JAK1 pathway inhibitors for the treatment of gastrointestinal diseases

Abstract

The present disclosure relates to JAK1 pathway inhibitors and their use in treating gastrointestinal diseases or disorders, such as ulcerative colitis.

Description

[Technical field]

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

[0002] Ulcerative colitis (UC) is the most common form of inflammatory bowel disease worldwide. It is a chronic, idiopathic, recurrent disease of the mucosa that typically involves the rectum and extends proximally to the colon, resulting in diffuse easy bleeding and erosion with bleeding. Although there is some correlation between the extent of disease and the severity of symptoms, the course of the disease is mild in many patients (Solberg et al., Scand. J. Gastroenterol. 2009;44:431-440). In most patients, the disease is characterized by periods of symptom flare-ups and remission, and patients may also experience disease prolongation over time.

[0003] Patients with UC typically experience recurrent episodes of rectal bleeding and diarrhea, often associated with crampy abdominal pain and tenesmus. Characteristic clinical features include diarrhea, rectal bleeding, mucus discharge, tenesmus, fecal urgency, and abdominal pain. Patients may also experience fatigue, fever, weight loss, and dehydration, especially in more severe cases. Mortality is generally not increased in UC, but the disease can present as a life-threatening fulminant colitis. Most patients follow a chronic intermittent course with periods of increased disease activity separated by periods of disease remission. After initial diagnosis, approximately half of patients will have active disease at any single time, and approximately 90% will have a disease course characterized by intermittent flares.

[0004] The incidence of UC in developing countries has been steadily increasing since the mid-20th century. The annual incidence of UC ranges from 1.2 to 20.3 cases per 100,000, with the highest incidence seen in Northern European and North American populations (Loftus et al. Gastroenterology 2004;126:1504-1517). Typical onset of UC occurs between the ages of 15 and 30 (Andres et al., Gastroenterol. Clin. North Am. 1999;28:255-281). Men and women appear to be affected in equal proportions. Westernized environments and lifestyles are recognized as risk factors for inflammatory bowel disease.

[0005] Current therapies for UC include mesalamine, glucocorticoids, thiopurines, and inhibitors of TNFα and α4β7 integrin. Many patients have no response or a non-sustainable response to these therapies.

[0006] Despite these treatment options, a significant proportion of UC patients still require colectomy due to refractory, severe fulminant disease, or in some cases, for cancer prevention. Patients with UC are often considered cured by rectal resection and anal-sparing proctocolectomy, but quality of life is poor and surgery can be associated with short- and long-term complications, including reduced fertility in women and the development of pouchitis.

[0007] Currently, current pharmacological therapies are unable to provide a cure for UC. The primary treatment goal is to induce and then maintain remission.

[0008] Thus, there is a need to develop new therapies for the treatment of gastrointestinal diseases or disorders, such as ulcerative colitis. The present application addresses this need and others. [Brief description of the drawings]

[0009] [Figure 1]1 shows the mean plasma concentration-time profiles for the mean individual maximum fecal concentrations following administration of Compound 1 at a single dose of 25 mg. [Diagram 2] 1 shows individual plasma concentration-time profiles of fecal concentrations following administration of a single 25 mg dose of Compound 1. [Diagram 3] 1 shows [14C]Compound 1 concentrations in colons from healthy and ulcerative colitis subjects after 1 hour of incubation. [Figure 4] 1 shows changes in IL-6 and TPO-induced STAT3 phosphorylation by Compound 1 treatment group in patients (PD-evaluable subjects) as described in Example 3. [Diagram 5] FIG. 1 shows the correlation between IL-6-stimulated inhibition of STAT3 phosphorylation in patients and efficacy measures (change from baseline in Static Physician's Global Assessment (sPGA) and Psoriasis Area and Severity Index (PASI)) as described in Example 3. [Figure 6A] 1 shows the change in IL-6-induced STAT3 phosphorylation on day 15 of cycle 1 in individuals as described in Example 3. [Figure 6B] 1 shows the change in TPO-induced STAT3 phosphorylation on day 15 of cycle 1 in individuals as described in Example 3. [Figure 7] Figure 7 shows that twice-daily Compound 1 treatment (30 mg / kg) reduces symptoms (Figure 7A), gross tissue abnormalities (Figure 7B), and histopathological evidence (Figures 7C-7D) in an IL-10 knockout mouse model of spontaneous colitis. Data represent mean + SEM, n=9-10 per treatment group. *p<0.05, ***p<0.001, ****p<0.0001. [Figure 8]Figure 8 shows that twice-daily Compound 1 treatment (30 mg / kg) reduces symptoms (Figure 8A), tissue damage (Figure 8B), and inflammatory swelling (Figure 8C) in a mouse model of oxazolone-induced colitis. Data represent mean + SEM, n=8 per treatment group. Non-parametric two-tailed Kruskal-Wallis with Dunn test for colitis disease and macroscopic assessment. Parametric two-tailed ANOVA with Holm-Sidak test for colon weight analysis *p<0.05, **p<0.01, ***p<0.0001. [Figure 9] Figure 9A shows that twice-daily Compound 1 treatment administered orally (Figure 9A) or via intracolonic injection (Figure 9B) significantly reduced disease severity in a TNBS-induced colitis model in mice. Data represent mean + SEM, n = 3-8 per treatment group. *p<0.05, **p<0.01. [Figure 10] Results show that twice-daily treatment with compound 1 at 30 mg / kg inhibited disease development in an IL-10 knockout (KO) mouse model of spontaneous colitis. Data represent mean + SEM, n = 9-10 per treatment group, p values ​​were calculated using Kaplan-Meier survival curve analysis. SEM, standard error of the mean. [Figure 11] Figure 11A shows that twice daily treatment with Compound 1 either orally (Figure 11A) or via intracolonic injection (Figure 11B) significantly reduced disease severity in a TNBS-induced colitis model in mice. High-dose oral (Figure 11C) and low-dose intracolonic (Figure 11D) achieved sustained drug exposure above the IC50 coverage. Data represent mean + SEM, n = 8 per treatment group. IBD, inflammatory bowel disease; SEM, standard error of the mean. *p<0.05, **p<0.01. [Figure 12A] 1 shows a volcano plot of differentially expressed genes in IL-10 KO mouse colon following oral administration of Compound 1 in a spontaneous colitis mouse model. [Figure 12B] Statistically significant differentially expressed genes in Compound 1 treated mice compared to the vehicle group in a spontaneous colitis mouse model. [Figure 13A] Figure 13 shows the results of systemic and localized intracolonic Compound 1 delivery in an oxazolone-induced mouse model of inflammatory bowel disease (IBD). Twice-daily Compound 1 treatment administered orally (Figure 13A, 13C) or intracolonically (Figure 13B, 13D) significantly improved stool consistency and reduced fecal occult blood scores in an oxazolone-induced colitis model in mice. Data represent mean + SEM, n = 8 per treatment group. SEM: standard error of the mean. **p<0.01, ***p<0.001, ****p<0.0001. [Figure 13B] Figure 13 shows the results of systemic and localized intracolonic Compound 1 delivery in an oxazolone-induced mouse model of inflammatory bowel disease (IBD). Twice-daily Compound 1 treatment administered orally (Figure 13A, 13C) or intracolonically (Figure 13B, 13D) significantly improved stool consistency and reduced fecal occult blood scores in an oxazolone-induced colitis model in mice. Data represent mean + SEM, n = 8 per treatment group. SEM: standard error of the mean. **p<0.01, ***p<0.001, ****p<0.0001. [Figure 13C] Figure 13 shows the results of systemic and localized intracolonic Compound 1 delivery in an oxazolone-induced mouse model of inflammatory bowel disease (IBD). Twice-daily Compound 1 treatment administered orally (Figure 13A, 13C) or intracolonically (Figure 13B, 13D) significantly improved stool consistency and reduced fecal occult blood scores in an oxazolone-induced colitis model in mice. Data represent mean + SEM, n = 8 per treatment group. SEM: standard error of the mean. **p<0.01, ***p<0.001, ****p<0.0001. [Figure 13D]Figure 13 shows the results of systemic and localized intracolonic Compound 1 delivery in an oxazolone-induced mouse model of inflammatory bowel disease (IBD). Twice-daily Compound 1 treatment administered orally (Figure 13A, 13C) or intracolonically (Figure 13B, 13D) significantly improved stool consistency and reduced fecal occult blood scores in an oxazolone-induced colitis model in mice. Data represent mean + SEM, n = 8 per treatment group. SEM: standard error of the mean. **p<0.01, ***p<0.001, ****p<0.0001. [Figure 14A] Representative images of colon shortening resulting from Compound 1 administered orally or directly into the colon compared to vehicle are shown. Oral (FIG. 14B) and intracolonic (FIG. 14D) Compound 1 treatment significantly improved colon shortening compared to the respective vehicle-treated controls (FIGS. 14A, 14C) in the oxazolone-induced mouse colitis model. Colon length data (FIG. 14E) are graphed as mean+SEM, n=8 per treatment group. SEM: standard error of the mean. ****p<0.0001. [Figure 14B] Representative images of colon shortening resulting from Compound 1 administered orally or directly into the colon compared to vehicle are shown. Oral (FIG. 14B) and intracolonic (FIG. 14D) Compound 1 treatment significantly improved colon shortening compared to the respective vehicle-treated controls (FIGS. 14A, 14C) in the oxazolone-induced mouse colitis model. Colon length data (FIG. 14E) are graphed as mean+SEM, n=8 per treatment group. SEM: standard error of the mean. ****p<0.0001. [Figure 14C] Representative images of colon shortening resulting from Compound 1 administered orally or directly into the colon compared to vehicle are shown. Oral (FIG. 14B) and intracolonic (FIG. 14D) Compound 1 treatment significantly improved colon shortening compared to the respective vehicle-treated controls (FIGS. 14A, 14C) in the oxazolone-induced mouse colitis model. Colon length data (FIG. 14E) are graphed as mean+SEM, n=8 per treatment group. SEM: standard error of the mean. ****p<0.0001. [Figure 14D]Representative images of colon shortening resulting from Compound 1 administered orally or directly into the colon compared to vehicle are shown. Oral (FIG. 14B) and intracolonic (FIG. 14D) Compound 1 treatment significantly improved colon shortening compared to the respective vehicle-treated controls (FIGS. 14A, 14C) in the oxazolone-induced mouse colitis model. Colon length data (FIG. 14E) are graphed as mean+SEM, n=8 per treatment group. SEM: standard error of the mean. ****p<0.0001. [Figure 14E] Representative images of colon shortening resulting from Compound 1 administered orally or directly into the colon compared to vehicle are shown. Oral (FIG. 14B) and intracolonic (FIG. 14D) Compound 1 treatment significantly improved colon shortening compared to the respective vehicle-treated controls (FIGS. 14A, 14C) in the oxazolone-induced mouse colitis model. Colon length data (FIG. 14E) are graphed as mean+SEM, n=8 per treatment group. SEM: standard error of the mean. ****p<0.0001. [Figure 15] Figure 15 shows that systemic Compound 1 delivery is associated with a significant protective effect on colon morphology in IL-10 KO mice. Figure 15A shows a representative colon, and Figure 15B shows a hematoxylin / eosin stained colon section. The white arrow in Figure 15B indicates the area of ​​mononuclear cell infiltration. 20x magnification, bar = 100 μm. Summary of the Invention

[0010] A method for treating a gastrointestinal disease or disorder in a subject in need thereof is provided herein, comprising administering to the subject a therapeutically effective amount of a JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof.

