JP7518900B2 - Use of JAK1 inhibitors for the treatment of cutaneous lupus erythematosus and lichen planus (LP) - Patent Application 20070233334 - Google Patents

Description

The present application provides methods for the treatment of cutaneous lupus erythematosus and/or lichen planus (LP) using compounds that modulate the activity of Janus kinase (JAK) 1.

Cutaneous lupus erythematosus (CLE) is an inflammatory autoimmune skin disease with heterogeneous subtypes that range from focal discoid plaques to severe widespread erythrosquamous lesions in affected patients (see, e.g., Kuhn & Landmann, J. Autoimmun. 2014, 48-49:14-19). The disease is characterized by a typical histological pattern called "interface dermatitis" (ID), consisting of colloid bodies and anti-epithelial cytotoxic lymphocytic infiltration at the epidermal-dermal junction, orchestrated by IFN-regulated inflammatory cytokines (see, e.g., Wenzel & Tuting, J. Invest. Dermatol. 2008, 128:2392-2402, and Wenzel et al, Br. J. Dermatol. 2007, 157:752-757).

CLE is multifactorial in origin and includes both genetic and environmental risk factors (see, e.g., Kunz et al, Exp. Dermatol. 2015, 24:510-515; Sinha & Dey-Rao, J. Investig. Dermatol. Symp. Proc., 2017, 18:S75-S80; and Szczech et al, Lupus, 2017, 26:791-807). UV radiation in particular promotes cell damage, apoptosis, and release of DNA-containing blebs. Clearance of cellular debris is thought to be impaired, with secondary necrosis (see, e.g., Kuhn et al, Arthritis Rheum. 2006, 54:939-950; Caricchio et al, The Journal of Immunology 2003, 171:5778-5786; and Mahajan et al, Front Immunol. 2016, 7). In some cases, such as familial pernio lupus, TREX1 gene mutations may be the underlying cause, leading to TREX1 exonuclease dysfunction and high accumulation of intracytoplasmic DNA (see, e.g., Gunther et al, J. Am. Acad. Dermatol. 2013, 69:e179-81; Grieves et al, Proc. Natl. Acad. Sci. U.S.A. 2015, 112:5117-5122; Gunther et al, Dermatology (Basel), 2009, 219:162-166; and Peschke et al, J. Invest. Dermatol. 2014, 134:1456-1459).

When nuclear components, such as endogenous nucleic acid motifs, are released outside the nucleus due to cell damage, they can be recognized as damage-associated molecular patterns (DAMPs) (see, e.g., Scholtissek et al, J. Invest. Dermatol. 2017, 137:1484-1492). In CLE, there is evidence that keratinocytes, particularly plasmacytoid dendritic cells (pDCs), respond inappropriately to DAMPs, with massive type I IFN production via TLR-dependent or TLR-independent pathways (e.g., Mustelin et al, Front. Immunol. 2019, 10:238; Wollenberg et al, J. Invest. Dermatol. 2002, 119:1096-1102; Wenzel et al, Arch. Dermatol. Res. 2009, 301:83-86; Yu et al, J. Autoimmun. 2013, 41:34-45; and Katayama et al, J. Immunol. 2013, 41:34-45). al, J. Invest. Dermatol. DOI:10.1016/j.jid. 2019.02.035). In an autocrine loop, IFN binds to the IFN-α/β receptor on keratinocytes, which activates the JAK/STAT pathway and expression of inflammatory mediators such as CXCL10, which is known to recruit CXCR3+ effector cells and pDCs to skin lesions (see, e.g., Wenzel et al, Arch. Dermatol. Res. 2009, 301:83-86, and Wenzel et al, J. Pathol. 2005, 205:435-442). These effector cells induce keratinocyte necroptosis (see, e.g., Lauffer et al, J. Invest. Dermatol. 2018, 138:1785-1794), cytokine release, and persistent "self-recruitment" as hallmarks of chronic inflammation (see, e.g., Meller et al, Arthritis Rheum. 2005, 52:1504-1516).

This cycle can have a strong impact on quality of life (see, e.g., Samotij et al, Postepy Dermatol. Aergol. 2018, 35:192-198, and Ogunsanya et al, Int. J. Womens Dermatol. 2018, 4:152-158). According to current guidelines, corticosteroids and antimalarials are established as first-line treatment for CLE. However, corticosteroids are limited by side effects and long-term treatment can be difficult, especially in patients with antimalarial resistance. Furthermore, there are no drugs approved specifically for CLE and only a few clinical trials have been conducted, especially due to clinical heterogeneity and the resulting difficulties in trial design (see, e.g., Chen et al, F1000Res. 2019, 8).

The American College of Rheumatology (ACR) has established a scheme of 11 clinical and laboratory criteria aimed at distinguishing systemic lupus erythematosus (SLE) from other autoimmune diseases. The ACR guidelines require the fulfillment of 4 of the 11 criteria for the diagnosis of SLE, but only 4 of the criteria (macular rash, discoid lesions, mucosal ulcers, and photosensitivity) are actually cutaneous, so the presence of cutaneous pathology is not essential for the diagnosis of SLE. Conversely, cutaneous lupus erythematosus (CLE) subtypes can occur in the absence of systemic disease (SLE), and CLE is 2-3 times more frequent than SLE (see, e.g., Tebbe & Orfanos, Lupus, 1997, 6(2):96-104).

Given these limitations, there is a medical need for new treatment options. This application addresses this and other needs.

The present application provides a method of treating a disease selected from cutaneous lupus erythematosus (CLE) and lichen planus (LP) in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a JAK1 selective inhibitor provided herein.

Figure 1 shows the functional role of JAK1 expression in skin lesions of CLE. Representative immunohistochemical sections (anti-phospho-JAK1 staining) from skin samples of patients with different inflammatory skin disorders (subacute cutaneous lupus erythematosus (SCLE, n=5), chronic discoid lupus erythematosus (CDLE, n=5), lichen planus (LP, n=6) and healthy controls (HC, n=5)) are shown (original magnification: ×200). 1 shows the functional role of JAK1 expression in skin lesions of CLE. From non-expression
Expression of pJAK1 is shown, scored semiquantitatively by scoring to strong expression (see, e.g., Wenzel et al. J. Pathol. 2005, 205:435-442). All bars represent mean ± SEM.
(Kruskal-Wallis test) is shown.
Figure 1 shows the functional role of JAK1 expression in CLE skin lesions.Up-regulation of expression of IFN-related genes in CLE lesional skin compared to healthy controls (HC) (2-fold, p<0.01;Welch's t-test). Figure 1 shows the functional role of JAK1 expression in CLE skin lesions. Figure 2 shows KEGG pathways that are upregulated in CLE. Classification of KEGG pathways was performed using Database for Annotation, Visualization and Integrated Discovery (DAVID ver. 6.8). P-values were generated with EASE Score. Count: number of genes that are upregulated in CLE by more than 2-fold within each KEGG pathway. Figure 1 shows the effect of pharmacological JAK1 inhibition on inflammatory cytokine expression and IFN-related gene regulation in cultured keratinocytes. Area intensity mean of anti-pJAK1 antibody staining in pixels is shown for stimulated HaCaT (eNA), stimulated and inhibitor-treated HaCaT (eNA + JAK1 inhibitor {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 (i.e., INCB039110); eNA + JAK1/2, inhibitor: ruxolitinib), and unstimulated HaCaT (C-medium). Measurements were performed using Fiji software. n=5 measurements per entity. All scatter plots are The results are shown (Mann-Whitney U test).
Figure 1 shows the effect of pharmacological JAK1 inhibition on inflammatory cytokine expression and IFN-related gene regulation in cultured keratinocytes. Representative immunofluorescence micrographs of HaCaT cells stimulated with eNA and either untreated (eNA) or inhibitor-treated (eNA+JAK1, inhibitor: INCB039110; eNA+JAK1/2, inhibitor: ruxolitinib) and unstimulated HaCaT cells (medium control) are shown. Anti-pJAK1 antibody staining is red, nuclear DAPI staining is blue, original magnification: 400x. Figure 1 shows the effect of pharmacological JAK1 inhibition on inflammatory cytokine expression and IFN-related gene regulation in cultured keratinocytes. Expression of genes typical of CLE in eNA-stimulated NHEKs (+, 4 samples) and expression downregulated by more than 2-fold (p<0.05;Welch's t-test) following treatment of stimulated NHEKs with JAK1-selective INCB039110 (JAK1; 4 samples). Figure 1 shows the effect of pharmacological JAK1 inhibition on inflammatory cytokine expression and IFN-related gene regulation in cultured keratinocytes. Figure 2 shows the effect of different JAK inhibitors on CXCL10 expression in stimulated HaCaT after 24 hours of incubation. Cells were treated with JAK1-selective INCB039110 (JAK1), JAK1/2-selective ruxolitinib (JAK1/2) and JAK3-selective FM-381 (JAK3). Measured by ELISA. All bars represent (Mann-Whitney U test) is shown.
Figure 1 shows the effect of pharmacological JAK1 inhibition on inflammatory cytokine expression and IFN-related gene regulation in cultured keratinocytes. Photomicrographs (original magnification: 400x) depicting CXCL10 expression in stimulated cells (eNA, n=3) and cells treated with a JAK1 inhibitor (INCB039110) after stimulation (eNA+JAK1, n=3) and the corresponding CXCL10 expression scores are shown. All bars represent the mean ± SEM. (Mann-Whitney U test) is shown.
Pharmacological JAK1 inhibition in vivo in a lupus-prone TREX1 ^−/− mouse model. Clinical findings of TREX1 ^−/− mice (n=8) subjected to UV stimulation (starting on day −3; 3×450 mJ/ ^cm2 for 115 sec) are shown at baseline (day 0, start of treatment) and 4 and 7 days after treatment with JAK1-selective INCB039110 or placebo. Figure 1 shows pharmacological JAK1 inhibition in vivo in a lupus-prone TREX1 ^-/- mouse model. Mouse-adapted modified CLE disease area and severity index (mouse RCLASI) mean scores ± SEM (n=4 mice per group). Figure 1 shows pharmacological JAK1 inhibition in vivo in a lupus-prone TREX1 ^-/- mouse model. Histological (H&E) and immunohistological (CD45) photomicrographs of lesional skin (original magnification: 400x) are shown. Corresponding CD45 expression inflammatory scores in lesional skin of placebo- and JAK1 inhibitor-treated mice. All bars represent mean ± SEM. (Mann-Whitney U test) is shown.

