Pyrazolopyrimidine aryl ether inhibitors of JAK kinases and their use

Abstract

Described herein are compounds and salts thereof useful as JAK kinase inhibitors. Also provided are pharmaceutical compositions comprising such JAK inhibitors and a pharmaceutically acceptable carrier, adjuvant, or vehicle, and methods for treating or lessening the severity of a disease or condition in a patient that responds to the inhibition of Janus kinase activity. JPEG2022537354000161.jpg45170

Description

Cross - reference to related applications

[0001] This application claims the priority of International Application No. PCT / CN2019 / 091710 filed on June 18, 2019 and U.S. Provisional Application No. 63 / 036,577 filed on June 9, 2020, and the disclosures of these are incorporated herein by reference.

Technical Field

[0002] The present invention relates to compounds that are inhibitors of Janus kinases such as JAK1 and JAK2, compositions containing these compounds, and methods of use including, but not limited to, the diagnosis or treatment of patients suffering from conditions responsive to the inhibition of JAK kinases.

Background Art

[0003] Cytokine pathways mediate a wide range of biological functions including many aspects of inflammation and immunity. Janus kinases (JAKs), including JAK1, JAK2, JAK3 and TYK2, are cytoplasmic protein kinases that associate with type I and type II cytokine receptors and regulate cytokine signaling. Engagement of a cytokine with its cognate receptor induces activation of the receptor - associated JAK, which in turn results in JAK - mediated tyrosine phosphorylation of signal - transducer and activator of transcription (STAT) proteins and ultimately activation of the transcriptional activity of specific gene sets (Schindler et al., 2007, J. Biol. Chem. 282:20059 - 63). JAK1, JAK2 and TYK2 exhibit a broad gene expression pattern, while JAK3 expression is limited to leukocytes. Cytokine receptors typically function as heterodimers, and as a result, more than one type of JAK kinase usually associates with the cytokine receptor complex. The specific JAKs that associate with different cytokine receptor complexes are often determined by genetic studies and supported by other experimental evidence. Exemplary therapeutic benefits of JAK enzyme inhibition are discussed, for example, in International Publication No. WO 2013 / 014567.

[0004] JAK1 was first identified in the screening of novel kinases (Wilks A.F., 1989, Proc. Natl. Acad. Sci. U.S.A. 86:1603-1607). Genetic and biochemical studies have shown that JAK1 is functionally and physically associated with type I interferon (e.g., IFN alpha), type II interferon (e.g., IFN gamma), and the IL-2 and IL-6 cytokine receptor complexes (Kisseleva et al., 2002, Gene 285:1-24; Levy et al., 2005, Nat. Rev. Mol. Cell Biol. 3:651-662; O’Shea et al., 2002, Cell, 109(suppl.):S121-S131). JAK1 knockout mice die during the perinatal period due to a defect in LIF receptor signaling (Kisseleva et al., 2002, Gene 285:1-24; O’Shea et al., 2002, Cell, 109(suppl.):S121-S131). Characterization of tissues derived from JAK1 knockout mice has demonstrated an important role for this kinase in the IFN, IL-10, IL-2 / IL-4, and IL-6 pathways. A humanized monoclonal antibody (tocilizumab) targeting the IL-6 pathway has been approved by the European Commission for the treatment of moderate to severe rheumatoid arthritis (Scheinecker et al., 2009, Nat. Rev. Drug Discov. 8:273-274).

[0005] CD4 T cells play an important role in the pathogenesis of asthma through the production of TH2 cytokines in the lung, including IL-4, IL-9, and IL-13 (Cohn et al., 2004, Annu. Rev. Immunol. 22:789-815). IL-4 and IL-13 induce increased mucus production, eosinophil recruitment to the lung, and increased IgE production (Kasaian et al., 2008, Biochem. Pharmacol. 76(2):147-155). IL-9 leads to mast cell activation, which exacerbates asthmatic symptoms (Kearley et al., 2011, Am. J. Resp. Crit. Care Med., 183(7):865-875). The IL-4Rα chain activates JAK1 and binds to either IL-4 or IL-13 when combined with the common γ chain or the IL-13Rα1 chain, respectively (Pernis et al., 2002, J. Clin. Invest. 109(10):1279-1283). The common γ chain can also bind to IL-9 in combination with IL-9Rα, and IL-9Rα similarly activates JAK1 (Demoulin et al., 1996, Mol. Cell Biol. 16(9):4710-4716). The common γ chain activates JAK3, but JAK1 is dominant over JAK3, and inhibition of JAK1 has been shown to be sufficient to inactivate signaling through the common gamma chain despite JAK3 activity (Haan et al., 2011, Chem. Biol. 18(3):314-323). Inhibition of IL-4, IL-13, and IL-9 signaling by blocking the JAK / STAT signaling pathway can alleviate asthmatic symptoms in preclinical models of lung inflammation (Mathew et al., 2001, J. Exp. Med. 193(9):1087-1096; Kudlacz et al., 2008, Eur. J. Pharmacol. 582(1-3):154-161).

[0006] Biochemical and genetic studies have shown an association of JAK2 with single-chain (e.g., EPO), IL-3, and interferon γ cytokine receptor families (Kisseleva et al., 2002, Gene 285:1-24; Levy et al., 2005, Nat. Rev. Mol. Cell Biol. 3:651-662; O’Shea et al., 2002, Cell, 109(suppl.):S121-S131). Consistent with this, JAK2 knockout mice die of anemia (O’Shea et al., 2002, Cell, 109(suppl.):S121-S131). Kinase activation mutations in JAK2 (e.g., JAK2 V617F) are associated with myeloproliferative disorders in humans. Furthermore, JAK2 associates with receptors for cytokines such as IL-5 and thymic stromal lymphopoietin (TSLP). IL-5 is an important cytokine responsible for eosinophil differentiation, growth, activation, survival, and mobilization to the airways (Pelaia et al., 2019, Front. Physiol., 10:1514; Stirling et al., 2001, Am. J. Respir. Crit. Care Med., 164:1403-9; Fulkerson and Rothenberg, 2013, Nat. Rev. Drug Discov., 12:117-9.; Varricchi and Canonica, 2016, Expert. Rev. Clin. Immunol., 12:903-5). Three monoclonal antibody drugs targeting either IL-5 (mepolizumab, reslizumab) or the α-chain of its receptor (benralizumab) are approved for the treatment of asthma with an eosinophilic phenotype. TSLP is an epithelial cell-derived cytokine that plays an important role in the regulation of type II immunity and acts as an alarmin upstream of TH2 cytokine production (Kitajima et al., 2011, Eur J Immunol., 41:1862-71). Tezepelumab is an antagonist antibody against TSLP. The results of a phase 2 trial have shown successful reduction of asthma exacerbation in both patients with and without a type 2 signature (Corren et al., 2017, 377:936-46).

[0007] JAK3 exclusively associates with the common gamma cytokine receptor chain present in the IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21 cytokine receptor complexes. JAK3 is important for the development and proliferation of lymphoid cells, and mutations in JAK3 result in severe combined immunodeficiency (SCID) (O’Shea et al., 2002, Cell, 109 (suppl.): S121-S131). Based on its role in lymphocyte regulation, JAK3 and JAK3-mediated pathways have been targeted for immunosuppressive indications such as transplant rejection and rheumatoid arthritis (Baslund et al., 2005, Arthritis & Rheumatism 52:2686-2692; Changelian et al., 2003, Science 302:875-878).

[0008] TYK2 associates with the type I interferon (e.g., IFNα), IL-6, IL-10, IL-12, and IL-23 cytokine receptor complexes (Kisseleva et al., 2002, Gene 285:1-24; Watford, W.T. & O’Shea, J.J., 2006, Immunity 25:695-697). Consistent with this, primary cells derived from TYK2-deficient humans are defective in type I interferon, IL-6, IL-10, IL-12, and IL-23 signaling. A fully human monoclonal antibody (ustekinumab) that targets the shared p40 subunit of the IL-12 and IL-23 cytokines was recently approved by the European Commission for the treatment of moderate to severe plaque psoriasis (Krueger et al., 2007, N. Engl. J. Med. 356:580-92; Reich et al., 2009, Nat. Rev. Drug Discov. 8:355-356). Furthermore, antibodies targeting the IL-12 and IL-23 pathways have been tested in clinical trials for the treatment of Crohn’s disease (Mannon et al., 2004, N. Engl. J. Med. 351:2069-79).

[0009] WO 2010 / 051549, WO 2011 / 003065, WO 2015 / 177326 and WO 2017 / 089390 describe certain pyrazolopyrimidine compounds reported to be useful as inhibitors of one or more Janus kinases. Data are presented for certain compounds that exhibit inhibition of JAK1 as well as JAK2, JAK3 and / or TYK2 kinases.

[0010] There remains a need for additional compounds that are inhibitors of Janus kinases. For example, compounds are needed that have useful efficacy as inhibitors of one or more Janus kinases (e.g., JAK1 and JAK2) in combination with other pharmacological properties required to achieve a useful therapeutic benefit. For example, compounds are needed that exhibit selectivity of one Janus kinase over other kinases (e.g., selectivity of JAK1 and / or JAK2 over other kinases such as leucine rich repeat kinase 2 (LRRK2)). Compounds are also needed that exhibit selectivity of one Janus kinase over other Janus kinases (e.g., selectivity of JAK1 and / or JAK2 over JAK3 and / or TYK2). Compounds that exhibit selectivity for both JAK1 and JAK2 over JAK3 and TYK2 may provide a therapeutic benefit in conditions that respond to inhibition of JAK1. Further, there is a current need for potent JAK1 inhibitors that have other properties (e.g., melting point, pK, solubility, etc.) required for formulation and administration by inhalation. Such compounds would be particularly useful for treating conditions such as asthma.

[0011] Accordingly, there is a need in the art for further or alternative treatments of JAK kinase-mediated conditions such as those described above. In particular, there is a need for JAK1 and JAK2 kinase inhibitors that are useful for inhaled delivery in the treatment of airway inflammatory conditions such as asthma.

[0012] Pyrazolopyrimidines that inhibit JAK kinases, such as pyrazolopyrimidines selected from compounds of formula (I), stereoisomers or salts thereof, such as pharmaceutically acceptable salts thereof, are provided herein. The JAK kinase can be JAK1, JAK2, or both.

Prior Art Documents

Patent Documents

[0013]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Non-Patent Documents

[0014]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Non-Patent Document 6

Non-Patent Document 7

Non-Patent Document 8

Non-Patent Document 9

Non-Patent Document 10

Non-Patent Document 11

Non-Patent Document 12

Non-Patent Document 13

Non-Patent Document 14

Non-Patent Document 15

Non-Patent Document 16

Non-Patent Document 17

Non-Patent Document 18

Non-Patent Document 19

Non-Patent Document 20

Non-Patent Document 21

Non-Patent Document 22

Non-Patent Document 23

Non-Patent Document 24

Non-Patent Document 25

Non-Patent Document 26

[0015] One embodiment is of formula (I): TIFF0007716993000001.tif53170[wherein, Ar is phenyl; 1,2,3,4-tetrahydroisoquinolinyl; pyrazolyl; pyridinyl; or pyridazinyl: R 1 , , k , 1 , , a , m , , , a , a , , a , , h , , 1 ,

[0015] , 3-6 , is hydrogen; C1-C6 alkyl; halo-C1-C6 alkyl; hydroxy-C1-C6 alkyl; -(CHR a ) h -het 1 ; -(CHR a ) k -NR a -het 1 ; or -(CHR a ) m -C 3-6 cycloalkyl, and the cycloalkyl moiety may be unsubstituted or Rd may be replaced one or two times; each R 2 is independently: C1-C6 alkyl; hydroxy-C1-C6 alkyl; halo-C1-C6 alkyl; C1-C6 alkoxy; C1-C6 alkoxy-C1-C6 alkyl; halo-C1-C6 alkoxy; halo-C1-C6 alkoxy-C1-C6 alkyl; C1-C6 alkyl-SO2-C1-C6 alkyl; hydroxyl; cyano; cyano-C1-C6 alkyl; halo; acetyl; -(CHR a ) p -het 2 ; -(CHR a ) q -NR b R c ; -(CHR a ) r -C(O)-NR b R c ; -(CHR a ) s -NR a -(CHR a ) s -C(O)-NR b R c ; or -(CHR a ) t -C 3-6 is cycloalkyl, and the cycloalkyl moiety may be unsubstituted or may be substituted one or two times with R e ; R 3 , R 4 and R 5 are each independently: hydrogen; or C1-C6 alkyl; each R a is independently: hydrogen; or C 1-6 alkyl; each R b is independently: hydrogen; C 1-6 alkyl; or hydroxy-C1-C6 alkyl; each R c is independently: hydrogen; C 1-6alkyl; hydroxy-C1-C6 alkyl; cyano-C1-C6 alkyl; C1-C6 alkoxy-C1-C6 alkyl; oxetanyl; 2-morpholinoethyl; 1-methyl-azetidin-3-yl; 2-(N,N-dimethylamino)-ethyl; hydroxycyclobutyl; or 3-(N,N-dimethylamino)-pyrrolidin-1-yl; -(CHR a ) u -C 3-6 is cycloalkyl, and the cycloalkyl moiety may be unsubstituted or substituted one or two times with R e ; or R b and R c may together with the nitrogen atom to which they are attached form het 3 ; each R d is independently: C1-C6 alkyl, hydroxy or halo; each R e is independently: C 1-6 alkyl; hydroxyl; cyano-C1-C6 alkyl; hydroxy-C1-C6 alkyl; morpholinyl; or -(CHR a ) v -NR g R h wherein R g and R h are each independently hydrogen or C 1-6 alkyl; h is 0 to 2; k is 0 to 2; m is 0 to 2; n is 0 to 2; p is 0 to 2; q is 0 to 2; r is 0 to 2; s is 0 to 2; t is 0 to 2; u is 0 to 2; v is 0 to 2; het 1is oxetanyl; tetrahydrofuranyl; tetrahydropyranyl; or pyrrolodinyl, each of which may be unsubstituted or substituted one or two times with R d and may be substituted one or two times with R het 2 is: azetidinyl; pyrrolidinyl; oxetanyl; piperidinyl; morpholinyl; piperazinyl; azepinyl; quinuclidinyl; or pyrazolyl, each of which may be unsubstituted or substituted one or two times with R e and may be substituted one or two times with R het 3 is: azetidinyl; pyrrolidinyl; piperidinyl; morpholinyl; piperazinyl; or azepinyl, each of which may be unsubstituted or substituted one or two times with R e and may be substituted one or two times with R] to provide a compound of or a pharmaceutically acceptable salt thereof.

[0016] Also provided is a pharmaceutical composition comprising a JAK inhibitor or a pharmaceutically acceptable salt thereof described herein and a pharmaceutically acceptable carrier, diluent or excipient.

[0017] Also provided is the use of a JAK inhibitor or a pharmaceutically acceptable salt thereof described herein in the treatment, for example, in the treatment of inflammatory diseases (such as asthma). Also provided is the use of a JAK inhibitor or a pharmaceutically acceptable salt thereof described herein for the preparation of a medicament for treating an inflammatory disease. Also provided is a method of preventing, treating or reducing the severity of a disease or condition responsive to inhibition of Janus kinase activity in a patient, comprising administering to the patient a therapeutically effective amount of a JAK inhibitor or a pharmaceutically acceptable salt thereof described herein.

[0018] The most well-validated cytokines in asthma (IL-4, IL-5, IL-9, IL-13, and TSLP) all signal through JAK1 and / or JAK2. The compounds of the present invention are active against both JAK1 and JAK2. Some of these compounds do not optimally have balanced co-activity against both JAK1 and JAK2, or have much higher activity against one of these kinases than the other, but rather have a slightly higher affinity for JAK1 than for JAK2. The subject compounds also have good selectivity against off-target kinases such as LRRK2 associated with lung toxicity.

[0019] Many compounds can show high affinity for both JAK1 and JAK2 in simple biochemical assays, but not all such compounds are effective in mediating the relevant cytokines associated with JAK1 and JAK2. The specific compounds of the present invention, in addition to being active against both JAK1 and JAK2, have been shown to be effective in mediating asthma-related cytokines associated with JAK1 and JAK2 in cell-based assays.

[0020] The compounds of the present invention also exhibit favorable pharmacokinetic (PK) properties in lung tissue and are useful for inhalation therapy. When administered via the inhalation route using techniques such as dry powder inhalation (DPI) or intranasal (IN) delivery, certain compounds unexpectedly show sustained retention within lung tissue and much lower concentrations in the systemic circulation. Such improved PK properties can advantageously result in lower dosage and less frequent dosing requirements for effective treatment. Certain compounds show unexpectedly improved solubility, which also provides improved efficacy in the lung. Specific compounds of the present invention also show an unexpected decrease in cytotoxicity compared to other JAK inhibitors. **Modes for Carrying Out the Invention**

[0021] Definitions "Halogen" or "halo" refers to F, Cl, Br or I. Further, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl, where one or more halogens replace the hydrogen(s) of an alkyl group.

[0022] The term "alkyl" refers to a saturated straight-chain or branched-chain monovalent hydrocarbon radical, and the alkyl radical may be optionally substituted. In one example, the alkyl radical has from 1 to 18 carbon atoms (C1-C 18 ) is. In other examples, the alkyl radical is C0-C6, C0-C5, C0-C3, C1-C 12 , C1-C 10,It is C1-C8, C1-C6, C1-C5, C1-C4, or C1-C3. C0 alkyl refers to a bond. Examples of alkyl groups include methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, -CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH3)3), 1-pentyl (n-pentyl, -CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3, 1-heptyl, and 1-octyl, etc.In some embodiments, examples of substituents for "optionally substituted alkyl" include F, Cl, Br, I, OH, SH, CN, NH2, NHCH3, N(CH3)2, NO2, N3, C(O)CH3, COOH, CO2CH3, methyl, ethyl, propyl, iso-propyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methanesulfonylamino, SO, SO2, phenyl, piperidinyl, piperidinyl and pyrimidinyl, and the alkyl, phenyl and heterocyclic moieties may be optionally substituted by, for example, one to four examples of substituents selected from the same list.

[0023] The term "alkenyl" refers to a straight or branched chain monovalent hydrocarbon radical having at least one unsaturated site, i.e., a carbon-carbon double bond, and the alkenyl radical may be optionally substituted and includes radicals having "cis" and "trans" orientations, or "E" and "Z" orientations. In one example, the alkenyl radical has 2 to 18 carbon atoms (C2-C 18 )). In other examples, the alkenyl radical is C2-C 12 , C2-C 10、It is C2-C8, C2-C6 or C2-C3. Examples include, but are not limited to, ethenyl or vinyl (-CH=CH2), prop-1-enyl (-CH=CHCH3), prop-2-enyl (-CH2CH=CH2), 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, but-1,3-dienyl, 2-methylbut-1,3-diene, hexa-1-enyl, hexa-2-enyl, hexa-3-enyl, hexa-4-enyl, and hexa-1,3-dienyl. In some embodiments, the substituents of "optionally substituted alkenyl" include F, Cl, Br, I, OH, SH, CN, NH2, NHCH3, N(CH3)2, NO2, N3, C(O)CH3, COOH, CO2CH3, methyl, ethyl, propyl, iso-propyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methanesulfonylamino, SO, SO2, phenyl, piperidinyl, 1 to 4 examples of piperidinyl and pyrimidinyl, and the alkyl, phenyl and heterocyclic moieties thereof may be optionally substituted by 1 to 4 examples of substituents selected from the same list.

[0024] The term "alkynyl" refers to a straight-chain or branched-chain monovalent hydrocarbon radical having at least one unsaturated site, i.e., a carbon-carbon triple bond, and the alkynyl radical may be optionally substituted. In one example, the alkynyl radical has 2 to 18 carbon atoms (C2-C 18 ) is. In another example, the alkynyl radical is C2-C 12 , C2-C 10、It is C2-C8, C2-C6 or C2-C3. Examples include, but are not limited to, ethynyl (-C≡CH), prop-1-ynyl (-C≡CCH3), prop-2-ynyl (propargyl, -CH2C≡CH), but-1-ynyl, but-2-ynyl, and but-3-ynyl. In some embodiments, the substituents of "optionally substituted alkynyl" include F, Cl, Br, I, OH, SH, CN, NH2, NHCH3, N(CH3)2, NO2, N3, C(O)CH3, COOH, CO2CH3, methyl, ethyl, propyl, iso-propyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methanesulfonylamino, SO, SO2, phenyl, piperidinyl, 1 to 4 examples of piperidinyl and pyrimidinyl, and the alkyl, phenyl and heterocyclic moieties thereof may be optionally substituted by 1 to 4 examples of substituents selected from the same list.

[0025] "Alkylene" means a saturated, branched, or straight-chain hydrocarbon group having two monovalent radical centers derived by removing two hydrogen atoms from the same or two different carbon atoms of the parent alkane. In one example, the divalent alkylene group has 1 to 18 carbon atoms (C1-C 18 )). In other examples, the divalent alkylene group is C0-C6, C0-C5, C0-C3, C1-C 12 , C1-C 10、 C1-C8, C1-C6, C1-C5, C1-C4, or C1-C3. The C0 alkylene group refers to a bond. Exemplary alkylene groups include (-CH2-), 1,1-ethyl (-CH(CH3)-), (1,2-ethyl (-CH2CH2-), 1,1-propyl (-CH(CH2CH3)-), 2,2-propyl (-C(CH3)2-), 1,2-propyl (-CH(CH3)CH2-), 1,3-propyl (-CH2CH2CH2-), 1,1-dimethylethane-1,2-yl (-C(CH3)2CH2-), 1,4-butyl (-CH2CH2CH2CH2-), etc.

[0026] The term "heteroalkyl" refers to a straight or branched chain monovalent hydrocarbon radical consisting of a specified number of carbon atoms, or up to 18 carbon atoms if not specified, and 1 to 5 heteroatoms selected from the group consisting of O, N, Si, and S, where the nitrogen and sulfur atoms can be optionally oxidized and the nitrogen heteroatoms can be optionally quaternized. In some embodiments, the heteroatoms are selected from O, N, and S, the nitrogen and sulfur atoms may be optionally oxidized, and the nitrogen heteroatoms may be optionally quaternized. The heteroatom(s) can be located at any internal position of the heteroalkyl group, including the position where the alkyl group is attached to the rest of the molecule (e.g., -O-CH2-CH3). Examples include -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -Si(CH3)3, and -CH2-CH=N-OCH3. For example, up to 2 heteroatoms may be consecutive, such as -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. The heteroalkyl group may be optionally substituted. In some embodiments, substituents for "optionally substituted heteroalkyl" include F, Cl, Br, I, OH, SH, CN, NH2, NHCH3, N(CH3)2, NO2, N3, C(O)CH3, COOH, CO2CH3, methyl, ethyl, propyl, iso-propyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methanesulfonylamino, SO, SO2, phenyl, piperidinyl, piperidinyl, and 1 to 4 examples of pyrimidinyl, and the alkyl, phenyl, and heterocyclic moieties thereof may be optionally substituted by 1 to 4 examples of substituents selected from the same list.

[0027] "Amino" means a primary (i.e., -NH2), secondary (i.e., -NRH), tertiary (i.e., -NRR), and quaternary (i.e., -N(+)RRR) amine, which is optionally substituted, wherein each R is the same or different and is selected from alkyl, cycloalkyl, aryl, and heterocyclyl, and the alkyl, cycloalkyl, aryl, and heterocyclyl groups are as defined herein. In many such embodiments, R is C1-C6 alkyl. Specific secondary and tertiary amines are alkylamine, dialkylamine, arylamine, diarylamine, aralkylamine, and dialkylamine, and the alkyl and aryl moieties may be optionally substituted. Specific secondary and tertiary amines are methylamine, ethylamine, propylamine, isopropylamine, phenylamine, benzylamine, dimethylamine, diethylamine, dipropylamine, and diisopropylamine. In some embodiments, the R groups of the quaternary amine are each independently an optionally substituted alkyl group.

[0028] "Aryl" means a carbocyclic aromatic group having a specified number of carbon atoms, or, when the number is not specified, up to 14 carbon atoms, whether or not fused to one or more groups. As an example, an aryl group having 6 to 14 carbon atoms can be mentioned. As another example, an aryl group having 6 to 10 carbon atoms can be mentioned. Examples of aryl groups include phenyl, naphthyl, biphenyl, phenanthrenyl, naphthacenyl, 1,2,3,4-tetrahydronaphthalenyl, 1H-indenyl, 2,3-dihydro-1H-indenyl, etc. (for example, Lang’s Handbook of Chemistry can be mentioned (Dean, J.A., ed.) 13 thSee ed. Table 7-2

[1985] . The specific aryl is phenyl. Substituted phenyl or substituted aryl is selected from groups such as F, Cl, Br, I, OH, SH, CN, NH2, NHCH3, N(CH3)2, NO2, N3, C(O)CH3, COOH, CO2CH3, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methanesulfonylamino, SO, SO2, phenyl, piperidinyl, piperidinyl and pyrimidinyl, etc., as specified herein (see the definition of "optionally substituted"), and means a phenyl group or aryl group substituted with 1, 2, 3, 4 or 5 substituents, for example 1 to 2, 1 to 3 or 1 to 4 substituents, and its alkyl, phenyl and heterocyclic moieties may be optionally substituted by 1 to 4 examples of substituents selected from the same list.Examples of the term "substituted phenyl" include mono- or di(halo)phenyl groups such as 2-chlorophenyl, 2-bromophenyl, 4-chlorophenyl, 2,6-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-bromophenyl, 3,4-dibromophenyl, 3-chloro-4-fluorophenyl, 2-fluorophenyl, 2,4-difluorophenyl, etc.; mono- or di(hydroxy)phenyl groups such as 4-hydroxyphenyl, 3-hydroxyphenyl, 2,4-dihydroxyphenyl, etc., and their protected hydroxy derivatives, etc.; nitrophenyl groups such as 3- or 4-nitrophenyl; cyanophenyl groups, such as 4-cyanophenyl; mono- or di(alkyl)phenyl groups such as 4-methylphenyl, 2,4-dimethylphenyl, 2-methylphenyl, 4-(isopropyl)phenyl, 4-ethylphenyl, 3-(n-propyl)phenyl, etc.; mono- or di(alkoxy)phenyl groups, such as 3,4-dimethoxyphenyl, 3-methoxy-4-benzyloxyphenyl, 3-ethoxyphenyl, 4-(isopropoxy)phenyl, 4-(t-butoxy)phenyl, 3-ethoxy-4-methoxyphenyl, etc.; 3- or 4-trifluoromethylphenyl; mono- or dicarboxyphenyl or (protected carboxy)phenyl groups, such as 4-carboxyphenyl, mono- or di(hydroxymethyl)phenyl or (protected hydroxymethyl)phenyl, such as 3-(protected hydroxymethyl)phenyl or 3,4-di(hydroxymethyl)phenyl; mono- or di(aminomethyl)phenyl or (protected aminomethyl)phenyl, such as 2-(aminomethyl)phenyl or 2,4-(protected aminomethyl)phenyl; or mono- or di(N-(methylsulfonylamino))phenyl, such as 3-(N-methylsulfonylamino))phenyl.Also, the term "substituted phenyl" refers to disubstituted phenyl groups having different substituents, such as 3-methyl-4-hydroxyphenyl, 3-chloro-4-hydroxyphenyl, 2-methoxy-4-bromophenyl, 4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl, 2-hydroxy-4-chlorophenyl, 2-chloro-5-difluoromethoxy, etc., and trisubstituted phenyl groups having different substituents, such as 3-methoxy-4-benzyloxy-6-methylsulfonylamino, 3-methoxy-4-benzyloxy-6-phenylsulfonylamino, and tetrasubstituted phenyl groups having different substituents, such as 3-methoxy-4-benzyloxy-5-methyl-6-phenylsulfonylamino. In some embodiments, the substituents of aryl such as phenyl include amides. For example, the aryl (e.g., phenyl) substituent can be -(CH2). 0-4 CONR’R’’, where R’ and R’’ are each independently, for example, hydrogen; unsubstituted C1-C6 alkyl; C1-C6 alkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; unsubstituted C1-C6 heteroalkyl; C1-C6 heteroalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; unsubstituted C6-C 10 aryl; C6-C substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, or NR’R’’ 10Aryl; unsubstituted 3- to 11-membered heterocyclyl (e.g., 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S, or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); and 3- to 11-membered heterocyclyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR’R’’ (e.g., 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S, or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); provided that R’ and R’’ may, in combination with the nitrogen atom, form a 3-, 4-, 5-, 6-, or 7-membered ring, the ring atoms of which are optionally substituted with N, O, or S, and the ring is optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR’R’’.