[0011] Provided herein are JAK1 pathway inhibitors for the treatment of a gastrointestinal disease or disorder in a subject in need thereof.

[0012] Provided herein is the use of a JAK1 pathway inhibitor for the manufacture of a medicament for use in treating a gastrointestinal disease or disorder in a subject in need thereof. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] The present invention provides, inter alia, a method for treating a gastrointestinal disease or disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof.

[0014] The methods described herein utilize JAK1 pathway inhibitors, particularly JAK1 selective inhibitors. JAK1 selective inhibitors are compounds that preferentially inhibit JAK1 activity over other Janus kinases. JAK1 plays a central role in several cytokine and growth factor signaling pathways that, when dysregulated, can result in or contribute to disease states. JAK1 has been shown to cooperate with other JAKs to mediate the signaling of several inflammatory cytokines associated with many inflammatory disorders, including ulcerative colitis (UC). Inhibition of JAK / STAT signaling by targeting multiple UC-associated cytokine pathways has the potential to simultaneously reduce inflammation, cell activation, and proliferation of key immune cells, and thus represents a promising therapeutic strategy for the treatment of UC. Tofacitinib for the treatment of UC was recently approved by the FDA. However, as a systemically acting pan-JAK inhibitor, tofacitinib therapy appears to carry an increased risk of immunosuppression (Sandborn et al., N. Engl. J. Med. 2017;376:1723-1736).

[0015] JAK1 pathway inhibitors, specifically compound 1 (i.e., {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, see Table 1), maximize colonic exposure while minimizing systemic exposure when administered in sustained release form and at lower doses than those used for systemic therapy (see, e.g., Example 1). As a result, the efficacy of JAK1 pathway inhibitors is expected to be mediated primarily through local, rather than systemic, JAK1 inhibition.

[0016] Furthermore, patients with gastrointestinal disorders may benefit from JAK1 inhibition, particularly selective JAK1 pathway inhibition. Selective inhibitors of JAK1 can be effective while avoiding the unnecessary and potentially undesirable effects of inhibiting other JAK kinases.

[0017] Accordingly, there is provided a method for treating a gastrointestinal-related disease or disorder in a subject, the method comprising administering to the subject a JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, wherein a maximum stool concentration of the JAK1 pathway inhibitor after administration of the JAK1 pathway inhibitor is about 25 nM or greater, and a maximum total plasma concentration (C max ) is less than about 450 nM.

[0018] The maximum fecal concentration can be determined by measuring the fecal concentration over a period of time following administration of the JAK1 pathway inhibitor (e.g., from 0 to about 48 hours following administration of the JAK1 pathway inhibitor) using, for example, liquid chromatography with tandem mass spectrometry (LC-MS / MS) analysis. Measuring the fecal concentration of Compound 1 can be performed by the methods described in Example C herein.

[0019] In some embodiments, the maximum stool concentration of the JAK1 pathway inhibitor is about 25 nM, about 30 nM, about 35 nM, about 40 nM, about 45 nM, about 50 nM, about 55 nM, about 60 nM, about 65 nM, about 70 nM, about 75 nM, about 80 nM, about 85 nM, about 90 nM, about 95 nM, or about 100 nM or more after administration of the JAK1 pathway inhibitor. In some embodiments, the maximum stool concentration of the JAK1 pathway inhibitor is about 50 nM or more after administration of the JAK1 pathway inhibitor. In some embodiments, the maximum stool concentration of the JAK1 pathway inhibitor is about 25 nM to 100 nM after administration of the JAK1 pathway inhibitor.

[0020] Maximum total plasma concentration (i.e., C max ) can be determined by measuring the plasma concentration using, for example, liquid chromatography with tandem mass spectrometry (LC-MS / MS) analysis over a period following administration of the JAK1 pathway inhibitor (e.g., 0 to about 48 hours following administration of the JAK1 pathway inhibitor). Measuring the plasma concentration of Compound 1 can be performed by the methods described in Example C herein.

[0021] In some embodiments, the maximum total plasma concentration of the JAK1 pathway inhibitor is about 450 nM, about 425 nM, about 400 nM, about 375 nM, about 350 nM, about 325 nM, about 300 nM, about 275 nM, about 250 nM, about 225 nM, about 200 nM, about 175 nM, about 150 nM, about 125 nM, about 100 nM, about 75 nM, or about 50 nM or less after administration of the JAK1 pathway inhibitor. In some embodiments, the maximum total plasma concentration of the JAK1 pathway inhibitor is about 150 nM or less after administration of the JAK1 pathway inhibitor. In some embodiments, the maximum total plasma concentration of the JAK1 pathway inhibitor is about 141 nM or less after administration of the JAK1 pathway inhibitor. In some embodiments, the maximum total plasma concentration of the JAK1 pathway inhibitor is about 100 nM or less after administration of the JAK1 pathway inhibitor. In some embodiments, the maximum total plasma concentration is between about 25 nM and 100 nM.

[0022] In some embodiments, the maximum unbound plasma concentration of the JAK1 pathway inhibitor is about 150 nM or less after administration of the JAK1 pathway inhibitor. The maximum unbound plasma concentration can be derived from the maximum total plasma concentration of the JAK1 pathway inhibitor (see, e.g., Example C) and in vitro protein binding, and can be determined by equilibrium dialysis. In some embodiments, the maximum unbound plasma concentration of the JAK1 pathway inhibitor is about 150 nM, about 125 nM, about 100 nM, about 75 nM, about 50 nM, or about 25 nM or less after administration of the JAK1 pathway inhibitor. In some embodiments, the maximum unbound plasma concentration of the JAK1 pathway inhibitor is about 100 nM or less after administration of the JAK1 pathway inhibitor.

[0023] In some embodiments, the maximum unbound plasma concentration of the JAK1 pathway inhibitor is about 50 nM or less following administration of the JAK1 pathway inhibitor.

[0024] In some embodiments, the ratio of maximum unbound plasma concentration to maximum stool concentration is about 6, about 5, about 4, about 3, about 2, or about 1 or less. In some embodiments, the ratio of maximum unbound plasma concentration to maximum stool concentration is about 2 or less. In some embodiments, the ratio of maximum unbound plasma concentration to maximum stool concentration is about 1 to about 6.

[0025] In some embodiments of the methods provided herein, the gastrointestinal-related disease or disorder is selected from ulcerative colitis, Crohn's disease, and celiac disease.

[0026] In some embodiments, the gastrointestinal disease is recurrent, refractory, or recurrent and refractory ulcerative colitis. In some embodiments, the subject has failed to respond to a previously administered treatment for ulcerative colitis. In other embodiments, the subject is intolerant to a previously administered treatment for ulcerative colitis. In some embodiments, the previously administered treatment is selected from (a) oral corticosteroids, (b) AZA or 6-MP, or (c) biological therapy such as infliximab or adalimumab.

[0027] I. JAK1 Pathway Inhibitors The methods described herein utilize a JAK1 pathway inhibitor. In some embodiments, the JAK1 pathway inhibitor is selective for JAK1 over JAK2, JAK3, and TYK2 (i.e., a JAK1 selective inhibitor). For example, a compound described herein, or a pharma- ceutically acceptable salt thereof, preferentially inhibits JAK1 over one or more of JAK2, JAK3, and TYK2. In some embodiments, the compound preferentially inhibits JAK1 over JAK2 (e.g., has a JAK2 / JAK1 IC of greater than 1). 50 In some embodiments, the compound or salt is about 10-fold more selective for JAK1 over JAK2. In some embodiments, the compound or salt has an IC 50 The antibody is 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 the affinity of the antibody to JAK1 (see, e.g., Example A).

[0028] In some embodiments, the JAK1 pathway inhibitor is a compound in Table 1, or a pharma- ceutically acceptable salt thereof. The compounds in Table 1 are selective JAK1 inhibitors (i.e., JAK1 pathway inhibitors that are 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. [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] [Table 1-11]

[0029] In some embodiments, the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof.

[0030] In some embodiments, the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate.

[0031] The synthesis and preparation of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile and its adipic acid salts can be found, for example, 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.

[0032] In some embodiments, the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide, or a pharma- ceutically acceptable salt thereof.

[0033] In some embodiments, the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide phosphate.

[0034] 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, which is incorporated herein by reference in its entirety.

[0035] In some embodiments, the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile, or a pharma- ceutically acceptable salt thereof.

[0036] In some embodiments, the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile monohydrate.

[0037] 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 anhydrous and monohydrate forms are described 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.

[0038] In some embodiments, the compounds in Table 1 are disclosed in U.S. Patent Publication Nos. 2011 / 0224190, filed March 9, 2011, 2014 / 0343030, filed May 16, 2014, 2014 / 0121198, filed October 31, 2013, 2014 / 0298334, filed May 21, 2010, 2011 / 0059951, filed August 31, 2010, 2011 / 0059951, filed November 1, 2011, and 2011 / 0062224. No. 2012 / 0149681, filed on May 8, 2012; 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.

[0039] In some embodiments, the JAK1 pathway inhibitor is a compound 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, or 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.

[0040] In some embodiments, the JAK1 pathway inhibitor is a compound of formula I [ka] or a pharma- ceutically acceptable salt thereof, X is N or CH; L is C(=O) or C(=O)NH; Each A is one or two independently selected R 1 phenyl, pyridinyl, or pyrimidinyl, optionally substituted with a group; Each R 1 is independently fluoro, or trifluoromethyl.

[0041] 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.

[0042] 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.

[0043] 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.

[0044] In some embodiments, the JAK1 pathway inhibitor is a compound of formula II [ka] or a pharma- ceutically acceptable salt thereof, R 2 is C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, or C 3-6 Cycloalkyl-C 1-3 is an alkyl group, 1-6 Alkyl, C 3-6 Cycloalkyl and C 3-6 Cycloalkyl-C 1-3 each alkyl is optionally substituted with 1, 2, or 3 substituents independently selected from fluoro, -CF3, and methyl; R 3 is H or methyl, R 4 is H, F, or Cl, R 5 is H or F, R 6 is H or F, R 7 is H or F, R 8 is H or methyl, R 9 is H or methyl, R 10 is H or methyl, R 11 is H or methyl.

[0045] 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.

[0046] In some embodiments, the JAK1 pathway inhibitor is a compound of formula III [ka] or a pharma- ceutically acceptable salt thereof, Cy 4 CN, OH, F, Cl, C 1-3 Alkyl, C 1-3 Haloalkyl, Cyano-C 1-3 Alkyl, HO-C 1-3 Alkyl, Amino, C 1-3 Alkylamino and di(C 1-3 a tetrahydro-2H-pyran ring optionally substituted with one or two groups independently selected from the group consisting of alkyl)amino; 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.

[0047] 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.

[0048] In some embodiments, the JAK1 pathway inhibitor is administered in a daily dose of about 1 mg to about 100 mg, about 3 mg to about 100 mg, about 5 mg to about 100 mg, about 10 mg to about 100 mg, about 10 mg to about 75 mg, or about 25 mg to about 75 mg on a free base basis.

[0049] In some embodiments, the JAK1 pathway inhibitor is administered in a daily dose of about 1 mg to about 100 mg, about 3 mg to about 100 mg, about 5 mg to about 100 mg, about 10 mg to about 100 mg, about 10 mg to about 75 mg, or about 25 mg to about 75 mg on a free base basis.

