the method further comprises administering prednisone to said subject. 6. The method of claim 1 or 2, wherein the method further comprises administering tocilizumab and a corticosteroid to said patient. 7. The method according to claim 1, where 4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof is administered to said subject as monotherapy. 8. The method of claim 2 wherein the adipic acid salt {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3 -d]pyrimidin-4-ul)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile is administered to said subject as monotherapy.