[0029] “Cycloalkyl” refers to a non-aromatic saturated or partially unsaturated hydrocarbon ring group, and the cycloalkyl group may be independently optionally substituted with one or more substituents described herein. In one example, the cycloalkyl group has 3 to 12 carbon atoms (C3-C 12 ). In another example, cycloalkyl is C3-C8, C3-C 10 or C5-C 10 . In another example, the cycloalkyl group as a monocyclic ring is C3-C8, C3-C6, or C5-C6. In another example, the cycloalkyl group as a bicyclic ring is 7-C 12 . In another example, the cycloalkyl group as a spiro system is C5-C 12It is. Examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopenta-1-enyl, 1-cyclopenta-2-enyl, 1-cyclopenta-3-enyl, cyclohexyl, perdeuterocyclohexyl, 1-cyclohexa-1-enyl, 1-cyclohexa-2-enyl, 1-cyclohexa-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. Exemplary arrangements of bicyclic cycloalkyls having 7 to 12 ring atoms include, but are not limited to, [4,4], [4,5], [5,5], [5,6], or [6,6] ring systems. Exemplary bridged bicyclic cycloalkyls include, but are not limited to, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, and bicyclo[3.2.2]nonane. Examples of spirocycloalkyls include spiro[2.2]pentane, spiro[2.3]hexane, spiro[2.4]heptane, spiro[2.5]octane, and spiro[4.5]decane. In some embodiments, substituents of "optionally substituted cycloalkyl" include, for example, F, Cl, Br, I, OH, SH, CN, NH2, NHCH3, N(CH3)2, NO2, N3, C(O)CH3, COOH, CO2CH3, methyl, ethyl, propyl, iso-propyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methanesulfonylamino, SO, SO2, phenyl, piperidinyl, 1 to 4 examples of piperidinyl and pyrimidinyl, and the alkyl, aryl, and heterocyclic moieties thereof may be optionally substituted by 1 to 4 examples of substituents selected from the same list. In some embodiments, the substituents of cycloalkyl include amides. For example, the cycloalkyl substituent is -(CH2) 0-4It can be CONR’R’’, and R’ and R’’ are each independently, for example, hydrogen; unsubstituted C1-C6 alkyl; C1-C6 alkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; unsubstituted C1-C6 heteroalkyl; C1-C6 heteroalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; unsubstituted C6-C 10 aryl; C6-C substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, or NR’R’’ 10 aryl; unsubstituted 3- to 11-membered heterocyclyl (for example, 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S); and 3- to 11-membered heterocyclyl (for example, 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S) substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; or, R’ and R’’ can combine with a nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring, the ring atoms are optionally substituted with N, O or S, and the ring is optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’.

[0030] "Complex ring group", "complex ring formula", "complex ring", "heterocyclyl" or "heterocycle" are used interchangeably and refer to any monocyclic, bicyclic, tricyclic or spiro ring system having 3 to 20 ring atoms (e.g., 3 to 10 ring atoms), a saturated or unsaturated aromatic (heteroaryl) or non-aromatic (e.g., heterocycloalkyl) ring system, where the ring atoms are carbon and at least one atom of the ring or ring system is a heteroatom selected from nitrogen, sulfur or oxygen. If any ring atom of the ring system is a heteroatom, the system is a heterocycle regardless of the point of attachment of the ring system to the remainder of the molecule. In one example, heterocyclyl contains 3 to 11 ring atoms ("members") and includes monocyclic, bicyclic, tricyclic, and spiro ring systems. Here, the ring atoms are carbon and at least one atom in the ring or ring system is a heteroatom selected from nitrogen, sulfur, or oxygen. In one example, heterocyclyl contains 1 to 4 heteroatoms. In one example, heterocyclyl contains 1 to 3 heteroatoms. In another example, heterocyclyl includes a 3- to 7-membered monocyclic ring having 1 to 2, 1 to 3, or 1 to 4 heteroatoms selected from nitrogen, sulfur, or oxygen. In another example, heterocyclyl includes a 4- to 6-membered monocyclic ring having 1 to 2, 1 to 3, or 1 to 4 heteroatoms selected from nitrogen, sulfur, or oxygen. In another example, heterocyclyl includes a 3-membered monocyclic ring. In another example, heterocyclyl includes a 4-membered monocyclic ring. In another example, heterocyclyl includes a 5- to 6-membered monocyclic ring, e.g., 5- to 6-membered heteroaryl. In another example, heterocyclyl includes 3- to 11-membered heterocycloalkyl, e.g., 4- to 11-membered heterocycloalkyl. In some embodiments, heterocycloalkyl contains at least one nitrogen. In one example, the heterocyclyl group contains 0 to 3 double bonds. Any nitrogen or sulfur heteroatom may optionally be oxidized (e.g., NO, SO, SO2), and any nitrogen heteroatom may optionally be quaternized (e.g., [NR4] + Cl - , [NR4] + OH -)). Exemplary complex rings include oxiranyl, aziridinyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, 1,2-dithietanyl, 1,3-dithietanyl, pyrrolidinyl, dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydrofuranyl, dihydrothienyl, tetrahydrothienyl, imidazolidinyl, piperidinyl, piperazinyl, isoquinolinyl, tetrahydroisoquinolinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, hexahydrothiopyranyl, hexahydropyrimidinyl, oxazinanil, thiazinanil, thioxanyl, homopiperazinyl, homopiperidinyl, azepanyl, oxepanyl, thiepanyl, oxazepinyl, oxazepanyl, diazepanyl, 1,4-diazepanyl, diazepinyl, thiazepinyl, thiazepanyl, tetrahydrothiopyranyl, oxazolidinyl, thiazolidinyl, isothiazolidinyl, 1,1-dioxoisothiazolidinonyl, oxazolidinonyl, imidazolidinonyl, 4,5,6,7-tetrahydro[2H]indazolyl, tetrahydrobenzimidazolyl, 4,5,6,7-tetrahydrobenzo[d]imidazolyl, 1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridinyl, thiazinyl, oxazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl, oxathiazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl, imidazolinyl, dihydropyrimidyl, tetrahydropyrimidyl, 1-pyrrolinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, thiapyranyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, pyrazolidinyl, dithianyl, dithiolanyl, pyrimidinonyl, pyrimidinedionyl, pyrimidine-2,4-dionyl, piperazinonyl, piperazinedionyl, pyrazolidinylimidazolinyl, 3-azabicyclo[3.1.0]hexanyl, 3,6-azabicyclo[3.1.1]heptanyl, 6-azabicyclo[3.1.1]heptanyl, 3-azabicyclo[3.1.1]heptanyl, 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 2-azabicyclo[3.2.1]octanyl, 8-azabicyclo[3.2.1]octanyl, 2-azabicyclo[2.2.2]octanyl, 8-azabicyclo[2.2.2]octanyl, 7-oxabicyclo[2.2.1]heptane, azaspiro[3.5]nonanyl, azaspiro[2.5]octanyl, azaspiro[4.5]decanyl, 1-azaspiro[4.5]decan-2-one (only), azaspiro[5.5]undecanyl, tetrahydroindolyl, octahydroindolyl, tetrahydroisoindolyl, tetrahydroindazolyl, 1,1-dioxohexahydrothiopyranyl. Examples of 5-membered heterocycles containing a sulfur atom or an oxygen atom and 1 to 3 nitrogen atoms include thiazolyl including thiazol-2-yl and thiazol-2-yl N-oxide, thiadiazolyl including 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl, oxazolyl such as oxazol-2-yl, and oxadiazolyl such as 1,3,4-oxadiazol-5-yl and 1,2,4-oxadiazol-5-yl. Examples of 5-membered heterocycles containing 2 to 4 nitrogen atoms include imidazolyl such as imidazol-2-yl; triazolyl such as 1,3,4-triazol-5-yl, 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl; and tetrazolyl such as 1H-tetrazol-5-yl. Examples of benzo-fused 5-membered heterocycles are benzoxazol-2-yl, benzthiazol-2-yl and benzimidazol-2-yl. Examples of 6-membered heterocycles include 1 to 3 nitrogen atoms and optionally a sulfur or oxygen atom, for example, pyridyl such as pyrid-2-yl, pyrid-3-yl and pyrid-4-yl; pyrimidyl such as pyrimid-2-yl and pyrimid-4-yl; 1,3,4-triazin-2-yl and 1,3,Triazinyl such as 5 - triazin - 4 - yl; pyridazinyl, especially pyridazin - 3 - yl and pyrazinyl. Pyridine N - oxide and pyridazine N - oxide, as well as pyridyl, pyrimido - 2 - yl, pyrimido - 4 - yl, pyridazinyl and 1,3,4 - triazin - 2 - yl groups are other exemplary heterocyclic groups. The heterocycle may be optionally substituted. For example, substituents of the "optionally substituted heterocycle" include F, Cl, Br, I, OH, SH, CN, NH2, NHCH3, N(CH3)2, NO2, N3, C(O)CH3, COOH, CO2CH3, methyl, ethyl, propyl, iso - propyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methanesulfonylamino, SO, SO2, phenyl, piperidinyl, piperidinyl and pyrimidinyl, with 1 to 4 examples thereof, and the alkyl, aryl and heterocyclic moieties thereof may be optionally substituted by 1 to 4 examples of substituents selected from the same list. In some embodiments, substituents of heterocyclic groups such as heteroaryl or heterocycloalkyl include amides. For example, the heterocyclic (e.g., heteroaryl or heterocycloalkyl) substituent can be -(CH2), 0-4 CONR’R’’, where R’ and R’’ are each independently, for example, hydrogen; unsubstituted C1 - C6 alkyl; C1 - C6 alkyl substituted with halogen, OH, CN, unsubstituted C1 - C6 alkyl, unsubstituted C1 - C6 alkoxy, oxo or NR’R’’; unsubstituted C1 - C6 heteroalkyl; C1 - C6 heteroalkyl substituted with halogen, OH, CN, unsubstituted C1 - C6 alkyl, unsubstituted C1 - C6 alkoxy, oxo or NR’R’’; unsubstituted C6 - C 10 aryl; C6 - C substituted with halogen, OH, CN, unsubstituted C1 - C6 alkyl, unsubstituted C1 - C6 alkoxy, or NR’R’’ 10Aryl; unsubstituted 3- to 11-membered heterocyclyl (e.g., 5- or 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); and 3- to 11-membered heterocyclyl (e.g., 5- or 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S) substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR’R’’; provided that R’ and R’’ may, in combination with the nitrogen atom, form a 3-, 4-, 5-, 6- or 7-membered ring, the ring atoms of which are optionally substituted with N, O or S, and the ring is optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR’R’’.

[0031] “Heteroaryl” refers to any monocyclic, bicyclic or tricyclic ring system in which at least one ring is a 5- or 6-membered aromatic ring containing 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, and in an exemplary embodiment, at least one heteroatom is nitrogen. For example, Lang’s Handbook of Chemistry (Dean, J.A., ed.) 13 thSee ed.Table 7-2

[1985] . This definition includes any bicyclic group in which any of the above heteroaryl rings is fused to an aryl ring and the aryl ring or heteroaryl ring is attached to the remainder of the molecule. In one embodiment, heteroaryl includes 5- to 6-membered monocyclic aromatic groups in which one or more ring atoms are nitrogen, sulfur, or oxygen. Exemplary heteroaryl groups include thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl, tetrazolo[1,5-b]pyridazinyl, imidazol[1,2-a]pyrimidinyl, and purinyl, and benzocondensed derivatives such as benzoxazolyl, benzofuryl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzimidazolyl, and indolyl. The heteroaryl group may be optionally substituted. In some embodiments, substituents of "optionally substituted heteroaryl" include one to four examples of F, Cl, Br, I, OH, SH, CN, NH2, NHCH3, N(CH3)2, NO2, N3, C(O)CH3, COOH, CO2CH3, methyl, ethyl, propyl, iso-propyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, trifluoromethyl, difluoromethyl, sulfonylamino, methanesulfonylamino, SO, SO2, phenyl, piperidinyl, piperidinyl, and pyrimidinyl, and the alkyl, phenyl, and heterocyclic moieties thereof may be optionally substituted by one to four examples of substituents selected from the same list. In some embodiments, the substituents of heteroaryl include amides. For example, the heteroaryl substituent is -(CH2) 0-4It can be CONR’R’’, and R’ and R’’ are each independently, for example, hydrogen; unsubstituted C1-C6 alkyl; C1-C6 alkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; unsubstituted C1-C6 heteroalkyl; C1-C6 heteroalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; unsubstituted C6-C 10 aryl; C6-C substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, or NR’R’’ 10 aryl; unsubstituted 3- to 11-membered heterocyclyl (for example, 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S); and 3- to 11-membered heterocyclyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’ (for example, 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S); or, R’ and R’’ can combine with a nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring, the ring atoms are optionally substituted with N, O or S, and the ring is optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’.

[0032] In certain embodiments, the heterocyclyl group is bonded to a carbon atom of the heterocyclyl group. For example, as a carbon-bonded heterocyclyl group, the 2, 3, 4, 5 or 6 positions of the pyridine ring, the 3, 4, 5 or 6 positions of the pyridazine ring, the 2, 4, 5 or 6 positions of the pyrimidine ring, the 2, 3, 5 or 6 positions of the pyrazine ring, furan, tetrahydrofuran, thiophene, thiophene, pyrrole, or the 2, 3, 4 or 5 positions of the tetrahydropyrrole ring, oxazole, imidazole, or the 2, 4 or 5 positions of the thiazole ring, isoxazole, pyrazole, or the 3, 4 or 5 positions of the isothiazole ring, the 2 or 3 positions of the aziridine ring, the 2, 3 or 4 positions of the azetidine ring, the 2, 3, 4, 5, 6, 7 or 8 positions of the quinoline ring, or the 1, 3, 4, 5, 6, 7 or 8 positions of the isoquinoline ring may be mentioned.

[0033] In certain embodiments, the heterocyclyl group is N-bonded. For example, as a nitrogen-bonded heterocyclyl or heteroaryl group, aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, the 1 position of 1H-indazole, the 2 position of isoindole or isoindoline, the 4 position of morpholine, and the 9 position of carbazole, or β-carboline may be mentioned.

[0034] The term "alkoxy" refers to a straight-chain or branched-chain monovalent radical represented by the formula -OR, where R is alkyl as defined herein. Examples of alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, mono-, di- and tri-fluoromethoxy and cyclopropoxy.

[0035] "Acyl" means a carbonyl containing a substituent represented by the formula -C(O)-R (wherein R is hydrogen, alkyl, cycloalkyl, aryl, or heterocyclyl, and alkyl, cycloalkyl, aryl, and heterocyclyl are as defined herein). Examples of acyl groups include alkanoyl (e.g., acetyl), aroyl (e.g., benzoyl), and heteroaroyl (e.g., pyridinoyl).

[0036] Unless otherwise specified, "optionally substituted" means that a group may be unsubstituted or may be substituted by one or more (e.g., 0, 1, 2, 3, 4, or 5 or more, or any range variable thereof) substituents (which may be the same or different) listed for that group. In one embodiment, an optionally substituted group has one substituent. In another embodiment, an optionally substituted group has two substituents. In another embodiment, an optionally substituted group has three substituents. In another embodiment, an optionally substituted group has four substituents. In another embodiment, an optionally substituted group has five substituents.

[0037] Any substituent of an alkyl radical, alone or as part of another substituent (e.g., alkoxy), and alkylenyl, alkenyl, alkynyl, heteroalkyl, heterocycloalkyl and cycloalkyl may also, each alone or as part of another substituent, be various groups such as those described herein, and halogen; oxo; CN; NO; N3; -OR'; perfluoro-C1-C4 alkoxy; unsubstituted C3-C7 cycloalkyl; C3-C7 cycloalkyl substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR'R''; unsubstituted C6-C 10 Aryl (e.g., phenyl); C6-C substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, or NR'R'' 10Aryl; unsubstituted 3- to 11-membered heterocyclyl (e.g., 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); 3- to 11-membered heterocyclyl substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’ (e.g., 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); -NR’R’’; -SR’; -SiR’R’’R’’’; -OC(O)R’; -C(O)R’; -CO2R’; -CONR’R’’; -OC(O)NR’R’’; -NR’’C(O)R’; -NR’’’C(O)NR’R’’; -NR’’C(O)2R’; -S(O)2R’; -S(O)2NR’R’’; -NR’S(O)2R’’; -NR’’’S(O)2NR’R’’; amidinyl; guanidyl; -(CH2) 1-4 -OR’; -(CH2) 1-4 -NR’R’’; -(CH2) 1-4 -SR’; -(CH2) 1-4 -SiR’R’’R’’’; -(CH2) 1-4 -OC(O)R’; -(CH2) 1-4 -C(O)R’; -(CH2) 1-4 -CO2R’; and -(CH2) 1-4 A group selected from the group consisting of CONR’R’’, or combinations thereof, which may be a number in the range of 0 to (2m’ + 1), where m’ is the total number of carbon atoms in each such radical. R’, R’’ and R’’’ are each independently, for example, hydrogen; unsubstituted C1-C6 alkyl; C1-C6 alkyl substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; unsubstituted C1-C6 heteroalkyl; C1-C6 heteroalkyl substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; unsubstituted C6-C 10Aryl; C6-C substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, or NR’R’’ 10 Aryl; unsubstituted 3- to 11-membered heterocyclyl (e.g., 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); and 3- to 11-membered heterocyclyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR’R’’ (e.g., 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); A group containing is meant. When R’ and R’’ are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring, where the ring atoms are optionally substituted with N, O, or S, and the ring is optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR’R’’. For example, -NR’R’’ means including 1-pyrrolidinyl and 4-morpholinyl.

[0038] Similarly, the optional substituents of the aryl group and heteroaryl group are various. In some embodiments, the substituents of the aryl group and heteroaryl group are halogen; CN; NO; N3; -OR’; perfluoro-C1-C4 alkoxy; unsubstituted C3-C7 cycloalkyl; C3-C7 cycloalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR’R’’; unsubstituted C6-C 10 Aryl (e.g., phenyl); C6-C substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, or NR’R’’ 10Aryl; unsubstituted 3- to 11-membered heterocyclyl (e.g., 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); 3- to 11-membered heterocyclyl substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’ (e.g., 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); -NR’R’’; -SR’; -SiR’R’’R’’’; -OC(O)R’; -C(O)R’; -CO2R’; -CONR’R’’; -OC(O)NR’R’’; -NR’’C(O)R’; -NR’’’C(O)NR’R’’; -NR’’C(O)2R’; -S(O)2R’; -S(O)2NR’R’’; -NR’S(O)2R’’; -NR’’’S(O)2NR’R’’; amidinyl; guanidyl; -(CH2) 1-4 -OR’; -(CH2) 1-4 -NR’R’’; -(CH2) 1-4 -SR’; -(CH2) 1-4 -SiR’R’’R’’’; -(CH2) 1-4 -OC(O)R’; -(CH2) 1-4 -C(O)R’; -(CH2) 1-4 -CO2R’; and -(CH2) 1-4 Selected from the group consisting of CONR’R’’, or combinations thereof, and is a number in the range of 0 to (2m’ + 1), where m’ is the total number of carbon atoms in each such radical. R’, R’’ and R’’’ are each independently, for example, hydrogen; unsubstituted C1-C6 alkyl; C1-C6 alkyl substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; unsubstituted C1-C6 heteroalkyl; C1-C6 heteroalkyl substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR’R’’; unsubstituted C6-C 10Aryl; C6-C substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, or NR’R’’ 10 Aryl; unsubstituted 3- to 11-membered heterocyclyl (e.g., 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); and 3- to 11-membered heterocyclyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR’R’’ (e.g., 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms selected from O, N, and S or 4- to 11-membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N, and S); A group containing is meant. When R’ and R’’ are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring, where the ring atoms are optionally substituted with N, O, or S, and the ring is optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR’R’’. For example, -NR’R’’ means including 1-pyrrolidinyl and 4-morpholinyl.

[0039] The term “oxo” refers to =O or (=O)2.

[0040] As used herein, the bond in the chemical structure intersects TIFF0007716993000002.tif7170 indicates the bonding point of an atom in the chemical structure where a wavy bond is attached to the rest of the molecule or the rest of the molecular fragment. In some embodiments, an asterisk together with an arrow is used like a wavy line to indicate the added point.

[0041] In certain embodiments, divalent groups are generally described without a specific bond structure. Unless otherwise specified, the general description is understood to mean including both bond structures. For example, the group R 1 -R 2 -R 3 In, the group R 2When described as -CH2C(O)-, unless otherwise specified, this group is R 1 -CH2C(O)-R 3 and R 1 -C(O)CH2-R 3 is understood to be bondable as both of them.

[0042] Terms such as "the compound(s) of the invention" and "the compound(s) of the present invention" include, unless otherwise specified, the compounds of formula (I) in this specification such as Compounds 1 to 18, which may also be called JAK inhibitors, including stereoisomers (including atropisomers), geometric isomers, tautomers, solvates, metabolites, isotopes, salts (e.g., pharmaceutically acceptable salts), and prodrugs thereof. In some embodiments, solvates, metabolites, isotopes or prodrugs, or any combination thereof are excluded.

[0043] The phrase "pharmaceutically acceptable" means molecular elements and compositions that do not produce side reactions, allergic reactions, or other side effects when appropriately administered to animals, such as humans.

[0044] The compounds of the present invention can be in the form of salts such as pharmaceutically acceptable salts. "Pharmaceutically acceptable salts" include both acid addition salts and base addition salts. "Pharmaceutically acceptable acid addition salts" mean salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, etc., which retain the biological effectiveness and properties of the free base and are salts that are biologically or otherwise desirable, and the organic acids can be selected from the classes of aliphatic, alicyclic, aromatic, araliphatic, heterocyclic, carboxylic acids, and sulfonic acids of organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, embonic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

[0045] Examples of the "pharmaceutically acceptable basic addition salts" include salts derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Specific basic addition salts are ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts. Examples of salts derived from pharmaceutically acceptable organic non-toxic bases include primary, secondary, and tertiary amines, naturally occurring substituted amines, cyclic amines, and substituted amines including basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, tromethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperizine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Specific organic non-toxic bases include isopropylamine, diethylamine, ethanolamine, tromethamine, dicyclohexylamine, choline, and caffeine.

[0046] In some embodiments, the salt is selected from hydrochloride, hydrobromide, trifluoroacetate, sulfate, phosphate, acetate, fumarate, maleate, tartrate, lactate, citrate, pyruvate, succinate, oxalate, methanesulfonate, p-toluenesulfonate, bisulfate, benzenesulfonate, ethanesulfonate, malonate, xinafoate, ascorbate, oleate, nicotinate, saccharinate, adipate, formate, glycolate, palmitate, L-lactate, D-lactate, aspartate, malate, L-tartrate, D-tartrate, stearate, phthalate (e.g., 2-phthalate or 3-phthalate), napadisylate (naphthalene-1,5-disulfonate, or naphthalene-1(sulfonic acid)-5-sulfonate), edisylate (ethane-1,2-disulfonate, or ethane-1-(sulfonic acid)-2-sulfonate), isethionate (2-hydroxyethylsulfonate), 2-mesitylenesulfonate, 2-naphthalenesulfonate, 2,5-dichlorobenzenesulfonate, D-mandelate, L-mandelate, cinnamate, benzoate, adipate, esilate, malonate, mesitylate (2-mesitylenesulfonate), naphthylate (2-naphthalenesulfonate), camsylate (camphor 10-sulfonate, e.g., (1S)-(+)-10-camphor-sulfonate), glutamate, glutarate, hippurate (2-(benzoylamino)acetate), orotate, xylate (p-xylene-2-sulfonate), and pamoate (2,2'-dihydroxy-1,1'-dinaphthylmethane-3,3'-dicarboxylate).

[0047] A "sterile" formulation is either sterile or contains no viable microorganisms and their spores.

[0048] "Stereoisomers" means compounds that have the same chemical structure but differ in the arrangement of atoms or groups in space. Examples of stereoisomers include diastereomers, enantiomers, conformational isomers, and the like.

[0049] "Chiral" refers to a molecule with the property that its mirror image partners cannot be superimposed, while the term "achiral" refers to a molecule that can be superimposed on its mirror image partners.

[0050] "Diastereomers" refer to stereoisomers that have two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectroscopic properties, or biological activity. A mixture of diastereomers can be separated under high-resolution analytical procedures such as electrophoresis and chromatography such as HPLC.

[0051] "Enantiomers" refer to two stereoisomers of a compound that are non-superimposable mirror images of each other.

[0052] The definitions and conventions of stereochemistry used in this specification generally follow S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., “Stereochemistry of Organic Compounds,” John Wiley & Sons, Inc., New York, 1994. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D and L, or R and S are used to denote the absolute configuration of the molecule around its chiral center. The prefixes d and l or (+) and (-) are used to denote the sign of rotation of plane-polarized light by the compound, and (-) or l means that the compound is levorotatory. A compound with the prefix (+) or d is dextrorotatory. In a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Also, a particular stereoisomer may be called an enantiomer, and a mixture of such isomers is often called a mixture of enantiomers. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur when there is no stereoselection or stereospecificity in a chemical reaction or process. The terms “racemic mixture” and “racemate” refer to an equimolar mixture of two enantiomeric species and are optically inactive.

[0053] The term “tautomer” or “tautomeric form” means structural isomers of different energies that are interconvertible by a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) involve interconversion by the migration of a proton, such as keto-enol and imine-enamine isomerization. Valence tautomers involve interconversion by the rearrangement of some bonding electrons.

[0054] The specific compounds of the present invention can exist not only in solvated forms including hydrated forms, but also in unsolvated forms. "Solvate" means a compound or complex of one or more solvent molecules and a compound of the present invention. Examples of solvents that form solvates include water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. Specific compounds of the present invention can exist in multiple crystalline or amorphous forms. Generally, all physical forms are intended to be within the scope of the present invention. The term "hydrate" refers to a complex in which the solvent molecule is water.

[0055] "Metabolite" means a product produced through metabolism in the body of a specified compound or a salt thereof. Such products can be brought about, for example, by oxidation, reduction, hydrolysis, amidation, amide decomposition, esterification, deesterification, enzymatic cleavage, etc. of the administered compound.

[0056] Metabolic products are typically identified by preparing a radiolabeled (e.g., 14 C or 3 H) isotope of the compound of the present invention, administering the isotope to animals such as rats, mice, guinea pigs, monkeys, etc., or humans at a detectable dose (e.g., more than about 0.5 mg / kg), allowing metabolism to occur for a sufficient time (typically about 30 seconds to 30 hours), and isolating the conversion product from urine, blood, or other biological samples. Such products are easily isolated because they are labeled (others are isolated by using antibodies that can bind to epitopes remaining in the metabolite). The structure of the metabolite is determined by conventional methods, such as MS, LC / MS, or NMR analysis. Generally, the analysis of metabolites is carried out in the same manner as conventional drug metabolism studies well-known to those skilled in the art. Metabolites are useful in diagnostic assays for the therapeutic administration of the compounds of the present invention, unless separately found in vivo.

[0057] "Subject", "individual", or "patient" is a vertebrate. In certain embodiments, the vertebrate is a mammal. Examples of mammals include, but are not limited to, livestock (such as cows), sports animals, pets (such as guinea pigs, cats, dogs, rabbits, and horses), primates, mice, and rats. In certain embodiments, the mammal is a human. In embodiments that include administering to a patient a JAK inhibitor or a pharmaceutically acceptable salt thereof, the patient may be in need thereof.

[0058] The term "Janus kinase" refers to JAK1, JAK2, JAK3, and TYK2 protein kinases. In some embodiments, the Janus kinase may be further defined as one of JAK1, JAK2, JAK3, or TYK2. In any embodiment, any one of JAK1, JAK2, JAK3, and TYK2 may be specifically excluded as a Janus kinase. In some embodiments, the Janus kinase is JAK1. In some embodiments, the Janus kinase is a combination of JAK1 and JAK2.

[0059] The terms "inhibit" and "reduce", or any variation of these terms, include any measurable reduction or complete inhibition to achieve the desired result. For example, there may be a decrease of about, up to about, or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or more, or any range variable thereof, a decrease in activity (e.g., JAK1 activity) compared to normal.

[0060] "Therapeutically effective amount" means an amount of a compound of the invention or a salt thereof (e.g., a pharmaceutically acceptable salt thereof) that (i) treats or prevents a particular disease, condition or disorder, or (ii) weakens, ameliorates or eliminates one or more symptoms of a particular disease, condition or disorder, and optionally (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition or disorder described herein. In some embodiments, a therapeutically effective amount is an amount sufficient to reduce or alleviate the symptoms of an autoimmune or inflammatory disease (e.g., asthma). In some embodiments, a therapeutically effective amount is an amount of a chemical entity described herein sufficient to significantly reduce the activity or number of B cells. In the case of cancer, a therapeutically effective amount of an agent can reduce the number of cancer cells, reduce tumor size, inhibit (i.e., slow to a low level and preferably stop) the invasion of cancer cells into peripheral organs, inhibit (i.e., slow to a low level and preferably stop) tumor metastasis, inhibit tumor growth to a low level, or reduce to a low level one or more symptoms associated with cancer. To the extent that an agent can prevent the growth of existing cancer cells or kill them, the agent can be cytostatic or cytotoxic. With regard to cancer therapy, efficacy can be measured, for example, by assessment of time to disease progression (TTP) or determination of response rate (RR).

[0061] "Treatment" (and variations such as "treat" or "treating") means a clinical intervention to alter the natural course of an individual or cell being treated and can be implemented for the prevention of, or during, a clinical condition. Desired effects of treatment include preventing the onset or recurrence of a disease, alleviating symptoms, reducing any direct or indirect pathological consequences of a disease, a stable (i.e., non-worsening) state of a disease, reducing the rate of disease progression, restoring or alleviating the state of a disease, extending survival compared to survival expected in the absence of treatment, and an improved remission or prognosis. In some embodiments, a compound of the invention or a salt thereof (e.g., a pharmaceutically acceptable salt thereof) is used to delay the onset of, or slow the progression of, a disease or disorder. Subjects in need of treatment include subjects already having a symptom or disorder, subjects having a tendency to have a symptom or disorder (e.g., due to a genetic mutation), or subjects in whom it is necessary to prevent a symptom or disorder.

[0062] "Inflammatory disorder" refers to any disease, disorder or syndrome in which an excessive or unregulated inflammatory response results in excessive inflammatory symptoms, host tissue damage or loss of tissue function. "Inflammatory disorder" also refers to a pathological condition mediated by the influx of leukocytes or neutrophil chemotaxis.

[0063] "Inflammation" refers to a local defensive response induced by tissue injury or destruction, which serves to destroy, dilute or remove (sequester) both the injurious agent and the injured tissue. Inflammation is particularly associated with the influx of leukocytes or neutrophil chemotaxis. Inflammation can result from infectious means such as infection by pathogenic organisms and viruses, as well as non-infectious means such as trauma or reperfusion after myocardial infarction or stroke, an immune response to foreign antigens, and an autoimmune response. Thus, inflammatory disorders suitable for treatment with a compound of the invention or a salt thereof (e.g., a pharmaceutically acceptable salt thereof) include disorders associated with the response of the specific defense system as well as the response of the non-specific defense system.