The present application provides, inter alia, a method of treating a disease selected from cutaneous lupus erythematosus (CLE) and lichen planus (LP) in a patient in need thereof, comprising administering a therapeutically effective amount of a JAK1 selective inhibitor, or a pharma- ceutically acceptable salt thereof. In some embodiments, the disease is cutaneous lupus erythematosus (CLE). In some embodiments, the cutaneous lupus erythematosus (CLE) is selected from acute cutaneous lupus erythematosus, subacute cutaneous lupus erythematosus, and chronic cutaneous lupus erythematosus. In some embodiments, the cutaneous lupus erythematosus (CLE) is acute cutaneous lupus erythematosus. In some embodiments, the cutaneous lupus erythematosus is selected from subacute cutaneous lupus erythematosus (SCLE) and chronic discoid lupus erythematosus (CDLE). In some embodiments, the cutaneous lupus erythematosus is subacute cutaneous lupus erythematosus (SCLE). In some embodiments, the cutaneous lupus erythematosus is chronic discoid lupus erythematosus (CDLE). In some embodiments, the disease is lichen planus (LP).

As used herein, "cutaneous lupus" refers to a form of the disease in which symptoms are limited to those affecting the skin. For example, a patient may be diagnosed with cutaneous lupus, but that does not mean that the patient has SLE, which affects multiple sites on the body. Similarly, if a patient has SLE, it does not necessarily mean that the patient has cutaneous lupus. To that end, the Cutaneous Lupus Area and Severity Index (CLASI) scoring system was devised to quantitatively measure disease activity and damage (see, e.g., Albrecht et al., J. Invest. Dermatol. 2005, Nov; 125(5):889-94). The index takes into account the morphology and anatomical location of the lesions and has subsequently been validated through reliability testing for both dermatologists and rheumatologists.

Cutaneous lupus is divided into several subtypes, including acute cutaneous lupus erythematosus, subacute cutaneous lupus erythematosus, and chronic cutaneous lupus erythematosus. Acute cutaneous LE typically presents in the second decade of life and is frequently associated with active SLE. The most typical form of acute cutaneous lupus is a malar rash in which the flat areas of the face become red and resemble a sunburn (see, e.g., Fabbri et al., Am. J. Clin. Dermatol. 2003, 4(7):449-65). These lesions are typically transient, sun-induced, and non-scarring. A malar rash is present in approximately 50% of SLE patients at the time of diagnosis, and clinical activity of the rash has been reported to parallel systemic disease activity (see, e.g., Rothfield et al., Clin. Dermatol.; 2006, Sep-Oct; 24(5):348-62).

Chronic cutaneous lupus includes discoid LE (DLE), profundus LE (LEP), chilblain LE (CHLE), and neoplastic LE (LET). In some embodiments, the chronic cutaneous lupus is selected from discoid erythematosus (DLE), profundus LE (LEP), chilblain LE (CHLE), and neoplastic LE (LET).

Discoid LE lesions are the most common lesion of chronic cutaneous lupus erythematosus (CCLE) and appear as round, discoid lesions. DLE occurs most frequently in women in their 30s and 40s. DLE patients generally have a more benign disease course compared to patients with other CLE subtypes, and most patients (approximately 90-95%) do not develop lupus (SLE) symptoms in other organ systems (see, e.g., Crowson & Magro, J. Cutan. Pathol. 2001, 28(1):1-23; Chong et al., Br. J. Dermatol. 2012, 166(1):29-35). Erosions usually appear on the scalp and face, but may also appear on other parts of the body. Discoid lupus erythematosus lesions are often red, scaly, and thickened. Lesions are usually not painful or pruritic. Histologic examination of active DLE lesions reveals hyperkeratosis, dilated, dense keratin-filled follicles, vacuolar degeneration of basal keratinocytes, and an intense inflammatory dermal infiltrate.

Profound LEP (LEP), or panniculitis, is characterized by painful, firm subcutaneous nodules that appear in areas of increased fat deposition, such as the upper and lower legs, face, and chest. LEP tends to have a chronic course characterized by remissions and relapses, eventually leaving atrophic scars (see, e.g., Fabbri et al., Am. J. Clin. Dermatol. 2003, 4(7):449-65). Histology shows lobular panniculitis with dense lymphocytic infiltrates.

Chilblain lupus pernio (CHLE) is a rare form of CCLE that resembles frostbite. Lesions appear as painful, purplish plaques and nodules at sites of exposure to cold. Central erosions or ulcerations may occur on acral surfaces such as fingers, toes, heels, nose, and ears. Chilblain lupus pernio occurs when there is a drop in temperature and can be difficult to distinguish from frostbite. Pathology shows epidermal atrophy, interface vacuolation, and perivascular mononuclear cell infiltrate. Approximately 80% of patients with CHLE do not develop SLE (see, e.g., Hedrich et al., Clin. Rheumatol. 2008, 27(8):949-54).

Tumor-associated lupus erythematosus is a subtype of CCLE that occurs preferentially in men, unlike SLE, which occurs predominantly in women, and is characterized by extreme photosensitivity and a benign prognosis. Clinically, these lesions appear on the face as well-defined, raised, smooth-surfaced, erythematous, edematous, urticarial, multicyclic plaques.

Predominantly in young to middle-aged women, subacute cutaneous lupus (SCLE) lesions are described as having a scaly, red, annular ("ring-shaped") erythema with central healing. Lesions may also be multicyclic, i.e., appearing as multiple rings clustered together. There are two morphological subtypes of SCLE: annular and papular-squamous. The annular variant is characterized by scaly, annular, erythematous plaques that tend to coalesce to produce multicyclic rows (see, e.g., Walling & Sontheimer, Am. J. Clin. Dermatol. 2009, 10(6):365-81). The papular-squamous variant may resemble eczema and, in some cases, pityriasis rubra (see, e.g., Caproni et al. Int. J. Dermatol. 2001, 40(1):59-62). SCLE lesions occur in exposed areas such as the upper chest (in a "V" distribution), upper back, and extensor surfaces of the arms and forearms. Approximately 10% of patients with SLE have SCLE lesions, and patients with systemic disease tend to have only mild manifestations in other organ compartments. Pathologic examination of SCLE lesions shows vacuolar degeneration of basal keratinocytes, dermal edema, hyperkeratosis, keratinous plugs, and sparse superficial inflammatory infiltrates (see, e.g., Fabbri et al., Am. J. Clin. Dermatol. 2003, 4(7):449-65).

The first evidence for a functional role of type I IFN in lupus erythematosus (i.e., LE) came from clinical observations of patients with carcinoid tumors who were treated with recombinant IFNα and developed SLE due to this treatment (see, e.g., Ronnblom et al, Acta Oncol. 1991, 30:537-540). This was supported by the findings of multiple sclerosis patients who developed CLE-like lesions at the injection site side after subcutaneous application of recombinant IFNβ (see, e.g., Arrue et al, J. Cutan. Pathol. 2007, 34, Suppl 1:18-21). Gene expression analyses in both SLE and CLE have revealed strong expression of type I IFN-related inflammatory cytokines in blood and skin that correlate with disease activity (see, e.g., Kuhn et al, Semin. Immunopathol. 2016, 38:97-112, and Mikita et al, J. Dermatol. 2011, 38:839-849). These findings are supported by data described herein demonstrating strong expression of IFN-regulated genes and immune pathways within CLE skin lesions (see, e.g., Figures 1A-1D).

Previous reports describing a strong IFN-signature prompted the development of anti-IFN-targeted therapeutic strategies in patients with LE. Anti-IFNα agents (e.g., sifalimumab, rontalizumab) were the first drugs tested in clinical trials. These agents reduced the circulating IFN signature but had limited effect on disease activity. Without being bound by theory, this could be due to the high diversity of different type I IFNs, including not only IFNα, but also IFNβ and IFNκ, which bind to the same receptor (see, e.g., Kaluunian et al, Ann. Rheum. Dis. 2016, 75:196-202; Merrill et al, Ann. Rheum. Dis. 2011, 70:1905-1913; and Sarkar et al, Ann. Rheum. Dis. 2018, 77:1653-1664). Therefore, targeting a common receptor is more effective, and the anti-IFNαβ receptor antibody anifrolumab significantly reduced CLASI skin scores in SLE patients in a recent clinical trial (see, e.g., Furie et al, Arthritis & Rheumatology, 2017, 69:376-386). Alternative strategies have focused on "indirect" inhibition of the IFN system by (i) targeting IFN-producing cells or (ii) targeting the intracellular IFN pathway. In the skin and blood, plasmacytoid DCs are considered the primary IFN-producing cells (see, e.g., Saadeh et al, Exp. Dermatol. 2016, 25:415-421), and pDC-specific antibodies BIIB059 and VIB7734 are currently in clinical trials, with initial results showing reduced CLASI activity scores (see, e.g., Furie et al, Ann. Rheum. Dis. 2017, 76(Suppl 2):857).