[0050] In some embodiments, the JAK1 pathway inhibitor is administered in a daily amount of about 10 mg to about 100 mg on a free base basis. Thus, in some embodiments, the selective JAK1 pathway inhibitor is administered in a daily amount of about 1 mg, about 3 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, or about 100 mg on a free base basis.

[0051] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 50 mg to about 100 mg.

[0052] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 25 mg to about 50 mg.

[0053] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 25 mg to about 75 mg.

[0054] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 1 mg.

[0055] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 2 mg.

[0056] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 2.5 mg.

[0057] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 3 mg.

[0058] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 5 mg.

[0059] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 10 mg.

[0060] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 15 mg.

[0061] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 25 mg.

[0062] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 30 mg.

[0063] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 50 mg.

[0064] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 100 mg.

[0065] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered once daily at a dose of about 25 mg.

[0066] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered at a daily dose of about 1 mg / kg to about 50 mg / kg (e.g., as a once or twice daily dose).

[0067] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered at a daily dose of about 3 mg / kg to about 30 mg / kg (e.g., as a once or twice daily dose).

[0068] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered as a dose of about 3 mg / kg twice daily (BID), for a total daily dose of about 6 mg / kg.

[0069] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered as a twice-daily (BID) intracolonic dose of about 3 mg / kg, for a total daily dose of about 6 mg / kg.

[0070] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered as a dose of about 30 mg / kg twice daily (BID), for a total daily dose of about 60 mg / kg.

[0071] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered as an oral dose of about 30 mg / kg twice daily (BID), for a total daily dose of about 60 mg / kg.

[0072] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered at a dose of about 25 mg twice daily, for a total daily dose of about 50 mg.

[0073] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered once daily at a dose of about 50 mg.

[0074] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered at a dose of about 50 mg twice daily, for a total daily dose of about 100 mg.

[0075] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered once daily at a dose of about 100 mg.

[0076] In some embodiments, the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered as one or more sustained release dosage forms, each comprising the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof.

[0077] Provided herein is a method for treating a gastrointestinal disorder in a subject, comprising administering to the subject a daily dose of about 25 mg to 100 mg of a JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, wherein the JAK1 pathway inhibitor, or a pharma-ceutically acceptable salt thereof, is administered in one or more sustained release dosage forms comprising the JAK1 pathway inhibitor, or a pharma-ceutically acceptable salt thereof.

[0078] The embodiments described herein are intended to be combined in any suitable combination as if the embodiments were multiple dependent claims (e.g., embodiments relating to selective JAK1 pathway inhibitors and dosages thereof, embodiments relating to maximum plasma concentrations (total or unbound), embodiments relating to any salt form of the compounds disclosed herein, embodiments relating to particular types of gastrointestinal-related disorders, and embodiments relating to compositions and / or administration may be combined in any combination).

[0079] Also provided herein are methods for treating a gastrointestinal disease selected from the group consisting of inflammatory bowel disorder, ulcerative colitis, spontaneous colitis, Crohn's disease, and celiac disease. In some embodiments, the gastrointestinal disease is selected from the group consisting of ulcerative colitis, Crohn's disease, and celiac disease.

[0080] In some embodiments, the gastrointestinal disorder is selected from the group consisting of inflammatory bowel disorder and spontaneous colitis.

[0081] In some embodiments, the gastrointestinal disease is spontaneous colitis.

[0082] For example, provided herein is a method for treating a gastrointestinal disease selected from the group consisting of ulcerative colitis, Crohn's disease, and celiac disease in a subject in need thereof, the method comprising administering to the subject {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof, wherein the {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, after administration of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof, The maximum fecal concentration 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 is greater than about 25 nM, and {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl) The maximum total plasma concentration 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}acetonyl following administration of -1H-pyrazol-1-yl]azetidin-3-yl}acetonyl, or a pharma- ceutically acceptable salt thereof, is less than about 150 nM.

[0083] Also provided herein is a method for treating a gastrointestinal disorder selected from the group consisting of ulcerative colitis, Crohn's disease, and celiac disease in a subject, the method comprising administering to a subject from about 25 mg to about 100 mg of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl} The method includes administering to a subject a once-daily dose of acetonitrile, or a pharma- ceutically acceptable salt thereof, wherein the dose comprises one or more sustained release dosage forms, each comprising {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-[7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof.

[0084] Also provided herein is a method for treating a gastrointestinal disorder selected from the group consisting of ulcerative colitis, Crohn's disease, and celiac disease in a subject, the method comprising administering about 25 mg of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetamide, on a free base basis. The method includes administering to a subject twice daily doses of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-[7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof, wherein the doses comprise one or more sustained release dosage forms, each comprising {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-[7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof.

[0085] Also provided herein is a method for treating a gastrointestinal disorder selected from the group consisting of ulcerative colitis, Crohn's disease, and celiac disease in a subject, the method comprising administering about 50 mg of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetamide, on a free base basis. The method includes administering to a subject twice daily doses of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-[7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof, wherein the doses comprise one or more sustained release dosage forms, each comprising {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-[7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof.

[0086] A sustained release dosage form of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof (Table 1, Compound 1) can be found in U.S. Publication No. 2015-0065484, filed August 6, 2014, which is incorporated herein by reference in its entirety. See also Example B below.

[0087] All possible combinations are not listed separately herein for the sake of brevity only.

[0088] The compounds described herein may be asymmetric (e.g., have one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise indicated. Compounds containing asymmetrically substituted carbon atoms may be isolated in optically active or racemic forms. Methods on how to prepare optically active forms from optically inactive starting materials are known in the art, such as by resolution of racemic mixtures or stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like, may also exist 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 the present invention are described and may be isolated as a mixture of isomers or as separated isomeric forms.

[0089] In some embodiments, the compounds have the (R) configuration. In some embodiments, the compounds have the (S) configuration.

[0090] The resolution of racemic mixtures of compounds can be carried out by any of a number of methods known in the art. An exemplary method includes fractional recrystallization using chiral resolving acids that are optically active salt-forming organic acids. Suitable resolving agents for fractional recrystallization methods 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 methods include stereoisomerically pure forms (e.g., S and R forms, or diastereomerically pure forms) of α-methylbenzylamine, 2-phenylglycinol, norephedrine, ephedrine, N-methylphedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, and the like.

[0091] Resolution of racemic mixtures can also be carried out by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). Suitable elution solvent compositions can be determined by one skilled in the art.

[0092] The compounds described herein also include tautomeric forms. Tautomeric forms result from the exchange of a single bond with an adjacent double bond, accompanied by the concomitant migration of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which protons can occupy more than one position of a heterocyclic ring system, such as 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomeric forms can be in equilibrium or sterically fixed in one form by appropriate substitution.

[0093] The compounds described herein may also include isotopically labeled compounds of the present disclosure. An "isotopically" or "radiolabeled" compound is a compound of the present disclosure in which one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occurring). Suitable radionuclides that can 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 131For example, one or more hydrogen atoms in the compounds of the present disclosure may be replaced by a deuterium atom (e.g., C of formula (I), (II), or (III)). 1-6 One or more hydrogen atoms of an alkyl group or of a compound of Table 1 can be optionally replaced with a deuterium atom, such as -CD3 replaced with -CH3).

[0094] The term "compound" as used herein is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted, unless the name indicates a specific stereoisomer. Compounds herein identified by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise specified.

[0095] In some embodiments, the compounds described herein, or salts thereof, are substantially isolated. By "substantially isolated" is meant at least partially or substantially separated from the environment in which the compound was formed or detected. Partial separation can include, for example, a composition enriched in the compounds described herein. Substantial separation can include 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.

[0096] All compounds and their pharma- ceutically acceptable salts may be found together with other substances such as water and solvents (e.g., hydrates and solvates) or may be isolated. When in the solid state, the compounds described herein and their salts may occur in various forms, for example, in the form of solvates, including hydrates. Because the compounds may be in any solid state form, such as polymorphs or solvates, unless otherwise expressly indicated, references herein to compounds and their salts should be understood to include any solid state form of the compounds.

[0097] The phrase "pharmacologically acceptable" is used herein to refer to those compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0098] The present invention also includes pharma- ceutically acceptable salts of the compounds described herein. The term "pharma- ceutically acceptable salts" refers to derivatives of the disclosed compounds, where the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharma- ceutically acceptable salts include, but are not limited to, mineral 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 invention include 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 invention 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, with non-aqueous media such as ether, ethyl acetate, alcohol (e.g., methanol, ethanol, isopropanol, or butanol) or acetonitrile (MeCN) being generally preferred. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th Ed., (Mack Publishing Company, Easton, 1985), p. 1418, Berge et al., J. Pharm. Sci., 1977, 66(1), 1-19, and Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (Wiley, 2002). In some embodiments, the compounds described herein include N-oxide forms.

[0099] The terms "individual," "patient," and "subject" are used interchangeably and refer to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, or primate, and most preferably a human.

[0100] The phrase "therapeutically effective amount" refers to an amount of an active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal, individual, or human that is being sought by a researcher, veterinarian, physician, or other clinician.

[0101] The term "treating" or "treatment" refers to one or more of: (1) inhibiting a disease, e.g., inhibiting a disease, condition, or disorder (i.e., halting further development of the pathology and / or symptoms) in an individual experiencing or displaying the pathology or symptoms of the disease, condition, or disorder, and (2) ameliorating a disease, e.g., improving a disease, condition, or disorder (i.e., reversing the pathology and / or symptoms) in an individual experiencing or displaying the pathology or symptoms of the disease, condition, or disorder, such as reducing the severity of the disease. In one embodiment, treating or treatment includes preventing a disease or reducing the risk of developing a disease, e.g., preventing or reducing the risk of developing a disease, condition, or disorder in an individual who may be predisposed to the disease, condition, or disorder, but has not yet experienced or displayed the pathology or symptoms of the disease.

[0102] The terms "for example" and "such as," as well as their grammatical equivalents, are understood to be followed by the phrase "and without limitation," unless expressly stated otherwise.

[0103] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0104] As used herein, the term "about" means "approximately" (eg, plus or minus about 10% of the indicated value).

[0105] Combination therapy The methods described herein can further include administering one or more additional therapeutic agents, including anti-inflammatory agents, steroids, immunosuppressants, or therapeutic antibodies.

[0106] For example, the methods described herein may be used in combination with current UC therapies such as oral mesalamine (5-ASA), oral corticosteroids, azathioprine (AZA), 6-mercaptopurine (6-MP), and methotrexate, infliximab, vedolizumab, mucosal addressin cell adhesion molecule (MADCAM1) inhibitors, and fecal transplants.

[0107] For example, oral 5-ASA (mesalamine, e.g., about 1600 mg / day to about 2400 mg / day) or sulfasalazine (maximum, e.g., about 1000 mg / day to 4000 mg / day) can be administered with a JAK1 pathway inhibitor for any of the methods described herein.

[0108] As another example, oral corticosteroids (e.g., from about 0.5 mg / day to about 60 mg / day of prednisone or an oral corticosteroid equivalent) can be administered with a JAK1 pathway inhibitor for any of the methods described herein.

[0109] As another example, about 50 mg / day to about 225 mg / day of azathioprine, up to, for example, about 30 mg / day to about 112.5 mg / day of 6-mercaptopurine, or up to, for example, about 25 mg of methotrexate may also be administered with a JAK1 pathway inhibitor for any of the methods described herein. In some embodiments, azathioprine is administered at about 50 mg / day to about 100 mg / day with a JAK1 pathway inhibitor for any of the methods described herein. In other embodiments, 6-mercaptopurine is administered at about 30 mg / day to about 50 mg / day with a JAK1 pathway inhibitor for any of the methods described herein.