[0064] The "specific defense system" refers to the components of the immune system that respond to the presence of a specific antigen. Examples of inflammation resulting from the response of the specific defense system include the classical response to foreign antigens, autoimmune diseases, and delayed-type hypersensitivity responses mediated by T cells. Chronic inflammatory diseases, rejection of solid transplant tissues and organs, such as kidney and bone marrow transplants, and graft-versus-host disease (GVHD) are further examples of inflammatory reactions of the specific defense system.

[0065] The term "nonspecific defense system" refers to inflammatory disorders mediated by white blood cells that do not develop immunological memory (e.g., granulocytes and macrophages). Examples of inflammation that results at least in part from the response of the nonspecific defense system include inflammation associated with conditions such as adult (acute) respiratory distress syndrome (ARDS) or multiple organ injury syndrome; reperfusion injury; acute glomerulonephritis; reactive arthritis; skin diseases with acute inflammatory components; other central nervous system inflammatory disorders such as acute pyogenic meningitis or stroke; thermal injury; inflammatory bowel disease; granulocyte transfusion-related syndrome; and cytokine-induced toxicity.

[0066] "Autoimmune disease" refers to any group of disorders in which tissue damage is associated with a humoral or cell-mediated response to components of the body itself. Non-limiting examples of autoimmune diseases include rheumatoid arthritis, lupus, and multiple sclerosis.

[0067] As used herein, "allergy disease" refers to any symptom, tissue damage, or loss of tissue function resulting from an allergy. As used herein, "arthritis disease" refers to any disease characterized by an inflammatory lesion of the joints resulting from various etiologies. As used herein, "dermatitis" refers to any of a large family of skin diseases characterized by inflammation of the skin resulting from various etiologies. As used herein, "transplant rejection" refers to any immune reaction against a transplanted tissue, such as an organ or cell (e.g., bone marrow), characterized by loss of function of the transplanted tissue and surrounding tissue, pain, swelling, leukocytosis, and thrombocytopenia. The treatment methods of the present invention include methods for treating disorders associated with inflammatory cell activation.

[0068] "Inflammatory cell activation" refers to the induction by stimuli of a proliferative cell response (including, but not limited to, cytokines, antigens or autoantibodies), the production of soluble mediators (including, but not limited to, cytokines, oxygen radicals, enzymes, prostaglandins, or vasoactive amines), or the cell surface expression of new or increased numbers of mediators (including, but not limited to, major histocompatibility antigens or cell adhesion molecules) in inflammatory cells (including, but not limited to, monocytes, macrophages, T lymphocytes, B lymphocytes, granulocytes (i.e., polymorphonuclear leukocytes such as neutrophils, basophils, eosinophils, etc.), mast cells, dendritic cells, Langerhans cells, and endothelial cells). One or a combination of these phenotypes in these cells can contribute to the initiation, persistence, or exacerbation of inflammatory disorders, as will be understood by those skilled in the art.

[0069] In some embodiments, inflammatory disorders that can be treated according to the methods of the present invention include, but are not limited to, asthma, rhinitis (e.g., allergic rhinitis), allergic airway syndrome, atopic dermatitis, bronchitis, rheumatoid arthritis, psoriasis, contact dermatitis, chronic obstructive pulmonary disease (COPD), and delayed-type hypersensitivity reactions.

[0070] The terms "cancer" and "cancerous", "neoplasm", and "tumor", and related terms refer to or describe physiological conditions in mammals typically characterized by unregulated cell growth. A "tumor" contains one or more cancer cells. Examples of cancer include carcinomas, sarcomas, blastomas, seminomas, glioblastomas, melanomas, leukemias, and malignancies of the bone marrow or lymphatic system. More specific examples of such cancers include squamous cell carcinoma (e.g., epidermal squamous cell carcinoma), and lung cancers (such as small cell lung cancer, non-small cell lung cancer ("NSCLC"), lung adenocarcinoma, and lung squamous cell carcinoma). Other cancers include skin cancer, keratoacanthoma, follicular carcinoma, hairy cell leukemia, oral cancer, pharyngeal (oral) cancer, lip cancer, tongue cancer, mouth cancer, salivary gland cancer, esophageal cancer, laryngeal cancer, hepatocellular carcinoma, gastric cancer, stomach cancer, gastrointestinal cancer, small intestine cancer, colorectal cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, liver cancer, breast cancer, colon cancer, rectal cancer, colorectal cancer, genitourinary cancer, biliary tract cancer, thyroid cancer, papillary cancer, liver cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer or renal carcinoma, prostate cancer, testicular cancer, vulvar cancer, peritoneal cancer, anal cancer, penile cancer, bone cancer, multiple myeloma, B-cell lymphoma, central nervous system, brain cancer, head and neck cancer, Hodgkin's disease, and related metastases. Examples of neoplastic disorders include myeloproliferative disorders such as polycythemia vera, essential thrombocythemia, myelofibrosis such as primary myelofibrosis, and chronic myelogenous leukemia (CML).

[0071] A "chemotherapeutic agent" is an agent useful in the treatment of a given disorder, such as cancer or an inflammatory disorder. Examples of chemotherapeutic agents are known in the art and include those described in U.S. Patent Application Publication No. 2010 / 0048557, which is incorporated herein by reference. Further, chemotherapeutic agents include any pharmaceutically acceptable salt, acid, or derivative of a chemotherapeutic agent, and combinations of two or more thereof.

[0072] "Package insert" is used to refer to the instructions customarily included in the commercial package of a therapeutic agent that contains information about the indications, usage, dosage, administration, contraindications, or warnings regarding its use.

[0073] Unless otherwise indicated, the structures shown in this specification also include compounds that differ only in that one or more isotope-enriched atoms are present. Exemplary isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 32 P, 33 P, 35 S, 18 F, 36 Cl, 123 I, and 125 I. Isotopically labeled compounds (e.g., 3 H and 14 C labeled compounds) can be useful in compound or substrate tissue distribution assays. Tritium labeling (i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes can be useful because of the ease of preparation and detection. Further, substitution with heavier isotopes, such as deuterium (i.e., 2 H), etc., can result in higher metabolic stability and, as a result, certain therapeutic advantages can be obtained (e.g., longer in vivo half-life or lower required dosage). In some embodiments, one or more hydrogen atoms are replaced with 2 H or 3 H, or one or more carbon atoms are replaced with 13 C or 14 C-enriched carbon. 15 O, 13 N, 11 C, and 18Positron-emitting isotopes such as F are useful in positron emission tomography (PET) studies to investigate substrate receptor occupancy. Isotope-labeled compounds can generally be prepared by replacing the non-isotope-labeled reagent with an isotope-labeled reagent by a procedure similar to the procedures described in the schemes or examples described herein.

[0074] It is specifically contemplated that any limitation discussed with respect to one embodiment of the invention may apply to any other embodiment of the invention. Further, any compound of the invention or a salt thereof (e.g., a pharmaceutically acceptable salt thereof) or composition can be used in any method of the invention, and any method of the invention can be used to produce or utilize any compound of the invention or a salt thereof (e.g., a pharmaceutically acceptable salt thereof) or composition.

[0075] The use of the term "or" means "and / or" unless explicitly indicated to refer only to alternatives or that the alternatives are mutually exclusive, in which case the disclosure supports a definition referring only to alternatives and "and / or".

[0076] Throughout this application, the term "about" is used to indicate that a value includes the standard deviation of error for the device or method used to measure that value.

[0077] As used herein, unless otherwise specified, "a" or "an" means one or more. As used herein, "another" means at least a second, or more.

[0078] The headings used herein are intended for organizational purposes only.

[0079] Inhibitor of Janus kinase One embodiment is of formula (I): TIFF0007716993000003.tif54170[wherein, Ar is phenyl; 1,2,3,4 - tetrahydroisoquinolinyl; pyrazolyl; pyridinyl; or pyridazinyl: R 1 is hydrogen; C1 - C6 alkyl; halo - C1 - C6 alkyl; hydroxy - C1 - C6 alkyl; -(CHR a ) h -het 1 ; -(CHR a ) k -NR a -het 1 ; or -(CHR a ) m -C 3-6 cycloalkyl, and the cycloalkyl moiety may be unsubstituted or may be substituted one or two times with R d ; each R 2 is independently: C1 - C6 alkyl; hydroxy - C1 - C6 alkyl; halo - C1 - C6 alkyl; C1 - C6 alkoxy; C1 - C6 alkoxy - C1 - C6 alkyl; halo - C1 - C6 alkoxy; halo - C1 - C6 alkoxy - C1 - C6 alkyl; C1 - C6 alkyl - SO2 - C1 - C6 alkyl; hydroxyl; cyano; cyano - C1 - C6 alkyl; halo; acetyl; -(CHR a ) p -het 2 ; -(CHR a ) q -NR b R c ; -(CHR a ) r -C(O)-NR b R c ; -(CHR a ) s -NR a -(CHR a ) s -C(O)-NR b R c [[ID=,66]]; or -(CHR a ) t -C 3-6 cycloalkyl, and the cycloalkyl moiety may be unsubstituted or may be substituted one or two times with R e ; R 3, R 4 and R 5 are each independently: hydrogen; or C1-C6 alkyl; each R a is independently: hydrogen; or C 1-6 alkyl; each R b is independently: hydrogen; C 1-6 alkyl; or hydroxy-C1-C6 alkyl; each R c is independently: hydrogen; C 1-6 alkyl; hydroxy-C1-C6 alkyl; cyano-C1-C6 alkyl; C1-C6 alkoxy-C1-C6 alkyl; oxetanyl; 2-morpholinoethyl; 1-methyl-azetidin-3-yl; 2-(N,N-dimethylamino)-ethyl; hydroxycyclobutyl; or 3-(N,N-dimethylamino)-pyrrolidin-1-yl; -(CHR a ) u -C 3-6 cycloalkyl, and the cycloalkyl moiety may be unsubstituted or may be substituted one or two times with R e ; or alternatively, R b and R c may together with the nitrogen atom to which they are attached form a het 3 ; each R d is independently: C1-C6 alkyl, hydroxy or halo; each R e is independently: C 1-6 alkyl; hydroxyl; cyano-C1-C6 alkyl; hydroxy-C1-C6 alkyl; morpholinyl; or -(CHR a ) v -NR g R h and R g and R h are each independently hydrogen or C 1-6 alkyl; h is from 0 to 2; k is from 0 to 2; m is from 0 to 2; n is from 0 to 2; p is from 0 to 2; q is from 0 to 2; r is from 0 to 2; s is from 0 to 2; t is from 0 to 2; u is from 0 to 2; v is from 0 to 2; het 1 is oxetanyl; tetrahydrofuranyl; tetrahydropyranyl; or pyrrolodinyl, each of which may be unsubstituted or substituted one or two times with R d ; het 2 is: azetidinyl; pyrrolidinyl; oxetanyl; piperidinyl; morpholinyl; piperazinyl; azepinyl; quinuclidinyl; or pyrazolyl, each of which may be unsubstituted or substituted one or two times with R e ; het 3 is: azetidinyl; pyrrolidinyl; piperidinyl; morpholinyl; piperazinyl; or azepinyl, each of which may be unsubstituted or substituted one or two times with R e To provide a compound of or a pharmaceutically acceptable salt thereof.

[0080] In certain embodiments, Ar is phenyl; or pyrazolyl.

[0081] In certain embodiments, Ar is phenyl.

[0082] In certain embodiments, Ar is pyrazolyl.

[0083] In certain embodiments, R 1 is hydrogen or C1-C6 alkyl.

[0084] In certain embodiments, R 1 is hydrogen. ​

[0085] In certain embodiments, R 1 is C1-C6 alkyl.

[0086] In certain embodiments, R 1 is methyl.

[0087] In certain embodiments, R 3 is hydrogen.

[0088] In certain embodiments, R 4 is hydrogen.

[0089] In certain embodiments, R 5 is hydrogen.

[0090] In certain embodiments, h is 0.

[0091] In certain embodiments, h is 1.

[0092] In certain embodiments, h is 2.

[0093] In certain embodiments, k is 0.

[0094] In certain embodiments, k is 1.

[0095] In certain embodiments, k is 2.

[0096] In certain embodiments, m is 0.

[0097] In certain embodiments, m is 1.

[0098] In certain embodiments, m is 2.

[0099] In certain embodiments, n is 0.

[0100] In certain embodiments, n is 1.

[0101] In certain embodiments, n is 2.

[0102] In certain embodiments, p is 0.

[0103] In certain embodiments, p is 1.

[0104] In certain embodiments, p is 2.

[0105] In certain embodiments, q is 0.

[0106] In certain embodiments, q is 1.

[0107] In certain embodiments, q is 2.

[0108] In certain embodiments, r is 0.

[0109] In certain embodiments, r is 1.

[0110] In certain embodiments, r is 2.

[0111] In certain embodiments, s is 0.

[0112] In certain embodiments, s is 1.

[0113] In certain embodiments, s is 2.

[0114] In certain embodiments, t is 0.

[0115] In certain embodiments, t is 1.

[0116] In certain embodiments, t is 2.

[0117] In certain embodiments, u is 0.

[0118] In certain embodiments, u is 1.

[0119] In certain embodiments, u is 2.

[0120] In certain embodiments, v is 0.

[0121] In certain embodiments, v is 1.

[0122] In certain embodiments, v is 2.

[0123] In some embodiments, each R 2 is independently selected from the following: TIFF0007716993000004.tif215170TIFF0007716993000005.tif80170.

[0124] In some embodiments, each R 2 is independently the following; TIFF0007716993000006.tif139170TIFF0007716993000007.tif137170.

[0125] In some embodiments, each R 2 is independently the following; TIFF0007716993000008.tif87170TIFF0007716993000009.tif184170.

[0126] In some embodiments, each R 2 is independently the following; TIFF0007716993000010.tif245170.

[0127] In some embodiments where n is 2, R 2One of them is selected from -CH2NH2; -OCF3; -CH2CN; -F; -CF3; -CHF2; -CH2CN; -OCF3; and TIFF0007716993000011.tif9170; and R 2 The other is as follows: TIFF0007716993000012.tif215170TIFF0007716993000013.tif67170.

[0128] In some embodiments where n is 1, R 2 is as follows: TIFF0007716993000014.tif163170TIFF0007716993000015.tif113170.

[0129] In some embodiments where n is 1, R 2 is as follows: TIFF0007716993000016.tif240170.

[0130] In certain embodiments, het 3 is selected from morpholinyl, azetidinyl, and piperazinyl.

[0131] In certain embodiments, the subject compound is of formula (II): TIFF0007716993000017.tif50170 where R 1 , R 2 and n are as defined herein.

[0132] In certain embodiments, the subject compound is of formula (III): TIFF0007716993000018.tif50170 where R 1 and R 2 are as defined herein.

[0133] In some embodiments, there is provided a compound selected from Table 1 below, or a salt thereof (e.g., a pharmaceutically acceptable salt) or a stereoisomer.

[0134] There is also provided a pharmaceutical composition comprising a JAK inhibitor described herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

[0135] There is also provided the use of a JAK inhibitor described herein or a pharmaceutically acceptable salt thereof in therapy, for example in the treatment of an inflammatory disease (e.g., asthma). There is also provided the use of a JAK inhibitor described herein or a pharmaceutically acceptable salt thereof for preparing a medicament for treating an inflammatory disease. There is also provided a method for preventing, treating or reducing the severity of a disease or condition responsive to inhibition of Janus kinase activity in a patient, comprising administering to the patient a therapeutically effective amount of a JAK inhibitor described herein or a pharmaceutically acceptable salt thereof.

[0136] In one embodiment, the disease or condition for treatment is cancer, polycythemia vera, essential thrombocythemia, myelofibrosis, chronic myeloid leukemia (CML), rheumatoid arthritis, inflammatory bowel syndrome, Crohn's disease, psoriasis, contact dermatitis or delayed type hypersensitivity reaction.

[0137] In one embodiment, there is provided the use of a JAK inhibitor described herein or a pharmaceutically acceptable salt thereof for the treatment of cancer, polycythemia vera, essential thrombocythemia, myelofibrosis, chronic myeloid leukemia (CML), rheumatoid arthritis, inflammatory bowel syndrome, Crohn's disease, psoriasis, contact dermatitis or delayed type hypersensitivity reaction.

[0138] In one embodiment, there is provided a composition formulated for administration by inhalation.

[0139] In one embodiment, there is provided a metered dose inhaler comprising a compound of the invention or a pharmaceutically acceptable salt thereof.

[0140] In one embodiment, the JAK inhibitor or a pharmaceutically acceptable salt thereof described herein is at least 5 times more potent as an inhibitor of JAK1 than as an inhibitor of LRRK2.

[0141] In one embodiment, the JAK inhibitor or a pharmaceutically acceptable salt thereof described herein is at least 10 times more potent as an inhibitor of JAK1 than as an inhibitor of LRRK2.

[0142] In one embodiment, there is provided a method for treating alopecia in a mammal, comprising administering to the mammal a JAK inhibitor or a pharmaceutically acceptable salt thereof described herein.

[0143] In one embodiment, there is provided the use of a JAK inhibitor or a pharmaceutically acceptable salt thereof described herein for the treatment of alopecia.

[0144] In one embodiment, there is provided the use of a JAK inhibitor or a pharmaceutically acceptable salt thereof described herein for preparing a medicament for treating alopecia in a mammal.

[0145] The compounds of the present invention may contain one or more asymmetric carbon atoms. Accordingly, the compounds can exist as diastereomers, enantiomers, or mixtures thereof. For the synthesis of the compounds, racemic compounds, diastereomers, or enantiomers can be used as starting materials or as intermediates. A mixture of specific diastereomeric compounds can be separated or enriched into one or more specific diastereomers by chromatography or crystallization methods. Similarly, enantiomeric mixtures can be separated or enantiomerically enriched using the same techniques or other techniques known in the art. Each of the asymmetric carbon or nitrogen atoms can be present in the R or S configuration, and both of these configurations are within the scope of the present invention.

[0146] In the structures shown in this specification, when the stereochemistry of any specific chiral atom is not specified, all stereoisomers are contemplated and included as compounds of the invention. When the stereochemistry is specified by a solid line wedge, or a dashed line representing a specific configuration, the stereoisomer is so specified and defined. Unless otherwise stated, when a solid line wedge, or a dashed line is used, relative stereochemistry is intended.

[0147] Another aspect includes prodrugs of the compounds described herein that are released and, for example, hydrolyzed to provide the compounds of the invention under physiological conditions and that include known amino protecting groups and carboxy protecting groups.

[0148] The term "prodrug" means a precursor or derivative form of a pharmaceutically active substance that is less active in patients compared to the parent drug and that can be activated by enzymatic or hydrolytic means or converted to a more active parent form. See, for example, Wilman, "Prodrugs in Cancer Chemotherapy", Biochemical Society Transactions, Vol. 14, pp. 375-382, 615th Meeting Belfast (1986) and Stella et al., "Prodrugs: A Chemical Approach to Targeted Drug Delivery", Directed Drug Delivery, Borchardt et al. (eds.), pp. 247-267, Humana Press (1985). Prodrugs include, but are not limited to, phosphate-containing prodrugs, thiophosphate-containing prodrugs, sulfate-containing prodrugs, peptide-containing prodrugs, D-amino acid-modified prodrugs, glycosylated prodrugs, β-lactam-containing prodrugs, optionally substituted phenoxyacetamide-containing prodrugs or optionally substituted phenylacetamide-containing prodrugs, and 5-fluorocytosine and 5-fluorouridine prodrugs.

[0149] Certain classes of prodrugs are compounds in which a nitrogen atom in an amino, amidino, aminoalkyleneamino, iminoalkyleneamino or guanidino group is substituted with a hydroxy group, an alkylcarbonyl (-CO-R) group, an alkoxycarbonyl (-CO-OR) or an acyloxyalkyl-alkoxycarbonyl (-CO-O-R-O-CO-R) group (wherein R is a monovalent or divalent group, such as alkyl, alkylene or aryl), or a compound substituted with the formula -C(O)-O-CP1P2-haloalkyl (wherein P1 and P2 are the same or different and are hydrogen, alkyl, alkoxy, cyano, halogen, alkyl or aryl). In certain embodiments, the nitrogen atom is one of the nitrogen atoms of the amidino group. The prodrug can be prepared by reacting the compound with an activated group, such as an acyl group, for example, by bonding a nitrogen atom in the compound to an exemplary carbonyl of the activated acyl group. Examples of activated carbonyl compounds are compounds containing a leaving group bonded to the carbonyl group, such as acyl halides, acylamines, acylpyridinium salts, acyl alkoxides, acyl phenoxides (such as p-nitrophenoxyacyl, dinitrophenoxyacyl, fluorophenoxyacyl, and difluorophenoxyacyl). The reaction is generally carried out at low temperatures, such as from -78 °C to about 50 °C, in an inert solvent. The reaction can also be carried out in the presence of an inorganic base, such as potassium carbonate or sodium bicarbonate, or an organic base, such as an amine including pyridine, trimethylamine, triethylamine, triethanolamine, etc.

[0150] Additional types of prodrugs are also included. For example, the free carboxyl group of the JAK inhibitors described herein can be derivatized as an amide or an alkyl ester. As another example, the compounds of the present invention containing a free hydroxy group can be derivatized as prodrugs by converting the hydroxy group to a group such as, but not limited to, a phosphate ester, a hemisuccinate, a dimethylaminoacetate, or a phosphoryloxymethyloxycarbonyl group, as outlined in Fleisher, D. et al., (1996) Improved oral drug delivery: solubility limitations overcome by the use of prodrugs (Advanced Drug Delivery Reviews, 19:115). Carbamate prodrugs of hydroxy and amino groups are also included, as well as carbonate prodrugs, sulfonic acid esters, and sulfate esters of hydroxy groups. Derivatization of hydroxy groups as (acyloxy)methyl and (acyloxy)ethyl ethers, where the acyl group can be an alkyl ester optionally substituted with groups including, but not limited to, ether, amine, and carboxylic acid functional groups, or where the acyl group is the above amino acid ester, is also included. This type of prodrug is described in J. Med. Chem., (1996), 39:10. As a more specific example, the hydrogen atom of an alcohol group is replaced with (C 1- C6) alkanoyloxymethyl, 1-((C 1- C6) alkanoyloxy)ethyl, 1-methyl-1((C 1- C6) alkanoyloxy)ethyl, (C 1- C6) alkoxycarbonyloxymethyl, N-(C 1- C6) alkoxycarbonylaminomethyl, succinoyl, (C 1- C6) alkanoyl, α-amino(C 1- C4) alkanoyl, arylacyl, and α-aminoacyl, or α-aminoacyl-α-aminoacyl (each α-aminoacyl group is a naturally occurring L-amino acid, P(O)(OH)2, -P(O)(O(C 1-Examples of replacement groups include groups such as (C6) alkyl) 2, or glycosyl (independently selected from groups resulting from removal of the hydroxyl group in the hemiacetal form of a carbohydrate).

[0151] A "leaving group" means a part of a first reactant in a chemical reaction that has been replaced from the first reactant in the chemical reaction. Examples of leaving groups include, but are not limited to, halogen atoms, alkoxy, and sulfonyloxy groups. Exemplary sulfonyloxy groups include alkylsulfonyloxy groups (e.g., methylsulfonyloxy (mesylate group) and trifluoromethylsulfonyloxy (triflate group)), and arylsulfonyloxy groups (e.g., p - toluenesulfonyloxy (tosylate group) and p - nitrosulfonyloxy (nosylate group)), but are not limited to these.

[0152] Synthesis of JANUS kinase inhibitor compounds Compounds can be synthesized by the synthetic routes described herein. In certain embodiments, in addition to or in light of the descriptions contained herein, processes well - known in the chemical art can be used. Starting materials are generally available from commercial sources such as Aldrich Chemicals (Milwaukee, Wis.), or can be readily prepared using methods well - known to those skilled in the art (e.g., Louis F. Fieser and Mary Fieser, Reagents for Organic Synthesis, v. 1 - 19, Wiley, N.Y. (1967 - 1999 ed.), Beilsteins Handbuch der organischen Chemie, 4, Aufl. ed. Springer - Verlag, Berlin, including supplements (also available via Beilstein online database)), or Comprehensive Heterocyclic Chemistry, Editors Katrizky and Rees, Pergamon Press, 1984).

[0153] The compounds can be prepared alone or as a library of compounds comprising at least two, for example 5 to 1,000 compounds, or 10 to 100 compounds. The library of compounds can be prepared by procedures known to those skilled in the art, by combinatorial "split and mix" approaches, or by multiple parallel synthesis using either solution-phase or solid-phase chemistry. Accordingly, according to a further aspect of the invention, there is provided a library of compounds comprising at least two compounds of the invention.

[0154] For illustrative purposes, the reaction scheme shown below provides a route for the synthesis of the compounds and key intermediates of the invention. For a more detailed description of the individual reaction steps, see the Examples section below. Those skilled in the art will understand that other synthetic routes are possible. Although some specific starting materials and reagents are described in the scheme and discussed below, other starting materials and reagents can be substituted to provide various derivatives or reaction conditions. Further, many of the compounds prepared by the methods described below can be further modified from the perspective of the present disclosure using conventional chemical phenomena well known to those skilled in the art.

[0155] In the preparation of the compounds of the invention, protection of the remote functionality of the intermediate (e.g., primary or secondary amine) may be required. The need for such protection will vary depending on the nature of the remote functional group and the conditions of the preparation method. Suitable amino protecting groups include acetyl, trifluoroacetyl, benzyl, phenylsulfonyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBz), and 9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protection can be readily determined by those skilled in the art. For a general description of protecting groups and their use, see T.W. Greene, "Protective Groups in Organic Synthesis", John Wiley & Sons, New York (1991).

[0156] Other transformations commonly used in the synthesis of the compounds of the present invention and which can be carried out using a variety of reagents and conditions include the following. (1) Formation of amides by reaction of carboxylic acids with amines. Such transformations can be achieved using a variety of reagents known to those skilled in the art, and a comprehensive review can be found in Tetrahedron, 2005, 61, 10827-10852. (2) The reaction of primary or secondary amines with aryl halides or pseudohalides, such as triflates, generally known as "Buchwald-Hartwig cross-coupling", can be achieved using a variety of catalysts, ligands and bases. A review of these methods is provided in Comprehensive Organic Name Reactions and Reagents, 2010, 575-581. (3) Palladium cross-coupling reactions between aryl halides and vinylboronic acids or boronic esters. This transformation is a type of "Suzuki-Miyaura cross-coupling", a reaction class that has been well studied in Chemical Reviews, 1995, 95(7), 2457-2483. (4) The hydrolysis of esters to give the corresponding carboxylic acids is well known to those skilled in the art and the conditions include the following: for methyl and ethyl esters, the use of strong aqueous bases such as lithium hydroxide, sodium hydroxide or potassium hydroxide or strong aqueous mineral acids such as HCl; for tert-butyl esters, hydrolysis is carried out using an acid, such as HCl in dioxane or trifluoroacetic acid (TFA) in dichloromethane (DCM).

[0157] Reaction Scheme 1 TIFF0007716993000019.tif126170

[0158] Reaction Scheme 1 illustrates the synthesis of the compound of formula I. The nitropyrazole compound 1 can be arylated with 4-bromo-1-(difluoromethoxy)-2-iodobenzene under palladium catalyst conditions to produce compound 2. The nitro group of compound 2 can be reduced under conditions such as iron chloride and ammonium chloride to produce aminoaniline 3. The commercially available pyrazolo[1,5-a]pyrimidine-3-carboxylic acid can be subjected to amide bond coupling in an organic solvent such as DMF using an organic base such as DIPEA and DMAP in the presence of a coupling reagent such as PyAOP to obtain compound 4. The compound of formula 6 can be synthesized by treating compound 4 with a suitably substituted hydroxyaryl compound 5 (such as phenol, pyrazole or pyridinol etc.) under Pd-catalyzed coupling conditions using a base such as cesium carbonate in a solvent such as toluene. The SEM protecting group of compound 6 can be removed using an acid such as HCl in a solvent such as 1,4-dioxane to obtain compound 7. Then, compound 7 can be N-alkylated with R 1 -X (wherein X is halo, for example bromo) to obtain compound 8.

[0159] It will be understood that when appropriate functional groups are present, compounds of various formulas or any intermediates used in their preparation can be further derivatized by one or more standard synthetic methods using condensation, substitution, oxidation, reduction or cleavage reactions. Specific substitution approaches include conventional alkylation, arylation, heteroarylation, acylation, sulfonylation, halogenation, nitration, formylation and coupling procedures.

[0160] In a further example, a primary amine group or a secondary amine group can be converted to an amide group (-NHCOR’ or -NRCOR’) by acylation. Acylation can be achieved by reaction with a suitable acid chloride in the presence of a base, such as triethylamine, in a suitable solvent, such as dichloromethane, or by reaction with a suitable carboxylic acid in the presence of a suitable coupling agent, such as HATU (O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate), in a suitable solvent, such as dichloromethane. Similarly, an amine group can be converted to a sulfonamide group (-NHSO2R’ or -NR’’SO2R’) by reacting it with a suitable sulfonyl chloride in the presence of a suitable base, such as triethylamine, in a suitable solvent, such as dichloromethane. A primary or secondary amine group can be converted to a urea group (-NHCONR’R’’ or -NRCONR’R’’) by reacting it with a suitable isocyanate in the presence of a suitable base, such as triethylamine, in a suitable solvent, such as dichloromethane.

[0161] An amine (-NH2) can be obtained by reducing a nitro (-NO2) group, for example, by catalytic hydrogenation using hydrogen in the presence of a metal catalyst, such as palladium on a carrier such as carbon, in a solvent such as ethyl acetate or an alcohol such as methanol. Alternatively, the conversion may be carried out by chemical reduction using a metal such as tin or iron in the presence of an acid such as hydrochloric acid.

