PYRAZOLOPYRIMIDINE SULFONE INHIBITORS OF JAK KINASES AND USES THEREOF - Patent application

Abstract

A compound of formula (I) TIFF2022537964000153.tif70170, R 1 , R 2 , R 3 , R 4 , R 5 and R 6 is as defined herein, and compounds useful as JAK kinase inhibitors, as well as salts thereof, are described herein. 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 responsive to inhibition of Janus kinase activity in a patient.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims priority to International Application No. PCT / CN2019 / 091712, filed June 18, 2019, and U.S. Provisional Application No. 63 / 035381, filed June 5, 2020, the disclosures of which are incorporated herein by reference.

[0002] (Technical field) The present invention relates to compounds that are inhibitors of Janus kinases (e.g., JAK1 and JAK2), as well as compositions comprising these compounds and methods of use, including but not limited to, the diagnosis or treatment of patients suffering from conditions that respond to the inhibition of JAK kinases. [Background technology]

[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. Binding of a cytokine to its cognate receptor triggers receptor-associated JAK activation, which results in JAK-mediated tyrosine phosphorylation of signal transducer and activator of transcription (STAT) proteins and eventual transcriptional activation of a specific set of genes (Schindler et al., 2007, J. Biol. Chem. 282: 20059-63). JAK1, JAK2, and TYK2 display a broad pattern of gene expression, while JAK3 expression is restricted to leukocytes. Cytokine receptors are typically functional as heterodimers, such that more than one type of JAK kinase is usually associated with a cytokine receptor complex. The specific JAKs that associate with different cytokine receptor complexes have in many cases been determined by genetic studies and corroborated by other experimental evidence. Examples of therapeutic benefit from inhibition of JAK enzymes are discussed, for example, in WO 2013 / 014567.

[0004] JAK1 was originally identified in a screening for novel kinases (Wilks AF, 1989, Proc. Natl. Acad. Sci. USA 86:1603-1607). Genetic and biochemical studies have shown that JAK1 functionally and physically associates with type I interferons (e.g., IFNα), type II interferons (e.g., IFNγ), and 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 perinatally due to defects 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 from JAK1 knockout mice demonstrated a key 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 a key role in the pathogenesis of asthma through the production of TH2 cytokines such as IL-4, IL-9 and IL-13 in the lungs (Cohn et al., 2004, Annu. Rev. Immunol. 22:789-815). IL-4 and IL-13 induce increased mucus production, recruitment of eosinophils to the lungs, and increased production of IgE (Kasaian et al., 2008, Biochem. Pharmacol. 76(2): 147-155). IL-9 leads to activation of mast cells, which exacerbates asthma symptoms (Kearley et al., 2011, Am. J. Resp. Crit. Care Med., 183(7): 865-875). The IL-4Rα chain activates JAK1 and binds 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 IL-9 when combined with IL-9Rα, which also activates JAK1 (Demoulin et al., 1996, Mol. Cell Biol. 16(9):4710-4716). While the common γ chain activates JAK3, it has been shown that JAK1 predominates over JAK3 and that inhibition of JAK1 is sufficient to inactivate signaling by the common γ chain despite the presence of 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 reduce asthma symptoms in preclinical pulmonary inflammation models (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 that JAK2 associates with the single chain (e.g., EPO), IL-3, and interferon gamma 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-activating mutations of JAK2 (e.g., JAK2 V617F) have been associated with myeloproliferative disorders in humans. In addition, JAK2 associates with receptors for cytokines such as IL-5 and thymic stromal lymphopoietin (TSLP). IL-5 is a key cytokine involved in eosinophil differentiation, growth, activation, survival, and recruitment 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 (Mepolizumab, Reslizumab, and Benralizumab) that target IL-5 or the α chain of its receptor have been approved for the treatment of asthma with an eosinophilic phenotype. TSLP is an epithelial cell-derived cytokine that plays a key role in regulating type II immunity and functions 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. Phase 2 results show that it successfully reduced asthma exacerbations in patients with and without a high type 2 signature (Corren et al., 2017, 377: 936-46).