The JAK family consists of four non-receptor tyrosine kinases (JAK1-3, TYK2) that transmit signals from growth factors and cytokines such as type I/III IFNs. JAK inhibitors were initially developed for the treatment of hematological disorders with well-defined JAK mutations and exhibit anticlonal activity (see, e.g., Morgan et al, Annu. Rev. Med. 2008, 59:213-222; and Pardanani et al, Leukemia 2008, 22:23-30), but they also produce significant immunosuppressive effects (see, e.g., Santos & Verstovsek, Expert Opin. Pharmacother. 2014, 15:1465-1473; Schwartz et al, Nat. Rev. Drug Discov. 2017, 16:843-862; Schwartz et al, Nat. Rev. Drug Discov. 2017, 16:843-862; Schwartz et al, Nat. Rev. Drug Discov. 2017, 16:843-862). al, Nat. Rev. Rheumatol. 2016,12:25-36; and Howell et al, Ann. Allergy Asthma Immunol. 2018,120:367-375).

Previous studies have demonstrated activation of the JAK/STAT pathway and expression of JAK proteins in CLE and related skin disorders (see, e.g., Scholtissek et al, J. Invest. Dermatol. 2017, 137:1484-1492; Alves de Medeiros et al, PLoS ONE 2016, 11:e0164080; and Wenzel et al, J. Am. Acad. Dermatol. 2008, 58:437-442). JAK inhibitors have been used to treat, for example, graft-versus-host disease (see, e.g., Spoerl et al, Blood, 2014, 123:3832-3842), dermatomyositis (see, e.g., Hornung et al, N. Engl. J. Med. 2014, 371:2537-2538, and Hornung et al, N. Engl. J. Med. 2015, 372:1274), and LE (see, e.g., Wenzel et al, J. Invest. Dermatol. 2016, 136:1281-1283, and Briand ... al, Ann. Rheum. Dis. 2019, 78:431-433), in addition to other inflammatory skin diseases (see, e.g., Banerjee et al, Drugs, 2017, 77:521-546; Welsch et al, Eur. J. Immunol. 2017, 47:1096-1107; and Shreberk-Hassidim et al, J. Am. Acad. Dermatol. 2017, 76:745-753.e19). In CLE, the JAK1/2 inhibitors ruxolitinib (see, e.g., Wenzel et al, J. Invest. Dermatol. 2016, 136:1281-1283; Briand et al, Ann. Rheum. Dis. 2019, 78:431-433; and Klaeschen et al, Exp. Dermatol. 2017, 26:728-730) and baricitinib (see, e.g., Zimmerman et al, JAMA Dermatol. DOI:10.1001/jamadermatol.2018.5077), as well as the JAK1/3 inhibitor tofacitinib (see, e.g., Konig et al. al, Ann. Rheum. Dis. 2017, 76:468-472) have been reported to be successful in treating patients, and these drugs significantly reduced the expression of chemokines typical of CLE in vitro (see, e.g., Klaeschen et al, Exp. Dermatol. 2017, 26:728-730, and Srivastava et al, Acta. Derm. Venereol. 2018, 98:772-775). One potential drawback of these drugs is the side effects, including anemia and thrombocytopenia, that may occur in some patients. The side effects are associated with inhibition of JAK2 and JAK3.

Of note, recent reports of double-blind, placebo-controlled trials for the treatment of SLE suggest that baricitinib was not effective compared to placebo (see, e.g., Wallace et al, Lancet, 2018, 392(10143):222-231, and Werth & Merrill, Br. J. Dermatol. 2019, 180(5):964-965). Baricitinib (2 mg or 4 mg) was administered once daily for 24 weeks. Eligible patients were aged 18 years or older, had a diagnosis of systemic lupus erythematosus (SLE), and had active disease affecting the skin or joints. The primary endpoint was the proportion of patients achieving resolution of arthritis or rash at week 24, as defined by the Systemic Lupus Erythematosus Disease Activity Index-2000 (SLEDAI-2K).

Exploratory endpoints assessed the Cutaneous Lupus Erythematosus Disease Area and Severity Index (CLASI) and showed no improvement in CLASI activity score with baricitinib. Unlike the SLEDAI, the CLASI scoring system is well-validated as a skin-specific endpoint for SLE and has been used successfully in several Phase II clinical trials for SLE and CLE. Recent reports also agree that the CLASI scoring system should be used as the cutaneous outcome for lupus testing for cutaneous reactivity (e.g., Albrecht J, et al., J. Invest. Dermatol. 2005;125:889-94; Klein et al., Arch. Dermatol. 2011,147:203-8; Furie et al., Arthritis Rheumatol. 2017,69:376-86; van Vollenhoven et al., The Lancet 2018,392:1330-9; Khamashta et al., J. Pharmacol. 2013,311:111-11; (see, e.g., E. et al., Ann. Rheumat. Dis. 2016, 75:1909-16, and Concha et al., J. Invest. Dermatol. 2018, https://doi.org/10.1016/j.jid.2018.08.017).

The methods provided herein utilize a compound or salt that is a JAK1 inhibitor (e.g., a JAK1 selective inhibitor). In some embodiments, the compound is selected from the following:
{1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-[4-fluoro-2-(trifluoromethyl)phenyl]piperidine-1-carboxamide,
[3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(1-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)azetidin-3-yl]acetonitrile,
4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide,
((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile,
3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,
3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,
4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile,
4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile,
[trans-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-3-(4-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperazin-1-yl)cyclobutyl]acetonitrile,
{trans-3-(4-{[4-[(3-hydroxyazetidin-1-yl)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
{trans-3-(4-{[4-{[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
{trans-3-(4-{[4-{[(2R)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile,
5-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide,
4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide,
5-{3-(cyanomethyl)-3-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide,
{1-(cis-4-{[6-(2-hydroxyethyl)-2-(trifluoromethyl)pyrimidin-4-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
{1-(cis-4-{[4-[(ethylamino)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
{1-(cis-4-{[4-(1-hydroxy-1-methylethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
{1-(cis-4-{[4-{[(3R)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
{1-(cis-4-{[4-{[(3S)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
{trans-3-(4-{[4-({[(1S)-2-hydroxy-1-methylethyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
{trans-3-(4-{[4-({[(2R)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
{trans-3-(4-{[4-({[(2S)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile and {trans-3-(4-{[4-(2-hydroxyethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
or a pharma- ceutically acceptable salt of any of the foregoing.

In some embodiments, the compounds or salts are selective for JAK1 over JAK2, JAK3, and TYK2. For example, some of the compounds described herein, or pharma- ceutically acceptable salts thereof, selectively inhibit JAK1 over one or more of JAK2, JAK3, and TYK2. JAK1 plays a central role in many cytokine and growth factor signaling pathways whose dysregulation can lead to or contribute to disease states. For example, IL-6 levels are elevated in rheumatoid arthritis, a disease in which IL-6 has been suggested to have deleterious effects (see, e.g., Fonesca, et al., Autoimmunity Reviews, 8:538-42, 2009). Because IL-6 signals, at least in part, through JAK1, IL-6 may indirectly inhibit JAK1, resulting in potential clinical benefit (see, e.g., Guschin, et al. Embo J 14:1421, 1995, and Smolen, et al. Lancet 371:987, 2008). In other autoimmune diseases and cancers, elevated systemic levels of inflammatory cytokines that activate JAK1 may also contribute to the disease and/or associated symptoms. Thus, patients with such diseases may benefit from inhibition of JAK1. Selective inhibitors of JAK1, as described herein, may be effective while avoiding the unnecessary and undesirable effects of inhibiting other JAK kinases.

In some embodiments, the compounds or salts selectively inhibit JAK1 over JAK2 (e.g., JAK2/JAK1 _IC50 ratio of greater than 1). In some embodiments, the compounds or salts provided herein are about 10-fold selective for JAK1 over JAK2. In some embodiments, the compounds or salts provided herein are about 3-fold, about 5-fold, about 10-fold, about 15-fold, or about 20-fold selective for JAK1 over JAK2, as calculated by measuring the _IC50 at 1 mM ATP (see Example 1).

In some embodiments, the JAK1 inhibitor is a compound of Table 1, or a pharma- ceutically acceptable salt thereof. The compounds of Table 1 are selective inhibitors of JAK1 (e.g., more selective than JAK2, JAK3, and TYK2). _IC50 values obtained by the method of Example 1 at 1 mM ATP are shown in Table 1.
+ means less than 10 nM (see Example 1 for assay conditions)
++ means 100 nM or less (see Example 1 for assay conditions)
+++ means 300 nM or less (see Example 1 for assay conditions)
Data for ^a enantiomer 1
^b Enantiomer 2 data

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

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

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.

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

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

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

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

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

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. US 2014/0343030, filed May 16, 2014, each of which is incorporated herein by reference in its entirety.

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

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

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.

In some embodiments, the compounds in Table 1 are selected from the group consisting of those 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, 2010/0298334, filed May 21, 2010, and 2010/030624, each of which is incorporated herein by reference in its entirety. It is prepared by the synthesis methods described in U.S. Patent Publication No. 11/0059951, U.S. Patent Publication No. 2012/0149681, filed November 18, 2011, 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.

In some embodiments, the JAK1 inhibitor is selected from the group consisting of those 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, 2010/0298334, filed May 21, 2010, and 2011/0026, filed August 31, 2010, each of which is incorporated herein by reference in its entirety. No. 59951, U.S. Patent Publication No. 2012/0149681 filed November 18, 2011, 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.

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

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

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

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.

In some embodiments, the JAK1 inhibitor is a compound of formula II
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 alkyl, each of which is optionally substituted with 1, ₂ , or 3 substituents independently selected from fluoro, _{—CF 3 ,} and methyl;
^R3 is H or methyl;
^R4 is H, F, or Cl;
^R5 is H or F;
^R6 is H or F;
^R7 is H or F;
^R8 is H or methyl;
^R9 is H or methyl;
R ¹⁰ is H or methyl;
R ¹¹ is H or methyl.

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.