[0162] In a further example, an amine (-CH2NH2) group can be obtained by reducing a nitrile (-CN) by catalytic hydrogenation using hydrogen, for example, in the presence of a metal catalyst, such as palladium on a carrier such as carbon or Raney nickel, in a solvent such as an ether, for example a cyclic ether such as tetrahydrofuran, at a suitable temperature, for example about -78 o °C to the reflux temperature of the solvent.

[0163] In a further example, an amine (-NH2) group can be obtained from a carboxylic acid group (-CO2H) by conversion to the corresponding acyl azide (-CON3), Curtius rearrangement and hydrolysis of the resulting isocyanate (-N=C=O).

[0164] An aldehyde group (-CHO) can be converted to an amine group (-CH2NR’R’’) by reductive amination using an amine and a boron hydride, such as sodium triacetoxyborohydride or sodium cyanoborohydride, in a solvent such as a halogenated hydrocarbon, for example dichloromethane, or an alcohol such as ethanol, in the presence of an acid such as acetic acid, near ambient temperature as required.

[0165] In a further example, an aldehyde group can be converted to an alkenyl group (-CH=CHR’) by use of a Wittig or Wadsworth-Emmons reaction using a suitable phosphorane or phosphonate under standard conditions known to those skilled in the art.

[0166] An aldehyde group can be obtained by reducing an ester group (-CO2Et etc.) or a nitrile (-CN) using diisobutylaluminum hydride in a suitable solvent such as toluene. Alternatively, an aldehyde group can be obtained by oxidation of an alcohol group using any suitable oxidizing agent known to those skilled in the art.

[0167] An ester group (-CO2R’) can be converted to the corresponding acid group (-CO2H) by acid or base-catalyzed hydrolysis, depending on the nature of R. When R is t-butyl, acid-catalyzed hydrolysis can be achieved, for example, by treatment with an organic acid such as trifluoroacetic acid in an aqueous solvent or by treatment with an inorganic acid such as hydrochloric acid in an aqueous solvent.

[0168] A carboxylic acid group (-CO2H) can be converted to an amide (CONHR’ or -CONR’R’’) by reaction with a suitable amine in the presence of a suitable coupling agent such as HATU in a suitable solvent such as dichloromethane.

[0169] In a further example, a carboxylic acid can be homologated by one carbon by conversion to the corresponding acid chloride (-COCl) followed by Arndt - Eistert synthesis (i.e., from -CO2H to -CH2CO2H).

[0170] In a further example, an -OH group can be generated by reduction from the corresponding ester (e.g., -CO2R’) or aldehyde (-CHO) using a complex metal hydride such as lithium aluminum hydride in, for example, diethyl ether or tetrahydrofuran, or sodium borohydride in a solvent such as methanol. Alternatively, an alcohol can be prepared by reduction of the corresponding acid (-CO2H) using lithium aluminum hydride in a solvent such as tetrahydrofuran, or borane in a solvent such as tetrahydrofuran.

[0171] An alcohol group can be converted to a leaving group such as a halogen atom or a sulfonyloxy group, such as an alkylsulfonyloxy, such as trifluoromethylsulfonyloxy or an arylsulfonyloxy, such as a p - toluenesulfonyloxy group, using conditions known to those skilled in the art. For example, an alcohol can be reacted with thionyl chloride in a halogenated hydrocarbon (e.g., dichloromethane) to obtain the corresponding chloride. A base (e.g., triethylamine) can also be used in the reaction.

[0172] In another example, an alcohol, phenol or amide group may be alkylated by coupling a phenol or amide with an alcohol in a solvent such as tetrahydrofuran in the presence of a phosphine, such as triphenylphosphine, and an activator, such as diethyl -, diisopropyl or dimethyl azodicarboxylate. Alternatively, alkylation can be achieved by deprotonating using a suitable base, such as sodium hydride, followed by addition of an alkylating agent, such as an alkyl halide.

[0173] The aromatic halogen substituents in the compound can be subjected to halogen-metal exchange by treatment with a base, such as a lithium base like n-butyl or t-butyl lithium, optionally at low temperature, e.g., about -78 o °C, in a solvent such as tetrahydrofuran, and then quenched with an electrophile to introduce the desired substituent. Thus, for example, a formyl group can be introduced by using N,N-dimethylformamide as the electrophile. Alternatively, the aromatic halogen substituent can be subjected to a metal (e.g., palladium or copper) catalyzed reaction to introduce, for example, an acid, ester, cyano, amide, aryl, heteroaryl, alkenyl, alkynyl, thio or amino substituent. Suitable procedures that can be used include those described by Heck, Suzuki, Stille, Buchwald or Hartwig.

[0174] The aromatic halogen substituent may also undergo nucleophilic substitution after reaction with a suitable nucleophile such as an amine or an alcohol. Advantageously, such a reaction can be carried out at high temperature in the presence of microwave irradiation.

[0175] Separation methods In each of the exemplary schemes, it may be advantageous to separate the reaction products from each other or from the starting materials. The desired product of each step or series of steps is separated or purified (hereinafter separated) to the desired degree of homogeneity by techniques common in the art. Typically, such separation includes multiphase extraction, crystallization or trituration from a solvent or solvent mixture, distillation, sublimation, or chromatography. Chromatography can involve any number of methods, including, for example, reverse and normal phase, size exclusion, ion exchange, supercritical fluid, high, medium, and low pressure liquid chromatography methods and apparatus, small scale analysis, simulated moving bed (SMB) and preparative thin or thick layer chromatography, and small scale thin layer and flash chromatography techniques.

[0176] Another class of separation methods involves treating the mixture with a selected reagent to bind to, or otherwise render separable, the desired product, unreacted starting materials, reaction by-products, etc. Such reagents include adsorbents or absorbents such as activated carbon, molecular sieves, ion exchange media, etc. Alternatively, the reagent may be an acid in the case of a basic substance, a base in the case of an acidic substance, a binding reagent such as an antibody, a binding protein, a selective chelating agent such as a crown ether, a liquid-liquid ion extraction reagent (LIX), etc.

[0177] The selection of an appropriate separation method depends on the nature of the materials involved. Examples of separation methods include boiling point and molecular weight in distillation and sublimation, presence or absence of polar functional groups in chromatography, stability of the materials in acidic and basic media in multiphase extraction, etc. One of ordinary skill in the art will apply the technique most likely to achieve the desired separation.

[0178] A mixture of diastereomers can be separated into their individual diastereoisomers based on their physicochemical differences by chromatography or by methods well known to one of ordinary skill in the art such as fractional recrystallization. Enantiomers can be separated by converting the enantiomer mixture to a diastereomer mixture by reaction with an appropriate optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers, and converting the individual diastereoisomers to the corresponding pure enantiomers (e.g., by hydrolysis). Also, some of the compounds of the present invention may be atropisomers (e.g., substituted biaryls) and are considered part of the invention. Enantiomers can also be separated by the use of a chiral HPLC column or supercritical fluid chromatography.

[0179] By resolving a racemic mixture using a method such as the formation of diastereomers using an optically active resolving agent, a single stereoisomer substantially free of its stereoisomers, for example an enantiomer, can be obtained (Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds, John Wiley & Sons, Inc., New York, 1994; Lochmuller, C. H., J. Chromatogr., 113(3):283 - 302(1975)). The racemic mixture of the chiral compounds of the present invention can be separated and isolated by any suitable method including the following. (1) Formation of an ionic diastereomeric salt with a chiral compound and separation by fractional recrystallization or other methods, (2) Formation of a diastereomeric compound with a chiral derivatizing reagent, separation of the diastereomers, and conversion to the pure stereoisomer, and (3) Separation of substantially pure or enriched stereoisomers under chiral conditions. See Drug Stereochemistry, Analytical Methods and Pharmacology, Irving W. Wainer, Ed., Marcel Dekker, Inc., New York (1993).

[0180] Diastereomeric salts can be formed by the reaction of enantiomerically pure chiral bases such as brucine, quinine, ephedrine, strychnine, α - methyl - β - phenylethylamine (amphetamine), etc. with asymmetric compounds having acidic functional groups such as carboxylic acids and sulfonic acids. The diastereomeric salts can be induced to separate by fractional recrystallization or ion chromatography. To separate the optical isomers of an amino compound, a chiral carboxylic acid such as camphorsulfonic acid, tartaric acid, mandelic acid, or lactic acid, or a sulfonic acid can be added to form a diastereomeric salt.

[0181] Alternatively, the substrate to be resolved is reacted with one enantiomer of a chiral compound to form a pair of diastereomers (Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds, John Wiley & Sons, Inc., New York, 1994, p. 322). Diastereomeric compounds can be formed by reacting an asymmetric compound with a chiral derivatizing reagent that is enantiomerically pure, such as a menthyl derivative, followed by separation of the diastereomers and hydrolysis to obtain the pure or enriched enantiomer. Methods for determining optical purity include preparing chiral esters of a racemic mixture, such as (-)-menthyl chloroformate in the presence of a base, or Mosher's ester, α-methoxy-α-(trifluoromethyl)phenylacetate (Jacob, J. Org. Chem. 47:4165 (1982)), and analyzing the NMR spectrum for the presence of the two atropisomeric enantiomers or diastereomers. Stable diastereomers of atropisomeric compounds can be separated and isolated by normal and reverse phase chromatography following a method for separating atropisomeric naphthyl-isoquinolines (International Publication No. WO 96 / 15111, incorporated herein by reference). By method (3), a racemic mixture of two enantiomers can be separated by chromatography using a chiral stationary phase (Chiral Liquid Chromatography, W.J. Lough, Ed., Chapman and Hall, New York, (1989); Okamoto, J. of Chromatogr. 513:375-378 (1990)). Enriched or purified enantiomers can be distinguished by methods used to distinguish other chiral molecules having asymmetric carbon atoms, such as optical rotation and circular dichroism. The absolute stereochemistry of the chiral center and enantiomers can be determined by X-ray crystallography.

[0182] Position isomers and intermediates for their synthesis can be observed by property evaluation methods such as NMR and analytical HPLC. For certain compounds with a sufficiently high energy barrier for interconversion, the E and Z isomers can be separated, for example, by preparative HPLC.

[0183] Pharmaceutical Compositions and Administration The compounds related to the present invention are JAK kinase inhibitors, such as JAK1 inhibitors, and are useful for the treatment of several diseases, such as inflammatory diseases like asthma.

[0184] Accordingly, another embodiment provides a pharmaceutical composition or medicament containing a compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient, and a method for preparing such a composition and medicament using the compound of the present invention.

[0185] In one example, the compound of the present invention or a pharmaceutically acceptable salt thereof can be formulated by mixing with a physiologically acceptable carrier, i.e., a carrier that is not toxic to the recipient at the dosages and concentrations used in the dosage forms of crude drugs, at an appropriate pH and desired purity at ambient temperature. The pH of the formulation mainly depends on the specific use and the concentration of the compound, but typically falls within a range of about 3 to about 8. In one example, the compound of the present invention or a pharmaceutically acceptable salt thereof is formulated at pH 5 in acetate buffer. In another embodiment, the compound of the present invention is sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation, or as an aqueous solution.

[0186] The composition is formulated, dosed, and administered in a manner consistent with good medical practice. Factors to be considered in this regard include the specific disorder being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of drug delivery, the method of administration, the dosing schedule, and other factors known to medical practitioners.

[0187] The specific dosage level for any particular patient will depend on a variety of factors including the activity of the specific compound being used, age, weight, general health, gender, diet, time of administration, route of administration, rate of excretion, drug combinations, and the severity of the particular disease being treated. Optimal dosage levels and dosing frequencies are determined by clinical trials as required in the pharmaceutical art. Generally, the daily dosage range for oral administration is from about 0.001 mg to about 100 mg per kg of human body weight, often from 0.01 mg to about 50 mg, for example 0.1 to 10 mg, in single or divided doses. Generally, the daily dosage range for inhalation administration is from about 0.1 μg to about 1 mg per kg of human body weight, preferably within the range of 0.1 μg to 50 μg per kg, in single or divided doses. In some cases, however, it may be necessary to use dosage amounts outside of these limits.

[0188] The compounds of the present invention or pharmaceutically acceptable salts thereof can be administered by any suitable method including oral administration, topical administration (including buccal and sublingual), rectal administration, intravaginal administration, transdermal administration, parenteral administration, subcutaneous administration, intraperitoneal administration, intralung administration, intradermal administration, intrathecal administration, inhalation and epidural as well as intranasal administration, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular administration, intravenous administration, intraarterial administration, intraperitoneal administration, or subcutaneous administration. In some embodiments, inhalation administration is used.

[0189] The compounds of the present invention or pharmaceutically acceptable salts thereof can be administered in any convenient dosage form, for example, tablets, powders, capsules, lozenges, granules, solutions, dispersions, suspensions, syrups, sprays, aerosols, suppositories, gels, emulsions, patches, etc. Such compositions can contain conventional ingredients in pharmaceutical preparations, for example, diluents (e.g., glucose, lactose or mannitol), carriers, pH adjusters, buffers, sweeteners, extenders, stabilizers, surfactants, wetting agents, lubricants, emulsifying agents, suspending agents, preservatives, antioxidants, opacifying agents, flow promoters, processing aids, colorants, fragrances, flavoring agents, other known additives and further active agents.

[0190] Suitable carriers and excipients are well-known to those skilled in the art and are described in detail, for example, in Ansel, Howard C., et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. For example, carriers include, as known to those skilled in the art, solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drugs, drug stabilizers, gels, binders, excipients, disintegrants, lubricants, sweetening agents, flavoring agents, dyes, such as similar materials and combinations thereof (see, for example, Remington’s Pharmaceutical Sciences, pp 1289-1329, 1990). Any conventional carrier is contemplated for use in a therapeutic or pharmaceutical composition, except when it is incompatible with the active ingredient. Exemplary excipients include dicalcium phosphate, mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, or combinations thereof. The pharmaceutical composition may contain different types of carriers or excipients depending on whether it is to be administered in solid, liquid, or aerosol form and whether it needs to be sterile for such a route of administration.

[0191] For example, tablets and capsules for oral administration may be in unit dosage forms and may contain conventional excipients such as binders, e.g., syrup, acacia, gelatin, sorbitol, tragacanth or polyvinylpyrrolidone; fillers, e.g., lactose, sugar, corn starch, calcium phosphate, sorbitol or glycine; tabletting lubricants, e.g., magnesium stearate, talc, polyethylene glycol or silica; disintegrants, e.g., potato starch, or acceptable wetting agents such as sodium lauryl sulfate. Tablets may be coated according to methods well known in the normal pharmaceutical practice. Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be provided as dry products for reconstitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, e.g., sorbitol, syrup, methyl cellulose, glucose syrup, gelatin hydrogenated edible fats; emulsifying agents, e.g., lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (which may include edible oils), e.g., almond oil, fractionated coconut oil, glycerin, propylene glycol, or oily esters such as ethyl alcohol; preservatives, e.g., methyl or propyl p-hydroxybenzoate or sorbic acid, and, if desired, conventional flavoring or coloring agents.

[0192] For topical application to the skin, the compounds can be formulated into creams, lotions or ointments. Cream or ointment formulations that can be used for the drug are conventional formulations well known in the art, such as those described in standard textbooks of pharmacy, e.g., the British Pharmacopoeia.

[0193] The compounds of the present invention or pharmaceutically acceptable salts thereof can also be formulated for inhalation, for example, as nasal sprays or dry powders or aerosol inhalers. For delivery by inhalation, the compounds are typically in particulate form, which can be prepared by various techniques including spray drying, freeze drying and micronization. Aerosol generation can be carried out, for example, using a pressure-driven jet nebulizer or ultrasonic nebulizer, such as by using a propellant-driven metered aerosol of the micronized compound from an inhalation capsule or other "dry powder" delivery system or administration without a propellant.

[0194] By way of example, the compositions of the present invention may be prepared as a suspension for delivery from a nebulizer or as an aerosol in a liquid propellant for use, for example, in a pressurized metered dose inhaler (PMDI). Propellants suitable for use in PMDIs are known to those skilled in the art and include CFC-12, HFA-134a, HFA-227, HCFC-22 (CCl2F2) and HFA-152 (CH4F2 and isobutane).

[0195] In some embodiments, the compositions of the present invention are in dry powder form for delivery using a dry powder inhaler (DPI). Many types of DPIs are known.

[0196] The fine particles for delivery by administration can be formulated with excipients that assist in delivery and release. For example, in dry powder formulations, the fine particles can be formulated with large carrier particles that assist the flow from the DPI to the lungs. Suitable carrier particles are known and include lactose particles. They can have, for example, an aerodynamic mass median diameter greater than 90 μm.

[0197] In the case of aerosol-based formulations, one example is as follows: Compound of the present invention * 24 mg / canister Lecithin, NF Liq. concentration 1.2 mg / canister Trichlorofluoromethane, NF 4.025 g / canister Dichlorodifluoromethane, NF 12.15 g / canister * or a pharmaceutically acceptable salt thereof.

[0198] The compounds of the invention or pharmaceutically acceptable salts thereof can be administered as described depending on the inhaler system used. In addition to the compound, the dosage form can further contain excipients as described above, or, for example, a propellant (e.g., Frigen in the case of a metered aerosol), a surfactant, an emulsifier, a stabilizer, a preservative, a flavoring, a filler (e.g., lactose in the case of a powder inhaler), or, if necessary, further active compounds.

[0199] For inhalation, a number of systems are available that can generate and administer an aerosol of optimal particle size using an inhalation technique suitable for the patient. In addition to the use of an adapter (spacer, expander) and an egg-shaped container (e.g., Nebulator®, Volumatic®), and an automatic device that releases a puff spray (Autohaler®), for metered aerosols, and in particular for powder inhalers, several technical solutions are available (e.g., Diskhaler®, Rotadisk®, Turbohaler® or inhalers as described in U.S. Patent No. 5,263,475, which is hereby incorporated by reference). Furthermore, the compounds of the invention or pharmaceutically acceptable salts thereof can be delivered in a multi-chamber device, thus enabling the delivery of combination agents.

[0200] The compound or a pharmaceutically acceptable salt thereof can also be administered parenterally in a sterile medium. Depending on the vehicle and concentration used, the compound can be suspended or dissolved in the vehicle. Advantageously, adjuvants such as local anesthetics, preservatives or buffers can be dissolved in the vehicle.

[0201] Targeted delivery of inhaled drugs The compounds of the present invention can be used for targeted inhalation delivery. Optimization of drugs for delivery to the lungs by topical (inhalation) administration has recently been investigated (Cooper, A.E. et al., Curr. Drug Metab. 2012, 13, 457-473).

[0202] Due to limitations of delivery devices, the dose of inhaled drugs may be limited in humans, which requires very potent molecules with good pulmonary pharmacokinetic properties. High potency against the target of interest is particularly important for inhaled drugs due to factors such as the limited amount of drug that can be delivered in a single puff from an inhaled drug and safety concerns associated with high aerosol loads in the lungs (e.g., cough or irritation). For example, in some embodiments, a Ki of about 0.5 nM or less in a JAK1 biochemical assay as described herein and an IC50 of about 20 nM or less in a JAK1-dependent cell-based assay as described herein may be desirable for an inhaled JAK1 inhibitor. In other embodiments, the predicted human dose of the compound of the present invention or a pharmaceutically acceptable salt thereof is at least one-half of the predicted human dose of a compound known in the art. Thus, in some embodiments, the compounds (or pharmaceutically acceptable salts thereof) described herein exhibit such potency values. The following procedures were used to evaluate the subject compounds for potential use as inhaled drugs.

[0203] IL13 Signaling. IL13 signaling is strongly involved in the etiology of asthma. IL13 is a cytokine that requires active JAK1 for signaling. Thus, inhibition of JAK1 inhibits IL13 signaling, which may provide benefits to asthma patients. Inhibition of IL13 signaling in animal models (e.g., mouse models) can predict future benefits for human asthma patients. Therefore, it may be beneficial for an inhaled JAK1 inhibitor to show suppression of IL13 signaling in an animal model. Methods for measuring such suppression are known in the art. For example, as discussed herein and known in the art, JAK1-dependent STAT6 phosphorylation is known to be a downstream consequence of IL13 stimulation. Thus, in some embodiments, the compounds (or pharmaceutically acceptable salts thereof) described herein show inhibition of pSTAT6 induction in the lung. To examine the pharmacodynamic effect on pSTAT6 levels, the compounds of the invention were co-administered with IL13 intranasally to female Balb / c mice. The compounds were formulated in 0.2% (v:v) Tween 80 in saline and mixed 1:1 (v:v) with IL13 immediately before administration. The intranasal dose was administered to lightly anesthetized (isoflurane) mice by pipetting a fixed volume (50 μL) directly into the nostrils to achieve the target dose levels (3 mg / kg, 1 mg / kg, 0.3 mg / kg, 0.1 mg / kg). At 0.25 hours after administration, blood samples (about 0.5 mL) were collected by cardiac puncture and plasma was generated by centrifugation (1500 g, 10 minutes, +4°C). The lungs were perfused with cold phosphate buffered saline (PBS), weighed, and snap frozen in liquid nitrogen. All samples were stored at about -80°C until analysis. Thawed lung samples were weighed and homogenized after adding 2 mL of HPLC grade water per gram of tissue using an Omni-Prep Bead Ruptor at 4°C. Plasma and lung samples were extracted by protein precipitation using three volumes of acetonitrile containing tolbutamide (50 ng / mL) and labetalol (25 ng / mL) as analytical internal standards.After vortex mixing and centrifugation at 3200 g for 30 minutes at 4 °C, the supernatant was appropriately diluted (e.g., 1:1 v:v) with HPLC-grade water in a 96-well plate. Representative aliquots of plasma and lung samples were assayed for the parent compound by LC-MS / MS against a series of matrix-matched calibrations and quality control standards. Standards were prepared by spiking test compounds into aliquots of control Balb / c mouse plasma or lung homogenate (2:1 in HPLC-grade water) and extracting as described for the experimental samples. The lung:plasma ratio was determined as the ratio of the mean lung concentration (μM) to the mean plasma concentration (μM) at the sampling time (0.25 h).

[0204] To measure pSTAT6 levels, mouse lungs were frozen at -80 °C until assay and homogenized in 0.6 ml of ice-cold cell lysis buffer (Cell Signalling Technologies, catalog number 9803S) supplemented with a cocktail of 1 mM PMSF and protease (Sigma Aldrich, catalog number P8340) and phosphatase (Sigma Aldrich, catalog numbers P5726 and P0044) inhibitors. Samples were centrifuged at 16060×g for 4 minutes at 4 °C to remove tissue debris and protein concentration of the homogenate determined using the Pierce BCA Protein Assay Kit (catalog number 23225). Samples were diluted to a protein concentration of 5 mg / ml in ice-cold distilled water and assayed for pSTAT6 levels by Meso Scale Discovery electrochemiluminescence immunoassay. Briefly, 5 μl / well of 150 μg / ml STAT6 capture antibody (R&D Systems, catalog number MAB 2169) was coated onto 96-well Meso Scale Discovery High Binding Plates (catalog number L15XB-3) and air-dried at room temperature for 5 hours. Plates were blocked by the addition of 150 μl / well of 30 mg / ml Meso Scale Discovery Blocker A (catalog number R93BA-4) and incubation for 2 hours at room temperature on a microplate shaker. The blocked plates were washed 4 times with Meso Scale Discovery TRIS wash buffer (catalog number R61TX-1) and then 50 μl / well of lung homogenate was transferred to achieve a protein load of 250 μg / well. The assay plates were incubated overnight at 4 °C, washed 4 times with TRIS wash buffer, and then 25 μl / well of 2.5 μg / ml sulfo-tag labeled pSTAT6 detection antibody (BD Pharmingen, catalog number 558241) was added for 2 hours at room temperature on a microplate shaker. The plates were washed 4 times with TRIS wash buffer and 150 μl / well of 1×Meso Scale Discovery Read Buffer T (catalog number R92TC-1) was added.The levels of pulmonary homogenate pSTAT6 were quantified by detection of electrochemiluminescence on a Meso Scale Discovery SECTOR S 600 instrument.

[0205] JAK and JAK2 inhibitor compounds that inhibit both JAK1 and JAK2 are potentially useful in the treatment of different types of asthma. Selectivity between JAK1 and JAK2 may also be important for inhaled JAK1 inhibitors. For example, GMCSF (granulocyte macrophage colony stimulating factor) is a cytokine that signals exclusively through JAK2. Neutralization of GMCSF activity is associated with pulmonary alveolar proteinosis (PAP) in the lung. However, submaximal JAK2 inhibition does not appear to be associated with PAP. Thus, even moderate JAK1 to JAK2 selectivity, or nearly equivalent inhibition of JAK1 and JAK2, may be beneficial in avoiding complete suppression of the GMCSF pathway and avoiding PAP. For example, in certain embodiments, compounds that are equipotent against JAK1 and JAK2 are desirable. In other embodiments, compounds having about 2- to 5-fold selectivity for JAK1 over JAK2 may be beneficial for inhaled JAK1 inhibitors. Thus, in some embodiments, the compounds (or pharmaceutically acceptable salts thereof) described herein exhibit such selectivity. Methods for measuring JAK1 and JAK2 selectivity are known in the art and information can also be found in the examples herein.

[0206] Kinase profiling. Further, it may be desirable for the inhaled JAK1 or JAK1 / JAK2 inhibitor to be selective for one or more other kinases, reducing the potential for toxicity resulting from off-target kinase pathway inhibition. Thus, it can also be beneficial for the inhaled JAK1 inhibitor to be selective for a broad range of non-JAK kinases, for example, in protocols available from ThermoFisher Scientific's SelectScreen™ Biochemical Kinase Profiling Service, Adapta™ Screening Protocol Assay Conditions (dated July 29, 2016), LanthaScreen™ Eu Kinase Binding Assay Screening Protocol and Assay Conditions (dated June 7, 2016), and / or Z’LYTE™ Screening Protocol and Assay Conditions (dated September 16, 2016). For example, the compounds of the present invention or pharmaceutically acceptable salts thereof exhibit at least 50-fold selectivity for JAK1 over a panel of non-JAK kinases. Thus, in some embodiments, the compounds (or pharmaceutically acceptable salts thereof) described herein exhibit such selectivity.

[0207] Cytotoxicity assay. Hepatotoxicity, general cytotoxicity, or cytotoxicity of unknown mechanism are undesirable features for potential drugs, including inhaled drugs. It can be beneficial for the inhaled JAK1 or JAK1 / JAK2 inhibitor to have low intrinsic cytotoxicity against various cell types. Typical cell types used to evaluate cytotoxicity include both primary cells such as human hepatocytes and established proliferating cell lines such as Jurkat and HEK-293. Thus, in some embodiments, the compounds (or pharmaceutically acceptable salts thereof) described herein exhibit such values. Methods for measuring cytotoxicity are known in the art. In some embodiments, the compounds described herein were tested as follows: (a) Jurkat and HEK293T cells were maintained at sub-confluent density in T175 flasks. The cells were seeded at 450 cells / 45 μl of medium in Greiner 384 well black / clear tissue culture treated plates (Greiner catalog number 781091). After dispensing the cells, the plates were equilibrated at room temperature for 30 minutes. After 30 minutes at room temperature, the cells were incubated overnight at 37 °C in a CO2 and humidity controlled incubator. The next day, the cells were treated with the compound diluted in 100% DMSO (final DMSO concentration on the cells = 0.5%) with a 10-point dose-response curve with a maximum concentration of 50 μM. The cells and the compound were then incubated for 72 hours overnight at 37 °C in a CO2 and humidity controlled incubator. After 72 hours of incubation, cell viability was measured for all wells using CellTiterGlo® (Promega catalog number G7572). After incubation at room temperature for 20 minutes, the plates were read on an EnVision® (Perkin Elmer Life Sciences) in luminescence mode. (b) Use of primary human hepatocytes: Test compounds were prepared as 10 mM solutions in DMSO. Further, positive controls such as chlorpromazine were prepared as 10 mM solutions in DMSO. Test compounds were typically evaluated using a 7-point dose-response curve with 2-fold dilutions. Typically, the maximum concentration tested was 50 - 100 μM. The highest concentration was typically determined by the solubility of the test compound. Cryopreserved primary human hepatocytes (BioreclamationIVT) (lot IZT) were thawed at 37 °C in InVitroGro™ HT Thaw Medium (BioreclamationIVT), pelleted, and resuspended. The viability of the hepatocytes was evaluated by trypan blue exclusion, and the cells were plated at a density of 13,000 cells / well in black-walled BioCoat™ Collagen 384-well plates (Corning BD) in InVitroGro™ CP Plating Medium supplemented with 1% Torpedo™ Antibiotic Mix (BioreclamationIVT) and 5% fetal bovine serum. The cells were incubated overnight for 18 hours (37 °C, 5% CO2) before treatment. After 18 hours of incubation, the plating medium was removed, and the hepatocytes were treated with the diluted compounds in InVitroGro™ HI Incubation Medium containing 1% Torpedo™ Antibiotic Mix and 1% DMSO (serum-free conditions). The hepatocytes were treated with test compounds at concentrations such as 0.78, 1.56, 3.12, 6.25, 12.5, 25, and 50 μM in a final volume of 50 μL. Positive controls (e.g., chlorpromazine) were typically included in the assay at the same concentrations as the test compounds. Additional cells were treated with 1% DMSO as a vehicle control. All treatments were performed for 48 hours (37 °C, 5% CO2), and each treatment condition was performed in triplicate. After 48 hours of compound treatment, the CellTiter-Glo® Cell Viability Assay (Promega) was used as an endpoint assay to measure the ATP content as a determination of cell viability. The assay was performed according to the manufacturer's instructions. Luminescence was measured with an EnVision™ Muliplate Reader (PerkinElmer, Waltham, MA, USA).The luminescence data were normalized against vehicle (1% DMSO) control wells. Inhibition curves and IC. 50 Estimates were generated by non-linear regression of log-transformed inhibitor concentration (7-point serial dilutions including vehicle) against normalized response, using a variable Hill slope, with upper and lower constraints fixed at constant values of 100 and 0, respectively (GraphPad Prism™, GraphPad Software, La Jolla, CA, USA).