[0110] As another example, a series of 2-10 mg / kg, e.g., 5 mg / kg, infliximab for induction and maintenance may be administered with a JAK1 pathway inhibitor for any of the methods described herein, in some embodiments, infliximab is administered at 5 mg / kg at weeks 0, 2, and 6, then every 8 weeks thereafter.

[0111] As another example, a dose of about 200 to about 400 mg, e.g., 300 mg, of vedolizumab may be administered with a JAK1 pathway inhibitor for any of the methods described herein. In some embodiments, vedolizumab is administered at weeks 0, 2, and 6, and then every 8 weeks thereafter.

[0112] When two or more pharmaceutical agents are administered to a subject, they may be administered simultaneously, sequentially, or in combination (eg, for three or more agents).

[0113] composition The compounds 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 by a variety of routes, depending on whether local or systemic treatment is indicated and on the area to be treated. Administration may be topical (to mucous membranes, including transdermal, epidermal, ophthalmic, and including intranasal, intravaginal, and rectal delivery), pulmonary (e.g., intratracheal or intranasal, by inhalation or insufflation of powders or aerosols, including by nebulizer), 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 may be in the form of a single bolus dose or may be, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners, and the like may be necessary or desirable. In some embodiments, administration is oral. In some embodiments, administration is colonic.

[0114] The pharmaceutical composition can contain the compound or its pharma- ceutically acceptable salt as active ingredient in combination with one or more pharma- ceutically acceptable carriers (excipients). In some embodiments, the composition is suitable for topical administration. When preparing the composition, 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 material 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 in a liquid medium), for example, ointment containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injection solutions, and sterile packaged powders.

[0115] When preparing formulation, active compound can be milled to provide suitable particle size before combining with other components.If active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh.If active compound is substantially water-soluble, particle size can be adjusted by milling to provide substantially uniform distribution in formulation, for example, about 40 mesh.

[0116] The compounds 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 invention can be prepared by processes known in the art, see, for example, WO2002 / 000196.

[0117] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose.The formulation can additionally include lubricants such as talc, magnesium stearate and mineral oil, wetting agents, emulsifying and suspending agents, preserving agents such as methylbenzoate and propylhydroxybenzoate, and sweeteners and flavoring agents.The composition of the present invention can be formulated to provide quick, sustained or delayed release of the active ingredient after administration to the patient by using procedures known in the art.

[0118] The components used to formulate pharmaceutical compositions are of high purity and substantially free of potentially harmful contaminants (e.g., at least National Food grade, generally at least analytical grade, and more typically at least pharmaceutical grade). Particularly for human consumption, the compositions are preferably manufactured or formulated under Good Manufacturing Practice regulations as defined in applicable regulations of the U.S. Food and Drug Administration. For example, suitable formulations can be sterile and / or substantially isotonic and / or fully compliant with all Good Manufacturing Practice regulations of the U.S. Food and Drug Administration.

[0119] The active compounds can be effective over a wide dosage range and are generally administered in a therapeutically 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 actual compound being administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.

[0120] The therapeutic dosage of a compound of the invention can vary according to, for example, the particular use for which the treatment is made, the manner 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 can vary depending on several factors, including dosage, chemical characteristics (e.g., hydrophobicity), and the route of administration.

[0121] 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 compound of the present invention. When referring to these preformulation compositions as homogenous, the active ingredient is typically dispersed evenly throughout the composition so that the composition can be easily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules. This solid preformulation is then subdivided into unit dosage forms of the type described above, for example, containing about 0.1 to about 1000 mg of the active ingredient of the present invention.

[0122] The tablet or pill of the present invention can be coated or otherwise compounded to provide a dosage form that provides the advantage of long-term action.For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope covering the former.The two components can be separated by an enteric layer, which functions to resist disintegration in the stomach and allows the inner component to pass intact into the duodenum or to be delayed in release.A variety of materials can be used for such enteric layers or coatings, including some polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol and cellulose acetate.

[0123] Liquid forms into which the compounds and compositions of the present invention may be incorporated for administration orally or by injection include aqueous solutions, suitably 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 vehicles.

[0124] The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of administration, such as prophylactic or therapeutic, the condition of the patient, the mode 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 disease state being treated, depending on factors such as the severity of the disease, the age, weight, and general condition of the patient, as well as the judgment of the attending clinician.

[0125] 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, and 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 understood that the use of some of the aforementioned excipients, carriers, or stabilizers will result in the formation of pharmaceutical salts.

[0126] kit The present application also includes useful pharmaceutical kits, including one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound, or any of their embodiments. Such kits may further include one or more of a variety of conventional pharmaceutical kit components, such as, for example, a container containing one or more pharma- ceutically acceptable carriers, additional containers, etc., as would be readily apparent to one of skill in the art. Instructions, either as an insert or as a label, indicating the amounts of components to be administered, guidelines for administration, and / or guidelines for mixing the components, may also be included in the kit. EXAMPLES

[0127] The present invention will be described in more detail with 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.

[0128] Example A: In vitro JAK kinase assay JAK1 pathway inhibitors that may be used for the treatment of 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 (aa837-1142), JAK2 (aa828-1132) and JAK3 (aa781-1124) 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 homogenous time-resolved fluorescence (HTRF). The IC of the compounds was determined by the following: 50is measured for each kinase in 40 μL reactions containing 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. IC 50 For measurements, the ATP concentration in the reaction is 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 in assay buffer (Perkin Elmer, Boston, Mass.). Binding to europium-labeled antibody occurred for 40 minutes and the HTRF signal was measured on a fusion plate reader (Perkin Elmer, Boston, Mass.). The compounds in Table 1 were tested in this assay and the IC 50 The values ​​are shown to have the following meanings:

[0129] Example B: Preparation of a sustained release formulation of Compound 1 Extended release tablets containing Compound 1 were prepared with excipients in the amounts shown in the table below. Protocol A was used for SR1 tablets, Protocol B was used for SR2 tablets, Protocol C was used for SR3 tablets and 25 mg SR tablets, and Protocol D was used for SR4 tablets. These procedures are disclosed in US Patent Publication No. 2015 / 0065484 and relate to extended release dosage forms of Compound 1.

[0130] Protocol A: Step 1. Screen compound 1 adipate salt, microcrystalline cellulose, hypromellose (Methocel K100 LV and Methocel K4M), and lactose monohydrate individually. Step 2. Transfer the screened material from step 1 into a suitable blender and mix. Step 3. Transfer the blend from step 2 into a suitable granulator and mix. Step 4. Add purified water while mixing. Step 5. Transfer the granules from step 4 into a suitable dryer and dry until the LOD is less than 3%. Step 6. Screen the granules from step 5. Step 7. Mix the screened magnesium stearate with the granules of Step 6 in a suitable blender. Step 8. Compress the final blend of Step 7 on a suitable rotary tablet press.

[0131] Protocol B: Step 1. Individually screen the adipate salt of the compound of formula I, microcrystalline cellulose, hypromellose and pregelatinized starch. Step 2. Transfer the screened material from step 1 into a suitable blender and mix. Step 3. Transfer the blend from step 2 into a suitable granulator and mix. Step 4. Add purified water while mixing. Step 5. Transfer the granules from step 4 into a suitable dryer and dry until the LOD is less than 3%. Step 6. Screen the granules from step 5. Step 7. Polyox, butylated hydroxytoluene and colloidal silicon dioxide were screened individually. Step 8. Transfer the granules from step 6 and the material from step 7 to a suitable blender and mix. Step 9. Add the screened magnesium stearate to the ingredients from Step 8 and continue blending. Step 10. Compress the final blend of Step 9 on a suitable rotary tablet press.

[0132] Protocol C: Step 1. Screen lactose monohydrate, the adipate salt of the compound of Formula I, microcrystalline cellulose, and hypromellose individually through a suitable screen. Step 2. Transfer the screened material from step 1 into a suitable blender and mix. Step 3. Transfer the blend from step 2 into a suitable granulator and mix. Step 4. Add purified water while mixing. Step 5. Screen the wet granules through a suitable screen. Step 6. Transfer the granules from step 5 into a suitable dryer and dry until the LOD is less than 3%. Step 7. Mill the granules from step 6. Step 8. Mix the screened magnesium stearate with the granules of Step 7 in a suitable blender. Step 9. Compress the final blend of Step 8 on a suitable rotary tablet press.

[0133] Protocol D: Step 1. Individually screen pregelatinized starch, the adipate salt of the compound of formula I, hypromellose, and a portion of the required microcrystalline cellulose through a suitable screen. Step 2. Transfer the screened material from step 1 into a suitable blender and mix. Step 3. Transfer the blend from step 2 into a suitable granulator and mix. Step 4. Add purified water while mixing. Step 5. Screen the wet granules through a suitable screen. Step 6. Transfer the granules from step 5 into a suitable dryer and dry until the LOD is less than 3%. Step 7. Mill the granules from step 6. Step 8. Screen the remaining portion of the microcrystalline cellulose and half of the sodium bicarbonate. Step 9. Transfer the milled granules from step 7 and the screened material from step 8 to a suitable blender and mix. Step 10. Screen the remaining portion of the sodium bicarbonate and mix with the Step 9 blend. Step 11. Screen the magnesium stearate and mix with the blend from step 10. Step 12. Compress the final blend of Step 11 on a suitable rotary tablet press. [Table 2] [Table 3] [Table 4] [Table 5] [Table 6]

[0134] Example C: Bioanalysis of Compound 1 in Plasma and Feces Two different assays can be used to understand the functional activity of JAK1 inhibition. The first one is a standard cell-based assay, while the other one uses whole blood. The former is performed using human peripheral blood mononuclear cells (PBMCs), and briefly, the cells are stimulated with IL-6 to increase JAK1 activity, measured via phosphorylated STAT3. As increasing concentrations of compound 1 are added, a corresponding decrease in phosphorylated STAT3 is observed. This assay is suitable for evaluating the JAK1 activity and / or inhibitory activity of compound 1 in samples lacking serum proteins, e.g., stool samples.

[0135] To evaluate the inhibitory activity of Compound 1 in serum-rich media, such as plasma or whole blood, an assay is performed using whole blood, and briefly, whole blood samples are stimulated with IL-6 and the level of phosphorylated STAT3 is determined. This assay can be performed either in vitro (human blood samples are spiked with Compound 1) or ex vivo (whole blood samples are collected from human subjects administered Compound 1).

[0136] I. Compound 1 in Human Plasma The method used to analyze compound 1 in human plasma has been validated. Briefly, 50 μL of human plasma samples are placed in a 96-well plate. An aliquot of 50 μL of internal standard (dissolved in 50:50 acetonitrile:water) is added, followed by an aliquot of 100 μL of 0.1 M NaHCO3. Then, 800 μL of methyl-t-butyl ether (MtBE) is added and the samples are covered and vortexed. After centrifugation, 700 μL of the MtBE layer is transferred to a clean 96-well plate. The samples are then dried under nitrogen at approximately 50° C. Then, an aliquot of 250 μL of reconstitution solution (acetonitrile:water, 50:50, v / v) is added to each sample. The plate is placed in an autosampler tray and injected into the LC-MS / MS for analysis. LC-MS / MS analysis is performed using an AB Sciex 4000 or Sciex 6500 QTRAP mass spectrometer coupled with an HPLC pump and autosampler. Chromatographic separation is achieved on a Waters T3 (50 mm x 2.1 mm) HPLC column using isocratic elution. The mass spectrometer is operated in positive ESI mode. Multiple reaction monitoring (MRM) transitions are m / z 554.1 -> 186.0 for compound 1 and m / z 558.1 -> 190.0 for the internal standard. Peak area integration is performed using the analysis software and concentrations are calculated in Watson LIMS. Concentrations are calculated using 10 concentration levels ranging from 5 nM to 5000 nM with weighted linear regression according to the following formula: y=ax+b(weighting factor=1 / x 2 ) where x=compound 1 concentration in nM, y=peak area ratio, a=slope, and b=intercept.