In some embodiments, the JAK1 inhibitor is a compound of formula III
or a pharma- ceutically acceptable salt thereof,
Cy ⁴ is a tetrahydro-2H-pyran ring, optionally substituted with one or two groups independently selected from CN, OH, F, Cl, C _1-3 alkyl, C _1-3 haloalkyl, CN-C _1-3 alkyl, HO-C _1-3 alkyl, amino, C _1-3 alkylamino, and di(C _1-3 alkyl)amino, wherein said C _1-3 alkyl and di(C _1-3 alkyl)amino are optionally substituted with one, two, or three substituents independently selected from F, Cl, C _1-3 alkylaminosulfonyl, and C _1-3 alkylsulfonyl;
^R12 is -CH2 _- OH, -CH( _CH3 )-OH, or _{-CH2 - NHSO2CH3} .

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

One or more constituent atoms of the compounds described herein can be replaced or substituted with an isotope of the atom at natural or non-natural abundance. In some embodiments, the compounds include at least one deuterium atom. In some embodiments, the compounds include two or more deuterium atoms. In some embodiments, the compounds include 1-2, 1-3, 1-4, 1-5, or 1-6 deuterium atoms. In some embodiments, all hydrogen atoms in the compounds can be replaced or substituted with deuterium atoms.

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

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

Thus, in some embodiments, a JAK1 inhibitor (e.g., a JAK1 selective inhibitor) is a compound in which one or more hydrogen atoms are replaced by deuterium atoms, or a pharma- ceutically acceptable salt thereof.

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

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

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

As used herein, the term "HO-C _1-3 -alkyl" refers to a group of formula -alkylene-OH, said alkylene group having 1 to 3 carbon atoms.

As used herein, the term "CN-C _1-3 alkyl" refers to a C _1-3 alkyl substituted by a cyano group.

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

As used herein, the term "di(C _1-3 -alkyl)amino" refers to a group of formula --N(alkyl) ₂ , wherein the two alkyl groups each independently have 1 to 3 carbon atoms.

As used herein, the term "C _1-3 alkylamino" refers to a group of formula -NH(alkyl), where the alkyl group has 1 to 3 carbon atoms.

As used herein, the term "di(C _1-3 alkyl)aminosulfonyl" refers to a group of formula -S(O) ₂ N(alkyl) ₂ , where each alkyl group independently has 1 to 3 carbon atoms.

As used herein, the term "C _1-3 alkylsulfonyl" refers to a group of formula -S(O) ₂ -alkyl, where the alkyl group has 1 to 3 carbon atoms.

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

As used herein, the term "C _nm haloalkyl," used alone or in combination with other terms, refers to a C _nm alkyl group having up to {2(n-m)+1} halogen atoms, which may be the same or different. In some embodiments, the halogen atoms are fluoro atoms. In some embodiments, the alkyl group has 1-6 or 1-3 carbon atoms. Examples of haloalkyl groups include CF ₃ , C ₂ F ₅ , CHF ₂ , CCl ₃ , CHCl ₂ , C ₂ Cl ₅ , and the like. In some embodiments, the haloalkyl group is a fluoroalkyl group.

As used herein, the term "C _1-3 fluoroalkyl" refers to a C _1-3 alkyl group which may be partially or fully substituted with fluoro atoms.

As used herein, the term "C _3-6 cycloalkyl", used alone or in combination with other terms, refers to a non-aromatic monocyclic hydrocarbon moiety having from 3 to 6 carbon atoms which may optionally contain one or more alkenylene groups as part of the ring structure. One or more ring-forming carbon atoms of the cycloalkyl group may be oxidized to form a carbonyl bond. Exemplary C _3-6 cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, and the like. In some embodiments, the cyclopropyl group is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

As used herein, the term "C _3-6 cycloalkyl-C _1-3 alkyl" refers to a group of formula -C _1-3 alkylene-C _3-6 cycloalkyl.

The compounds described herein may be asymmetric (e.g., having 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 for preparing optically active forms from optically inactive starting materials are known in the art, for example, by resolution of racemic mixtures or stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like may also be present in the compounds described herein, and all such stable isomers are contemplated in this application. Cis and trans geometric isomers of the compounds of this application are described and may be isolated as a mixture of isomers or as separated isomeric forms. In some embodiments, the compounds have the (R) configuration. In some embodiments, the compounds have the (S) configuration.

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

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

The compounds described herein also include tautomeric forms. Tautomeric forms arise from the simultaneous migration of a proton along with the interchange of a single bond with an adjacent double bond. Tautomeric forms include prototropic tautomers, which are isomeric protonation states having 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 a proton can occupy more than one position in a heterocyclic 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 may be in equilibrium or sterically locked into one form by appropriate substitution. For example, it will be recognized that two tautomers can form in the pyrazole ring below.
The claims are intended to cover both tautomers.

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.

In some embodiments, the compounds described herein or salts thereof are substantially isolated. "Substantially isolated" means that the compound is at least partially or substantially separated from the environment in which it was formed or detected. Partial separation can include, for example, compositions enriched in the compounds described herein. Substantial separation can include compositions that contain 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 salts thereof are routine in the art.

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

As used herein, the terms "ambient temperature" and "room temperature" or "rt" are understood in the art and generally refer to a temperature, e.g., a reaction temperature, that is close to the temperature of the room in which the reaction is carried out, e.g., from about 20°C to about 30°C.

The present application also includes pharma- ceutically acceptable salts of the compounds described herein. As used herein, "pharma- ceutically acceptable salts" refers to derivatives of the disclosed compounds, in which the parent compound has been 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, inorganic or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, and the like. The pharma- ceutically acceptable salts of the present application include conventional non-toxic salts of the parent compound, for example, formed from non-toxic inorganic or organic acids. The pharma- ceutically acceptable salts of the present application can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. In general, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or in a mixture of both, generally in non-aqueous media such as ether, ethyl acetate, alcohol (e.g., methanol, ethanol, isopropanol, or butanol) or acetonitrile (ACN). Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

As used herein, the term "contacting" refers to combining the indicated moieties together in an in vitro system or in an in vivo system. For example, "contacting" a JAK with a compound of the invention includes administering a compound of the present application to an individual or patient, such as a human, having a JAK, as well as, for example, introducing a compound of the invention into a sample containing a cell preparation or purified preparation containing a JAK.

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

In some embodiments, the inhibitor is administered in a therapeutically effective amount. As used herein, the phrase "therapeutically effective amount" refers to an amount of an active compound or pharmaceutical agent that elicits the biological or pharmaceutical response that a researcher, veterinarian, physician, or other clinician is seeking in a tissue, system, animal, individual, or human.

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

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

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

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

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

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

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

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

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

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

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

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

In some embodiments, the method further comprises administering an additional therapeutic agent selected from antithymocyte globulin, recombinant human granulocyte colony stimulating factor (GCSF), granulocyte monocytic CSF (GM-CSF), erythropoietin stimulating factor (ESA), and cyclosporine.

In some embodiments, the method further comprises administering to the patient an additional JAK inhibitor. In some embodiments, the additional JAK inhibitor is selected from a JAK2 inhibitor (e.g., a selective JAK2 inhibitor), a JAK1/JAK2 inhibitor, a JAK3 inhibitor (e.g., a selective JAK3 inhibitor), and a JAK1/JAK3 inhibitor, or a pharma- ceutically acceptable salt of any of the foregoing.

In some embodiments, the additional JAK inhibitor is a JAK1/JAK2 inhibitor. In some embodiments, the JAK1/JAK2 inhibitor is selective for JAK1 and JAK2 over JAK3 and TYK2. In some embodiments, the JAK1/JAK2 inhibitor is 3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, or a pharmaceutically acceptable salt thereof. In some embodiments, the JAK1/JAK2 inhibitor is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile (ruxolitinib), or a pharmaceutically acceptable salt thereof. Ruxolitinib has an _IC50 of less than 10 nM at 1 mM ATP for JAK1 and JAK2. 3-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile and ruxolitinib can be made by the procedure (Example 67) described in US Pat. No. 7,598,257, filed Dec. 12, 2006, which is incorporated by reference in its entirety. In some embodiments, the JAK1/JAK2 inhibitor is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile phosphate. The phosphate can be made as described in US Pat. No. 8,722,693, which is incorporated by reference in its entirety. In some embodiments, the JAK1/JAK2 inhibitor is barcitinib, or a pharma- ceutically acceptable salt thereof. In some embodiments, the JAK1/JAK2 inhibitor is barcitinib.

In some embodiments, the additional JAK1 inhibitor is a JAK1/JAK3 inhibitor. In some embodiments, the JAK1/JAK3 inhibitor is tofacitinib, or a pharma- ceutically acceptable salt thereof. In some embodiments, the JAK1/JAK3 inhibitor is tofacitinib.

In some embodiments, the JAK1/JAK2 inhibitor may be an isotopically labeled compound, or a pharma- ceutically acceptable salt thereof. An "isotopically" or "radioactively labeled" compound is a compound in which one or more atoms have been replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occurring). Suitable radionuclides that may be incorporated into the compounds of the present disclosure include, but are not limited to, ^2H (also written as deuterium, D), ^3H (also written as tritium, T), ^11C , ^13C , ^14C , ^13N , ^15N , ^15O , ^17O , ^18O , ^18F , ^35S , ^36Cl , ^82Br , ^75Br , ^76Br , ^77Br , ^123I , ^124I , ^125I , and ^131I . For example, one or more hydrogen atoms in the compounds of the present disclosure may be replaced by a deuterium atom, e.g., _-CH3 is replaced with _-CD3 ).

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

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

In some embodiments, the JAK1/JAK2 inhibitor is a compound of formula IV
or a pharma- ceutically acceptable salt thereof,
R ¹ is selected from H and D;
each ^R2 is independently selected from H and D, provided that each ^R2 attached to the common carbon is the same;
each ^R3 is independently selected from H and D, provided that each ^R3 attached to the common carbon is the same;
^R4 is selected from H and D;
each ^R5 is the same and is selected from H and D;
R ⁶ , R ⁷ , and R ⁸ are each independently selected from H and D, with the proviso that when R ¹ is H, each R ² and each R ³ are H, R ⁴ is H, and R ⁶ , R ⁷ , and R ⁸ are each H, then each R ⁵ is D.