[0208] hERG inhibition. Inhibition of the hERG (human ether-a-go-go related gene) potassium channel can lead to long QT syndrome and cardiac arrhythmias. Although the plasma levels of inhaled JAK1 or JAK1 / JAK2 inhibitors are expected to be low, pulmonary deposited compounds exiting the lung into the bloodstream via pulmonary absorption circulate directly to the heart. Thus, the local cardiac concentration of an inhaled JAK1 inhibitor can transiently be higher than the total plasma level, especially immediately after dosing. Therefore, it can be beneficial to minimize hERG inhibition by an inhaled JAK1 inhibitor. For example, in some embodiments, an hERG IC50 greater than 30-fold the Cmax of free drug plasma is preferred. Thus, in some embodiments, the compounds of the invention (or pharmaceutically acceptable salts thereof) exhibit minimized hERG inhibition under the following conditions: (a) Using hERG 2pt automated patch clamp conditions, the in vitro effect of the compound on hERG expressed in mammalian cells was examined and evaluated at room temperature using the QPatch HT® (Sophion Bioscience A / S, Denmark), an automated parallel patch clamp system. In some cases, the compound was tested at only one or two concentrations, such as 1 or 10 μM. In other cases, a broader concentration-response relationship was established to enable estimation of the IC50. For example, the test compound concentrations were selected to span a range of approximately 10-90% inhibition in semi-logarithmic increments. Each test article concentration was tested in two or more cells (n≥2). The exposure period to each test article concentration was at least 3 minutes; and / or (b) In the example of International Publication No. 2014 / 074775, it is described in the protocol of "Effect on Cloned hERG Potassium Channels Expressed in Mammalian Cells" (ChanTest™, Charles River Company), with the following modifications: Cells stably expressing hERG were held at -80 mV. The onset and steady-state inhibition of hERG potassium current by the compound were measured using a pulse pattern with a constant amplitude (conditioning prepulse: +20 mV for 1 second; repolarization test ramp to -90 mV (-0.5 V / s) repeated at 5-second intervals). Each recording was terminated with the final application of the highest concentration of the reference substance E-4021 (500 nM) (Charles River Company). The remaining uninhibited current was digitally subtracted from the data offline to determine the potency of the test substance against hERG inhibition.

[0209] CYP (Cytochrome P450) inhibition assay. CYP inhibition may not be a desirable feature for inhaled JAK1 or JAK1 / JAK2 inhibitors. For example, reversible or time-dependent CYP inhibitors can cause undesirable increases in their own plasma levels or the plasma levels of other co-administered drugs (drug-drug interactions). Additionally, time-dependent CYP inhibition can sometimes be caused by the in vivo conversion of the parent drug to reactive metabolites. Such reactive metabolites can covalently modify proteins and potentially lead to toxicity. Therefore, minimizing reversible and time-dependent CYP inhibition can be beneficial for inhaled JAK1 inhibitors. Thus, in some embodiments, the compounds of the present invention (or pharmaceutically acceptable salts thereof) exhibit minimal or no reversible and / or time-dependent CYP inhibition. Methods for measuring CYP inhibition are known in the art. The CYP inhibition of the compounds described herein was evaluated over a concentration range of 0.16 - 10 μM using pooled (n = 150) human liver microsomes (Corning, Tewksbury, MA) using a previously reported method (Halladay et al., Drug Metab. Lett. 2011, 5, 220 - 230). The incubation period and protein concentration were dependent on the CYP isoform and the probe substrate / metabolite being evaluated. The following substrates / metabolites, along with the incubation time and protein concentration for each CYP, were used: CYP1A2, phenacetin / acetaminophen, 30 minutes, 0.03 mg / ml protein; CYP2C9, warfarin / 7-hydroxywarfarin, 30 minutes, 0.2 mg / ml protein; CYP2C19, mephenytoin / 4-hydroxymephenytoin, 40 minutes, 0.2 mg / ml protein; CYP2D6, dextromethorphan / dextrorphan, 10 minutes, 0.03 mg / ml protein; CYP3A4, midazolam / 1-hydroxymidazolam, 10 minutes, 0.03 mg / ml protein, and, CYP3A4 testosterone / 6-hydroxytestosterone, 10 minutes, 0.06 mg / ml protein. These conditions were previously determined to be linear rates of formation for CYP-specific metabolites.All reactions were initiated with 1 mM NADPH and terminated by the addition of 0.1% formic acid in acetonitrile containing an appropriate stable-labeled internal standard. Samples were analyzed by LC-MS / MS.

[0210] Mouse lung tissue binding. A high binding fraction or percentage of JAK1 / JAK2 inhibitors to lung tissue may be undesirable as it may reduce the amount of free drug available to inhibit JAK1 or JAK2.

[0211] (a) Tissue binding experiments were performed in triplicate (n = 3) using disposable RED plates according to the standard protocol. First, individual drugs were spiked into tissue homogenate (pH 7.4) to achieve a final concentration of 1 μM, and then 300 μL of the drug-tissue homogenate mixture was transferred to the donor wells of a RED plate pre-filled with 500 μL of phosphate buffered saline (133 mM) on the receiver well. The RED plate was sealed with a gas-permeable membrane and placed in a 37 °C shaking incubator (450 rpm, VWR Symphony™) containing 5% CO2 for 6 h. At the end of incubation, an aliquot of 30 μL of the sample was removed from the RED device and an equal volume of matrix equalized with tissue homogenate or buffer was obtained, and then the resulting sample was immediately quenched with ice-cold acetonitrile (sample: acetonitrile 1:4) containing either propranolol or labetalol as the internal standard. After shaking at 500 rpm for 15 min on a Thermo Scientific Compact Digital MicroPlate Shaker, all samples were then subjected to centrifugation at 3700 rpm for 15 min (Beckman Coulter Allegra X 12R) to remove plasma proteins. The supernatant was then recovered and diluted with an equal volume of water prior to LC-MS / MS analysis.

[0212] (b) In another procedure, the extent of lung tissue binding of the test compound to mouse lung homogenate can also be determined by equilibrium dialysis using a Pierce RED (rapid equilibrium dialysis) apparatus (Fisher Scientific 89811 and 89809). A 10 mM solution of the compound in DMSO was prepared and diluted to 1 mM with DMSO. This 1 mM aliquot (4 μL) was added to the lung homogenate (dilution factor 1:9, lung tissue: potassium phosphate buffer (0.05 M, pH 7.4)) to obtain a final compound incubation concentration of 5 μM containing a solvent that occupies 0.5% (v / v) of the final incubation volume.

[0213] For each assay, the percentage of bound lung tissue was determined in triplicate. Lung homogenate (200 μL) was filled in triplicate on one side of the RED apparatus insert, and 350 μL of potassium phosphate buffer was filled on the other side. The RED apparatus was sealed and incubated at 37 °C (about 150 rpm) for about 4 hours on an orbital shaker.

[0214] After incubation, aliquots of the lung homogenate (8 μL) and the dialysate (72 μL) were matrix-matched prior to analysis (lung homogenate containing 72 μL of phosphate buffer, dialysate containing 8 μL of lung homogenate). 160 μL of acetonitrile containing an internal standard was added to precipitate the protein from the samples. For the evaluation of mass balance, the same matrix matching and protein precipitation procedures were performed on the lung homogenate aliquot sampled at the start of the experiment (t = 0 minute sample). The quenched samples were centrifuged (4000 rpm, 30 minutes, 4 °C), and the resulting supernatant was diluted with water (3:1 (v / v), supernatant: water), and the samples were analyzed for the parent compound by liquid chromatography mass spectrometry assay.

[0215] The unbound fraction (fu) in the lung homogenate was determined from the ratio of the dialysate to the homogenate peak area, corrected to account for the lung homogenate dilution (D), and the total lung tissue binding could be estimated using the following equation: Undiluted fu = (1 / D) / [((1 / apparent fu)-1)+(1 / D)] Corrected binding fraction (%) = (1 - undiluted fu) * 100

[0216] Kinetic solubility. To reduce the amount of undissolved particulate matter in the lungs, good water solubility of the JAK1 / JAK2 inhibitor for inhaled delivery may be desirable. In one procedure for measuring kinetic solubility, 4 μL of a 10 mM DMSO stock solution of the test compound is added to 196 μL of pH 7.4 phosphate-buffered saline in a Millipore Multiscreen® 96-well filter plate to obtain a test concentration of 200 μM containing 2% residual DMSO. The filter plate is sealed with an aluminum-sealing film and shaken at room temperature for 24 hours, then the mixture is vacuum filtered into a clean 96-well plate. The filtrate sample is diluted two-fold using pH 7.4 phosphate-buffered saline and then 5 μL of the resulting solution is analyzed by ultra-high performance liquid chromatography (UHPLC) using chemiluminescent nitrogen detection (CLND) and ultraviolet (UV) detection at a wavelength of 254 nm. The sample concentration is typically quantified by the CLND intensity related to the number of nitrogens in the compound. UV detection is used primarily to confirm sample purity, except in rare cases where the test compound contains no nitrogen. In those cases, a compound-specific calibration curve is collected based on UV absorbance. This curve is then used to determine the sample concentration.

[0217] Lipophilicity: Lipophilicity is generally related to the solubility, absorption, tissue penetration, protein binding, distribution, and ADME and PK properties of potential drugs. Therefore, the calculated logP (cLogP), which is the logarithm of the partition coefficient of the compound between n-octanol and water (i.e., log(concentration of the compound in n-octanol / concentration of the compound in water)), can be an important consideration for JAK1 / JAK inhibitors for inhaled delivery.

[0218] Liver microsomal stability. To minimize the systemic exposure of inhaled JAK1 / JAK2 inhibitors, it may be beneficial to optimize rapid metabolism in the liver. The liver microsomal stability assay was performed on a BioCel 1200 liquid handling workstation (Agilent Technologies, Santa Clara, CA). The compound (1.0 μM) was incubated at 37 °C for 5 minutes in 100 μL of reaction mixture containing 100 mM phosphate buffer (pH 7.4), 0.5 mg / mL liver microsomes and 1 mM NADPH. At different time intervals (0, 20, 40 and 60 minutes), aliquots of 20 μL of the reaction mixture were removed and mixed with 4 volumes of acetonitrile (ACN) containing 0.1 μM propranolol as internal standard to stop the metabolic reaction. The samples were then centrifuged at 3250 xg for 40 minutes to remove the precipitated protein. The supernatant was then transferred to a new 96-well plate, diluted 2-fold using deionized water, and then subjected to LC-MS / MS analysis using an ABI Sciex 5500 QTRAP® mass spectrometer (Applied Biosystems, Foster City, CA) combined with an Agilent 1260 HPLC (Agilent Technologies, Santa Clara, CA). The residual rate was calculated using the peak area ratio of the test compound to the internal standard at different time points compared to the control (T = 0 minutes). See B. Williamson, C. Wilson, G. Dagnell, RJ Riley. Harmonised high throughput microsomal stability assay. J. Pharmacol. Toxicol. Methods. 2017;84:31-36.

[0219] Solid state properties. For compounds intended to be delivered by dry powder inhalation, it is also necessary to be able to produce a crystalline form of the compound that can be micronized to a size of 1 - 5 μm. Particle size is an important determinant of lung deposition of inhaled compounds. Particles with a diameter less than 5 microns (μm) are typically defined as respirable. Particles with a diameter greater than 5 μm are more likely to deposit in the oropharynx and correspondingly less likely to deposit in the lungs. Also, fine particles with a diameter less than 1 μm are more likely to remain suspended in the air than larger particles and correspondingly more likely to be exhaled from the lungs. Thus, a particle diameter of 1 - 5 μm can be beneficial for inhaled pharmaceuticals whose site of action is the lung. Typical methods used to measure particle size include laser diffraction and cascade impaction. Typical values used to define particle size include · D10, D50, and D90. These are measurements of particle diameter indicating that 10%, 50%, or 90% of the sample, respectively, is below that value. For example, a D50 of 3 μm indicates that 50% of the sample is less than 3 μm in size. · Mass median aerodynamic diameter (MMAD). MMAD is the diameter at which 50% of the particles are larger and 50% are smaller, by mass. MMAD is a measure of central tendency. · Geometric standard deviation (GSD). GSD is a measure of the spread of the aerodynamic particle size distribution, i.e., the magnitude of the dispersion from the MMAD.

[0220] Common formulations for inhaled pharmaceuticals are dry powder formulations that include an active pharmaceutical ingredient (API) blended with a carrier such as lactose, with or without additional additives such as magnesium stearate. For such formulations, it can be beneficial for the API itself to have properties that enable it to be milled to a respirable particle size of 1 - 5 μm. Particle aggregation should be avoided, which can be measured by methods known in the art, such as examining the D90 value under different pressure conditions. Thus, in some embodiments, the compounds (or pharmaceutically acceptable salts thereof) of the present invention can be prepared at such respirable particle sizes with little or no aggregation.

[0221] Regarding crystallinity, for some formulations of inhaled drugs containing lactose blends, it is important that a specific crystalline form of the API is used. Crystallinity and crystal form can affect many parameters related to inhaled drugs, including, but not limited to, chemical and aerodynamic stability over time, compatibility with components of inhaled formulations such as lactose, hygroscopicity, lung retention, and lung irritation. Thus, a stable and reproducible crystal form can be beneficial for inhaled drugs. Further, the techniques used to mill the compound to the desired particle size are often energy-intensive and can convert a low melting point crystal form to another crystal form or render it fully or partially amorphous. Crystal forms with melting points below 150 °C may not be suitable for milling, and crystal forms with melting points below 100 °C are highly likely not to be suitable for milling. Thus, it can be beneficial for inhaled pharmaceuticals to have a melting point of at least above 100 °C, ideally above 150 °C. Thus, in some embodiments, the compounds (or pharmaceutically acceptable salts thereof) described herein exhibit such properties.

[0222] Furthermore, minimizing the molecular weight can help reduce the effective dose of the inhaled JAK1 inhibitor. A lower molecular weight results in a correspondingly higher number of molecules per unit mass of the active pharmaceutical ingredient (API). Thus, it can be beneficial to find an inhaled JAK1 inhibitor of the minimum molecular weight that retains all other desirable properties of the inhaled drug.

[0223] Finally, the compound needs to maintain a sufficient concentration in the lungs over a given period so that it can exert a pharmacological effect for the desired period and have a low systemic exposure for pharmacological targets where systemic inhibition of the target is not desirable. Since the lungs have essentially high permeability to both large molecules (proteins, peptides) and small molecules with a short pulmonary half-life, it may be necessary to attenuate the lung absorption rate by modifying one or more characteristics of the compound: minimizing membrane permeability, optimizing pKa, cLogP, solubility, dissolution rate, or introducing a degree of basicity (e.g., introducing an amine) into the compound to enhance binding to phospholipid-rich lung tissue or through sequestration in acidic intracellular compartments such as lysosomes (pH 5). Methods for measuring such characteristics are known in the art.

[0224] Thus, in some embodiments, the compounds (or pharmaceutically acceptable salts thereof) of the invention favorably exhibit one or more of the above characteristics. Further, in some embodiments, the compounds of the invention advantageously exhibit one or more of these characteristics as compared to compounds known in the art - this can particularly apply to compounds in the art intended as oral drugs versus inhalation. For example, compounds with rapid oral absorption typically are not retained in the lungs upon inhalation.

[0225] Methods of Treatment and Use of Janus Kinase Inhibitors The compounds of the present invention or pharmaceutically acceptable salts thereof inhibit the activities of Janus kinases such as JAK1 kinase. For example, the compound or its pharmaceutically acceptable salt inhibits the phosphorylation of signal transducer and activator of transcription (STAT) by JAK1 kinase and STAT-mediated cytokine production. The compounds of the present invention are useful for inhibiting JAK1 kinase activity in cells via cytokine pathways such as the IL-6, IL-15, IL-7, IL-2, IL-4, IL-9, IL-10, IL-13, IL-21, G-CSF, IFNα, IFNβ or IFNγ pathways. Thus, in one embodiment, there is provided a method of inhibiting the Janus kinase activity (e.g., JAK1 activity) of a cell by contacting the cell with a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0226] The compounds can be used for the treatment of immunological disorders caused by abnormal IL-6, IL-15, IL-7, IL-2, IL-4, IL9, IL-10, IL-13, IL-21, G-CSF, IFNα, IFNβ or IFNγ cytokine signaling.

[0227] Thus, one embodiment comprises a compound of the present invention or a pharmaceutically acceptable salt thereof for use in therapy.

[0228] In some embodiments, there is provided the use of a compound of the present invention or a pharmaceutically acceptable salt thereof in the treatment of inflammatory diseases. There is further provided the use of a compound of the present invention or a pharmaceutically acceptable salt thereof for preparing a medicament for treating an inflammatory disease such as asthma. There is also provided a compound of the present invention or a pharmaceutically acceptable salt thereof for use in the treatment of inflammatory diseases such as asthma.

[0229] Another embodiment includes a method of preventing, treating, or reducing the severity of a disease or condition such as asthma that responds to the inhibition of Janus kinase activity, such as JAK1 kinase activity, in a patient. The method may include administering to the patient a therapeutically effective amount of a compound of the invention or a pharmaceutically acceptable salt thereof. In one embodiment, the disease or condition that responds to the inhibition of a Janus kinase such as JAK1 kinase is asthma.

[0230] In one embodiment, the disease or condition is cancer, stroke, diabetes, hepatomegaly, cardiovascular disease, multiple sclerosis, Alzheimer's disease, cystic fibrosis, viral disease, autoimmune disease, atherosclerosis, restenosis, psoriasis, rheumatoid arthritis, inflammatory bowel disease, asthma, allergic disorder, inflammation, neuropathy, hormone-related disease, conditions associated with organ transplantation (e.g., transplant rejection), immunodeficiency disorder, destructive bone disorder, proliferative disorder, infectious disease, conditions associated with cell death, thrombin-induced platelet aggregation, liver disease, pathological immune states involving T cell activation, CNS disorder, or myeloproliferative disorder.

[0231] In one embodiment, the inflammatory disease is rheumatoid arthritis, psoriasis, asthma, inflammatory bowel disease, contact dermatitis, or delayed-type hypersensitivity. In one embodiment, the autoimmune disease is rheumatoid arthritis, lupus, or multiple sclerosis.

[0232] In another embodiment, the compound of the invention or a pharmaceutically acceptable salt thereof can be used to treat lung diseases such as fibrotic lung disease or interstitial lung disease (e.g., interstitial pneumonia). In some embodiments, the compound of the invention or a pharmaceutically acceptable salt thereof is idiopathic pulmonary fibrosis (IPF), systemic sclerosis interstitial lung disease (SSc-ILD), nonspecific interstitial pneumonia (NSIP), rheumatoid arthritis-associated interstitial lung disease (RA-ILD), sarcoidosis, hypersensitivity pneumonitis, or ILD secondary to connective tissue disease beyond scleroderma (e.g., polymyositis, dermatomyositis, rheumatoid arthritis, systemic lupus erythematosus (SLE), or mixed connective tissue disease).

[0233] In one embodiment, the cancer is breast, ovary, cervix, prostate, testis, penis, urogenital tract, seminoma, esophagus, larynx, stomach, gastric, gastrointestinal, skin, keratoacanthoma, follicular carcinoma, melanoma, lung, small cell lung carcinoma, non-small cell lung carcinoma (NSCLC), lung adenocarcinoma, squamous cell carcinoma of the lung, colon, pancreas, thyroid, papilla, bladder, liver, bile duct, kidney, bone, myeloid disorder, lymphoid disorder, hairy cell, oral and pharynx (mouth), lips, tongue, oral cavity, salivary gland, pharynx, small intestine, colon, rectum, anus, kidney, prostate, vulva, thyroid, large intestine, endometrium, uterus, brain, central nervous system, peritoneal cancer, hepatocellular carcinoma, head cancer, neck cancer, Hodgkin or leukemia.

[0234] In one embodiment, the disease is a myeloproliferative disorder. In one embodiment, the myeloproliferative disorder is polycythemia vera, essential thrombocythemia, myelofibrosis or chronic myelogenous leukemia (CML).

[0235] Another embodiment includes the use of a compound of the invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating a disease described herein (e.g., an inflammatory disorder, an immunological disorder or cancer). In one embodiment, the invention provides a method of treating a disease or condition described herein (e.g., an inflammatory disorder, an immunological disorder or cancer) by targeting the inhibition of a JAK kinase, such as JAK1.

[0236] Combination therapy The compound can be used alone or in combination with other agents for treatment. The second or further (e.g., third) compound of the pharmaceutical composition or dosing regimen typically has complementary activity to the compound of the invention so as not to adversely affect each other. Such agents are suitably combined and present in amounts effective for the intended purpose. The compounds may be administered together in a unitary pharmaceutical composition or separately, and if administered separately, this may occur simultaneously or sequentially, and such sequential administration may be at short or long intervals between administrations.

[0237] For example, for the prevention or treatment of inflammatory diseases such as asthma, other compounds can be combined with the compounds of the present invention or pharmaceutically acceptable salts thereof. Therapeutic agents suitable for combination therapy include, but are not limited to: adenosine A2A receptor antagonists; anti-infectives; non-steroidal glucocorticoid receptor (GR receptor) agonists; antioxidants; □2 adrenergic receptor agonists; CCR1 antagonists; chemokine antagonists (not CCR1); corticosteroids; CRTh2 antagonists; DP1 antagonists; formyl peptide receptor antagonists; histone deacetylase activators; chloride channel hCLCA1 blockers; epithelial sodium channel blockers (ENAC blockers; intercellular adhesion molecule 1 blockers (ICAM blockers); IKK2 inhibitors; JNK inhibitors; transient receptor potential ankyrin 1 (TRPA1) inhibitors; Bruton's tyrosine kinase (BTK) inhibitors (e.g., fenebrutinib); spleen tyrosine kinase (SYK) inhibitors; tryptase-beta antibodies; ST2 receptor antibodies (e.g., AMG282); cyclooxygenase inhibitors (COX inhibitors); lipoxygenase inhibitors; leukotriene receptor antagonists; dual 2 adrenergic receptor agonists / M3 receptor antagonists (MABA compounds); MEK-1 inhibitors; myeloperoxidase inhibitors (MPO inhibitors); muscarinic antagonists; p38 MAPK inhibitors; phosphodiesterase PDE4 inhibitors; phosphatidylinositol 3 kinase delta inhibitors (PI3 kinase delta inhibitors); phosphatidylinositol 3 kinase inhibitors (PI3 kinase inhibitors); peroxisome proliferator-activated receptor agonists (PPAR agonists); protease inhibitors; retinoic acid receptor modulators (RAR□ modulators); statins; thromboxane antagonists; TLR7 receptor agonists; or vasodilators.

[0238] Furthermore, the compound of the present invention or a pharmaceutically acceptable salt thereof may be combined with the following: (1) corticosteroids such as alclometasone dipropionate, amcinafal, beclomethasone dipropionate, budesonide, butixocort propionate, biclonide, clobetasol propionate, desisobutyryl ciclesonide, dexamethasone, etiprednol dicloacetate, fluocinolone acetonide, fluticasone furoate, fluticasone propionate, loteprednol etabonate (topical), or mometasone furoate; (2) β2 - adrenergic receptor agonists such as salbutamol, albuterol, terbutaline, fenoterol, bitolterol, carbuterol, clenbuterol, pirbuterol, rimiterol, terbutaline, tretoquinol, tulobuterol, and long - acting β2 - adrenergic receptor agonists such as metaproterenol, isoproterenol, isoprenaline, salmeterol, indacaterol, formoterol (including formoterol fumarate), arformoterol, carmoterol, abediterol, vilanterol trifenatate, or olodaterol; (3) combination products of corticosteroids / long - acting β2 agonists such as salmeterol / fluticasone propionate (also sold as Advair®, Seretide®), formoterol / budesonide (Symbicort®), formoterol / fluticasone propionate (Flutiform®), formoterol / ciclesonide, formoterol / mometasone furoate, indacaterol / mometasone furoate, vilanterol trifenatate / fluticasone furoate (BREO ELLIPTA), or arformoterol / ciclesonide; (4) anticholinergic drugs, for example, muscarinic - 3 (M3) receptor antagonists such as ipratropium bromide, tiotropium bromide, acridinium bromide (LAS - 34273), glycopyrronium bromide, or umeclidinium bromide.(5) Combinations of M3 - anti - cholinergic / β2 - adrenergic receptor agonists such as vilanterol / umeclidinium (Anoro® Ellipta®), olodaterol / thiotropium bromide, glycopyrronium bromide / indacaterol (also sold as Ultibro®, Xoterna®), fenoterol hydrobromide / ipratropium bromide (Berodual®), albuterol sulfate / ipratropium bromide (Combivent®), formoterol fumarate / glycopyrrolate, or acridinium bromide / formoterol; (6) Dual - pharmacology M3 - anti - cholinergic / β2 - adrenergic receptor agonists, for example, bitolterol succinate, AZD - 2115 or LAS - 190792; (7) Leukotriene modulators, for example, leukotriene antagonists such as montelukast, zafirlukast or pranlukast, or leukotriene biosynthesis inhibitors such as zileuton, or LTB4 antagonists such as amlexanox, or FLAP inhibitors such as fibrafenapone, GSK - 2190915; (8) Phosphodiesterase - IV (PDE - IV) inhibitors (oral or inhaled), for example roflumilast, cilomilast, oglemilast, rolipram, tetomilast, AVE - 8112, levamisole, CHF 6001; (9) Anti - histamine drugs, for example, selective histamine - 1 (H1) receptor antagonists such as fexofenadine, cetirizine, loratadine or astemizole, or dual H1 / H3 receptor antagonists such as GSK 835726 or GSK 1004723; (10) Cough suppressants such as codeine or dextromethorphan; (11) Mucolytics, for example, N - acetylcysteine or erdosteine; (12) Expectorant / mucokinetic agents, for example, ambroxol, hypertonic solutions (e.g., saline or mannitol) or surfactants; (13) Peptide mucolytics, for example, recombinant human deoxyribonuclease I (dornase - α and rhDNase) or heli cidin; (14) Antibiotics, for example, azithromycin, tobramycin or aztreonam; (15) Non - selective COX - 1 / COX - 2 inhibitors, for example, ibuprofen or ketoprofen;(16) COX-2 inhibitors, such as celecoxib and rofecoxib; (17) VLA-4 antagonists such as those described in International Publication No. WO 97 / 03094 and International Publication No. WO 97 / 02289, each incorporated herein by reference; (18) TACE inhibitors and TNF-α inhibitors, such as anti-TNF monoclonal antibodies such as Remicade® and CDP-870, and TNF receptor immunoglobulin molecules such as Enbrel®; (19) inhibitors of matrix metalloproteases (such as MMP-12); (20) human neutrophil elastase inhibitors such as BAY-85-8501, or those described in International Publication No. WO 2005 / 026124, International Publication No. WO 2003 / 053930, and International Publication No. WO 2006 / 082412, each incorporated herein by reference; (21) A2b antagonists such as those described in International Publication No. WO 2002 / 42298, incorporated herein by reference; (22) modulators of chemokine receptor function, such as antagonists of CCR3 and CCR8; (23) compounds that modulate the action of other prostanoid receptors, such as thromboxane A2 antagonists; DP1 antagonists such as laropiprant or asapiprant; CRTH2 antagonists such as OC000459, fevipiprant, ADC 3680, or ARRY 502; (24) PPAR agonists including PPARα agonists (such as fenofibrate), PPARδ agonists, and PPARγ agonists (such as pioglitazone, rosiglitazone, and balaglitazone); (25) methylxanthines such as theophylline or aminophylline, and combinations of methylxanthines / corticosteroids such as theophylline / budesonide, theophylline / fluticasone propionate, theophylline / ciclesonide, theophylline / mometasone furoate, and theophylline / beclomethasone dipropionate; (26) A2a agonists such as those described in European Patent No. EP 1052264 and European Patent No. EP 1241176; (27) CXCR2 antagonists or IL-8 antagonists such as AZD-5069, AZD-4721, or danirixin;(28) IL-R signaling modulators such as kineret and ACZ 885; (29) MCP-1 antagonists such as ABN-912; (30) p38 MAPK inhibitors, such as BCT197, JNJ49095397, rosmapimod or PH-797804; (31) TLR7 receptor agonists, such as AZD 8848; (32) PI3 kinase inhibitors such as RV1729 or GSK2269557 (nemiralisib); (33) three-component mixtures such as TRELEGY ELLIPTA (fluticasone furoate, umeclidinium bromide, and vilanterol); or, (34) small molecule inhibitors of TRPA1, BTK or SYK.

[0239] In some embodiments, the compounds of the invention or pharmaceutically acceptable salts thereof can be used in combination with one or more additional drugs, such as anti-proliferative agents, anti-cancer agents, cell growth inhibitors, cytotoxic agents, anti-inflammatory agents or chemotherapeutic agents, such as those disclosed in US Patent Application Publication No. 2010 / 0048557, which is incorporated herein by reference. The compounds of the invention or pharmaceutically acceptable salts thereof can also be used in combination with radiation therapy or surgery, as is known in the art.

[0240] Combinations of any of the foregoing with the compounds of the invention or pharmaceutically acceptable salts thereof are specifically contemplated.

[0241] Article of manufacture Another embodiment includes an article of manufacture (e.g., a kit) for treating a disease or disorder responsive to inhibition of a Janus kinase, such as JAK1 kinase. The kit can include (a) a first pharmaceutical composition comprising a compound of the invention or a pharmaceutically acceptable salt thereof, and (b) instructions for use, and can include: In another embodiment, the kit can (c) further include a second pharmaceutical composition, such as a pharmaceutical composition comprising an agent for such treatment, such as an agent for treating an inflammatory disorder or a chemotherapeutic agent.