[0137] The lower limit of quantification is 5 nM and the calibration curve ranges from 5 nM to 5000 nM for compound 1 in human plasma.

[0138] II. Compound I in human feces The method used to analyze Compound 1 in human stool is a qualified method. Human stool samples are collected at the clinical site in a 1:1 homogenate [1 part water (mL): 1 part stool (g)]. Prior to sample analysis, additional water is added to the sample homogenate to achieve a final stool to water ratio of 1:19 for the calibration standards and QC samples. The final homogenate is processed and analyzed along with the calibration standards and QC samples.

[0139] For human homogenate analysis, briefly, 100 μL of stool homogenate (blank, QC and study samples) is placed in a test tube. An aliquot of 20 μL of internal standard is added and mixed, followed by an aliquot of 200 μL of 0.1 M NaHCO3 and vortexing. Then, 2 mL of MtBE is added and the sample is vortexed. After centrifugation, the MtBE layer is transferred to a clean test tube. The samples are then dried at about 40° C. under nitrogen. An aliquot of 1 mL of reconstitution solution (acetonitrile:water, 50:50, v / v) is then added to each sample and vortexed. 10 μL of sample is then diluted with 3 mL of reconstitution solution in a clean test tube. Samples are transferred to an autosampler vial and 10 μL is injected into the LC-MS / MS for analysis. LC-MS / MS analysis is performed using an AB Sciex API 4000 or API 4000 QTrap mass spectrometer coupled with an HPLC pump and autosampler. Chromatographic separation is achieved on an Agilent Eclipse Plus C8 50×4.6 mm, 5 μm HPLC column using gradient elution. The mass spectrometer is operated in positive ESI mode. MRM transitions are m / z 554.3→186.2 for compound 1 and m / z 558.4→190.2 for the internal standard. Peak area integration is performed using the analysis software and concentrations are calculated in Watson LIMS. Concentrations in human stool homogenates are calculated using eight concentration levels ranging from 1 μg / g to 300 μg / g (1.8 μM to 542 μM) with weighted linear regression according to the following formula: y=ax+b(weighting factor=1 / x 2 ) where x=compound 1 concentration in μg / g in human stool homogenate, y=peak area ratio, a=slope, and b=intercept.

[0140] Example 1: Dosing Strategy for Compound I, a Selective JAK1 Inhibitor, for the Treatment of Ulcerative Colitis Compound 1 is a JAK1 inhibitor currently in development for oncology and autoimmune diseases. Clinical and ex vivo studies were conducted to understand the colonic losses that are important for ulcerative colitis (UC).

[0141] Methods: Compound 1 concentrations in plasma and feces (colon surrogates) were determined after a single sustained release 25 mg oral dose (see, e.g., Example B, 25 mg SR composition). Compound 1 concentrations in plasma after a single 100 mg dose were also determined in a separate study (see, e.g., Example C for measuring Compound 1 concentrations in plasma). Ex vivo studies: Colon tissue samples from healthy and UC subjects (2 / group) were mounted on vertical Ussing diffusion chambers. [ 14 [C] Compound 1 was applied to the upper side of the chamber at 100 and 1000 nM and incubated for 1 h. Samples were collected from the donor and receiver sides for determination of Compound 1 concentration. Colon tissue was snap frozen for quantitative autoradiography.

[0142] Results: Compound 1 is delivered as a sustained release formulation, with 27.1% of the dose being eliminated as unchanged Compound 1 in the stool (see, e.g., Example C for measuring the concentration of Compound 1 in the stool). After a single 25 mg dose of Compound 1, 8 of 12 patients demonstrated in vitro IC 50 The maximum fecal concentrations were greater than 0.01 (i.e., 58 nM) (Figures 1 and 2). Maximum fecal PK mean (SD), GM = 93.4 nM (41.4 nM), 85.5 nM, where PK is pharmacokinetic, SD is standard deviation, and GM is geometric mean. Maximum fecal concentrations were taken directly from observed fecal data, e.g., fecal concentrations collected from 0-24 hours.

[0143] Systemic concentrations were calculated as IC for JAK1 inhibition in whole blood for either dose. 50 Below the mean (SD) C max = 18.9 (7.46) nM for 25 mg and 84.4 (45.8) nM for 100 mg (Figure 1 and table below). [Table 7]

[0144] Standard non-compartmental pharmacokinetic methods were used to analyze Compound 1 plasma concentrations. max (maximum plasma concentration) and T max (the time when maximum plasma concentration occurs) was taken directly from the observed plasma concentration data. z ) was estimated using log-linear regression of the concentration data during the terminal elimination phase, and t 1 / 2 ln(2) / λ z The AUC all is defined as the area under the plasma concentration-time curve from time 0 to the last observation, calculated using the linear trapezoidal rule for increasing concentrations and the logarithmic trapezoidal rule for decreasing concentrations. AUC 0-inf AUC 0-t +C t / λ z Calculated as AUC 0-t was defined as the area under the plasma concentration-time curve from time 0 to the last measurable concentration (also calculated using the linear up / log down trapezoidal rule), C t is the last measurable concentration; Cl / F is the apparent clearance, Dose / AUC 0-inf Vz / F is calculated as dose / (λ z *AUC 0-inf ) is the apparent volume of distribution based on the terminal phase calculated as

[0145] Ex vivo, no Compound 1-associated radioactivity was detected from the receiver side. Compound 1 penetrated the mucosal layer in a concentration-dependent manner, and to a lesser extent the submucosal layer (see Example 2).

[0146] Summary: A dose range of about 25 mg to about 100 mg BID (twice daily) or about 25 mg to about 200 mg QD (once daily) is recommended for studies in UC patients to maximize colonic exposure while minimizing the potential for systemic exposure.

[0147] Example 2: Microautoradiography (MARG) and quantitative autoradioluminography (QARL) of colons from healthy subjects with ulcerative colitis 14 C] Tissue penetration and distribution analysis of compound 1 I. Purpose The aim of this study was to determine the expression of α-amyloid in colon samples collected from healthy and ulcerative colitis (UC) human subjects using quantitative autoradioluminography (QARL) and microautoradiography (MARG). 14 The aim of this study was to determine the tissue distribution of [C]Compound 1-associated radioactivity.

[0148] II. Materials and Methods A. Sample submission Small pieces of colon samples from two healthy subjects and two subjects with UC (total of eight samples) were provided by Analytical Biological Services Inc. (Wilmington, Del.) and stored at -70°C until use.

[0149] B. Medication Formulations Dosage formulations, i.e., 100 nM and 1000 nM, were prepared on the day of the experiment for all tissues in the study. 14 C] Compound 1 (1.06 mg) was dissolved in dimethyl sulfoxide (DMSO; 1.514 mL) to produce a 1 mM stock solution (0.7 mg / mL). The stock solution (20 μL) was diluted with Krebs-Ringer bicarbonate (KRB) buffer (20 mL) to reach a final concentration of 1000 nM. The 1000 nM dosing formulation (2 mL) was diluted with KRB buffer (18 mL) to reach a final concentration of 100 nM. The pH of both dosing formulations was approximately 5.5.

[0150] Dosing formulations were analyzed prior to incubation to determine radioactivity concentration and homogeneity. 100 μL aliquots were taken from the top, middle, and bottom of the formulation container, each weighed and diluted to 10 mL with DMSO for radioactivity analysis. Triplicate aliquots of each 10 mL dilution were analyzed by liquid scintillation counting (LSC).

[0151] C. Incubation and Sample Recovery Intestinal tissue permeation studies were performed using a vertical Ussing diffusion chamber system (Harvard Apparatus, Holliston, MA) for healthy tissue. Frozen tissue was thawed to ambient temperature, rinsed with pre-warmed KRB buffer used for dosing formulation, and then gently placed on the apparatus. Permeation was performed from mucosa to serosa for 1 hour at 37°C with the test article in KRB buffer added to the mucosal side. The receiver side of the Ussing chamber containing blank KRB buffer was agitated with air bubbles from an aeration device. Due to limited sample availability, UC tissue was mounted (mucosal side up) on one end of a polypropylene tube with both ends cut open, and the serosal side was placed in blank KRB buffer in a vial with a stir bar. KRB containing the test article was added to the tube, thus exposing the mucosal side of the tissue to the test article during incubation. After 1 h incubation at 37 °C, samples (100-500 μL) were collected from both the donor and receiver sides and then transferred to 1.5 mL tubes for assessment of permeation. Tissue samples were gently removed from the chambers (healthy tissue) or tubes (UC tissue) and flash frozen for approximately 30 s in liquid nitrogen-cooled isopentane. Individual frozen healthy and UC colon samples were embedded in cryogel medium and the larger healthy tissue sample was split in half for primary and secondary samples.

[0152] D. Sample Analysis [ 14 [C] Compound 1 concentrations in both the donor and receiver sides were analyzed by LSC. The lower limit of quantification (LLOQ) was determined as twice the background (21 dpm).

[0153] Tissue samples were mounted for sectioning in a manner that would allow the tissue to be sectioned in cross section, from the mucosal layer to the serosal layer represented in each section.

[0154] Samples were cryosectioned at 40 μm (for QARL) and 6-8 μm (for MARG) at approximately -20°C and collected onto glass microscope slides by thaw mounting followed by heat fixing on a slide warmer. Approximately three tissue sections were obtained from each sample for QARL. After QARL sectioning, approximately ten sets of three sections / slide were obtained for MARG.

[0155] E.Q.A.R.L. Slides with 40 μm sections were mounted on cardboard backing, covered with plastic wrap, and co-exposed to phosphorus contrast screens with [14C]spiked blood calibration standards (10 concentrations in triplicate ranging from 0.00030 μCi / g to 7.72 μCi / g). The contrast plates, sections, and calibration standards were placed in light-tight exposure cassettes in copper-lined lead storage cabinets for 4 days of exposure at room temperature. The contrast plates were scanned using a Typhoon FLA 9500 image acquisition system (GE Healthcare, Sunnyvale, CA), and the resulting images were stored on a dedicated QPS computer server. Images generated by the [14C]spiked blood calibration standards were used to generate image calibration curves using image analysis software (Microcomputer Imaging Device (MCID Image Analysis System, Interfocus Imaging, Cambridge, Linton, UK)).

[0156] F.MARG All tissue sections were thaw-mounted on subbed glass microscope slides that had been pre-coated with photographic emulsion in the dark and heat-fixed on a slide warmer. Slides were then placed in a black slide box containing desiccant. The slide box was taped with black electrical tape and placed in a lead-lined container at 4°C. Slides were exposed to photographic emulsion for 72 hours, 1 week, 10 days, 2 weeks, 4 weeks, 6 weeks, and 8 weeks. Slides were developed using Kodak D19 Replacement developer and Kodak fixer. Slides were stained with hematoxylin and eosin. Examination and digital photomicrographs of representative results were obtained using a digital camera mounted on an Olympus BX51 microscope. Location of radioactivity is visualized on the slides as small black grains of silver precipitate generated from the emulsion exposed to the radioactive test article. Observations and conclusions are based on evaluation of all samples. Conclusions regarding quantitative tissue concentrations cannot be made using MARG images.