In some embodiments, the JAK1 and/or JAK2 inhibitor is a compound of formula IV selected from compounds 100-130 in the table below, where R ⁶ , R ⁷ , and R ⁸ are each H, or a pharmaceutically acceptable salt thereof. In some embodiments, the JAK1/JAK2 inhibitor is a compound of formula IV selected from compounds 200-231 in the table below, where R ⁶ , R ⁷ , and R ⁸ are each D, or a pharmaceutically acceptable salt thereof.

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

In some embodiments, the additional JAK inhibitor is selected from baricitinib, tofacitinib, oclacitinib, filgotinib, gandotinib, restortinib, momelotinib, bacritinib, PF-04965842, upadacitinib, peficitinib, fedratinib, cucurbitacin I, ATI-501 (Aclaris), ATI-502 (Aclaris), JTE-052 (i.e., delgocitinib; Leo Pharma and Japan Tobacco), and CHZ868.

For the treatment of cutaneous lupus erythematosus (CLE), additional agents such as, but not limited to, anti-inflammatory agents, immunosuppressants, PI3Kδ inhibitors, mTor inhibitors, Bcr-Abl inhibitors, Flt-3 inhibitors, RAF inhibitors, and FAK kinase inhibitors (e.g., those described in WO 2006/056399, which is incorporated herein by reference in its entirety), or other agents, can be used in combination with the JAK1 inhibitors described herein. One or more additional agents can be administered to the patient simultaneously or sequentially.

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

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

Examples of suitable RAF inhibitors include compounds and pharma- ceutically acceptable salts thereof, such as those disclosed in WO 00/09495 and WO 05/028444, both of which are incorporated herein by reference in their entireties.

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

In some embodiments, one or more of the JAK1 inhibitors described herein can be used in combination with one or more other kinase inhibitors, e.g., imatinib, particularly to treat patients who are resistant to imatinib or other kinase inhibitors.

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

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

In some embodiments, the additional therapeutic agent is Dehydrex™ (Holles Labs), Civamide (Opko), sodium hyaluronate (Vismed, Lantibio/TRB Chemedia), cyclosporine (ST-603, Sirion Therapeutics), ARG101(T) (testosterone, Argentis), AGR1012(P) (Argentis), ecabet sodium (Senju-Ista), gefarnate (Santen), 15-(s)-hydroxyeicosatetraenoic acid (15(S)-HETE), cevilemine, doxycycline (ALTY-0501, Alacrity), minocycline, iDestrin™ (NP50301, Nascent Pharmaceuticals), cyclosporine A (Nova 22007, Novagali), oxytetracycline (duramycin, MOLI 1901, Lantibio), CF101 (2S,3S,4R,5R)-3,4-dihydroxy-5-[6-[(3-iodophenyl)methylamino]purin-9-yl]-N-methyl-oxolane-2-carbamyl, Can-Fite Biopharma), voclosporin (LX212 or LX214, Lux Biosciences), ARG103 (Agentis), RX-10045 (synthetic resolvin analog, Resolvyx), DYN15 (Dyanmis Therapeutics), Rivoglitazone (DE011, Daiichi Sanko), TB4 (RegeneRx), OPH-01 (Ophtalmis Monaco), PCS101 (Pericor Science), REV1-31 (Evolutec), Lacritin (Senju), Rebamipide (Otsuka-Novartis), OT-551 (Othera), PAI-2 (University of Pennsylvania and Temple) The agent is selected from: pilocarpine, tacrolimus, pimecrolimus (AMS981, Novartis), loteprednol etabonate, rituximab, diquafosol tetrasodium (INS365, Inspire), KLS-0611 (Kissei Pharmaceuticals), dehydroepiandrosterone, anakinra, efalizumab, sodium mycophenolate, etanercept (Embrel®), hydroxychloroquine, NGX267 (TorreyPines Therapeutics), Actemra, gemcitabine, oxaliplatin, L-asparaginase, or thalidomide.

In some embodiments, the additional therapeutic agent is an antiangiogenic agent, a cholinergic agent, a TRP-1 receptor modulator, a calcium channel blocker, a mucin secretagogue, a MUC1 stimulator, a calcineurin inhibitor, a corticosteroid, a P2Y2 receptor agonist, a muscarinic receptor agonist, or an mTOR inhibitor. In some embodiments, the additional therapeutic agent is a tetracycline derivative (e.g., minocycline or doxycline). In some embodiments, the additional therapeutic agent binds to FKBP12.

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

In some embodiments, the additional therapeutic agent is selected from antibiotics, antivirals, antifungals, anesthetics, anti-inflammatory agents (e.g., steroidal and nonsteroidal anti-inflammatory agents), and anti-allergy agents. Examples of suitable agents include aminoglycosides, such as amikacin, gentamicin, tobramycin, streptomycin, netilmicin, and kanamycin; fluoroquinolones, such as ciprofloxacin, norfloxacin, ofloxacin, trovafloxacin, lomefloxacin, levofloxacin, and enoxacin; naphthyridines; sulfonamides; polymyxins; chloramphenicol; neomycin; paramomycin; colistimethate; bacitracin; vancomycin; cyclosporine ... These include, but are not limited to, mycin; tetracyclines; rifampicin and its derivatives ("rifampin"); cycloserine; beta-lactams; cephalosporins; amphotericin; fluconazole; flucytosine; natamycin; miconazole; ketoconazole; corticosteroids; diclofenac; flurbiprofen; ketorolac; suprofen; cromolyn; lodoxamide; levocabastine; naphazoline; antazoline; pheniramine; or azalide antibiotics.

Pharmaceutical Formulations and Dosage Forms When used as pharmaceuticals, the JAK1 inhibitors provided herein can be administered in the form of pharmaceutical compositions. These compositions can be prepared in a manner well known in the pharmaceutical art and can be administered by various routes depending on whether local or systemic treatment is desired and on the area to be treated. Administration can be topical (including intradermal, transdermal, epidermal, ophthalmic, and mucosal delivery, including intranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, such as by a nebulizer; intratracheal or intranasal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal intramuscular or injection or infusion; or intracranial, e.g., intrathecal or intraventricular administration. Parenteral administration can be in the form of a single bolus dose or can be, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration include transdermal patches, skin patches, solutions, suspensions, foams, 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, the composition is formulated for topical administration by transdermal patches, skin patches, solutions, suspensions, gels, creams, ointments, lotions, sprays, foams, liquids, drops, suppositories, and powders. In some embodiments, the composition is formulated for topical administration by transdermal patches. In some embodiments, the composition is formulated for topical administration by skin patches. In some embodiments, the composition is formulated as a foam (e.g., for topical administration).

Topical medications that can penetrate the skin barrier and limit systemic effects are particularly important for treating skin disorders such as cutaneous lupus erythematosus (CLE).

Topical (cutaneous/intradermal) formulations are typically solutions, suspensions, gels, creams, ointments, lotions, sprays, and foams. Preferred topical formulations should be physically and chemically stable, should not cause skin irritation, and should deliver the active agent (e.g., a selective JAK1 inhibitor, or a pharma- ceutically acceptable salt thereof, as described herein) to the appropriate layers of the skin at a concentration that provides a therapeutic response with limited systemic exposure.

In some embodiments, administration is topical and consists of a formulation having one or more pharma- ceutically (e.g., dermatologically) acceptable excipients. Examples of dermatologically acceptable excipients include, but are not limited to, pH adjusters, chelating agents, preservatives, cosolvents, membrane permeability enhancers, humectants, thickening agents, gelling agents, viscosity enhancers, surfactants, propellants, flavoring agents, coloring agents, or any combination or mixture thereof. In some embodiments, the topical formulation is administered locally to the patient (e.g., administered to the site of a lesion).

In some embodiments, the pH adjusting agent is selected from an acid, an acid salt, a base, a basic salt, and a buffer, or any mixture thereof. Exemplary acids include, but are not limited to, lactic acid, acetic acid, citric acid, and benzoic acid, and salts thereof. Exemplary bases include, but are not limited to, trolamine, tromethamine, and salts thereof. Exemplary buffers include, but are not limited to, citrate/citric acid, acetate/acetic acid, edetate/edetic acid, lactate/lactic acid, and the like.

In some embodiments, the chelating agent is a single additive. In some embodiments, the chelating agent is a mixture of two or more chelating agents. Exemplary chelating agents include, but are not limited to, ethylenediaminetetraacetic acid (EDTA), or salts thereof. In some embodiments, the chelating agent comprises a mixture of a chelating agent and an antioxidant, where the chelating agent and the antioxidant prevent, minimize, or reduce oxidative degradation reactions in the composition. Exemplary antioxidants include, but are not limited to, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), tocopherol, and propyl gallate.

In some embodiments, the composition includes one or more preservatives. In some embodiments, the composition includes a mixture of two or more preservatives. In some embodiments, the composition includes 1-5 preservatives. Exemplary preservatives include, but are not limited to, benzyl alcohol, phenonyexthanol, methylparaben, ethylparaben, propylparaben, butylparaben, and imidazolidinyl urea.

In some embodiments, the composition includes one or more co-solvents. In some embodiments, the composition includes a mixture of two or more co-solvents. In some embodiments, the composition includes 1-5 co-solvents. Exemplary solvents include, but are not limited to, water, propylene glycol, diethylene glycol monoethyl ether, dimethyl isosorbide, ethyl alcohol, isopropyl alcohol, benzyl alcohol, propanediol, propylene glycol, polyethylene glycol (e.g., polyethylene glycol 200, polyethylene glycol 300, polyethylene glycol 400, etc.). In some embodiments, the solvent is a water-insoluble agent. Exemplary water-insoluble agents include, but are not limited to, diethyl sebacate, diisopropyl adipate, isopropyl myristate, isopropyl palmitate, and medium chain triglycerides.