[0007] JAK3 associates only with the cytokine receptor gamma chain, which is common to IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21 cytokine receptor complexes. JAK3 is important for lymphocyte growth and proliferation, and mutations in JAK3 lead to severe combined immunodeficiency (SCID) (O'Shea et al., 2002, Cell, 109 (suppl.): S121-S131). Based on its role in regulating lymphocytes, 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 type I interferons (e.g., IFNα), IL-6, IL-10, IL-12 and IL-23 cytokine receptor complexes (Kisseleva et al., 2002, Gene 285:1-24; Watford, WT & O'Shea, JJ, 2006, Immunity 25:695-697). Consistent with this, primary cells derived from humans lacking TYK2 are defective in signaling type I interferons, IL-6, IL-10, IL-12 and IL-23. A fully human monoclonal antibody (ustekinumab) targeting the p40 subunit shared by IL-12 and IL-23 cytokines has recently been approved by the European Commission for the treatment of moderate to severe chronic plaque psoriasis (Krueger et al., 2007, N. Engl. J. Med. 356:580-92; Reich et al., 2009, Nat. Rev. Drug Discov. 8:355-356). In addition, antibodies targeting the IL-12 and IL-23 pathways are undergoing 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 discuss certain pyrazolopyrimidine compounds that are reported to be useful as inhibitors of one or more Janus kinases, where data are presented showing that certain specific compounds inhibit JAK1 as well as JAK2, JAK3 and / or TYK2 kinases.

[0010] Currently, there is still a need for further compounds as inhibitors of Janus kinases. There is a need for compounds as inhibitors of one or more Janus kinases (e.g., JAK1 and JAK2) that have useful potency, for example, in combination with other pharmacological properties, to provide useful therapeutic benefits. For example, there is a need for potent compounds that exhibit selectivity for one Janus kinase over other kinases in general (e.g., selectivity for JAK1 and / or JAK2 over other kinases, such as leucine-rich repeat kinase 2 (LRRK2)). There is also a need for potent compounds that exhibit selectivity for one Janus kinase over other Janus kinases (e.g., selectivity for JAK1 and / or JAK2 over JAK3 and / or TYK2). Compounds that exhibit selectivity for JAK1 and JAK2 over JAK3 and TYK2 in conditions that respond to inhibition of JAK1 could provide therapeutic benefits. Moreover, there is currently a need for potent JAK1 inhibitors that possess other necessary properties (e.g., melting point, pK, solubility, etc.) for formulation and administration by inhalation. Such compounds would be particularly useful for treating conditions such as asthma.

[0011] Thus, there is a need for further or alternative technologies for the treatment of conditions mediated by JAK kinases (e.g., as described above), in particular for JAK1 and JAK2 kinase inhibitors that can be used for inhaled delivery in the treatment of indications of airway inflammation (e.g., asthma). Summary of the Invention

[0012] The present invention provides a pyrazolopyrimidine that inhibits a JAK kinase, e.g., selected from the compounds of formula (I), a stereoisomer thereof, or a salt thereof, such as a pharma- ceutically acceptable salt thereof. The JAK kinase can be JAK1, JAK2, or both.