[0157] G. Data Analysis All response curves determined for image analysis calibration were fitted to a weighted first-order polynomial linear equation (1 / MDC / mm 2 ) was used to generate the fit. A numerical estimate of the goodness of fit was given by the relative error, with absolute values ​​for the relative error of each calibration standard being less than or equal to 0.250 as accepted.

[0158] Standard curve calculation: Response (MDC / mm 2 ) = a1 × concentration (density standard in μCi / g) + a0 Where: Density standard = concentration in μCi / g MDC / mm2=Molecular Dynamic Count / area of ​​tissue a1=slope a0=y intercept The relative error for each standard was calculated using the standard curve according to the following:

number

[0159] The LLOQ was determined as 3 times the average background for each panel. Ten target areas were sampled to determine the average value for each panel.

[0160] LLOQ for healthy tissue = 3 x (0.00111) = 0.0033 μCi / g LLOQ for UC tissue = 3 x (0.00106) = 0.0032 μCi / g

[0161] Tissue concentration data were obtained using a profile image analysis sampling technique.

[0162] Profile imaging involved collecting density data at regular intervals (50 μm) across the image of each section using a ribbon-type sampling area provided by using the MCID "Profile" function. Density data was obtained continuously throughout the section and corresponded to the labeled layers of each sample.

[0163] III. Results A. Medication Formulation Analysis The concentrations of radioactivity in the dosing formulations averaged 4.53 and 48.7 nCi / mL (80.7 and 868 nM) for the pre-dose aliquots on the day of dosing. The coefficients of variation for the analysis of triplicate aliquots of the formulations, each analyzed in triplicate, were 1.5% and 0.7%, respectively, indicating that the formulations were homogeneous (Table, below). [Table 8]

[0164] The radiopurity of the formulations pre- and post-dose was greater than 96%.

[0165] B. Transmission research [ in colons after 1-h incubation from healthy and UC subjects 14 C] The permeation results of Compound 1 are listed in the table below (where INCB039110 is Compound 1). [Table 9]

[0166] The medial side of the tissue in subject 2 in group 2 broke while attempting to tightly wrap one end of the polypropylene tube with the tissue, possibly due to loss of extensibility from the UC condition. The permeation results of subject 2 in group 4 were not determined due to leakage during the 1 hour incubation, but this tissue sample was used for QARL and MARG. All [ 14 C] Compound 1 concentrations were below the LLOQ.

[0167] C. Autoradiography Analysis QARL A summary of the individual sample concentration profile data through the sample layers is plotted in FIG. 3 and listed in the table below (in the table, INCB039110 is compound 1). [Table 10]

[0168] The recovered peaks represented the variability of the colonic tissue layers. 14 C] Compound 1 was distributed primarily to the mucosal layer but was detected throughout the submucosa (five of seven tissues).

[0169] MARG No MARG reaction was observed in the first set of slides (72 hour samples), and subsequent slides developed a reaction that reached stationary at 4-8 weeks. The relative concentrations of drug-derived radioactivity were consistent across samples, drug concentrations, and tissue layers between healthy and UC conditions. The highest concentrations were present in the villi and associated crypts across all samples, followed by the submucosa. Little or no radioactivity was observed in the muscle layer. Outside the muscle layer was in background.

[0170] Example 3: IL-6-mediated STAT3 phosphorylation and JAK1 Interleukin-6 (IL-6) signals through the common gp130 receptor and the specific IL-6Rα coreceptor to activate the Janus kinase (JAK) signal transduction and activator of transcription (STAT) signaling pathways (Heinrich et al. The Biochemical journal. 2003;374:1-20). Ulcerative colitis biopsies have identified IL-6 as the predominant cytokine within inflamed areas of the intestine, and its concentrations correlate with Mayo endoscopic scores (see Bernado et al., 2012). Aberrant inflammatory IL-6 / STAT3 pathway activation has been described in peripheral blood mononuclear cells (PBMCs) from rheumatoid arthritis patients (RA) (Isomaki, P et al. Rheumatology, Volume 54, Issue 6, 1 June 2015, 1103-1113), and anti-IL-6 therapy demonstrates remarkable clinical efficacy (Expert Rev Clin Immunol. 2017 Jun; 13(6): 535-551, J Dermatolog Treat. 2018 Sep; 29(6): 569-578). The pathogenesis of plaque psoriasis (Ps) is driven by IL-23-mediated T helper 17 (Th17) / IL-17 inflammation (see). IL-6 plays a key role in promoting STAT3-dependent induction of the IL-23 receptor, which in turn is essential for endowing Th17 cells with full effector function (Zhou et al. Nat. Immunol. 2007;8:967-974, Hirota et al. J. Exp. Med. 2007;204:41-47, Calautti et al. Int J Mol Sci. 2018 Jan;19(1):171). Inhibition of signaling through the JAK / STAT pathway can be measured indirectly in cytokine-promoted cell-based assays. Assessment of phosphorylated STATs is often measured in response to JAK1 stimulation using recombinant human IL-6.

[0171] The systemic effects of Compound 1 have been studied in the autoimmune diseases RA and Ps. Inhibition of STAT3 phosphorylation after stimulation with IL-6, a marker of JAK1 inhibition, and TPO, a marker of JAK2 inhibition, was measured in both studies. In patients with Ps, doses of 100mg QD, 200mg QD, 200mg BID, and 600mg QD were studied. In response to ex vivo IL-6 stimulation, there was Compound 1 concentration-dependent inhibition of pSTAT3. However, in response to TPO, there was no significant inhibition of pSTAT3 at doses of 100mg QD, 200mg QD, and 200mg BID (Figure 4). There was also a dose-dependent response in the primary efficacy measure mean change from baseline sPGA at day 28 (see table below). [Table 11]

[0172] The 200 mg BID (p=0.053) and 600 mg QD (p=0.003) doses demonstrated clinically meaningful changes from baseline, whereas the 100 mg or 200 mg QD doses did not and were not statistically different from placebo (p=0.270, p=0.118, respectively). There is good correlation between pharmacokinetic markers of ex vivo IL-6-stimulated STAT3 inhibition and efficacy endpoints (Figure 5). No neutropenia was observed, consistent with the observation that no significant inhibition of JAK2 (as determined by TPO-stimulated pSTAT3 inhibition) was observed at doses of 100 mg QD, 200 mg QD, and 200 mg BID, and neutropenia and other cytopenias are believed to be a specific consequence of JAK2 inhibition inducing myelosuppression (Bissonnette R et al J Dermatolog Treat,2016 27(4)332-338; Mascarenhas et al.Haematolgica 2017 102(2):327-335.).

[0173] In RA patients, doses of 100 mg QD and BID, 200 mg BID, 300 mg QD, 400 mg BID, and 600 mg QD were studied, and again a general trend of dose-dependent inhibition of IL-6-induced pSTAT3 was observed (Figures 6A and 6B). A general trend of increasing TPO-induced pSTAT3 inhibition was also observed. However, the greatest inhibition appeared to be observed after 200 mg BID dosing. Also of note is that the 100 mg QD dose had less TPO-induced pSTAT3 inhibition than placebo. In this study, there were some cases of decreased ANC, but no dose-dependent trends were observed. With regard to efficacy, no dose-dependent trends were evident across the dose range, except for a statistically significant difference in ACR20, ACR50, and ACR70 responses between Compound 1 and placebo for the 600 mg QD treatment group at the 84th visit (primary endpoint visit).

[0174] Taken together, clinical data from RA and Ps patients suggest that a 100 mg QD dose has minimal systemic effects based on the pooled safety, efficacy, and biomarker data. Daily exposure following 50 mg BID is expected to be less than 100 mg QD given that Compound 1 exhibits supra-linear PK with respect to dose.

[0175] Example 4: A Phase 2, Double-Blind, Dose-Ranging, Placebo-Controlled Study with Open-Label Extension to Evaluate the Safety and Efficacy of Compound 1 in Moderate to Severe Ulcerative Colitis I. Purpose This study will evaluate the safety and efficacy of oral Compound 1 in participants with moderate to severe active UC. Compound 1 will be administered in an SR formulation. Oral bioavailability of Compound 1 in humans is moderate, with approximately 30% of an administered dose being excreted intact as the parent compound in the stool. Inhibition of IL-6-stimulated phosphorylation of STAT3 is a measure of JAK1 inhibition. A dose of 50 mg BID Compound 1 demonstrated an in vitro IC50-fold increase in inhibition of IL-6-stimulated phosphorylation of STAT3 in PBMCs (58 nM). 50This is expected to result in fecal concentrations (approximately 200 nM) that exceed the ex vivo whole blood IC value. However, the corresponding plasma concentrations associated with this dose are low and 50 C well below the value of 141 nM max value (51 nM). As a result, the efficacy of Compound 1 is expected to be mediated primarily through local, rather than systemic, inhibition of JAK1.

[0176] As a selective and locally acting JAK1 inhibitor, Compound 1 may possess the anti-inflammatory properties seen with other JAK inhibitors without the associated risks of anemia or neutropenia. Given the favorable safety profile of Compound 1 in selected dose ranges, concurrent use of immunosuppressive UC therapies (AZA, 6-MP, and methotrexate) would be tolerated.

[0177] II. Overall design Approximately 206 participants will be enrolled overall for 12 weeks in Part A (n=30) and Part B (n=176). Both Parts A and B are randomized, double-blind, placebo-controlled, and parallel design.

[0178] In Part A, 30 participants will be randomly assigned to receive 50 mg BID or placebo in a 2:1 distribution ratio. Part A participants will complete an overnight in-clinic visit at week 4. This visit will obtain a 24-hour stool sample for stool drug concentration analysis and serial blood samples for PK analysis of plasma drug concentrations. In addition to undergoing endoscopy at baseline and week 12, Part A participants (only) will undergo endoscopy at week 4. Part A is intended to establish proof of mechanism at 50 mg BID in a 2:1 ratio, and Part B is intended to evaluate the clinical efficacy of a range of doses of 25 to 100 mg total daily dose given either QD or BID. The dose regimen used in Part B will be selected after Part A. Participants who complete either Part A or Part B and all associated study procedures, including endoscopy at week 12, will be eligible to enter the corresponding 40-week OLE period of the study.

[0179] In Part B, 176 participants will be randomized in a 1:1:1:1 ratio to one of three dose levels of Compound 1 tablets or placebo. In addition to placebo, doses included in Part B are 25 mg BID, 50 mg BID, and 100 mg QD. The dose regimen for Part B will be confirmed at the conclusion of Part A (within the total daily dose range of 25-100 mg administered QD or BID). Part B participants will undergo endoscopy at baseline and week 12. In addition, a total of 24 Part B participants (6 from each treatment group) will complete an overnight in-clinic visit at week 4. This visit will obtain 24-hour stool collections for stool drug concentrations and serial blood samples for PK analysis.

[0180] Background stable therapy for UC in both Part A and Part B must not be changed during the screening and double-blind treatment periods until the Week 12 evaluation is completed.