In some embodiments, the composition comprises one or more membrane permeability enhancers. In some embodiments, the composition comprises a mixture of two or more membrane permeability enhancers. In some embodiments, the composition comprises 1-5 membrane permeability enhancers. A membrane permeability enhancer can act as both a solvent and a membrane permeability enhancer. Exemplary membrane permeability enhancers include, but are not limited to, fatty acids, fatty acid esters, fatty alcohols, pyrrolidones, sulfoxides, alcohols, diols and polyols, or any mixtures thereof. In some embodiments, a co-solvent provided herein is a membrane permeability enhancer.

In some embodiments, the composition includes one or more thickening agents, gelling agents, or thickening agents. In some embodiments, the composition includes a mixture of two or more thickening agents, gelling agents, or thickening agents. In some embodiments, the composition includes 1-5 thickening agents, gelling agents, or thickening agents. Exemplary thickening agents, gelling agents, or thickening agents include, but are not limited to, cellulose derivatives (e.g., hydroxyethyl cellulose (HEC), carboxymethyl cellulose, hydroxypropyl cellulose (HPC), and hydroxypropyl methyl cellulose (HPMC), and polyvinylpyrrolidone (PVP).

A surfactant is a compound that reduces the surface tension between two liquids or between a liquid and a solid. The surfactant may be a mixture of two or more surfactants. Exemplary surfactants include, but are not limited to, ethoxylated fatty alcohol ethers (e.g., steareth-2, steareth-10, steareth-20, ceteareth-2, ceteareth-10, etc.), PEG esters (e.g., PEG-4 dilaurate, PEG-20 stearate, etc.), glyceryl esters or derivatives thereof (e.g., glyceryl dioleate, glyceryl stearate, etc.), polymeric ethers (e.g., poloxamer 124, poloxamer 181, poloxamer 182, etc.), sorbitan derivatives (e.g., polysorbate 80, sorbitan monostearate, etc.), fatty alcohols (e.g., cetyl alcohol, stearyl alcohol, cetearyl alcohol, etc.), and emulsifying waxes (e.g., emulsifying wax NF, a mixture of cetearyl alcohol and polysorbate 60, etc.).

In some embodiments, administration is topical to the skin.

In some embodiments, administration is oral.

In some embodiments, administration of a topical composition comprising a selective JAK1 inhibitor provided herein to a patient results in a systemic exposure at steady state ( _Cmax ) of less than about 5% of the applied dose, e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%, etc. In some embodiments, administration of a topical composition comprising a selective JAK1 inhibitor provided herein to a patient results in a systemic exposure at steady state ( _Cmax ) of less than about 1% of the applied dose.

The present invention also includes pharmaceutical compositions containing as an active ingredient a JAK1 inhibitor provided herein or a pharma- ceutically acceptable salt thereof in combination with one or more pharma- ceutically acceptable carriers (additives). In some embodiments, the compositions are suitable for topical administration. In making the compositions of the present invention, the active ingredient is typically mixed with an additive, diluted by an additive, or enclosed in a carrier, such as, for example, a capsule, sachet, paper or other container form. When used as a diluent, an additive can be a solid, semi-solid, or liquid substance that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (either as solid or liquid media), ointments, e.g., containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

In preparing the formulation, the active compound can be milled to provide the appropriate particle size prior to combining with the other ingredients. If the active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh. If the active compound is substantially water soluble, the particle size can be adjusted by milling to provide a substantially uniform distribution in the formulation, e.g., about 40 mesh.

The JAK1 inhibitors provided herein may 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 JAK1 inhibitors can be prepared by methods known in the art (see, e.g., International Application No. WO 2002/000196).

Some examples of suitable additives include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulation may additionally include lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl benzoate and propyl hydroxybenzoate; sweetening agents; and flavoring agents. The compositions of the present invention may be formulated to provide immediate, sustained, or delayed release of the active ingredient after administration to the patient by methods known in the art.

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

In some embodiments, the composition is a sustained release composition comprising at least one compound described herein, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier. In some embodiments, the composition comprises at least one compound described herein, or a pharma- ceutically acceptable salt thereof, and at least one component selected from microcrystalline cellulose, lactose monohydrate, hydroxypropyl methylcellulose, and polyethylene oxide. In some embodiments, the composition comprises at least one compound described herein, or a pharma- ceutically acceptable salt thereof, and microcrystalline cellulose, lactose monohydrate, and hydroxypropyl methylcellulose. In some embodiments, the composition comprises at least one compound described herein, or a pharma- ceutically acceptable salt thereof, and microcrystalline cellulose, lactose monohydrate, and polyethylene oxide. In some embodiments, the composition further comprises magnesium stearate or silicon dioxide. In some embodiments, the microcrystalline cellulose is Avicel PH102™. In some embodiments, the lactose monohydrate is Fast-flo 316™. In some embodiments, the hydroxypropyl methylcellulose is hydroxypropyl methylcellulose 2208 K4M (e.g., Methocel K4 M Premier™) and/or hydroxypropyl methylcellulose 2208 K100LV (e.g., Methocel K00LV™). In some embodiments, the polyethylene oxide is polyethylene oxide WSR 1105 (e.g., Polyox WSR 1105™).

In some embodiments, a wet granulation process is used to prepare the composition. In some embodiments, a dry granulation process is used to prepare the composition.

The compositions can be formulated in unit dosage form, with each dosage containing about 1 to about 1,000 mg, about 1 mg to about 100 mg, about 1 mg to about 50 mg, and about 1 mg to about 10 mg of active ingredient. Preferably, the dosage is about 1 mg to about 50 mg or about 1 mg to about 10 mg of active ingredient. In some embodiments, each dosage contains about 10 mg of active ingredient. In some embodiments, each dosage contains about 50 mg of active ingredient. In some embodiments, each dosage contains about 25 mg of active ingredient. The term "unit dosage form" refers to a physically discrete unit suitable as a unitary dosage for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with suitable pharmaceutical excipients.

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

The active compounds may be effective over a wide dosage range and are generally administered in a pharma- ceutical effective amount. However, it will be understood that the amount of compound actually administered will usually be determined by the physician according to the relevant circumstances, e.g., the condition to be treated, the selected route of administration, the compound actually administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, etc.

To prepare solid compositions such as tablets, the primary active ingredient is mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogenous mixture of the compounds of the present application. When these preformulation compositions are referred to as homogenous, the active ingredient is typically dispersed evenly throughout the composition such that the composition can be readily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules. This solid preformulation is then subdivided into unit dosage forms of the type described above, containing, for example, from about 0.1 to about 1000 mg of the active ingredient of the present application (e.g., a JAK1 inhibitor provided herein).

The tablets or pills of the present application can be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill can comprise an inner dose and an outer dose component, the latter in the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permit 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 a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.

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

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

Topical (e.g., intradermal) administration offers the advantages of treating skin disorders locally, minimizing potential adverse events associated with systemic exposure, and allowing treatment to be easily discontinued, if necessary. Additionally, some topically applied formulations, such as creams, ointments, and gels, have the advantage of additives that can act as emollients or occlusive agents, enhancing patient well-being and compliance during treatment. Other routes of administration, such as oral, parenteral, and inhalation, can result in systemic drug concentrations that significantly exceed therapeutic efficacy, increased potential for adverse events, drug-drug interactions, and generation of active/toxic metabolites, which can lead to treatment discontinuation and poor patient compliance.

Topical formulations intended for dermal delivery are typically solutions, suspensions, gels, creams, ointments, lotions, sprays, and foams and may contain one or more conventional carriers as described herein. The formulation composition should be prepared with the objective of delivering the active ingredient to the appropriate layer(s) of the skin, minimizing systemic exposure, and preventing skin irritation. In addition, the pharmaceutical composition should be physically and chemically stable. Depending on the dosage form selected, one or more additional additives as described herein may be required, such as pH adjusters, chelating agents, preservatives, cosolvents, membrane permeability enhancers, humectants, thickeners, gelling agents, viscosity enhancers, surfactants, propellants, flavoring agents, colorants, or any combination or mixture thereof.

In some embodiments, the topical formulation may contain one or more conventional carriers as described herein. In some embodiments, the ointment may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, and the like. The carrier composition of the cream may be based on water combined with glycerol and one or more other ingredients, for example, glycerol monostearate, PEG-glycerol monostearate, and cetylstearyl alcohol. Gels may be formulated using isopropyl alcohol and water, preferably in combination with other ingredients, such as, for example, glycerol, hydroxyethylcellulose, and the like. In some embodiments, the topical formulation contains at least about 0.1%, at least about 0.25%, at least about 0.5%, at least about 1%, at least about 2%, or at least about 5% by weight of a compound of the present invention. The topical formulation may be suitably packaged in a tube, for example, a 100 g tube, and is optionally accompanied by instructions for the treatment of cutaneous lupus erythematosus (CLE).

The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of the administration, such as prophylaxis or treatment, the condition of the patient, the method of administration, etc. In therapeutic applications, the compositions may be administered to a patient already suffering from cutaneous lupus erythematosus (CLE) in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose will vary depending on the disease state being treated and at the discretion of the attending physician depending on factors such as the severity of the disease, the age, weight, and general condition of the patient.

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 for use as is or lyophilized, the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the compound preparations is typically 3-11, more preferably 5-9, and most preferably 7-8. It will be appreciated that the use of certain of the aforementioned additives, carriers, or stabilizers may result in the formation of pharmaceutical salts.

The therapeutic dosage of the compounds of the present application may vary according to, for example, the particular application for which the treatment is made, the method of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of a compound of the present invention in a pharmaceutical composition may vary depending on a number of factors, including the dosage amount, chemical characteristics (e.g., hydrophobicity), and the route of administration. For example, the compounds of the present invention may be provided in an aqueous physiological buffer solution containing about 0.1 to about 10 w/v % of the compound for parenteral administration. Some typical dosage ranges are about 1 μg/kg to about 1 g/kg of body weight per day. In some embodiments, the dosage range is about 0.01 mg/kg to about 100 mg/kg of body weight per day. The dosage may vary depending on such variables as the type and extent of progression of the disease or disorder, the general health of the particular patient, the relative biological effectiveness of the selected compound, the formulation of excipients, and its route of administration. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

The compositions of the present invention may further include one or more additional pharmaceutical agents, examples of which are listed hereinbefore.