[0242] In one embodiment, the instructions describe the simultaneous, sequential, or separate administration of the first and second pharmaceutical compositions to a patient in need thereof.

[0243] In one embodiment, the first and second compositions are contained in separate containers. In another embodiment, the first and second compositions are contained in the same container.

[0244] Containers for use include, for example, bottles, vials, syringes, blister packs, etc. The container may be formed from various materials such as glass or plastic. The container contains a compound of the invention or a pharmaceutically acceptable salt thereof effective in treating the condition and may have a sterile access port (e.g., the container can be an intravenous fluid bag or a vial with a stopper penetrable by a hypodermic needle). The label or package insert indicates that the compound is used to treat a selected condition such as asthma or cancer. In one embodiment, the label or package insert indicates that the compound can be used to treat a disorder. Further, the label or package insert may indicate that the patient to be treated has a disorder characterized by hyperactive or irregular Janus kinase activity, e.g., hyperactive or irregular JAK1 activity. The label or package insert may also indicate that the compound can be used to treat other disorders.

[0245] Alternatively or additionally, the kit may further comprise a second (or third) container containing a pharmaceutically acceptable buffer such as bacteriostatic water for injection (BWFI), phosphate buffered saline, Ringer's solution, or dextrose solution. Other materials desirable from a commercial and user perspective may be further included, including other buffers, diluents, filters, needles, and syringes.

[0246] To illustrate the present invention, the following examples are included. However, it should be understood that these examples are not intended to limit the present invention, but only to propose a method of implementing the present invention. Those skilled in the art will recognize that the described chemical reactions can be readily adapted to prepare other compounds of the present invention, and that alternative methods for preparing the compounds are within the scope of the present invention. For example, the synthesis of non-exemplary compounds according to the present invention can be carried out by making obvious changes to those skilled in the art, such as appropriately protecting intervening groups, using other suitable reagents known in the art other than those described, or making routine changes to the reaction conditions. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the present invention.

Example

[0247] Details of General Experiments Unless otherwise specified, all solvents and commercially available reagents were used as received. When the product was purified by chromatography on silica, this was done using either a glass column manually packed with silica gel (Kieselgel 60, 220 - 440 mesh, 35 - 75 μm) or an Isolute® SPE Si II cartridge. An "Isolute SPE Si cartridge" refers to a packed polypropylene column containing unbonded activated silica with irregular particles having an average size of 50 μm and a porosity of nominally 60 Å. When an Isolute® SCX-2 cartridge was used, an "Isolute® SCX-2 cartridge" refers to a packed polypropylene column containing an end-capped propylsulfonic acid functionalized silica strong cation exchange adsorbent.

[0248] LCMS Conditions Method A Experiments were carried out using a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3 mm Shim-Pack XR-ODS, particle size 2.2 μm), and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000020.jpg40170

[0249] Method B Experiments were carried out using a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3 mm Shim-Pack XR-ODS, particle size 2.2 μm), and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000021.jpg46170

[0250] Method C Experiments were carried out using a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3 mm Shim-Pack XR-ODS, particle size 2.2 μm), and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000022.jpg40170

[0251] Method D Experiments were carried out using a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3 mm Shim-Pack XR-ODS, particle size 2.2 μm), and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000023.jpg46170

[0252] Method E Experiments were carried out on a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3 mm Shim-Pack XR-ODS, particle size 2.2 μm), and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000024.jpg46170

[0253] Method F Experiments were carried out on a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3 mm Shim-Pack XR-ODS, particle size 2.2 μm), and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000025.jpg46170

[0254] Method G Experiments were carried out on a SHIMADZU 20A HPLC equipped with a C18 reversed-phase column (50×2.1 mm Ascentis Express C18, particle size 2.7 μm), and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000026.jpg40170

[0255] Method H Experiments were carried out on a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3 mm Shim-Pack XR-ODS, particle size 2.2 μm), and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000027.jpg39170

[0256] Method I Poroshell HPH-C 18, experiments were carried out on a SHIMADZU 20A HPLC equipped with a column (50×3 mm, particle size 2.7 μm), and elution was performed with solvent A: water / 5 mM NH4HCO3; solvent B: acetonitrile. Gradient: JPEG0007716993000028.jpg38170

[0257] Method J C18 reverse-phase column (50×3 mm Kinetex XB-C 18 , particle size 2.6 μm), experiments were carried out on a SHIMADZU LCMS-2020, and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000029.jpg40170

[0258] Method K C18 reverse-phase column (50×3 mm Shim-Pack XR-ODS, particle size 2.2 μm), experiments were carried out on a SHIMADZU LCMS-2020, and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000030.jpg40170

[0259] Method L C18 reverse-phase column (50×2.1 mm Kinetex XB-C 18 100A, particle size 2.6 μm), experiments were carried out on a SHIMADZU LCMS-2020, and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000031.jpg40170

[0260] Method M C18 reverse-phase column (30×2.1 mm Kinetex C18-100A, particle size 1.7 μm), experiments were carried out on a SHIMADZU LCMS-2020, and elution was performed with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000032.jpg42170

[0261] Method N Experiments were conducted using a SHIMADZU LCMS - 2020 equipped with a C18 reverse - phase column (50×3.0 mm Poroshell HPH - C18, particle size 2.7 μm), and elution was performed with Solvent A: water + 5 mM ammonium bicarbonate; Solvent B: acetonitrile. Gradient: JPEG0007716993000033.jpg40170

[0262] Method O Experiments were conducted using a SHIMADZU LCMS - 2020 equipped with a C18 reverse - phase column (50×3.0 mm Titank C18, particle size 3.0 μm), and elution was performed with Solvent A: water + 5 mM ammonium bicarbonate; Solvent B: acetonitrile. Gradient: JPEG0007716993000034.jpg39170

[0263] Method P Experiments were conducted using a SHIMADZU LCMS - 2020 equipped with a C18 reverse - phase column (30×2.1 mm Halo C18, particle size 2.0 μm), and elution was performed with Solvent A: water + 0.05% trifluoroacetic acid; Solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000035.jpg40170

[0264] Method Q Experiments were conducted using a SHIMADZU LCMS - 2020 equipped with a C18 reverse - phase column (50×3.0 mm YMC - Triart C18, particle size 2.5 μm), and elution was performed with Solvent A: water + 0.1% formic acid; Solvent B: acetonitrile + 0.1% formic acid. Gradient: JPEG0007716993000036.jpg40170

[0265] Method R Experiments were conducted using a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3 mm Shim-Pack XR-ODS, particle size 2.2 μm), and elution was performed with Solvent A: water + 0.05% trifluoroacetic acid; Solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000037.jpg40170

[0266] Method S Experiments were conducted using a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3.0 mm Poroshell HPH-C18, particle size 2.7 μm), and elution was performed with Solvent A: water + 5 mM ammonium bicarbonate; Solvent B: acetonitrile. Gradient: JPEG0007716993000038.jpg46170

[0267] Method T Experiments were conducted using a SHIMADZU 20A HPLC equipped with a C18 reversed-phase column (50×2.1 mm Ascentis Express C18, particle size 2.7 μm), and elution was performed with Solvent A: water + 0.05% trifluoroacetic acid; Solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000039.jpg40170

[0268] Method U Experiments were conducted using a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3 mm Shim-Pack XR-ODS, particle size 2.2 μm), and elution was performed with Solvent A: water + 0.05% trifluoroacetic acid; Solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient: JPEG0007716993000040.jpg46170

[0269] Method V Experiments were conducted using a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (50×3.0 mm Poroshell HPH-C18, particle size 2.7 μm), and elution was performed with Solvent A: water + 5 mM ammonium bicarbonate; Solvent B: acetonitrile. Gradient: JPEG0007716993000041.jpg47170

[0270] Method W Experiments were carried out on a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (Waters Acquity BEH, 50×2.1 mm, particle size 1.7 μm), and elution was performed with solvent A: water + 0.1% formic acid; solvent B: acetonitrile + 0.1% formic acid. Gradient: JPEG0007716993000042.jpg40170

[0271] Method X Experiments were conducted on an Agilent 1290 UHPLC connected to an Agilent MSD (6140) mass spectrometer using ESI as the ionization source. For LC separation, a Phenomenex XB-C18, 1.7 μm, 50×2.1 mm column was used at a flow rate of 0.4 ml / min. Mobile phase A was water with 0.1% formic acid, and mobile phase B was acetonitrile with 0.1% formic acid. The gradient started at 2% B for 7 minutes, ended at 98% B, and was maintained at 98% B for 1.5 minutes after 1.5 minutes of equilibration. The LC column temperature was 40°C. UV absorbance was collected at 220 nm and 254 nm, and full-scan mass spectrometry was applied to all experiments.

[0272] Method Y Experiments were carried out on a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (Gemini-NX, 50×3.0 mm, particle size 3.0 μm), and elution was performed with solvent A: water + 5 mM NH4HCO3; solvent B: acetonitrile + 5 mM NH4HCO3. Gradient: JPEG0007716993000043.jpg40170

[0273] Method Z Experiments were carried out on a SHIMADZU LCMS-2020 equipped with a C18 reversed-phase column (Gemini-NX, 50×3.0 mm, particle size 3.0 μm), and elution was performed with solvent A: water + 5 mM NH4HCO3; solvent B: acetonitrile + 5 mM NH4HCO3. Gradient: JPEG0007716993000044.jpg40170

[0274] List of common abbreviations ACN Acetonitrile Brine Saturated aqueous sodium chloride solution CH3OD Methanol-d CDCl3 Chloroform-d DCM Dichloromethane DIEA or DIPEA Diisopropylethylamine DMA Dimethylacetamide DMAP 4-Dimethylaminopyridine DMF Dimethylformamide DMSO Dimethyl sulfoxide DMSO-d6 Dimethyl sulfoxide-d6 EDC or EDCI 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide EtOAc Ethyl acetate EtOH Ethanol FA Formic acid HOAc Acetic acid g Gram h Hour HATU (O-(7-Azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate) HCl Hydrochloric acid HOBt Hydroxybenzotriazole HPLC High performance liquid chromatography IMS Industrial methylated spirit L Liter LCMS Liquid chromatography mass spectrometry LiHMDS or LHMDS Lithium hexamethyldisilazide MDAP Mass-directed automated purification MeCN Acetonitrile MeOH Methanol min Minute mg Milligram mL Milliliter NMR Nuclear Magnetic Resonance Spectroscopy Pd2(dba)3.CHCl3 Tris(dibenzylideneacetone)dipalladium(0)-chloroform adduct PE Petroleum Ether Preparative HPLC Preparative High Performance Liquid Chromatography SCX-2 Strong Cation Exchange TBAF Tetra-n-butylammonium fluoride THF Tetrahydrofuran TFA Trifluoroacetic acid Xantphos 4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene

[0275] The representative compounds in Table 1 below were prepared using the same procedures as those described in the schemes and examples of this specification. The absolute stereochemistry of each of the following compounds may not be shown. Therefore, structures representing a single stereoisomer may appear more than twice each. JPEG0007716993000045.jpg 241170 JPEG0007716993000046.jpg 228170 JPEG0007716993000047.jpg 234170 JPEG0007716993000048.jpg 229170 JPEG0007716993000049.jpg 224170 JPEG0007716993000050.jpg 231170 JPEG0007716993000051.jpg 233170 JPEG0007716993000052.jpg 223170 JPEG0007716993000053.jpg 229170 JPEG0007716993000054.jpg 233170 JPEG0007716993000055.jpg 234170 JPEG0007716993000056.jpg 225170 JPEG0007716993000057.jpg 222170 JPEG0007716993000058.jpg 225170 JPEG0007716993000059.jpg 235170 JPEG0007716993000060.jpg 221170 JPEG0007716993000061.jpg 221170 JPEG0007716993000062.jpg 228170 JPEG0007716993000063.jpg 221170 JPEG0007716993000064.jpg 228170 JPEG0007716993000065.jpg 246170 JPEG0007716993000066.jpg 214170 JPEG0007716993000067.jpg 217170 JPEG0007716993000068.jpg 220170 JPEG0007716993000069.jpg 215170 JPEG0007716993000070.jpg 224170 JPEG0007716993000071.jpg 230170 JPEG0007716993000072.jpg 224170 JPEG0007716993000073.jpg 225170 JPEG0007716993000074.jpg 215170 JPEG0007716993000075.jpg 221170 JPEG0007716993000076.jpg 224170 JPEG0007716993000077.jpg216170JPEG0007716993000078.jpg223170JPEG0007716993000079.jpg223170JPEG0007716993000080.jpg219170JPEG0007716993000081.jpg213170JPEG0007716993000082.jpg225170JPEG0007716993000083.jpg233170JPEG0007716993000084.jpg223170JPEG0007716993000085.jpg255170JPEG0007716993000086.jpg214170JPEG0007716993000087.jpg220170JPEG0007716993000088.jpg223170JPEG0007716993000089.jpg213170JPEG0007716993000090.jpg214170JPEG0007716993000091.jpg214170JPEG0007716993000092.jpg232170JPEG0007716993000093.jpg255170JPEG0007716993000094.jpg239170JPEG0007716993000095.jpg228170JPEG0007716993000096.jpg223170JPEG0007716993000097.jpg230170JPEG0007716993000098.jpg124170.

[0276] Intermediate 1 TIFF0007716993000099.tif64170N-(5-(5-Bromo-2-(difluoromethoxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0277] Step 1: Synthesis of 4-bromo-1-(difluoromethoxy)-2-iodobenzene A solution of N,N-dimethylformamide (2000 mL) and water (500 mL) containing 2-bromo-2-iodophenol (282 g, 943 mmol) was added with sodium 2-chloro-2,2-difluoroacetate (216 g, 1.42 mol) and cesium carbonate (617 g, 1.89 mol). The reaction vessel was equipped with a gas outlet for CO2 release. The resulting mixture was stirred at 120 °C overnight, cooled to room temperature, and poured into ice water (3000 mL). The resulting solution was extracted with ethyl acetate (3 x 1500 mL), and the organic layers were combined. The ethyl acetate extract was washed with brine (1000 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel eluting with ethyl acetate / petroleum ether (1 / 10) to give 300 g (91%) of 4-bromo-1-(difluoromethoxy)-2-iodobenzene as a yellow oil. 1 1H NMR (300 MHz, CDCl3) δ 7.96 (dd, J = 5.7 Hz, 2.4 Hz, 1H), 7.45 (dd, J = 8.7 Hz, 2.4 Hz, 1H), 7.03 (d, J = 8.7 Hz, 1H), 6.39 (t, J = 72.9 Hz, 1H).

[0278] Step 2: Synthesis of 5-[5-bromo-2-(difluoromethoxy)phenyl]-4-nitro-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazole A solution of 331704-nitro-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazole (100 g, 411 mmol) in anhydrous THF (1000 mL) was added dropwise to a solution of LiHMDS (490 mL, 1.0 mol / L in THF) with stirring at -70 °C under nitrogen. The resulting solution was stirred at -50 °C for 1 hour and then cooled to -70 °C. ZnCl2 (500 mL, 0.7 mol / L in THF) was added dropwise at -70 °C. The resulting solution was warmed to room temperature and stirred at room temperature for 1 hour. To the mixture were added 4-bromo-1-(difluoromethoxy)-2-iodobenzene (150 g, 860 mmol) and Pd(PPh3)4 (24.0 g, 20.8 mmol). The resulting solution was heated at reflux temperature overnight, cooled to room temperature, and concentrated under reduced pressure. This reaction on this scale was repeated once more, and the crude products from the two runs were combined for purification. The residue was purified by flash chromatography on silica gel eluting with ethyl acetate / petroleum ether (1 / 20). Appropriate fractions were combined and concentrated under reduced pressure. Thereby, 300 g (79%) of 5-[5-bromo-2-(difluoromethoxy)phenyl]-4-nitro-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazole was obtained as an overall pale yellow solid. 1 H NMR (300 MHz, CDCl3) δ 8.27 (s, 1H), 7.68 (dd, J = 8.7, 2.4 Hz, 1H), 7.62 (d, J = 2.4 Hz, 1H), 7.19 (d, J = 8.4 Hz, 1H), 6.39 (t, J = 72.5 Hz, 1H), 5.44 - 5.19 (m, 2H), 3.72 - 3.54 (m, 2H), 0.94 - 0.89 (m, 2H), 0.02 (s, 9H).

[0279] Step 3: Synthesis of 5-(5-bromo-2-(difluoromethoxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-amine A solution of ethanol (2000 mL) and water (200 mL) containing (50.1 g, 108 mmol) of 331705-(5-bromo-2-(difluoromethoxy)phenyl)-4-nitro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole was added to iron powder (60.1 g, 1.07 mol) and NH4Cl (28.0 g, 0.523 mol). The reaction mixture was stirred at reflux temperature for 3 hours under nitrogen. The solid was filtered and washed with ethanol (100 mL). The filtrate was concentrated under reduced pressure. The residue was dissolved in 3000 mL of ethyl acetate. The ethyl acetate solution was washed with 1×500 mL of brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 50.1 g of crude 5-(5-bromo-2-(difluoromethoxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-amine as a yellow oil. The crude product was used in the next step without further purification. LC / MS (method G, ESI): [M+H] + = 434.②, R T = 0.93 min.

[0280] Step 4: Synthesis of N-(5-(5-bromo-2-(difluoromethoxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide To a solution of (5-bromo-2-(difluoromethoxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-amine (50.1 g, 115 mmol) in DMA (1500 mL) were added pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (32.1 g, 196.0 mmol), PyAOP (102 g, 196 mmol), DMAP (1.41 g, 11.0 mmol) and DIPEA (44.1 g, 0.341 mol). The resulting solution was stirred at 60 °C in an oil bath for 3 h and then cooled to room temperature. The reaction mixture was then partitioned between water / ice (2000 mL) and ethyl acetate (2000 mL). The aqueous phase was extracted with ethyl acetate (2×). The combined organic layers were washed with brine (1000 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel eluting with ethyl acetate / petroleum ether (4:1). The appropriate fractions were combined and concentrated under reduced pressure. Water (150 mL) was added to the residue and the mixture was stirred in water at room temperature for 1 h. The solid was collected by filtration, air-dried to give 60.1 g (91%) of N-(5-(5-bromo-2-(difluoromethoxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide as a pale yellow solid. LC / MS (method G, ESI): [M+H] + = 579.1 and 581.1, R T = 1.10 min. 1 H NMR (300 MHz, CDCl3) δ 9.62 (s, 1H), 8.80 (dd, J = 6.9, 1.7 Hz, 1H), 8.73 (s, 1H), 8.53 (dd, J = 4.2, 1.7 Hz, 1H), 8.38 (s, 1H), 7.79 (d, J = 2.4 Hz, 1H), 7.67 (dd, J = 8.8, 2.5 Hz, 1H), 7.29 (d, J = 1.4 Hz, 1H), 7.00 (dd, J = 6.9, 4.2 Hz, 1H), 6.43 (t, J = 72.6 Hz, 1H), 5.53 - 5.27 (m, 2H), 3.73 - 3.50 (m, 2H), 0.88 (ddd, J = 9.5, 6.4, 4.4 Hz, 2H), 0.00 (s, 9H).

[0281] Intermediate 2 TIFF0007716993000104.tif51170N-[3-[5-Bromo-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0282] N-[5-[5-Bromo-2-(difluoromethoxy)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 2, 5.00 g, 8.63 mmol) was treated with HCl / dioxane (150 mL, 4 M) at room temperature overnight. The resulting mixture was concentrated under reduced pressure. Thereby, 3.80 g of N-[3-[5-Bromo-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a yellow solid. The purity of the intermediate was sufficient for use in the next step without further purification. LC / MS (Method I, ESI): [M+H] + = 449.0, R T = 1.02 min. 1 H NMR (400 MHz, CD3OD) δ 9.11 (dd, J = 6.8, 1.6 Hz, 1H), 8.67 - 8.64 (m, 2H), 8.32 (s, 1H), 7.80 (d, J = 2.4 Hz, 1H), 7.72 (dd, J = 8.8, 2.4 Hz, 1H), 7.37 (d, J = 8.8 Hz, 1H), 7.23 (dd, J = 7.0, 4.2 Hz, 1H), 6.81 (t, J = 73.2 Hz, 1H).

[0283] Intermediate 3 TIFF0007716993000105.tif52170N-[3-[5-Bromo-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0284] In dichloromethane (200 mL), to a solution of N-[5-[5-bromo-2-(difluoromethoxy)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 1, 10.1 g, 17.3 mmol) was added Me3OBF4 (2.81 g, 18.9 mmol) at room temperature. The resulting solution was stirred at room temperature for 2 hours. Then, 10 mL of EtOH was added to the reaction mixture, and the reaction mixture was stirred for 1 hour. To this solution was added 5.0 mL of HCl (concentrated), and the mixture was stirred for 1 hour. The resulting mixture was concentrated under vacuum. The pH value of the solution was adjusted to 8 using sodium bicarbonate (20%). The resulting solution was extracted with 3 × 300 mL of ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under high vacuum. The residue was applied to a silica gel column eluting with ethyl acetate / petroleum ether (80%) to give 5.5 g (69%) of N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a pale yellow solid. 1 H NMR (400 MHz, CDC13): δ (ppm) 9.86 (s, 1H), 8.80 (dd, J = 7.0, 1.6 Hz, 1H), 8.74 (s, 1H), 8.60 (dd, J = 4.2, 1.6 Hz, 1H), 8.32 (s, 1H), 7.85 (d, J = 2.4 Hz, 1H), 7.58 (dd, J = 8.4, 2.4 Hz, 1H), 7.24 (d, J = 8.8 Hz, 1H), 7.02 (dd, J = 7.0, 4.2 Hz, 1H), 6.49 (t, J = 74.0 Hz, 1H), 4.01 (s, 3H).

[0285] Intermediate 4 TIFF0007716993000106.tif51170N-(3-(2-(Difluoromethoxy)-5-hydroxyphenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0286] Synthesis of Project 1: N-[3-[2-(Difluoromethoxy)-5-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007716993000107.tif51170 To a 30 mL sealed tube purged and maintained under an inert atmosphere of nitrogen, N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 3, 1000 mg, 2.16 mmol), 4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (824 mg, 3.25 mmol, 1.50 equiv), Pd(dppf)Cl2-CH2Cl2 (176 mg, 0.216 mmol, 0.100 equiv), DPPF (119 mg, 0.215 mmol, 0.100 equiv), potassium acetate (636 mg, 6.48 mmol, 3.00 equiv), and dioxane (18 mL) were added. The resulting mixture was stirred at 110 °C overnight and then concentrated under vacuum. The residue was purified by silica gel column chromatography eluting with ethyl acetate / petroleum ether (60:40). This gave 965 mg (88%) of N-[3-[2-(difluoromethoxy)-5-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a pale yellow solid. LC / MS (Method H, ESI): [M+H] + = 511.2, R T = 1.31 min.

[0287] Synthesis of Project 2: N-(3-(2-(Difluoromethoxy)-5-hydroxyphenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007716993000108.tif53170100 mL round-bottomed flask was charged with N-[3-[2-(difluoromethoxy)-5-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (965 mg, 1.89 mmol), tetrahydrofuran (10 mL), and H2O2 (0.5 mL, 21.5 mmol). The resulting solution was stirred at room temperature overnight and then concentrated under vacuum. As a result, 680 mg (90%) of N-[3-[2-(difluoromethoxy)-5-hydroxyphenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a gray solid. LC / MS (method H, ESI): [M+H] + = 401.1, R T = 1.04 min.

[0288] Example Example 1 TIFF0007716993000109.tif55170N-(3-(2-(Difluoromethoxy)-5-phenoxyphenyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0289] Step 1: Synthesis of N-(5-(2-(Difluoromethoxy)-5-phenoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide In a 10 mL round-bottom flask purged and maintained under an inert nitrogen atmosphere, N-(5-(5-bromo-2-(difluoromethoxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 1, 100 mg, 0.173 mmol), phenol (32.5 mg, 0.345 mmol, 2.00 eq), [PdCl(allyl)]2 (6.31 mg, 0.017 mmol, 0.10 eq), RockPhos (16.2 mg, 0.035 mmol, 0.20 eq), cesium carbonate (84.3 mg, 0.26 mmol, 1.50 eq), and toluene (2 mL) were added. The resulting solution was stirred at 100 °C in an oil bath for 14 h. The resulting mixture was concentrated under vacuum. The residue was applied to a silica gel column using ethyl acetate / petroleum ether (1:1). Thereby, 100 mg (98%) of N-(5-(2-(difluoromethoxy)-5-phenoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a yellow solid. LC / MS (Method J, ESI): [M+H] + = 593.3, R T = 1.35 min.

[0290] Step 2: Synthesis of N-(3-(2-(difluoromethoxy)-5-phenoxyphenyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide. TIFF0007716993000111.tif5517025 mL round-bottom flask was charged with N-(5-(2-(difluoromethoxy)-5-phenoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.169 mmol), trifluoroacetic acid (2 mL) and dichloromethane (8 mL). The resulting solution was stirred at room temperature for 30 minutes and concentrated under vacuum. The crude product was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, C18 silica gel; mobile phase, CH3CN:H2O = 30:70, increasing to CH3CN:H2O = 70:30 within 14 minutes; detector, UV 254 nm. Thereby, 28.5 mg (37%) of N-(3-(2-(difluoromethoxy)-5-phenoxyphenyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a pale yellow solid. LC / MS (method G, ESI): [M+H] + = 463.2, R T = 0.86 min. 1 1H NMR (300 MHz, CD3OD) δ 9.14 - 9.12 (m, 1H), 8.69 - 8.65 (m, 2H), 8.28 - 8.26 (m, 1H), 7.45 - 7.42 (m, 1H), 7.33 - 7.04 (m, 8H), 6.72 (t, J = 73.7 Hz, 1H).

[0291] Example 133 TIFF0007716993000112.tif54170N-(3-(2-(difluoromethoxy)-5-((5-((dimethylamino)methyl)pyridin-3-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide N-[3-[5-Bromo-2-(difluoromethoxy)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 3, 300 mg, 0.648 mmol), 5-[(dimethylamino)methyl]pyridin-3-ol (197 mg, 1.30 mmol), [PdCl(allyl)]2 (9.48 mg, 0.026 mmol), RockPhos (30.4 mg, 0.065 mmol), cesium carbonate (422 mg, 1.30 mmol), and toluene (13 mL) were added to a 100 mL round-bottom flask and degassed with nitrogen for 5 minutes. The reaction mixture was stirred at 100 °C overnight and then concentrated under vacuum. The crude product was purified by reverse-phase HPLC to give the title (39.0 mg, 0.071 mmol, 10.9% yield) product as a white solid. LC / MS (Method W, ESI): [M+H] + = 535.2, R T = 1.04 min. 1 H NMR (400 MHz, DMSO-d6) δ 9.73 (s, 1H), 9.35 (dd, J = 7.0, 1.6 Hz, 1H), 8.80 - 8.51 (m, 2H), 8.41 - 8.09 (m, 3H), 7.47 (d, J = 8.9 Hz, 1H), 7.40 - 6.96 (m, 5H), 3.89 (s, 3H), 3.39 (s, 2H), 2.10 (s, 6H).

[0292] Example 157 TIFF0007716993000113.tif55170N-(3-(2-(Difluoromethoxy)-5-((1,2,3,4-tetrahydroisoquinolin-7-yl)oxy)phenyl)-1-(oxetan-3-yl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0293] Step 1: Synthesis of N-(3-(5-Bromo-2-(difluoromethoxy)phenyl)-1-(oxetan-3-yl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide In a 5417025 mL round-bottom flask, N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 2, 200 mg, 0.445 mmol), N,N-dimethylformamide (8 mL), cesium carbonate (403 mg, 1.24 mmol, 3.0 equivalents), and 3-iodooxetane (97.3 mg, 0.529 mmol, 1.2 equivalents) were added. The resulting solution was stirred at 60 °C for 4 hours. The resulting mixture was concentrated under vacuum. The residue was poured onto a silica gel column equipped with dichloromethane / methanol (10:1). Thereby, 187 mg (83%) of N-(3-(5-bromo-2-(difluoromethoxy)phenyl)-1-(oxetan-3-yl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a yellow solid. LC / MS (Method J, ESI): [M+H] + = 505.1 and 507.1, R T = 1.03 minutes.

[0294] Step 2: Synthesis of tert-butyl 7-(4-(difluoromethoxy)-3-(1-(oxetan-3-yl)-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenoxy)-3,4-dihydroisoquinoline-2(1H)-carboxylate In a sealed tube of 5417030 mL of TIFF0007716993000115.tif, N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-(oxetan-3-yl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (187 mg, 0.37 mmol), toluene (10 mL), Pd2(allyl)2Cl2 (6.8 mg, 0.019 mmol, 0.050 equiv), t-BuBrettPhos (17.9 mg, 0.037 mmol, 0.10 equiv), cesium carbonate (145 mg, 0.45 mmol, 1.2 equiv), tert-butyl 7-hydroxy-1,2,3,4-tetrahydroisoquinoline-2-carboxylate (111 mg, 0.45 mmol, 1.2 equiv) were added. The resulting solution was stirred at 100 °C overnight. The resulting mixture was concentrated under vacuum. The residue was applied to a silica gel column using ethyl acetate / petroleum ether (1:1). Thereby, 125 mg (50%) of tert-butyl 7-(4-(difluoromethoxy)-3-(1-(oxetan-3-yl)-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenoxy)-3,4-dihydroisoquinoline-2(1H)-carboxylate was obtained as a yellow solid. LC / MS (method H, ESI): [M+H] + = 674.4, R T = 1.40 min.