[0181] Participants requiring initiation of a new therapy for UC during this period should undergo endoscopy and discontinue the study with appropriate standard of care treatment given at the investigator's discretion. After endoscopy at week 12, the daily corticosteroid dose may be increased or decreased at the investigator's discretion. In addition to the analyses of week 12 data from Parts A and B, there are three additional interim analyses planned for the study: 1. The first interim analysis will be performed when the 15 participants randomized to Part A have week 4 data available. The unblinded PK / PD team will assess systemic exposure and perform exploratory biomarker analysis to confirm whether Compound 1 has an effect on the JAK / STAT signaling pathway. 2. A second interim analysis will be performed when these 15 participants reach week 12. In addition to considering the PK / PD results, the study may be terminated if there is insufficient evidence of efficacy demonstrated in this section. 3. The third interim analysis will be performed after 88 participants randomized to Part B have week 12 data available. The study may be terminated if there is insufficient evidence of efficacy.

[0182] At the conclusion of Part A, the SRC (composed of members of the sponsor's research team) will conduct a final analysis of Part A to review all safety and PD data on an unblinded basis to determine whether to proceed to Part B or terminate the study. The choice of dose regimen for Part B will be informed by this analysis of the data. The Part B dose regimen will be either QD or BID with a total daily dose of 25 to 100 mg. In addition, the dose regimens in the OLE period of Parts A and B may be modified by the sponsor's research team based on the Part A results). The dose in the Part A OLE is 50 mg BID. The dose in the Part B OLE period may be modified later within the same dose range (total daily dose of 25 mg to 100 mg).

[0183] When 88 participants complete Week 12 of Part B, the DMC may recommend continuing the study (details regarding the results of the current safety analysis will not be available before the next scheduled analysis) or may recommend stopping the study (based on lack of efficacy or any safety findings). The DMC may also recommend regarding an OLE dose modification for Part B.

[0184] The final analysis of the double-blind period will be conducted when all Part B participants have completed Week 12.

[0185] The final study analysis will occur after all participants have completed the OLE period of the study, including the 30-day follow-up period. III. Research treatment [Table 12]

[0186] IV. Effectiveness Evaluation Definitions of efficacy endpoints based on the Mayo score are defined below and will be used throughout the protocol. A. List of efficacy endpoint definitions based on Mayo score [Table 13]

[0187] B. Endoscopy Endoscopy (preferably colonoscopy) is required at baseline and week 12. In addition, endoscopy (colonoscopy or flexible sigmoidoscopy at the discretion of the institution) is required at week 4 for all Part A participants only. This procedure will be performed to establish a three-component Mayo score, including mMES, where any bleeding tendency results in a score of at least 2 (Food and Drug Administration. Guidance for Industry: Ulcerative Colitis: Clinical Trial Endpoints. 2016. https: / / www.fda.gov / downloads / Drugs / GuidanceComplianceRegulatoryInformation / Guidances / UCM515143.pdf). The duration of time between endoscopy and scheduled visits should not exceed 14 days. Endoscopy also allows pinch biopsies to evaluate PD effects in mucosal tissue.

[0188] A trained endoscopist should perform the endoscopies. If possible, the same endoscopist should perform the endoscopies at all visits. All results will be read and adjudicated centrally as described in the study manual.

[0189] Histologic evaluation of biopsy specimens obtained during endoscopy may also be reviewed by a trained pathologist, as described in a separate protocol.

[0190] C. Inflammatory Bowel Disease Questionnaire (IBDQ) The IBDQ is a psychometrically validated patient-reported outcome instrument to measure disease-specific quality of life in participants with inflammatory bowel disease, including UC. The IBDQ contains 32 items, grouped into four dimensions, and scored as follows: ·Defecation symptoms: 10~70. ·Systemic symptoms: 5~35. Emotional function: 12-84 · Social functioning: 5-35.

[0191] Total IBDQ scores range from 32 to 224. For the total score and for each domain, higher scores indicate better quality of life. A score of at least 170 corresponds to clinical remission, and an increase of at least 16 points is considered to indicate a clinically meaningful improvement.

[0192] The IBDQ will be assessed at baseline and at each designated study visit.

[0193] D.3 Component Mayo Score The three-component Mayo score will be used to measure UC disease activity in this study. The three-component Mayo score (Mayo score without PGA ranges from 0 to 9 points) consists of three subscores, each rated from 0 to 3, with higher scores indicating more severe disease: Stool frequency (0-3) ·Rectal bleeding (0~3) mMES(0-3)

[0194] A three-component Mayo score will be determined based on incorporating endoscopic results as assessed by a central reviewer at baseline and each designated study visit. In the event that central endoscopic results are missing, endoscopic subscores as determined by the investigator will be used in the calculation.

[0195] The three-component Mayo score will be calculated using stool frequency and rectal bleeding data from the most recent 3 days of available data prior to the visit. Data collected from the following periods will not be included in this calculation: - Days when medication for constipation or diarrhea was administered On days of procedures or preparation for procedures that may affect stool frequency or blood content (e.g., enemas, other laxatives, clear liquid diets). - 48 hours after use of antimotility drugs (e.g. loperamide). - 48 hours after endoscopy.

[0196] E. Doctor's Overall Evaluation The PGA will be calculated separately from the three-component Mayo score. The PGA recognizes three criteria: Participants' daily recall of abdominal discomfort, and The general health of the participant, and Other observations of the Participant, such as a physical examination and the Participant's performance status. The PGA Standards will be scored as follows: ·0=normal 1 = mild illness 2 = moderate disease 3 = severe illness PGA will be assessed at baseline and at each designated study visit.

[0197] V. Pharmacokinetic Evaluation A. Blood and Stool Sample Collection At the PK visits (Weeks 2 and 12), participants must abstain from study drug prior to arriving at the study site. A pre-dose PK sample should be collected. Following collection of the pre-dose PK sample, Compound 1 will be administered and subsequent regular samples will be collected from the participant. The dates and times of blood collection for PK analysis, the last dose of study drug, and the last two meals prior to blood draw (e.g., dinner the night before and breakfast that morning) will be recorded.

[0198] At week 4, all participants in Part A and a subset of Part B participants (n=approximately 24) will complete an overnight inpatient clinic visit. At this visit, participants will collect a 24-hour stool sample to determine Compound 1 stool concentrations in the stool and serial blood samples will be obtained for analysis of plasma drug concentrations (see, e.g., Example C). An endoscopy (colonoscopy or flexible sigmoidoscopy) will be performed before participants are discharged from the CRU and evaluated by a central reviewer. [Table 14]

[0199] VI. Objectives and Endpoints [Table 15]

[0200] Example 5. Preclinical mouse model of spontaneous colitis The interleukin-10 (IL-10) knockout (KO) mouse model is the IL-10 KO mouse, BALB / cAnNTac-Il10 em7Tac Colitis in IL-10 KO mice spontaneously develops colitis, reflecting the multifactorial nature of inflammatory bowel diseases (IBD), such as ulcerative colitis and Crohn's disease. Colitis in IL-10 KO mice is mediated by CD4 + It results from an aberrant response of T helper 1-like T cells and excessive secretion of pro-inflammatory cytokines that signal through the Janus kinase / signal transducer and activator of transcription (JAK / STAT) pathway. Compound 1 is a potent JAK1 inhibitor with 22- to >500-fold selectivity over JAK2, JAK3, and TYK2 and is currently being investigated as monotherapy in clinical trials for moderate to severe ulcerative colitis.

[0201] Female IL-10 homozygous knockout mice on BALB / c strain background were provided by Taconic (USA). From 6 weeks of age, compound 1 and vehicle (10 mL / kg) were administered by oral gavage twice a day. Diarrhea was quantified on a 0-3 rating scale (0=normal, 1=soft but still formed, 2=very soft, 3=diarrhea). Mice were euthanized by CO2 asphyxiation and colon length and weight were measured. Histopathology was scored on a 0-10 scale based on the following criteria: Lymphocytes infiltrate the mucosa and gut-associated lymphoid tissue located in the lamina propria / submucosa, mucosal erosions / ulcers, and transmural inflammation. Body weight, stool consistency, fecal occult blood, and rectal bleeding were scored. The incidence of rectal prolapse was recorded.

[0202] Significant improvements were observed in terms of total disease burden, as shown in FIG. 7A, and the development of rectal prolapse as a marker of severe disease, as shown in FIG. 7B. Ex vivo, colonic tissues of mice treated with Compound 1 were characterized by reduced histopathology, as shown in FIG. 7C-7D. Oral twice daily administration of Compound 1 at 30 mg / kg significantly (p<0.001) delayed the onset of colitis and modulated disease-associated weight loss. Cumulative clinical disease scores were significantly (p<0.0001) reduced in animals treated with Compound 1 compared to vehicle controls. The incidence of rectal prolapse was also significantly (p<0.01) lower. Compound 1 administration resulted in a significant (p<0.01) reduction in colonic structural pathology. Lymphocyte infiltration and transmural inflammation were also significantly (p<0.01) reduced in mice treated with Compound 1 versus vehicle controls. As shown in Figure 10, Compound 1 was also found to ameliorate spontaneous colitis in an IL-10 KO mouse model, as evidenced by significantly slower disease onset, and Compound 1 treatment also resulted in differential gene expression profiles in the colon of IL-10 KO mice compared to vehicle controls, as shown in Figures 12A-12B. As shown in Figures 15A-15B, systemic Compound 1 delivery was associated with a significant protective effect on colon morphology in IL-10 KO mice.

[0203] These data suggest that Compound 1 may be useful as a therapeutic agent for the treatment of IBD (eg, spontaneous colitis).

[0204] Example 6. Experimentally induced inflammatory bowel disease in a mouse model Inflammatory bowel diseases (IBD), such as ulcerative colitis and Crohn's disease, are a group of idiopathic chronic and relapsing inflammatory conditions that arise from a complex interaction between the immune system and tissues of the gastrointestinal tract. Multiple cytokines and growth factors in the pathogenesis of IBD signal through the Janus kinase / signal transduction and activator of transcription pathway.

[0205] A preclinical model of IBD was established in BALB / c mice by intracolonic injection of 2,4,6-trinitrobenzenesulfonic acid (TNBS) or 4-ethoxymethylene-2-phenyl-2-oxazolin-5-one (oxazolone) to induce an immune response, as described below. Body weight, stool consistency, and stool blood were scored. Additional readouts included colon weight-to-length ratio and histological evaluation. Blood was collected for pharmacokinetic analysis.

[0206] Mouse oxazolone-induced colitis model Male BALB / c mice were purchased commercially (Charles River Laboratories). On day 0, mice were sensitized by applying oxazolone (150 μL, 3% in acetone / olive oil, 4:1 v / v) to their pre-shaved rostral dorsal area. Animals were resensitized with oxazolone on day 5. Mice were fasted before intrarectal oxazolone challenge. Distal colitis was induced by intracolonic instillation of oxazolone solution (1 mg in 0.1 mL of 40% ethanol), after which animals were maintained in a vertical position for 30 seconds to ensure that the solution remained in the colon. Sham-control mice received 0.1 mL of 40% ethanol alone. Compound 1 and vehicle (10 mL / kg) were administered by oral gavage twice daily. Diarrhea was quantified on a 0-3 rating scale (0=normal, 1=soft but still forming, 2=very soft, 3=diarrhea). Fecal occult blood was detected using SY occult blood paper (Shih-Yung Medical Instruments, Taiwan) on a 0-3 scale (0=negative, 1=positive, 2=visible bloodstain, 3=rectal bleeding). On day 8, mice were euthanized by CO2 asphyxiation and colon length and weight were measured. Furthermore, when the abdominal cavity was opened, adhesion between the colon and other organs was noted as the presence of colonic ulcers after removal and weighing of each colon. Macroscopic scoring was performed on a 0-12 scale as shown in Table A. Normalized colon weight represents the increase in tissue relative to sham control mice. [Table 16]

[0207] Intrarectal administration of oxazolone in an ethanol vehicle induces direct tissue damage and induces an immune response that leads to mucosal inflammation, epithelial microulceration, and histopathological changes in the distal colon reminiscent of human ulcerative colitis (see, e.g., Kojima et al, J. Pharmacol. Sci. 2004, 96(3):307-313). The latter inflammatory phase is driven by the production of Th2 cytokines, such as IL-4, IL-5, and IL-13 secretion (see, e.g., Randhawa et al, J. Physiol. Pharmacol. 2014, 18(4):279-288).