Kits The present application also includes pharmaceutical kits useful, for example, in the treatment and/or prevention of cutaneous lupus erythematosus (CLE), which include one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a JAK1 inhibitor as described herein. Such kits may further include one or more of a variety of conventional pharmaceutical kit components as needed, such as one or more containers with pharma- ceutically acceptable carriers, additional containers, etc., as would be readily apparent to one of skill in the art. Instructions 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, as an insert or label.

The present invention will now be described in further detail with reference to specific examples. The following examples are provided for illustrative purposes and are not intended to limit the invention in any manner. Those of skill in the art will readily recognize a variety of non-critical parameters that can be changed or modified to yield essentially the same results.

All statistical analyses of in vitro experiments were performed with GraphPad prism software (version 7) using Kruskal-Wallis test and Mann-Whitney U test. Gene expression was analyzed with Partek Flow genomic analysis software and Subio Platform software v1.22.5266 using Welch's t-test. Confidence intervals were determined at 95%. P<0.05 was considered "significant" ( ^* ) and p<0.01 "highly significant" ( ^** ). KEGG pathways were mapped to differentially expressed genes using DAVID v6.8 (Database for Annotation, Visualization and Integrated Discovery).

Example 1. In Vitro JAK Kinase Assays JAK1 inhibitors that can be used to treat cytokine-related diseases or disorders are tested for inhibitory activity of JAK targets according to the following in vitro assay described in Park et al., Analytical Biochemistry 1999, 269, 94-104. The catalytic domains of human JAK1 (amino acids 837-1142), JAK2 (amino acids 828-1132), and JAK3 (amino acids 781-1124) with N-terminal His tags are expressed using baculovirus in insect cells and purified. The catalytic activity of JAK1, JAK2, or JAK3 was assayed by measuring phosphorylation of a biotinylated peptide. The phosphorylated peptide was detected by homogeneous time-resolved fluorescence (HTRF). The IC50 of the compounds is measured for each kinase in 40 microL reactions containing enzyme, ATP, and 500 nM peptide in ₅₀ mM Tris (pH 7.8) buffer containing 100 mM NaCl, 5 mM DTT, and 0.1 mg/mL (0.01%) BSA. For 1 mM _IC50 measurements, the ATP concentration in the reaction is 1 mM. The reactions are carried out at room temperature for 1 hour and then stopped with 20 μL of 45 mM EDTA, 300 nM SA-APC, 6 nM Eu-Py20 assay buffer (Perkin Elmer, Boston, Mass.). Binding to europium-labeled antibodies was carried out for 40 minutes and the HTRF signal was measured on a Fusion plate reader (Perkin Elmer, Boston, Mass.). The compounds in Table 1 were tested in this assay and shown to have the IC ₅₀ values in Table 1.

Example 2. Activated JAK1 is strongly expressed in human CLE skin lesions To investigate the specific role of JAK1-mediated signaling in CLE, the expression of phosphorylated JAK1 (pJAK1) in lesional skin (SCLE and CDLE subsets) was compared with lichen planus (LP) and healthy controls. All punch biopsies of different inflammatory skin disorders (N=34) were obtained from active skin lesions for diagnostic purposes. In healthy controls (N=9), biopsies were obtained from unaffected skin obtained at plastic surgery. Skin samples were fixed overnight in 4% formalin or frozen nitrogen and proceeded to immunohistochemical staining or RNA isolation. RNA was processed by the Next Generation Sequencing (NGS) Core Facility of the Medical Faculty of the University of Bonn using the QuantSeq 3'-mRNA Library Prep Kit by Lexogen. RNA sequencing was performed using an Illumina HiSeq 2500 (50 cycles of standard 3' RNA sequencing).

In CLE skin lesions, pJAK1 expression was significantly increased in keratinocytes from the basal to granular layers and in dermal infiltrating immune cells. We also observed that pJAK1 was significantly increased in lichen planus, an autoimmune disease that shares common histological features with CLE.

Example 2. JAK/STAT-associated innate inflammatory pathways are significantly activated in human CLE skin. H&E staining was performed on lesional skin samples from CLE patients to confirm the clinical diagnosis in each case by an experienced dermatopathologist. Immunohistochemical staining was performed using REAL™ Detection Systems with specific antibodies against pJAK (ABIN196869, antibodies-online), CXCL10 (ab9807, Cambridge, UK), MxA (M143, Haller, Freiburg, Germany) and CD45 (550539, BD, New Jersey) using Fast Red (Agilent, Santa Clara, USA) for chromatin. Expression was assessed using the following:
(Wenzel et al. J. Pathol. 2005, 205:435-442) and semiquantitatively recorded. Immunofluorescence analysis of JAK1 phosphorylation detected by anti-rabbit Rhodamine Red-X (711-295-152; Jackson ImmunoResearch, Baltimore, MD, USA) and DAPI (D9542, Sigma-Aldrich) was performed using a high-resolution microscope (Axio Observer Z1, Zeiss, Germany).

Expression analysis in CLE lesion skin revealed significant activation of genes associated with both innate and adaptive immune pathways compared to healthy controls. In particular, genes for LE-associated proinflammatory chemokines (CXCL10, 9, 11) and other IFN-regulated proteins (OASL, OAS2, Mx1) as well as key drivers in cell death and B cell activation (CXCR3, CASP10, AIM2, TRAIL, BLyS) were highly expressed. As JAK/STAT signaling is a key regulator of inflammatory gene transcription, gene expression of STAT1 in LE lesions was also significantly increased, as shown in Figure 1C. Innate immune pathways upregulated corresponding to individual genes included the JAK/STAT pathway and associated cytokine-/chemokine signaling pathways as well as upstream TLR-dependent and TLR-independent DAMP recognition pathways, as shown in Figure 1D.

Example 3. INCB039110 Significantly Inhibits JAK1 Phosphorylation in Cultured Immortalized Human Keratinocytes To analyze the functional principles and effects of JAK inhibition, the following established in vitro model of CLE was used: Immortalized keratinocytes (HaCaT) were obtained from CLS Cell Lines Service GmbH, Eppelheim, Germany, normal human epidermal keratinocytes (NHEK, FC-0025) and human epidermal equivalents (epiCS, CS-1001) were obtained from CellSystems, Troisdorf, Germany. These cell lines were cultured according to the manufacturer's protocols. Cultured keratinocytes were stimulated with endogenous nucleic acids (eNA, 1.25 μg/mL) isolated from unstimulated keratinocytes using the "Genomic DNA from tissue" kit (Machery-Nagel, Düren, Germany). Lipofectamine 2000 (Invitrogen, Carlsbad, USA) served as the transfection reagent (2.5 μL/mL). INCB039110, and ruxolitinib (Selleckchem, Eching, Germany) were added to a final concentration of 1 μM, and JAK3-selective FM-381 was used (100 nM) as recommended (see, e.g., Forster et al, Cell Chem. Biol. 2016, 23:1335-1340). All experiments were performed in biological triplicates. Enzyme-linked immunosorbent assay for human CXCL10 (DY266-05 R&D systems) was performed using DuoSet Ancillary Reagent Kit 2 (DY008 R&D systems) according to the provided protocol, measured by Synergy HT Multi-Detection Multiplate Reader (BioTek, Winooski, VT, USA) and read with Gen5 software (version 1.11.5).

JAK1 phosphorylation was strongly enhanced in immortalized keratinocytes (HaCaT) after stimulation with eNA, consistent with the findings in CLE skin lesions described above (see Examples 2 and 3). As shown in Figures 2A and 2B, JAK1-specific INCB039110 and JAK1/2-specific ruxolitinib significantly reduced JAK1 activation in stimulated cells.

Example 4. Pharmacological JAK1 inhibition blocks the expression of inflammatory cytokines and pathway molecules typical of CLE in vitro To investigate the efficacy of pharmacological JAK1 inhibition, in vitro analysis was performed in three different CLE models: (i) NHEK cells, (ii) HaCaT cells, and (iii) 3D epidermal equivalents. As shown in Figure 2C, pharmacological JAK1 inhibition induced a significant downregulation of genes encoding key drivers of innate inflammatory pathways such as IFN-regulated chemokines (CXCL10, CXCL11), cell death (TRAIL, AIM2, TREX1) and crosstalk to adaptive immune cells (BlyS) in primary keratinocytes (NHEK) compared to untreated eNA inflammatory cells. The relevant downregulated KEGG pathways are shown in Table A. These results were confirmed in HaCaT cells, where both JAK1 and JAK1/2 inhibitors significantly reduced CXCL10 protein expression.
^a KEGG pathways were classified using Database for Annotation, Visualization and Integrated Discovery (DAVID ver. 6.8). P-values were generated with EASE Score. Count: Number of genes downregulated by more than 2-fold by INCB039110 in NHEK within each KEGG pathway.

The in vitro data disclosed herein demonstrate that JAK1 selective inhibitors are as potent as JAK1/2 inhibitors in suppressing CLE typical cytokines and prevent expression of genes encoding proinflammatory chemokines (CXCL10, 11), lymphocyte activating factor (BLyS) (see e.g., Wenzel et al, Exp. Dermatol. 2018, 27:95-97) and cell death promoters (TRAIL (see e.g., Zahn et al, Br. J. Dermatol. 2011, 165:1118-1123), AIM2, caspase 10, TREX1), as shown in Figure 2D. Interestingly, inhibition of JAK3 did not result in a reduction in CXCL10 expression, a central mediator of the "interface dermatitis" typical of CLE, as shown in Figure 2D. Without being bound by theory, this may explain the early failure of the JAK3/SYK blocker R333 in CLE clinical trials (see, e.g., Presto et al, Br. J. Dermatol. 2018, 178:1308-1314). In 3D epidermal equivalents, exposure of a JAK1-selective inhibitor to stimulatory epiCS revealed a significant reduction in CXCL10 protein expression, consistent with the findings above, as shown in FIG. 2E.