[0295] Step 3: Synthesis of N-(3-(2-(difluoromethoxy)-5-((1,2,3,4-tetrahydroisoquinolin-7-yl)oxy)phenyl)-1-(oxetan-3-yl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007716993000116.tif5417025 mL round-bottom flask was charged with tert-butyl 7-(4-(difluoromethoxy)-3-(1-(oxetan-3-yl)-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenoxy)-3,4-dihydroisoquinoline-2(1H)-carboxylate (150 mg, 0.223 mmol), hydrogen chloride (4 M in dioxane, 1 mL), and 1,4-dioxane (4 mL). The resulting solution was stirred at room temperature for 1 hour. The resulting mixture was concentrated under vacuum. This afforded 120 mg (94%) of N-(3-(2-(difluoromethoxy)-5-((1,2,3,4-tetrahydroisoquinolin-7-yl)oxy)phenyl)-1-(oxetan-3-yl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. LC / MS (Method A, ESI): [M+H] + = 574.3, R T = 1.30 min. 1 H NMR (300 MHz, CD3OD) δ 9.11 (dd, J = 7.2, 1.8 Hz, 1H), 8.67 - 8.64 (m, 2H), 8.41 (s, 1H), 7.40 (d, J = 8.8 Hz, 1H), 7.25 - 7.21 (m, 2H), 7.17 - 7.14 (m, 1H), 7.08 - 6.51 (m, 3H), 5.62 - 5.57 (m, 1H), 5.12 - 5.08 (m, 4H), 3.89 (s, 2H), 3.10 - 3.06 (m, 2H), 2.82 - 2.80 (m, 2H).

[0296] Example 128 TIFF0007716993000117.tif54170N-(3-(2-(difluoromethoxy)-5-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0297] Process 1: N-(3-(2-(Difluoromethoxy)-5-((1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007716993000118.tif49170N2, N-(3-(5-Bromo-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide i (Intermediate 3, 56 g, 121 mmol), 1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-ol (35.0 g, 163 mmol), ditert-butyl-[6-methoxy-3-methyl-2-(2,4,6-triisopropylphenyl)phenyl]phosphane (5.78 g, 12.3 mmol) and allyl(chloro)palladium (2.30 g, 6.29 mmol) in toluene (1.40 L) were added K2CO3 (50.0 g, 361 mmol) at 25 °C. The resulting solution was stirred at 100 °C for 3 hours. The resulting mixture was concentrated under vacuum and diluted with water (500 mL). The resulting solution was extracted with ethyl acetate (3 × 400 mL) and the organic layers were combined. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash chromatography on silica gel eluting with ethyl acetate / petroleum ether (75%) to give N-(3-(2-(Difluoromethoxy)-5-((1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (150 g, crude) as a green solid. LC / MS (Method G, ESI): [M+H] + = 597.2, R T = 1.19 min.

[0298] Process 2: N-(3-(5-((1H-Pyrazol-4-yl)oxy)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide A solution of methanol (450 mL) containing TIFF0007716993000119.tif49170N-(3-(2-(difluoromethoxy)-5-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (45 g, 75.4 mmol) was added with HCl (230 g, 6.29 mol, 225 mL). The reaction mixture was heated to 60 °C and stirred for 30 minutes. This procedure was repeated two more times on the same scale, and then the three reaction mixtures were combined for work-up. The mixture was cooled to 25 °C and concentrated in vacuo. The crude product was dissolved in THF (1.50 L), and the solution was adjusted to pH 8 with sodium bicarbonate. The resulting solid was filtered from the solution and dissolved in THF. Ethyl acetate was added, and the resulting precipitate was isolated by vacuum filtration (76.5 g). The filtrate was concentrated in vacuo, dissolved in THF, and precipitated with EtOAc. A second portion (4.95 g) of the desired product was obtained by vacuum filtration. The filtrate was concentrated, and the residue was purified by preparative HPLC (column: Phenomenex luna c18 250 mm*100 mm*10 um; mobile phase: [water (0.1% TFA)-ACN]; B%: 20%-50%, 23 minutes) to obtain a third portion (13.8 g) of the desired product. In total, N-(3-(5-((1H-pyrazol-4-yl)oxy)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (95.3 g, 65.6%) was obtained as an off-white solid. 1 H NMR: DMSO 400 MHz δ 12.83 (s, 1H), 9.74 (s, 1H), 9.35 - 9.33 (m, 1H), 8.68 - 8.66 (m, 2H), 8.27 - 7.81 (s, 1H), 7.48 - 7.41 (s, 1H), 7.38 - 7.31 (s, 1H), 7.29 - 7.28 (m, 1H), 7.17 - 6.90 (m, 4H). LC / MS (method I, ESI): [M+H] + = 467.3, R T = 0.91 min.

[0299] Step 3: tert-Butyl 3-((4-(4-(Difluoromethoxy)-3-(1-methyl-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenoxy)-1H-pyrazol-1-yl)methyl)-3-hydroxyazetidine-1-carboxylate TIFF0007716993000120.tif54170 A solution of methanol (12 mL), N-(3-(5-((1H-pyrazol-4-yl)oxy)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (850 mg, 1.82 mmol), and tert-butyl 1-oxa-5-azaspiro[2.3]hexane-5-carboxylate (675 mg, 3.65 mmol) was added with DIEA (706 mg, 5.47 mmol) at room temperature. The resulting solution was stirred at 80 °C overnight. The obtained residue was purified by reverse-phase chromatography (acetonitrile 0 - 50 / 0.05% aqueous NH4HCO3 solution) to give tert-butyl 3-((4-(4-(difluoromethoxy)-3-(1-methyl-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenoxy)-1H-pyrazol-1-yl)methyl)-3-hydroxyazetidine-1-carboxylate (837 mg, 1.28 mmol, 70.5% yield) as a yellow solid. LC / MS (Method H, ESI): [M+H] + = 652.3, R T = 1.20 min.

[0300] Step 4: N-(3-(2-(Difluoromethoxy)-5-((1-((3-hydroxyazetidin-3-yl)methyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007716993000121.tif54170tert-Butyl 3-((4-(4-(difluoromethoxy)-3-(1-methyl-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenoxy)-1H-pyrazol-1-yl)methyl)-3-hydroxyazetidine-1-carboxylate (737 mg, 1.13 mmol) in a solution, hexafluoro-2-propanol (5.0 mL, 1.13 mmol) containing trifluoroacetic acid (84 mg, 0.73 mmol) was added at room temperature. The resulting solution was stirred at 30 °C for 2 days. The obtained residue was purified by reverse-phase chromatography (acetonitrile 0 - 40 / 0.05% aqueous NH4HCO3) to give N-(3-(2-(difluoromethoxy)-5-((1-((3-hydroxyazetidin-3-yl)methyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (1.11 g) as a white solid. LC / MS (method N, ESI): [M+H] + = 552.3, R T = 1.11 min. 1 1H NMR (300 MHz, DMSO-d6) δ 9.75 (s, 1H), 9.35 (dd, J = 6.9, 1.5 Hz, 1H), 8.67 - 8.65 (m, 2H), 8.27 (s, 1H), 7.70 (s, 1H), 7.41 - 6.84 (m, 6H), 5.67 (s, 1H), 3.91 (s, 3H), 3.34 - 3.17 (m, 5H).

[0301] Step 5: N-(3-(2-(Difluoromethoxy)-5-((1-((3-hydroxy-1-methylazetidin-3-yl)methyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007716993000122.tif54170N-(3-(2-(Difluoromethoxy)-5-((1-((3-hydroxyazetidin-3-yl)methyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (950 mg, 1.72 mmol) was added with a solution of HCHO / H2O (1.79 g, 1.72 mmol) in methanol (24 mL) at room temperature. The resulting solution was stirred for 2 h, then sodium triacetoxyborohydride (5.04 g, 1.72 mmol) was added. The reaction mixture was stirred at room temperature for 2 h and concentrated in vacuo. The crude product was purified by reverse phase chromatography (acetonitrile 0 - 45 / 0.1% aqueous NH4HCO3) to give the desired product as a white solid (864.1 mg). The solid was recrystallized from cyclohexane / isopropyl alcohol = 3 / 1 (60 mL) stirred at 55 °C for 2 h and then at room temperature for 3 days. Filtration gave N-(3-(2-(difluoromethoxy)-5-((1-((3-hydroxy-1-methylazetidin-3-yl)methyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (836.6 mg, 1.4793 mmol, 85.9% yield) as a white solid. LC / MS (method A, ESI): [M+H] + = 566.3, R T = 1.21 min. 1 H NMR (400 MHz, DMSO-d6) δ 9.74 (s, 1H), 9.34 (dd, J = 7.2, 1.6 Hz, 1H), 8.67 - 8.65 (m, 2H), 8.27 (s, 1H), 7.70 (s, 1H), 7.41 - 6.90 (m, 6H), 5.60 (s, 1H), 4.22 (s, 2H), 3.90 (s, 3H), 3.37 - 3.31 (m, 2H), 2.75 - 2.73 (m, 2H), 2.20 (s, 3H).

[0302] Examples 32 and 33 TIFF0007716993000123.tif49170(N-[3-[2-(Difluoromethoxy)-5-[3-[(3R)-morpholin-3-yl]phenoxy]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide and (N-[3-[2-(Difluoromethoxy)-5-[3-[(3S)-morpholin-3-yl]phenoxy]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0303] Step 1. Synthesis of tert-butyl 3-(3-(4-(difluoromethoxy)-3-(1-methyl-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamide)-1H-pyrazol-3-yl)phenoxy)phenyl)morpholine-4-carboxylate TIFF0007716993000124.tif50170To a 30 mL sealed tube purged and maintained under an inert atmosphere of nitrogen were added toluene (10 mL), N-[3-[2-(difluoromethoxy)-5-hydroxyphenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 4, 400 mg, 1.00 mmol), tert-butyl 3-(3-bromophenyl)morpholine-4-carboxylate (410 mg, 1.2 mmol, 1.20 equivalents), [PdCl(allyl)]2 (7.31 mg, 0.020 mmol, 0.02 equivalents), RockPhos (18.7 mg, 0.040 mmol, 0.040 equivalents), and cesium carbonate (651 mg, 2.0 mmol, 2.0 equivalents). The resulting solution was stirred at 90 °C in an oil bath for 14 hours. The resulting mixture was concentrated under vacuum. The residue was poured onto a silica gel column equipped with dichloromethane / methanol (10:1). Thereby, 200 mg (30%) of tert-butyl 3-[3-[4-(difluoromethoxy)-3-(1-methyl-4-[pyrazolo[1,5-a]pyrimidine-3-amide]-1H-pyrazol-3-yl)phenoxy]phenyl]morpholine-4-carboxylic acid was obtained as a pale yellow solid.

[0304] Process 2: Synthesis of (N-[3-[2-(Difluoromethoxy)-5-[3-[(3R)-Morpholin-3-yl]phenoxy]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide) and (N-[3-[2-(Difluoromethoxy)-5-[3-[(3S)-Morpholin-3-yl]phenoxy]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide) 4917025 mL of a round-bottom flask was charged with hydrogen chloride / dioxane (4 M, 3 mL) and tert-butyl 3-[3-[4-(difluoromethoxy)-3-(1-methyl-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-3-yl)phenoxy]phenyl]morpholine-4-carboxylate (200 mg, 0.302 mmol). The resulting solution was stirred at room temperature for 2 hours. The resulting mixture was concentrated under vacuum. The crude product was purified by preparative HPLC using the following conditions (2#-Analytical HPLC-SHIMADZU (HPLC-10)): column, XBridge Prep C18 OBD column, 19*150 mm 5um 13nm; mobile phase, water containing 10 mmol of NH4HCO3 and ACN (24.0% ACN up to 42.0% in 8 minutes); detector, UV 220 nm. The racemic product was purified by chiral-preparative HPLC using the following conditions (Preparative HPLC-009): column, CHIRALPAK-AD-H-SL001, 20*250 mm; mobile phase, Hex and IPA (holding 50.0% IPA in 50 minutes); detector, UV 254 / 220 nm. Thereby, arbitrarily assigning the stereochemistry, (N-[3-[2-(difluoromethoxy)-5-[3-[(3R)-morpholin-3-yl]phenoxy]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (peak 1, 11.4 mg, yield 7%) was obtained as a white solid, and (N-[3-[2-(difluoromethoxy)-5-[3-[(3S)-morpholin-3-yl]phenoxy]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (peak 2, 8.4 mg, yield 5%) was obtained as a white solid. 11H NMR (300 MHz, DMSO-d6) δ 9.74 (s, 1H), 9.35 (dd, J = 6.9, 1.5 Hz, 1H), 8.68 - 8.66 (m, 2H), 8.26 (s, 1H), 7.45 - 6.90 (m, 9H), 3.89 (s, 3H), 3.76 - 3.68 (m, 3H), 3.32 (m, 1H), 3.09 (m, 1H), 2.82 - 2.80 (m, 2H), 1.24 (m, 2H), 0.88 - 0.83 (m, 1H). LC / MS (Method E, ESI): [M+H] + = 562.3, R T = 2.73 min.

[0305] Example 73 TIFF0007716993000126.tif53170N-(3-(2-(Difluoromethoxy)-5-((1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0306] Step 1: Synthesis of N-(3-(5-((1-(2-bromoethyl)-1H-pyrazol-4-yl)oxy)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide In a 5217030 mL sealed tube, N-(3-(5-((1H-pyrazol-4-yl)oxy)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (200 mg, 0.429 mmol), N,N-dimethylformamide (15 mL), cesium carbonate (700 mg, 2.15 mmol, 5.00 equivalents), and 1,2-dibromoethane (1.6 g, 8.52 mmol, 20.000 equivalents) were added. The resulting solution was stirred at 60 °C in an oil bath for 3 hours. The resulting mixture was concentrated under vacuum. The residue was applied to a silica gel column using ethyl acetate / petroleum ether (100% EA). Thereby, 180 mg (73%) of N-(3-(5-((1-(2-bromoethyl)-1H-pyrazol-4-yl)oxy)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a yellow oil. LC / MS (method J, ESI): [M+H] + = 573.2 and 575.2, R T = 1.00 minute.

[0307] Step 2: Synthesis of N-(3-(2-(difluoromethoxy)-5-((1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide Into a 30 mL sealed tube were added N-[3-(5-[[1-(2-bromoethyl)-1H-pyrazol-4-yl]oxy]-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (180 mg, 0.314 mmol, 1.000 equivalent), CH3CN (14 mL, 266 mmol), DIEA (203 mg, 1.57 mmol, 5.00 equivalents), and dimethylamine hydrochloride (76.5 mg, 0.94 mmol, 3.00 equivalents). The resulting solution was stirred in an oil bath at 70 °C for 3 hours. The resulting mixture was concentrated under vacuum. The residue was loaded onto a silica gel column with dichloromethane / methanol (85:15). The crude product (120 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, silica gel; mobile phase, ACN / H2O (10 mmol NH4HCO3) = 15%, increasing to 37% within 8 minutes; detector, UV 254 nm. Thereby, 34.9 mg (21%) of N-(3-(2-(difluoromethoxy)-5-((1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a pale yellow solid. LC / MS (method T, ESI): [M+H] + = 538.3, R T = 0.94 min. 1 H NMR (300 MHz, DMSO-d6) δ 9.74 (s, 1H), 9.35 (dd, J = 6.9, 1.5 Hz, 1H), 8.67 - 8.66 (m, 2H), 8.27 (s, 1H), 7.82 (s, 1H), 7.41 - 7.38 (m, 2H), 7.33 - 6.84 (m, 4H), 4.14 - 4.10 (m, 2H), 3.90 (s, 3H), 2.63 - 2.59 (m, 2H), 2.13 (s, 6H).

[0308] Example 124 TIFF0007716993000128.tif52170N-(3-(2-(Difluoromethoxy)-5-((1-(2-hydroxy-2-methylpropyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide To an 8 mL sealed tube were added N-[3-[2-(difluoromethoxy)-5-(1H-pyrazol-4-yloxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (80 mg, 0.172 mmol), DIEA (90 mg, 0.70 mmol, 4.06 equiv), methanol (5 mL), and 2,2-dimethyloxirane (25 mg, 0.35 mmol, 2.02 equiv). The resulting solution was stirred at 80 °C in an oil bath for 20 h. The resulting mixture was concentrated under vacuum. The residue was applied to a silica gel column eluting with dichloromethane / methanol (5% MeOH). Thereby, 33.3 mg (36%) of N-(3-(2-(difluoromethoxy)-5-((1-(2-hydroxy-2-methylpropyl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a white solid. LC / MS (Method A, ESI): [M+H] + = 539.3, R T = 1.70 min. 1 H NMR (400 MHz, DMSO-d6) δ 9.75 (s, 1H), 9.35 (dd, J = 7.2, 1.6 Hz, 1H), 8.67 - 8.66 (m, 2H), 8.28 (s, 1H), 7.71 (s, 1H), 7.41 - 7.39 (m, 2H), 7.30 - 7.27 (m, 1H), 7.18 - 6.90 (m, 3H), 4.68 (s, 1H), 3.96 (s, 2H), 3.90 (s, 3H), 1.05 (s, 6H).

[0309] Example 117 TIFF0007716993000129.tif52170N-(3-(5-((1-(3-(Cyanomethyl)-1-methylazetidin-3-yl)-1H-pyrazol-4-yl)oxy)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide Into a 10 mL round bottom flask purged and maintained under an inert atmosphere of nitrogen, N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 3, 100 mg, 0.216 mmol), 2-[3-(4-hydroxy-1H-pyrazol-1-yl)-1-methylazetidin-3-yl]acetonitrile (40 mg, 0.21 mmol, 0.96 eq), Pd2(allyl)2Cl2 (3 mg, 0.008 mmol, 0.038 eq), t-BuBrettPhos (8 mg, 0.017 mmol, 0.076 eq), cesium carbonate (60 mg, 0.18 mmol, 0.85 eq), and toluene (5 mL) were added. The resulting solution was stirred at 80 °C overnight. The resulting mixture was concentrated under vacuum. The residue was loaded onto a silica gel column with dichloromethane / methanol (90 / 10). The collected fractions were combined and concentrated under vacuum. The crude product was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, silica gel; mobile phase, H2O(NH4HCO3) / CH3CN = 90 / 10, increasing to 50 / 50 within 10 minutes; detector, UV 254 nm. Thereby, 16.4 mg (13%) of N-(3-(5-((1-(3-(cyanomethyl)-1-methylazetidin-3-yl)-1H-pyrazol-4-yl)oxy)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as an off-white solid. LC / MS (Method A, ESI): [M+H] + = 575.3, R T = 1.28 minutes. 11H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 9.36 (dd, J = 6.8, 1.6 Hz, 1H), 8.68 - 8.67 (m, 2H), 8.28 (s, 1H), 8.16 (s, 1H), 7.60 (s, 1H), 7.41 (d, J = Hz, 1H), 7.31 - 6.93 (m, 4H), 3.92 (s, 3H), 3.58 - 3.56 (m, 2H), 3.49 - 3.47 (m, 2H), 3.43 (s, 2H).

[0310] Example 203 TIFF0007716993000130.tif52170N-(3-(2-(Difluoromethoxy)-5-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0311] Step 1. Synthesis of tert-butyl 4-(4-(4-(difluoromethoxy)-3-(1-methyl-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenoxy)-1H-pyrazol-1-yl)piperidine-1-carboxylate TIFF0007716993000131.tif Under 51170 nitrogen, a solution of N-[3-[2-(difluoromethoxy)-5-(1H-pyrazol-4-yloxy)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100, 0.210 mmol), tert-butyl 4-iodopiperidine-1-carboxylate (334 mg, 1.07 mmol) and cesium carbonate (209 mg, 0.640 mmol) in N,N-dimethylformamide (10 mL) was added at room temperature. The resulting solution was stirred at 120 °C overnight. The crude reaction mixture was filtered through Celite®. The organic layer was diluted with water (100 mL). The resulting solution was extracted with EA (100 * 3 mL), and the organic layers were combined. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The resulting residue was purified by reverse-phase chromatography (acetonitrile 0 - 56 / 0.05% aqueous NH4HCO3 solution) to obtain tert-butyl 4-(4-(4-(difluoromethoxy)-3-(1-methyl-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenoxy)-1H-pyrazol-1-yl)piperidine-1-carboxylate (96.4 mg) as a white solid. LC / MS (method H, ESI): [M+H] + = 467.2, R T = 1.09 min.

[0312] Step 2. Synthesis of N-(3-(2-(difluoromethoxy)-5-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide A solution of tert-butyl 4-[4-[4-(difluoromethoxy)-3-[1-methyl-4-(pyrazolo[1,5-a]pyrimidin-3-carbonylamino)pyrazol-3-yl]phenoxy]pyrazol-1-yl]piperidine-1-carboxylate (96.4 mg, 0.150 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1.0 mL, 0.150 mmol) at room temperature. The resulting solution was stirred at room temperature for 4 hours and concentrated under vacuum. The crude product was used without further purification.

[0313] Step 3. Synthesis of N-(3-(2-(difluoromethoxy)-5-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide In 52170 methanol (6 mL), a solution of N-[3-[2-(difluoromethoxy)-5-[1-(4-piperidyl)pyrazol-4-yl]oxy-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (150 mg, 0.270 mmol) and HCHO / H2O (360 mg, 12.0 mmol) was added at RT. The resulting solution was stirred at room temperature for 2 hours. Then, sodium triacetoxyborohydride (1.1 mg, 0.01 mmol) was added and stirred at room temperature for 2 hours. The reaction mixture was concentrated under vacuum. The obtained residue was purified by reverse-phase chromatography; column: XBridge Prep OBD C18 column, 19*250 mm, 5 um; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: EtOH; flow rate: 25 mL / min; gradient: from 40B to 64B over 10 minutes to give N-(3-(2-(difluoromethoxy)-5-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (47.2 mg, 0.084 mmol, yield 30.6%) as a white solid. LC / MS (method A, ESI): [M+H] + = 564.3, R T = 1.23 minutes. 1 H NMR (400 MHz, DMSO-d6) δ 9.73 (s, 1H), 9.34 (dd, J = 7.2, 1.6 Hz, 1H), 8.66 - 8.65 (m, 2H), 8.27 (s, 1H), 7.87 (s, 1H), 7.41 - 7.40 (m, 2H), 7.28 (dd, J = 7.2, 4.4 Hz, 1H), 7.17 - 6.89 (m, 3H), 4.04 - 3.98 (m, 1H), 3.90 (s, 3H), 2.83 - 2.80 (m, 2H), 2.18 (s, 3H), 2.03 - 1.87 (m, 6H).

[0314] Example 79 TIFF0007716993000134.tif52170N-(3-(2-(Difluoromethoxy)-5-(3-(3-hydroxy-1-methylazetidin-3-yl)phenoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0315] Step 1: Synthesis of tert-butyl 3-(3-(4-(difluoromethoxy)-3-(1-methyl-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenoxy)phenyl)-3-hydroxyazetidine-1-carboxylate TIFF0007716993000135.tif53170Into a 30 mL sealed tube purged and maintained under an inert atmosphere of nitrogen, N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 3, 462 mg, 1.00 mmol), tert-butyl 3-hydroxy-3-(3-hydroxyphenyl)azetidine-1-carboxylate (549 mg, 2.07 mmol, 2.07 equiv), [PdCl(allyl)]2 (41.1 mg, 0.112 mmol, 0.113 equiv), t-BuBrettPhos (98.1 mg, 0.202 mmol, 0.203 equiv), cesium carbonate (395 mg, 1.21 mmol, 1.22 equiv), and toluene (15 mL) were added. The resulting solution was stirred at 100 °C in an oil bath for 12 h. The reaction mixture was brought to room temperature and the solid was removed by filtration. The filtrate was concentrated under vacuum. The residue was applied to a silica gel column using ethyl acetate / petroleum ether (10:1). Thereby, tert-butyl 3-(3-(4-(difluoromethoxy)-3-(1-methyl-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenoxy)phenyl)-3-hydroxyazetidine-1-carboxylate was obtained as a yellow-green solid. LC / MS (Method I, ESI): [M+H] + = 648.3, R T = 1.18 min.

[0316] Process 2: Synthesis of N-(3-(2-(Difluoromethoxy)-5-(3-(3-hydroxyazetidin-3-yl)phenoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide To a 50 mL round-bottom flask, tert-butyl 3-[3-[4-(difluoromethoxy)-3-(1-methyl-4-[pyrazolo[1,5-a]pyrimidine-3-amide]-1H-pyrazol-3-yl)phenoxy]phenyl]-3-hydroxyazetidine-1-carboxylate (1.15 g, 1.78 mmol), dichloromethane (20 mL), and trifluoroacetic acid (4 mL) were added. The resulting solution was stirred at room temperature for 2 hours and concentrated under vacuum. Thereby, 1.21 g (crude) of N-[3-[2-(difluoromethoxy)-5-[3-(3-hydroxyazetidin-3-yl)phenoxy]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a solid and used without further purification. LC / MS (Method G, ESI): [M+H] + = 548.3, R T = 0.69 min.

[0317] Process 3: Synthesis of N-(3-(2-(Difluoromethoxy)-5-(3-(3-hydroxy-1-methylazetidin-3-yl)phenoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007716993000137.tifTo a 5217050 mL round-bottom flask, N-[3-[2-(difluoromethoxy)-5-[3(3-hydroxyazetidin-3-yl)phenoxy]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (1.21 g, 2.21 mmol, 1.00 equiv), formaldehyde (0.75 mL, 30.2 mmol, 13.7 equiv), and NaBH(OAc)3 (937 mg, 4.42 mmol, 2.00 equiv) in dichloromethane (20 mL) were added. The resulting solution was stirred at room temperature for 18 h. The resulting mixture was concentrated under vacuum. The residue was applied to a silica gel column using tetrahydrofuran / MeOH (10 / 1). Thereby, 0.68 g (55%) of N-(3-(2-(difluoromethoxy)-5-(3-(3-hydroxy-1-methylazetidin-3-yl)phenoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a white solid. LC / MS (method H, ESI): [M+H] + =562.3,R T =0.69 min. 1 H NMR (400 MHz, CD3OD) δ 9.13 - 9.11 (m, 1H), 8.67 - 8.65 (m, 2H), 8.25 (s, 1H), 7.44 - 7.36 (m, 3H), 7.28 - 7.20 (m, 4H), 7.02 - 6.97 (m, 1H), 6.73 (t, J = 74 Hz, 1H), 3.98 (s, 3H), 3.73 (d, J = 9.2 Hz, 2H), 3.51 - 3.50 (m, 2H).

[0318] The liquid chromatography mass spectrometry (LCMS) method, retention time, and m / z of each compound in Table 1 are shown in Table 2. JPEG0007716993000138.jpg255170JPEG0007716993000139.jpg255170JPEG0007716993000140.jpg255170JPEG0007716993000141.jpg255170JPEG0007716993000142.jpg149170

[0319] Assay Test agent The test agent sample was prepared as a 10 mM solution in dimethyl sulfoxide (DMSO) and stored in the dark at room temperature before use.

[0320] JAK1 and JAK2 biochemical assay The in vitro biochemical assay quantifies JAK-catalyzed phosphorylation of a synthetic peptide detected using a LabChip® EZ Reader II microfluidic mobility shift device (PerkinElmer; Waltham, MA). The substrate peptide Y-1B has the sequence 5-FAM-VALVDGYFRLTT-NH2. Y-1B is fluorescently labeled with 5-FAM (5-carboxyfluorescein) at the N-terminus and contains a single tyrosine residue (Y) that can be phosphorylated by JAK activity. The substrate peptide stock is prepared at 5 mM in DMSO. The purified recombinant human JAK1 kinase domain protein (residues 854 - 1154) was expressed in insect cells and procured from Proteros Biostructures GmbH (Martinsried, Germany). The recombinant human JAK2 kinase domain protein (residues 812 - 1132) was expressed in insect cells and purified at Genentech, Inc. (South San Francisco, CA).

[0321] The kinase reaction mixture contained 100 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer (pH 7.2), 10 mM magnesium chloride, 0.015% Brij® 35, 4 mM dithiothreitol, 1.5 μM Y-1B peptide substrate, 25 μM adenosine triphosphate (ATP), 1 nM total JAK1 or 0.2 nM total JAK2, and up to 1000 nM test compound in a final concentration of 2% (volume to volume [v / v]) DMSO. In each titration experiment, the test compound was tested in duplicate at each of 12 concentrations. The blank reaction contained ATP, peptide, and DMSO but no JAK or test compound, while the non-inhibited control reaction contained ATP, peptide, JAK, and DMSO but no test compound.

[0322] A peptide + ATP mixture (24 μL) was added to 1 μL of the test compound (or DMSO alone) in DMSO. The reaction was initiated by adding 25 μL of JAK enzyme to the inhibitor / peptide / ATP mixture before thoroughly mixing the resulting solution. The reactions were incubated at room temperature (22 °C - 23 °C) in a 384-well plate at a final volume of 50 μL / well. After a 30-minute incubation, the reaction was stopped by adding 25 μL of 150 mM ethylenediaminetetraacetic acid in 100 mM HEPES buffer (pH 7.2) containing 0.015% Brij 35 to each well.

[0323] In each reaction, the remaining Y-1B substrate and the generated phospho-peptide product were separated using an EZ Reader II device. Electrophoretic separation of the product molecule from the substrate molecule was achieved at an operating pressure of -1.3 psi using downstream and upstream voltages of -500 and -2600 V, respectively. The 5-FAM group present in both the substrate peptide and the product peptide was excited at 488 nm, the fluorescence was detected at 530 nm, and the peak height was reported.

[0324] Data analysis The degree (or percentage) of conversion of the substrate to the product was calculated from the corresponding peak heights of the electrophoretogram using the HTS Well Analyzer software, version 5.2 (PerkinElmer) and the following equation (Equation 1).