[0208] Daily Compound 1 treatment (30 mg / kg BID) was effective in accelerating recovery from diarrhea and rectal bleeding as shown in Figure 8A, improving gross histopathology as shown in Figure 8B, and reducing normalized colon weight as a surrogate readout of inflammatory swelling as shown in Figure 8C. These data are consistent with published results demonstrating that tofacitinib inhibits oxazolone-induced colitis (see, e.g., Beattie et al, J. Inflamm. (Lond). 2017, 44:28), and suggest that a significant proportion of the anti-inflammatory efficacy is driven by JAK1 inhibition. In addition, twice-daily Compound 1 treatment (oral or intracolonic) significantly improved stool consistency and reduced fecal occult blood scores compared to vehicle-treated controls (see Figures 13A-13D), and Compound 1 treatment significantly improved colon shortening compared to the respective vehicle-treated controls (see Figures 14A-14E).

[0209] TNBS-induced colitis model Male BALB / c mice were purchased (Charles River Laboratories) and distal colitis was induced by intracolonic instillation of TNBS (2,4,6-trinitrobenzenesulfonic acid solution, 1 mg in 0.1 mL of 50% ethanol). Compound 1 treatment was administered at 30 mg / kg by oral gavage (PO) or 3 mg / kg by intracolonic injection (IC) twice daily (BID). Diarrhea was quantified on a 0-3 rating scale (0=normal, 1=soft but still forming, 2=very soft, 3=diarrhea) on days 3-5 after TNBS sensitization.

[0210] As shown in Figure 9A, oral Compound 1 treatment significantly reduced diarrhea compared to vehicle-treated animals. This data is consistent with the oxazolone-induced model data shown in Figure 8A. Low-dose Compound 1 treatment delivered directly to the colon was also highly effective in enhancing disease recovery, as shown in Figure 9B. For example, in the oxazolone model, Compound 1 PO BID at 30 mg / kg showed a significant (p<0.05) reduction in colon shortening (see Figures 14A-14B and 14E) and weight gain. Compound 1 IC BID at 3 mg / kg also significantly (p<0.05) reduced colon shortening (see Figures 14C-14E).

[0211] Oral dosing (PO) of intracolonic doses of Compound 1 at 30 mg / kg or 3 mg / kg twice daily significantly (p<0.05) improved stool consistency compared to controls. In addition, a significant (p<0.05) reduction in stool blood score was achieved at 3 mg / kg IC BID. Furthermore, both routes of administration (oral, IC) resulted in significant (p<0.05) improvements in stool consistency and stool blood score. Compound 1 IC BID at 3 mg / kg improved total colonic gross damage. Intracolonic dosing of Compound 1 significantly improved JAK1 IC 50 While maintaining a systemic drug exposure of less than 100 mg / kg, the study achieved comparable inhibition of experimental IBD. Taken together, these data suggest that compound 1 may be useful as a therapeutic agent for the treatment of IBD.

[0212] Intrarectal administration of haptenizing agents (TNBS) renders colonic proteins immunogenic to the host immune system, thereby initiating a T helper (Th)1-mediated immune response characterized by infiltration of the lamina propria with CD4+ T cells, neutrophils, and macrophages. Compound 1 was administered orally at 30 mg / kg or directly to the colon at 3 mg / kg to determine whether localized JAK1 inhibition is effective. Consistent with the oxazolone model, oral compound 1 accelerated disease score recovery compared to vehicle-treated animals, as shown in Figure 11A. Low-dose compound 1 administered directly to the colon appeared to induce recovery more rapidly and mediate a greater therapeutic response, as shown in Figure 11B.

[0213] In further studies, quantification of circulating and tissue drug concentrations clearly distinguished local versus systemic JAK1 target inhibition. Oral dosing resulted in peak circulating drug levels of approximately 11 μM, which was similar to colonic concentrations, as shown in FIG. 11C. In contrast, localized Compound 1 delivery was characterized by a minimal peak systemic concentration of approximately 0.04 μM, but sustained exposure of 0.45 μM or greater in colonic tissue, as shown in FIG. 11D. Thus, strategic targeting or release of JAK1 inhibitors in inflamed gastrointestinal tissues could potentially achieve an improved benefit-risk profile.

[0214] Low doses of Compound 1 administered directly to the site of intestinal inflammation were highly effective in TNBS-induced colitis, and this therapeutic response was independent of systemic JAK1 inhibition, as Compound 1 plasma concentrations were minimal. This data strongly supports the rationale that localized JAK inhibition may be sufficient to achieve a therapeutic response, thereby avoiding the need for systemic immunosuppression. Without wishing to be bound by theory, these data also suggest that JAK1 is the predominant mechanism by which pathogens are activated.

[0215] 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, including all patents, patent applications, and publications cited in this application, is hereby incorporated by reference in its entirety. The present application also includes the following aspects. [Aspect 1] A method for treating a gastrointestinal disease or disorder in a subject, comprising administering to the subject a JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, wherein a maximum stool concentration of the JAK1 pathway inhibitor after administration of the JAK1 pathway inhibitor is greater than about 25 nM and a maximum total plasma concentration of the JAK1 pathway inhibitor after administration of the JAK1 pathway inhibitor is less than about 150 nM. [Aspect 2] The method of embodiment 1, wherein the JAK1 pathway inhibitor is selective for JAK1 over JAK2, JAK3, and Tyk2. [Aspect 3] The method of embodiment 1 or 2, wherein said gastrointestinal disease or disorder is ulcerative colitis (UC). [Aspect 4] The method of embodiment 1 or 2, wherein the gastrointestinal disease or disorder is Crohn's disease. [Aspect 5] 3. The method of claim 1 or 2, wherein the gastrointestinal disease or disorder is celiac disease. [Aspect 6] The method of embodiment 1 or 2, wherein said gastrointestinal disease or disorder is spontaneous colitis. [Aspect 7] The method of embodiment 1, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 50 mg to about 100 mg. [Aspect 8] The method of embodiment 1, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 25 mg to about 75 mg. [Aspect 9] The method of embodiment 1, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 25 mg. [Aspect 10] The method of embodiment 1, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 50 mg. [Aspect 11] The method of embodiment 1, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in a daily dose of about 100 mg. [Aspect 12] The method of embodiment 1, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered once daily at a dosage of about 25 mg. [Aspect 13] The method of embodiment 1, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered at a dose of about 25 mg twice daily, for a total daily dose of about 50 mg. [Aspect 14] The method of embodiment 1, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered once daily at a dosage of about 50 mg. [Aspect 15] The method of embodiment 1, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered at a dose of about 50 mg twice daily, for a total daily dose of about 100 mg. [Aspect 16] The method of embodiment 1, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered once daily at a dosage of about 100 mg. [Aspect 17] The method of any one of aspects 1-16, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered as one or more sustained release dosage forms, each comprising the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof. [Aspect 18] 18. The method of any one of aspects 1-17, wherein the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof. [Aspect 19] 18. The method of any one of aspects 1-17, wherein the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate. [Aspect 20] The method of any one of aspects 1-19, wherein the maximum stool concentration of the JAK1 pathway inhibitor is greater than about 50 nM following administration of the JAK1 pathway inhibitor. [Aspect 21] 21. The method of any one of aspects 1-20, wherein the maximum total plasma concentration of the JAK1 pathway inhibitor is less than about 150 nM following administration of the JAK1 pathway inhibitor. [Aspect 22] 22. The method of any one of aspects 1-21, wherein the maximum total plasma concentration of the JAK1 pathway inhibitor is less than about 141 nM following administration of the JAK1 pathway inhibitor. [Aspect 23] A method for treating a gastrointestinal disorder in a subject, comprising administering to the subject a daily dose of about 25 mg to 100 mg of a JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is administered in one or more sustained release dosage forms comprising the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof. [Aspect 24] 1. A method for treating a gastrointestinal disease selected from the group consisting of ulcerative colitis, Crohn's disease and celiac disease in a subject in need thereof, comprising administering to the subject {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof; a maximum stool concentration 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 after administration of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof, is greater than about 25 nM; The method, wherein the maximum total plasma concentration of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof, after administration is less than about 150 nM. [Aspect 25] 1. A method for treating a gastrointestinal disorder selected from the group consisting of ulcerative colitis, Crohn's disease, and celiac disease in a subject, comprising administering to a subject from about 25 mg to about 100 mg of {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, on a free base basis; administering to the subject a once-daily dose of a pharmaceutically acceptable salt thereof, wherein the dose comprises one or more sustained release dosage forms each comprising {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-[7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharmaceutically acceptable salt thereof.

Claims

1. A medicament for treating ulcerative colitis in a subject, the medicament comprising {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof; a maximum fecal concentration of the {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, of greater than 25 nM; the maximum total plasma concentration of the {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, is less than 150 nM; The said medicine.

2. 2. The medicament of claim 1, wherein the {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, is administered in a daily dose of 50 mg to 100 mg.

3. 2. The medicament of claim 1, wherein the {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, is administered in a daily dose of 25 mg to 75 mg.

4. 2. The medicament of claim 1, wherein the {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, is administered in a daily dose of 25 mg.

5. 2. The medicament of claim 1, wherein the {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, is administered in a daily dose of 50 mg.

6. 2. The medicament of claim 1, wherein the {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, is administered in a daily dose of 100 mg.

7. The pharmaceutical composition of claim 1, wherein the {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, is administered at a dose of 25 mg once a day.

8. 2. The medicament of claim 1, wherein the {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, is administered at a dose of 25 mg twice daily for a total daily dose of 50 mg.

9. The pharmaceutical composition of claim 1, wherein the {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, is administered at a dose of 50 mg once a day.

10. 2. The medicament of claim 1, wherein the {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, is administered at a dose of 50 mg twice daily for a total daily dose of 100 mg.

11. The pharmaceutical composition of claim 1, wherein the {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, is administered at a dose of 100 mg once a day.

12. The pharmaceutical composition according to any one of claims 1 to 11, wherein the {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, is administered as one or more sustained release dosage forms, each comprising the {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.

13. The pharmaceutical agent according to any one of claims 1 to 12, wherein the {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 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 adipic acid salt.

14. The pharmaceutical agent according to any one of claims 1 to 13, wherein the maximum fecal concentration of the {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 is greater than 50 nM after administration of the {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.

15. The pharmaceutical agent according to any one of claims 1 to 14, wherein the maximum total plasma concentration of the {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 is less than 141 nM after administration of the {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.

16. A medicament for treating ulcerative colitis in a subject, the medicament comprising {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof; {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, is administered in a once daily dose of 25 mg to 100 mg on a free base basis; the doses comprise one or more sustained release dosage forms each comprising {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-[7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharma- ceutically acceptable salt thereof; The said medicine.

17. The medicine according to claim 16, wherein the {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 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 adipic acid salt.

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