Example 5. In vivo topical application of INCB039110 ameliorates CLE-like lesions in lupus-prone TREX1 ^−/− mice TREX1 ^{−/− mice (established on a C57BL/6J background; Cancer Research Institute, London, UK) were bred and housed under specific pathogen-free conditions at the Animal Core Facility of UKB Bonn (HET, Bonn, Germany). TREX1 −/ −} mice (n=8) were shaved on the back and treated with 0.2% DNFB (1-fluoro-2,4-dinitrobenzol, Sigma Aldrich). After 4 days, UV irradiation was performed using a UV801KL (Waldmann, Villingen-Schwenningen, Germany) starting with 450 mJ/ ^cm2 UVB for 115 seconds per day for 3 consecutive days. 1% INCB039110 or vehicle dissolved in DMSO and olive oil (50 μL/mouse) was applied topically for 7 days. Mice were photographed daily and weighed every 2 days.

TREX1 ^-/- mice spontaneously developed CLE-like erythrosquamous partially ulcerated skin lesions at a certain age, which were enhanced after challenge with UVB. As shown in Figures 3A and 3B, topical treatment with the JAK1-specific INCB039110 for 7 days resulted in a sustained improvement in lesional skin with respect to erythema, induration, scaling, and size, and a significant reduction in the Lupus-Skin-Activity-Score (adapted CLASI score) compared to placebo-treated mice. Furthermore, distinct histological features such as epidermal thickness and infiltrating dermal immune cells were significantly improved by JAK1 inhibition, as shown in Figure 3C.

The data disclosed in Examples 2-5 demonstrate that JAK1-specific inhibition significantly reduces the expression of inflammatory cytokines typical of CLE in vitro and in vivo. Topical application of JAK1-selective inhibitors was highly effective in treating CLE-like lesions in lupus-prone mice, supporting their potential use in human CLE.

In addition to those described herein, various modifications of the present invention will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited within this disclosure, including all patents, patent applications, and publications, is hereby incorporated by reference in its entirety.
The present application also includes the following aspects.
[Aspect 1]
1. A method of treating a disease selected from cutaneous lupus erythematosus (CLE) and lichen planus (LP) in a patient in need thereof, 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,
4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-[4-fluoro-2-(trifluoromethyl)phenyl]piperidine-1-carboxamide,
[3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(1-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)azetidin-3-yl]acetonitrile,
4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide,
((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile,
3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,
3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,
4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile,
4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile,
[trans-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-3-(4-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperazin-1-yl)cyclobutyl]acetonitrile,
{trans-3-(4-{[4-[(3-hydroxyazetidin-1-yl)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
{trans-3-(4-{[4-{[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
{trans-3-(4-{[4-{[(2R)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile,
5-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide,
4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide,
5-{3-(cyanomethyl)-3-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide,
{1-(cis-4-{[6-(2-hydroxyethyl)-2-(trifluoromethyl)pyrimidin-4-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
{1-(cis-4-{[4-[(ethylamino)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
{1-(cis-4-{[4-(1-hydroxy-1-methylethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
{1-(cis-4-{[4-{[(3R)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
{1-(cis-4-{[4-{[(3S)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,
{trans-3-(4-{[4-({[(1S)-2-hydroxy-1-methylethyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
{trans-3-(4-{[4-({[(2R)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile {trans-3-(4-{[4-({[(2S)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile, and
{trans-3-(4-{[4-(2-hydroxyethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile,
or a pharma- ceutically acceptable salt of any of the foregoing, in a therapeutically effective amount to said patient.
[Aspect 2]
The method of aspect 1, wherein the JAK1 selective inhibitor, or a pharma- ceutically acceptable salt thereof, is selective for JAK1 over JAK2, JAK3, and TYK2.
[Aspect 3]
3. The method of claim 1 or 2, wherein the JAK1 selective 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 4]
The method of claim 3, wherein the pharma- ceutically acceptable salt is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate.
[Aspect 5]
3. The method of claim 1 or 2, wherein the JAK1 selective 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.
[Aspect 6]
The method of claim 5, wherein the pharma- ceutically acceptable salt is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide phosphate.
[Aspect 7]
3. The method of claim 1 or 2, wherein the JAK1 selective 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.
[Aspect 8]
The method of any one of the preceding claims, wherein the JAK1 selective inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile monohydrate.
[Aspect 9]
Aspect 9. The method of any one of aspects 1-8, further comprising administering to said patient an additional therapeutic agent.
[Aspect 10]
The method of aspect 9, wherein said additional therapeutic agent is selected from a JAK1/JAK2 inhibitor, a JAK1/JAK3 inhibitor, a TYK2 inhibitor, or a pharma- ceutically acceptable salt of any of the foregoing.
[Aspect 11]
The method of aspect 10, wherein the additional therapeutic agent is a JAK1/JAK2 inhibitor, or a pharma- ceutically acceptable salt thereof.
[Aspect 12]
12. The method of aspect 10 or 11, wherein the JAK1/JAK2 inhibitor, or a pharma- ceutically acceptable salt thereof, is selective for JAK1 and JAK2 over JAK3 and TYK2.
[Aspect 13]
The method of any one of aspects 10-12, wherein said JAK1/JAK2 inhibitor is ruxolitinib, or a pharma- ceutically acceptable salt thereof.
[Aspect 14]
The method of aspect 13, wherein said method comprises local administration of said ruxolitinib, or a pharma- ceutically acceptable salt thereof, to said patient.
[Aspect 15]
The method of aspect 13, wherein the method comprises oral administration of the ruxolitinib, or a pharma- ceutically acceptable salt thereof, to the patient.
[Aspect 16]
16. The method of any one of aspects 13 or 15, wherein the pharma- ceutically acceptable salt is ruxolitinib phosphate.
[Aspect 17]
The method of any one of aspects 10-12, wherein said JAK1/JAK2 inhibitor is ruxolitinib, or a pharma- ceutically acceptable salt thereof, wherein one or more hydrogen atoms are replaced by deuterium atoms.
[Aspect 18]
The method of aspect 10, wherein the additional therapeutic agent is a JAK1/JAK3 inhibitor, or a pharma- ceutically acceptable salt thereof.
[Aspect 19]
The method of aspect 10 or 18, wherein the JAK1/JAK3 inhibitor is tofacitinib, or a pharma- ceutically acceptable salt thereof.
[Aspect 20]
Aspect 20. The method of any one of aspects 1-19, wherein said method comprises local administration of said JAK1 selective inhibitor to said patient.
[Aspect 21]
Aspect 20. The method of any one of aspects 1-19, wherein said method comprises oral administration of said JAK1 selective inhibitor to said patient.
[Aspect 22]
22. The method according to any one of aspects 1 to 21, wherein said disease is cutaneous lupus erythematosus (CLE).
[Aspect 23]
23. The method of aspect 22, wherein said cutaneous lupus erythematosus (CLE) is selected from subacute cutaneous lupus erythematosus (SCLE) and chronic discoid lupus erythematosus (CDLE).
[Aspect 24]
22. The method according to any one of aspects 1 to 21, wherein said disease is Lichen planus (LP).

Claims (16)

A pharmaceutical for treating cutaneous lupus erythematosus (CLE ) , comprising a JAK1 selective inhibitor , wherein the JAK1 selective 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 . The pharmaceutical agent according to claim 1, wherein the JAK1 selective 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. The pharmaceutical composition of claim 1 or 2 , which is administered together with an additional therapeutic agent. The pharmaceutical composition of claim 3 , wherein the additional therapeutic agent is selected from a JAK1/JAK2 inhibitor, a JAK1/JAK3 inhibitor, a TYK2 inhibitor, or a pharma- ceutically acceptable salt of any of the foregoing. The pharmaceutical composition of claim 4 , wherein the additional therapeutic agent is a JAK1/JAK2 inhibitor, or a pharma- ceutically acceptable salt thereof. The pharmaceutical agent according to claim 4 or 5 , wherein the JAK1/JAK2 inhibitor, or a pharma- ceutically acceptable salt thereof, is selective for JAK1 and JAK2 over JAK3 and TYK2. The pharmaceutical agent according to any one of claims 4 to 6 , wherein the JAK1/JAK2 inhibitor is ruxolitinib or a pharma- ceutically acceptable salt thereof. The method of claim 7 , wherein the ruxolitinib, or a pharma- ceutically acceptable salt thereof, is administered locally. The method of claim 7 , wherein the ruxolitinib, or a pharma- ceutically acceptable salt thereof, is administered orally. The pharmaceutical composition according to claim 7 or 9 , wherein the pharma- ceutically acceptable salt of ruxolitinib is ruxolitinib phosphate. The pharmaceutical agent according to any one of claims 4 to 6, wherein the JAK1/JAK2 inhibitor is ruxolitinib, or a pharma- ceutically acceptable salt thereof, in which one or more hydrogen atoms are replaced by deuterium atoms. The pharmaceutical composition of claim 4 , wherein the additional therapeutic agent is a JAK1/JAK3 inhibitor, or a pharma- ceutically acceptable salt thereof. The pharmaceutical agent according to claim 4 or 12 , wherein the JAK1/JAK3 inhibitor is tofacitinib, or a pharma- ceutically acceptable salt thereof. The pharmaceutical composition according to any one of claims 1 to 13 , which is for topical administration. The pharmaceutical composition according to any one of claims 1 to 13 , which is for oral administration. The pharmaceutical composition according to any one of claims 1 to 15 , wherein the cutaneous lupus erythematosus (CLE) is selected from subacute cutaneous lupus erythematosus (SCLE) and chronic discoid lupus erythematosus (CDLE).