[0325] Equation 1 % Conversion = [P ÷ (S + P)] × 100 Wherein S and P represent the peak heights of the substrate and product, respectively. After subtracting any baseline signal from blank wells without JAK from the signals of all test wells, the % conversion data was converted to fractional activity as shown in Equation 2, where v i and v o are the % conversions in the presence and absence of the test compound, respectively. The % conversion observed in a non-inhibited control reaction containing JAK and DMSO vehicle but no test compound was defined to have a fractional activity = 1 (in the absence of inhibitor, v i = v o ), while blank wells without JAK were defined to have a fractional activity = 0. The fractional activity was plotted against the test compound concentration and the data was fit to a quadratic equation (see Equation 2) for apparent inhibition constant (K i app ) (Williams JW, Morrison JF. The kinetics of reversible tight-binding inhibition. Methods Enzymol 1979;63:437-67.) using XLfit software (IDBS; Guildford, United Kingdom) and used to calculate the fractional activity and K i app .

[0326] Equation 2 JPEG0007716993000143.jpg28170 Wherein, [E] T and [I] Tis the total concentration of the active enzyme (initial estimated values of 0.15 nM for JAK1 and 0.048 nM for JAK2) and the inhibitor (variable parameter), respectively. Finally, by applying the competitive inhibition relationship, K i app to K i was calculated (Equation 3).

[0327] Equation 3 JPEG0007716993000144.jpg13170where [ATP] is the concentration of ATP = 25 μM, and K m app is the apparent ATP Michaelis constant = 32.1 μM for JAK1 and K m app = 11.7 μM for JAK2. By applying Equation 2 for tight-binding to account for inhibitor depletion and Equation 3 for competitive-inhibition relationship, the sensitivity of the assay can be expanded at least to the calculated K values of 0.008 nM for JAK1 and 0.0015 nM for JAK2 i or less.

[0328] Kinase Selectivity The in vitro kinase selectivity of the test agent was evaluated at a concentration of 1 μM in a panel of recombinant human kinase activity and binding assays including cytoplasmic tyrosine kinases, receptor tyrosine kinases, serine / threonine kinases, and lipid kinases (SelectScreen® Kinase Profiling Services, ThermoFisher Scientific, Madison, WI). The kinase activity assays measure peptide phosphorylation (Z’-LYTE®) or ADP production (Adapta®), while the binding assays monitor displacement of an ATP-site binding probe (LanthaScreen®). The ATP concentration used in the activity assays is typically the apparent Michaelis constant (K m app) was within 2-fold of the value, although the competitive binding tracer concentration used in the binding assay was generally within 3-fold of the experimentally determined dissociation constant (K d ) value. Inhibitors were tested in duplicate against each kinase and the mean percent inhibition values were reported. For kinases inhibited near or above 50% at the initial 1-μM test concentration, a 10-point inhibitor titration was performed using the same assay to determine the inhibitor concentration (IC 50 ) that caused 50% inhibition. The total JAK1 concentration used in this assay panel was 75 nM. If 100% of the 75 nM JAK1 protein was catalytically active, the limit of JAK1 inhibitor sensitivity from the vendor's JAK1 assay would theoretically be an IC 50 value of 37.5 nM (half of the total enzyme concentration). However, the SelectScreen® JAK1 assay was much lower than 37.5 nM and generated JAK1 IC 50 values for several inhibitors that were consistent with our in-house determinations. Thus, the active JAK1 enzyme concentration in the SelectScreen® assay should be much lower than the 75 nM total nominal JAK1 protein concentration used in the assay, and the observed sensitivity of this assay is much better than the theoretical sensitivity IC 50 limit of 37.5 nM.

[0329] Data Analysis To fit the data to a concentration-kinase inhibition plot, SelectScreen® Kinase Profiling Services used XLfit software (IDBS), model number 205 (sigmoid concentration-response model), a 4-parameter logistic fit model described by Equation 4.

[0330] Equation 4

[0001] y = A + {(B - A) ÷ [1 + (C ÷ x) D} where x is the inhibitor concentration, y is the observed percent inhibition, A is the minimum y-value, B is the maximum y-value, and C is the IC 50where the value, and D is the Hill slope. In certain cases, a 3-parameter logistic fit was used. For example, if the plateau of the curve at infinitely low inhibitor concentration did not fit between -20% and 20% inhibition, the lower plateau was set to 0% inhibition, while if the plateau of the curve at infinite inhibitor concentration did not fit between 70% and 130% inhibition, the upper plateau was set to 100% inhibition.

[0331] TF-1 Cell Line Phospho-STAT JAK1 and JAK2 Pathway Selectivity Assay TF-1 human erythroleukemia cells (ATCC®; Manassas, VA; catalog number CRL-2003™) were grown in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 2 ng / mL granulocyte-macrophage colony-stimulating factor, 1× non-essential amino acids (NEAA), and 1 mM sodium pyruvate. The day before the assay, the cultures were transferred to Opti-MEM®, 1× NEAA, 1 mM sodium pyruvate, and 0.5% charcoal-stripped FBS (starvation medium). An inhibitor stock solution (5 mM in DMSO) was serially diluted 1:2 in DMSO to generate a 10-point concentration titration (500× test concentration), which was further diluted by 50-fold dilution in assay medium (RPMI containing 1× NEAA and 1 mM sodium pyruvate) to generate a 10× concentration titration (in 2% DMSO). Cells (300,000 cells / well in 35 μL of assay medium) were seeded into 384-well Greiner plates. The diluted inhibitor at 10× concentration (5 μL) was added to the cells, and the plates were incubated at 37 °C for 30 min in a humidified incubator. The cells were, for each individual lot, as previously determined for each EC 90Stimulated with human recombinant cytokines at the indicated concentrations. For the phosphorylated signal transducer and activator of transcription 6 (P-STAT6) TF-1+ interleukin-13 (IL-13) assay, 10 μL of 250 ng / mL IL-13 (R&D Systems; Minneapolis, MN) was added to the cells, which were then incubated at 37 °C for 10 minutes. For the P-STAT5 TF-1+ erythropoietin (EPO) assay, 10 μL of 110 IU / mL EPO (Gibco Life Technologies, catalog number PHC2054) was added to the cells, which were then incubated at 37 °C for 30 minutes. For both assays, after incubation, 5 μL of ice-cold 10× cell lysis buffer (Cell Signaling Technologies; Danvers, MA; catalog number 9803S) containing 1 mM phenylmethylsulfonyl fluoride (PMSF) was added to the cells. The assay plates were frozen at -80 °C for at least 1 hour. In the IL-13 assay, goat anti-rabbit (GAR) plates (Meso Scale Discovery [MSD]; Rockville, MD; catalog number MSD L21RA-1) were coated with rabbit anti-human total STAT6 antibody (Cell Signaling Technologies; catalog number 9362S), and the cell lysates in the coated plates were incubated overnight at 4 °C. P-STAT6 was then measured by detecting with mouse anti-P-STAT6 (Tyr641) clone 16E12 antibody (MilliporeSigma; Burlington, MA; catalog number 05-590; custom-labeled by MSD using a SULFO-tag) using standard MSD plate processing, washing, and detection protocols. In the EPO assay, P-STAT5 was detected using a phospho-STAT5a,b Whole Cell Lysate kit (MSD; catalog number K150IGD-1). The electrochemiluminescence (ECL) signal of the wells was read using a MESO SECTOR S600 (MSD) reader.

[0332] Data analysis Subtract the mean ECL value of the negative controls (cytokine-stimulated cells and cells treated with 20 μM control inhibitor) from the ECL values of all wells, determine the ratio of the control to the ECL value of the test compound wells relative to the mean ECL value of the positive controls (cytokine-stimulated cells and cells treated with DMSO), and use the four-parameter logistic fit model shown in Equation 4 to determine the IC 50 of the test compound, thereby performing data analysis.

[0333] P-STAT6 BEAS-2B + IL-13 Cell Assay To study the effect of JAK1 inhibitors in cell lines related to the cell biology of human asthma, an IL-13-stimulated STAT6 phosphorylation assay in the human bronchial epithelial BEAS-2B cell line was developed.

[0334] BEAS-2B cells (ATCC® CRL-9609™) were grown in bronchial epithelial growth medium (BEGM) (Lonza catalog number CC-3170; Walkersville, MD; or PromoCell catalog number C-21060; Heidelberg, Germany). A test compound stock solution (0.5 mM in DMSO) was serially diluted 1:2 in DMSO to create a 10-point concentration curve (500× test concentration), which was further diluted by a 50-fold dilution step in BEGM to create a 10× concentration curve (in 2% DMSO). Cells were seeded at 100,000 cells / well in 200 μL of BEGM in a 96-well plate and incubated at 37 °C in a humidified incubator for 48 hours. The medium was aspirated from the cells and replaced with 70 μL of fresh BEGM. Diluted test compound (10 μL; or 2% DMSO in assay medium) was added to the cells, and the plate was incubated at 37 °C in a humidified incubator for 1 hour. Then, 20 μL of 250 ng / mL human recombinant IL-13 (Bio Techne catalog number 213-ILB) was added to the cells and incubated at 37 °C for 15 minutes. The medium was aspirated from the cells, and 60 μL of ice-cold 1× cell lysis buffer (Cell Signaling Technologies; catalog number 9803S) containing 1 mM PMSF was added to the cells. The assay plate was incubated at -80 °C for at least 1 hour. P-STAT6 was measured by coating a GAR plate (MSD; catalog number L45RA-1) with rabbit anti-human total STAT6 antibody (Cell Signaling Technologies; catalog number 9362S), incubating the cell lysates in the coated plate overnight at 4 °C, and then detecting with mouse anti-phospho-STAT6 (Tyr641) clone 16E12 antibody (Millipore; catalog number 05-590, custom-labeled by MSD using SULFO-tag) using standard MSD plate processing, washing, and detection protocols. The plates were read on a MESO SECTOR S600.

[0335] Data analysis Data analysis was performed by subtracting the negative control values from all wells and determining the percentage of the control using the positive control values. IC 50 was determined using the 4-parameter logistic fit model shown in Equation 4.

[0336] P-STAT6 BEAS-2B + IL-13 Cell Assay Using Inhibitor Washout (WO) An inhibitor washout (WO) assay in the human bronchial epithelial BEAS-2B cell line was developed to evaluate the ability of JAK1 inhibitors to inhibit STAT6 phosphorylation stimulated by IL-13 and retain the ability to remove free unbound inhibitor after cell washing. Retention of inhibitory activity after washout of the inhibitor is consistent with persistent binding of the inhibitor to the JAK1 protein and / or retention of inhibitor molecules within the cell after washout.

[0337] Similar to the standard BEAS-2B cell assay (see above), BEAS-2B cells were grown in bronchial epithelial growth medium (BEGM). A test compound stock solution (0.5 mM in DMSO) was serially diluted 1:2 in DMSO to create a 10-point concentration curve (500× test concentration), which was further diluted by a 50-fold dilution step in BEGM to create a 10× concentration curve (in 2% DMSO). Cells were seeded at 100,000 cells / well in 200 μL of BEGM in a 96-well plate and incubated at 37 °C in a humidified incubator for 48 hours. The medium was aspirated from the cells and replaced with 70 μL of fresh BEGM. The diluted test compound (10 μL; or 2% DMSO in assay medium) was added to the cells, and the plate was incubated at 37 °C in a humidified incubator for 1 hour. The medium was aspirated from the cells and replaced with 80 μL of fresh BEGM to wash out the inhibitor from the cells, and then the cell plate was incubated at 37 °C in a humidified incubator for 10 minutes. This washout procedure was repeated two more times. After the third wash step, the cell plate was returned to the 37 °C humidified incubator and incubated for 1 hour. Then, 20 μL of 250 ng / mL of IL-13 was added to the cells and incubated at 37 °C for 15 minutes. The medium was aspirated from the cells, and 60 μL of ice-cold 1× cell lysis buffer (Cell Signaling Technologies; catalog number 9803S) containing 1 mM of PMSF was added to the cells. The assay plate was incubated at -80 °C for at least 1 hour. P-STAT6 was measured by coating a GAR plate (MSD; catalog number L45RA-1) with rabbit anti-human total STAT6 antibody (Cell Signaling Technologies; catalog number 9362S), incubating the cell lysates in the coated plate overnight at 4 °C, and then detecting with mouse anti-phospho-STAT6 (Tyr641) clone 16E12 antibody (Millipore; catalog number 05-590, custom-labeled by MSD using SULFO-tag) using standard MSD plate processing, washing, and detection protocols. The plate was read on a MESO SECTOR S600.

[0338] Data analysis Data analysis was performed by subtracting the negative control values from all wells and determining the percentage of the control using the positive control values. IC 50 was determined using the four-parameter logistic fit model shown in Equation 4.

[0339] Cell cytotoxicity assay A549 (ATCC® CCL-185™), Jurkat clone E6-1 (ATCC® TIB-152™) and HEK-293T (ATCC® CRL-1573™) cells maintained at sub-confluent density in T175 flasks were used. Cells in the exponential growth phase were seeded into Greiner 384-well black / clear tissue culture treated plates (Greiner catalog number 781091) (450 cells in 45 μL of medium). After dispensing the cells, the plates were equilibrated at room temperature for 30 minutes, and then the cell plates were placed in a 37 °C CO2 and humidity controlled incubator overnight. The next day, the cells were treated with the test agent diluted in 100% DMSO (0.5% final DMSO concentration on the cells) at 10 dilutions and a maximum concentration of 50 μM. The cells and compounds were then incubated in a 37 °C CO2 and humidity controlled incubator for 72 hours, after which cell viability was measured by adding CellTiter-Glo® (Promega G7572) reagent to all wells. The plates were incubated at room temperature for 20 minutes and then the luminescence of the wells was read on an EnVision plate reader (Perkin Elmer Life Sciences).

[0340] Data from the enzyme assays of the compounds in Table 1 are shown in Table 3. JPEG0007716993000145.jpg236170JPEG0007716993000146.jpg247170JPEG0007716993000147.jpg248170JPEG0007716993000148.jpg248170JPEG0007716993000149.jpg248170JPEG0007716993000150.jpg134170

[0341] Animal model Mouse house dust mite (HDM) model Female C57BL / 6J mice at 7 - 8 weeks of age were purchased from Jackson West. The mice were immunized on days 0 and 14 by intraperitoneal administration of house dust mite (HDM, D. Pteronyssinus, purchased from Greer Laboratories, normalized to 0.918 μg of DerP1 per mouse) mixed with 2 mg of alum (Thermo Scientific) diluted in sterile PBS. On days 21 and 24, the mice were challenged with HDM (re - normalized to 0.918 μg of DerP1 content) in PBS administered by intratracheal inhalation. Before each inhaled HDM challenge (and in a subset of the group also on days 22 and 23), the animals received the test compound by nasal - only inhalation (using a Wright dust delivery device and a dry powder inhaler from Electro - Medical Measurement Systems (EMMS) containing 4 - layer / 24 - port or 2 - layer / 12 - port, directed flow, nasal - only inhalation tower), ending 1 hour before the challenge. Control animals received air - only nasal - only inhalation. 24 hours after the final treatment, the mice were retro - orbitally bled for plasma PK and then euthanized by CO2 inhalation. After euthanasia, BAL fluid was collected for total cell count (by FACS using a known amount of spike - in reference beads) and differential cell count (by Wright Giemsa - stained cytospin). The lungs and spleens were collected, weighed, and frozen for PK. There were 5 or 6 animals per group.

[0342] Furthermore, to verify the pulmonary delivery dose, the test compound is administered to the PK satellite groups of three naive animals by nasal-only inhalation for 1 or 4 consecutive days. Immediately after the final inhalation administration, retro-orbital blood collection is performed on the PK satellite animals for plasma PK, and then the animals are euthanized by CO2 inhalation. The lungs and spleens are collected and weighed for PK analysis.

[0343] Rat OVA model Six-week-old male Brown Norway rats from Charles River-Kingston. The rats are immunized on day 0 by intraperitoneal administration of 150 μg of OVA (Sigma) mixed with 40 mg of alum (Thermo Scientific) diluted in sterile PBS. Twenty-eight days after sensitization, the rats are challenged for 3 consecutive days for 30 minutes with 2% OVA in aerosolized PBS via a nebulizer. Prior to each OVA challenge, the animals receive the JAK1 / JAK2 test compound by nasal-only inhalation (using a Wright dust feeder and a dry powder inhaler manufactured by Electro-Medical Measurement Systems (EMMS) with 4 layers and 24 ports, directed flow, nasal-only inhalation tower) and end 1 hour before the challenge. Control animals receive either orally administered MCT buffer or nasal-only inhalation of air only. Twenty-four hours after the final treatment, the rats are euthanized by CO2 inhalation. Blood is collected from the abdominal aorta for plasma PK and whole blood FACS analysis. After euthanasia, BAL fluid is collected for total cell count (by FACS using a known amount of spike-in reference beads) and differential cell count (by Wright Giemsa stained cytospin). The lungs are collected, weighed, and frozen for PK. The spleens are weighed and cut in half for PK and FACS analysis. Blood and spleen samples are analyzed by FACS for total cell count and % NK cells (CD161a positive). There are 6 animals per group, excluding the naive control group which contains 5 animals.

[0344] Furthermore, to verify the pulmonary delivery dose, the JAK1 / JAK2 test compound was administered to the PK satellite groups of 3 naive animals by nasal-only inhalation for 1 day or 3 days, respectively. Immediately after the final inhalation administration, the PK satellite animals were euthanized by CO2 inhalation. Blood was collected from the abdominal aorta for plasma PK. The lungs and spleens were collected and weighed for PK analysis.

[0345] The plasma levels and lung levels of the test compound and their ratios are determined in the following manner. BALB / c mice from Charles River Laboratories are used in the assay. The test compound is individually formulated in 0.2% Tween 80 in saline, and the dosing solution is introduced into the trachea of the mice by oral aspiration. At various time points after dosing (typically 0.167, 2, 6, 24 hours), blood samples are removed by cardiac puncture, and the intact lungs are excised from the mice. The blood samples are centrifuged at approximately 12,000 rpm for 4 minutes at 4°C (Eppendorf centrifuge, 5804R) to recover the plasma. The lungs are dried with pads, weighed, and homogenized in sterile water at a 1:3 dilution. The plasma levels and lung levels of the test compound are determined by LC-MS analysis against analytical standards constructed on a standard curve in the test matrix. The lung-to-plasma ratio is determined as the ratio of the lung AUC in μg hr / g units to the plasma AUC in μg hr / mL units, where AUC is conventionally defined as the area under the curve of the test compound concentration versus time.

[0346] Pharmacokinetics in Plasma and Lungs of Mice The pharmacokinetics of the compound are determined in female Balb / c mice after administration of a target dose of 0.3 mg / kg formulated in 0.2% Tween 80 in saline by single intranasal (IN) bolus solution / suspension administration. Female Balb / c mice, 7 - 8 weeks of age, can be purchased from Charles River. The mice are housed under specific pathogen-free conditions until used in the study.

[0347] Do not fast the animals before administration. Blood samples are collected from three animals at each of the time points of 0.083, 2, 7, and 24 hours after administration under anesthesia (intraperitoneal injection of pentobarbital) via cardiac puncture into EDTA-coated microtainers. The blood samples are centrifuged (1500 g, 10 minutes at 4 °C) to separate the plasma. The plasma samples are frozen at approximately -80 °C. After intranasal administration and before lung perfusion, the spleen is removed, weighed, and snap-frozen. After confirmation of death, the lungs of the administered animals are perfused with chilled PBS to remove residual blood from the pulmonary vasculature. The lungs are then excised and weighed (all weights are recorded). All tissue samples are frozen by immersion in liquid nitrogen. The tissue samples are stored frozen until analysis (at approximately -80 °C).

[0348] Before PK analysis, thawed tissue samples (spleen and lung) are weighed and homogenized using an Omni-Prep Bead Ruptor (Omni Inc., Kennesaw, GA) at 4 °C after adding 4 mL of HPLC-grade water per gram of tissue. Plasma and tissue homogenate samples are extracted using protein precipitation with four volumes of acetonitrile containing tolbutamide (200 ng / mL) or labetalol (100 ng / mL) as the internal standard. The samples are mixed and centrifuged at 3200 g and 4 °C for 30 minutes to remove the precipitated protein, and the supernatant is appropriately diluted (e.g., 1:1, v / v) with HPLC-grade water in a 96-well plate. Representative aliquots of plasma, spleen, and lung samples are assayed for compound concentration by LC-MS / MS in positive ion mode using a Waters Xevo TQ-S (Waters, Elstree, UK) against matrix-matched calibration curves and quality control standards. The standards are prepared by adding the compound to aliquots of control plasma, spleen, and lung tissue homogenates and are extracted as described for the experimental samples. The assay limit of detection is 0.168 mg / mL to 4000 ng / mL in all matrices.

[0349] Concentrations below the lower limit of quantification (LLOQ) are treated as 0 for the calculation of the mean and SD. The mean concentration measured in the samples is used to construct a semi-logarithmic concentration-time curve profile. Pharmacokinetic (PK) analysis is performed using the non-compartmental method in Biobook (E-Workbook IDBS).

[0350] Mouse model of Alternaria alternata-induced eosinophilic inflammation in the lung Airway eosinophilia is a prominent feature of human asthma. Alternaria alternata is a fungal aeroallergen that can exacerbate human asthma and induce eosinophilic inflammation in the lungs of mice (Havaux et al., Clin Exp Immunol. 2005, 139(2):179-88). In mice, it has been demonstrated that Alternaria indirectly activates tissue-resident type 2 innate lymphoid cells in the lung, which release JAK-dependent cytokines (e.g., IL-5 and IL-13) in response to (e.g., IL-2 and IL-7) and regulate eosinophilic inflammation (Bartemes et al., J Immunol. 2012, 188(3):1503-13).

[0351] Taconic male C57 mice, 7 - 9 weeks old, are used in the study. On the day of the study, the animals are lightly anesthetized with isoflurane and either the vehicle or the test compound is administered by oropharyngeal aspiration. The animals are placed in the recumbent position after administration and, after monitoring for complete recovery from anesthesia, are returned to their home cages. One hour later, the animals are anesthetized again briefly and challenged with either the vehicle or the Alternaria extract via oropharyngeal aspiration, then monitored for recovery from anesthesia and returned to their home cages. Forty-eight hours after Alternaria administration, bronchoalveolar lavage fluid (BALF) is collected and eosinophils in the BALF are counted using an Advia 120 Hematology System (Siemens).

[0352] Compound activity in the model is demonstrated by a decrease in the level of eosinophils present in the BALF of treated animals at 48 hours compared to control animals treated with vehicle and challenged with Alternaria. Data are presented as percent inhibition of the BALF eosinophil response in vehicle-treated animals challenged with Alternaria. To calculate percent inhibition, the number of BALF eosinophils for each condition is converted to the mean percent of BALF eosinophils in vehicle-treated animals challenged with Alternaria and subtracted from 100%.

Claims

1. Formula (I) [wherein, Ar is phenyl; 1,2,3,4-tetrahydroisoquinolinyl; pyrazolyl; pyridinyl; or pyridazinyl: R １ is hydrogen; C １ -C ６ -alkyl; halo-C １ -C ６ -alkyl; hydroxy-C １ -C ６ -alkyl; -(CHR ａ ) ｈ -het １ ; -(CHR ａ ) ｋ -NR ａ -het １ ; or -(CHR ａ ) ｍ -C ３－６ -cycloalkyl, and the cycloalkyl moiety may be unsubstituted or substituted one or two times with R ｄ ; Each R ２ is independently: C １ -C ６ -alkyl; hydroxy-C １ -C ６ -alkyl; halo-C １ -C ６ -alkyl; C １ -C ６ -alkoxy; C １ -C ６ -alkoxy-C １ -C ６ -alkyl; halo-C １ -C ６ -alkoxy; halo-C １ -C ６ -alkoxy-C １ -C ６ -alkyl; C １ -C ６ -alkyl-SO ２ -C １ -C ６ -alkyl; hydroxyl; cyano; cyano-C １ -C ６ -alkyl; halo; acetyl; -(CHR ａ ) ｐ -het ２ ; -(CHR ａ ) ｑ -NR ｂ R ｃ ; -(CHR ａ ) ｒ -C(O)-NR ｂ R ｃ ; -(CHR ａ ) ｓ -NR ａ -(CHR ａ ) ｓ -C(O)-NR ｂ R ｃ ; or -(CHR ａ ) ｔ -C ３－６ -cycloalkyl, and the cycloalkyl moiety may be unsubstituted or substituted one or two times with R ｅ ; R ３ , R ４ and R ５ are each independently: hydrogen; or C １ -C ６ alkyl; Each R ａ is independently: hydrogen; or C １－６ alkyl; Each R ｂ is independently: hydrogen; C １－６ alkyl; or hydroxy-C １ -C ６ alkyl; Each R ｃ is independently: hydrogen; C １－６ alkyl; hydroxy-C １ -C ６ alkyl; cyano-C １ -C ６ alkyl; C １ -C ６ alkoxy-C １ -C ６ alkyl; oxetanyl; 2-morpholinoethyl; 1-methyl-azetidin-3-yl; 2-(N,N-dimethylamino)-ethyl; hydroxycyclobutyl; or 3-(N,N-dimethylamino)-pyrrolidin-1-yl; -(CHR ａ ) ｕ -C ３－６ cycloalkyl, wherein the cycloalkyl moiety may be unsubstituted or may be substituted one or two times with R ｅ ; Or, R ｂ and R ｃ may together with the nitrogen atom to which they are attached form het ３ ; Each R ｄ is independently: C1-C ６ alkyl, hydroxy or halo; Each R ｅ is independently: C １－６ alkyl; hydroxyl; cyano-C １ -C ６ alkyl; hydroxy-C １ -C ６ alkyl; morpholinyl; or -(CHR ａ ) ｖ -NR ｇ R ｈ wherein, R ｇ and R ｈ are each independently hydrogen or C １－６ alkyl; h is from 0 to 2; k is from 0 to 2; m is from 0 to 2; n is from 0 to 2; p is from 0 to 2; q is from 0 to 2; r is from 0 to 2; s is from 0 to 2; t is from 0 to 2; u is from 0 to 2; v is from 0 to 2; het １ is oxetanyl; tetrahydrofuranyl; tetrahydropyranyl; or pyrrolidinyl; each of which may be unsubstituted or substituted one or two times with R ｄ and may be substituted one or two times with R; het ２ is: azetidinyl; pyrrolidinyl; oxetanyl; piperidinyl; morpholinyl; piperazinyl; azepinyl; quinuclidinyl; or pyrazolyl; each of which may be unsubstituted or may be substituted one or two times with R ｅ and may be substituted one or two times with R; het ３ is: azetidinyl; pyrrolidinyl; piperidinyl; morpholinyl; piperazinyl; or azepinyl; each of which may be unsubstituted or may be substituted one or two times with R ｅ ​ a compound of or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein Ar is phenyl; or pyrazolyl.

3. The compound according to claim 1 or 2, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein Ar is phenyl.

4. The compound according to claim 1 or 2, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein Ar is pyrazolyl.

5. R １ is hydrogen or C １ -C ６ -alkyl, the compound according to claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof.

6. R １ The compound according to any one of claims 1 to 5, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R is methyl.

7. The compound according to any one of claims 1 to 6, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein n is 0.

8. The compound according to any one of claims 1 to 6, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein n is 1.

9. The compound according to any one of claims 1 to 6, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein n is 2.

10. Each R ２ is independently as follows: The compound according to any one of claims 1 to 9, or a stereoisomer or pharmaceutically acceptable salt thereof, selected from;

11. Each R ２ is independently as follows: The compound according to any one of claims 1 to 9, or a stereoisomer or pharmaceutically acceptable salt thereof, selected from;

12. where n is 1 and R ２ is as follows: The compound according to any one of claims 1 to 9, or a stereoisomer or pharmaceutically acceptable salt thereof, selected from;

13. The compound according to claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein the compound is a compound of formula (II):

14. The compound according to claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein the compound is a compound of formula (III):

15. A medicament for preventing, treating or reducing the severity of a disease or condition responsive to inhibition of Janus kinase activity in a patient, the medicament comprising a therapeutically effective amount of the compound according to any one of claims 1 to 14 or a stereoisomer or pharmaceutically acceptable salt thereof.

16. ​ ​ The medicament according to claim 15, wherein the disease or symptom is cancer, stroke, diabetes, hepatomegaly, cardiovascular disease, multiple sclerosis, Alzheimer's disease, cystic fibrosis, viral disease, autoimmune disease, atherosclerosis, restenosis, psoriasis, rheumatoid arthritis, inflammatory bowel disease, asthma, allergic disorder, inflammation, neuropathy, hormone-related disease, symptoms related to organ transplantation (e.g., transplant rejection), immunodeficiency disorder, destructive bone disorder, proliferative disorder, infectious disease, symptoms related to cell death, thrombin-induced platelet aggregation, liver disease, pathological immune state accompanied by T cell activation, CNS disorder or myeloproliferative disorder.

17. A pharmaceutical composition comprising a compound according to any one of claims 1 to 14, or a stereoisomer or pharmaceutically acceptable salt thereof, and comprising fine particles of the compound suitable for inhalation delivery.

18. The pharmaceutical composition according to claim 17, wherein the fine particles are prepared by spray drying, freeze drying or micronization.

19. (a) A first pharmaceutical composition comprising a compound according to any one of claims 1 to 14, or a stereoisomer or pharmaceutically acceptable salt thereof; (b) Instructions for use; A kit comprising the same.

20. The kit according to claim 19, further comprising a second pharmaceutical composition comprising an agent or chemotherapeutic agent for the treatment of inflammatory disorders.

21. A medicament comprising a compound according to any one of claims 1 to 14, or a stereoisomer or pharmaceutically acceptable salt thereof, for the treatment of inflammatory diseases.

22. The medicament according to claim 21, wherein the inflammatory disease is asthma.

23. Use of a compound according to any one of claims 1 to 14, or a stereoisomer or pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment of inflammatory diseases.

24. The use according to claim 23, wherein the inflammatory disease is asthma.

25. The compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, selected from the group consisting of

Images