[0013] In one embodiment, the compound of formula (I): TIFF0007657740000001.tif69170 or a pharma- ceutically acceptable salt thereof is provided, During the ceremony, R 1 is C 1-6 Alkyl, Cyano-C 1-6 Alkyl, C 1-6 Alkoxy-(CO)-, -(CHR a ) m -NR b R c , or -(CHR a ) n -het 1 and R 2 is C 1-6 Alkyl, Hydroxy-C 1-6 Alkyl, halo-C 1-6 Alkyl, C 1-6 Alkoxy-C 1-6 Alkyl, C 3-6 Cycloalkyl, -(CHR a ) p -NR b R c , het 2 , -(CHR a ) q -het 3 , or even unsubstituted R d phenyl, optionally substituted one or two times by R 3 is hydrogen, amino, or C 1-6 is alkyl, R 4 is hydrogen or C 1-6 is alkyl, R 5 is hydrogen, or C 1-6 is alkyl, R 6 is hydrogen, C 1-6 alkyl or R 2 and R 6 may form a six-membered ring together with the atoms to which they are attached, m is 2 to 3; n is 0 to 2, p is 0 to 2; Each Ra are independently hydrogen or C 1-6 is alkyl, Each R b are independently hydrogen or C 1-6 is alkyl, Each R c are independently hydrogen or C 1-6 is alkyl, het 1 R is tetrahydrofuranyl, azetidinyl, or pyrrolidinyl, each of which may be unsubstituted or e may be substituted once or twice by het 2 is pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, or optionally partially saturated isoquinolinyl, each of which may be unsubstituted or f may be substituted once or twice by het 3 is azetidinyl, pyrrolidinyl, oxetanyl, or piperidinyl, each of which may be unsubstituted or g may be substituted once by Each R d is independently 1-6 Alkyl, Hydroxy, C 1-6 Alkoxy-C 1-6 Alkyl, -(CHR a ) q -NR b R c or phenyl; Each R e is independently 1-6 alkyl, or oxo; Each R f is independently 1-6 Alkyl, Hydroxy-C 1-6 Alkyl, oxo, -(CHR a ) r -NR b R c , -(CHR a ) s -het 4 and Each R g is independently 1-6alkyl, or acetyl; q is 1 to 2; r is 2 to 3; s is 2 to 3; het 4 is azetidin-1-yl, 1-methyl-azetidin-3-yl, quinuclidinyl, 1-methyl-pyrrolidin-2-yl, or 4-methylpiperazin-1-yl.

[0014] Also provided is a pharmaceutical composition comprising a JAK inhibitor described herein, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier, diluent, or excipient.

[0015] Also provided is the use of a JAK inhibitor described herein, or a pharma- ceutically acceptable salt thereof, in therapy, for example, in the treatment of an inflammatory disease (e.g., asthma).Also provided is the use of a JAK inhibitor described herein, or a pharma- ceutically acceptable salt thereof, for the preparation of a medicament for the treatment of 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 described herein, or a pharma- ceutically acceptable salt thereof.

[0016] The most effective 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 have activity against JAK1 and JAK2. Some of these compounds have optimally balanced co-activity against JAK1 and JAK2, or have slightly higher affinity for JAK1 than JAK2, rather than one of these kinases having much higher activity than the other. The compounds of the present invention also have good selectivity against off-target kinases, such as LRRK2, which is associated with pulmonary toxicity.

[0017] Although many compounds may show high affinity to JAK1 and JAK2 in simple biochemical assays, not all such compounds are effective in regulating important cytokines associated with JAK1 and JAK2.Certain compounds of the present invention, in addition to having activity against JAK1 and JAK2, are also shown to be effective in regulating asthma-related cytokines associated with JAK1 and JAK2 in cell-based assays.

[0018] The compounds of the present invention also show favorable pharmacokinetic (PK) properties in lung tissue, and are useful in inhalation therapy.When administered via inhalation route, for example using techniques such as dry powder inhalation (DPI) or intranasal (IN) delivery, certain compounds unexpectedly show very low concentrations in systemic circulation while being persistently retained within lung tissue.Such improved PK properties advantageously require lower dosages and less frequent administration to be effective therapeutics.Certain compounds show unexpected improved solubility, which also improves efficacy in lungs.Certain compounds of the present invention also show unexpectedly low cytotoxicity compared to other JAK inhibitors. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] definition "Halogen" or "halo" refers to F, Cl, Br, or I. Additionally, terms such as "haloalkyl," are meant to include monohaloalkyl and polyhaloalkyl, where one or more halogens replace a hydrogen on the alkyl group.

[0020] The term "alkyl" refers to a saturated linear or branched monovalent hydrocarbon group, which may be optionally substituted. In one example, an alkyl group is an alkyl group having 1 to 18 carbon atoms (C1-C 18 In other examples, the alkyl 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. "C0 alkyl" means a bond. Examples of alkyl groups are 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(C H3)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 (-CH2CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH 3)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), 1-heptyl, and 1-octyl.In some embodiments, substituents for "optionally substituted alkyl" include 1 to 4 of 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, piperazinyl, and pyrimidinyl, where the alkyl, phenyl, and heterocycle portions can be optionally substituted, e.g., with 1 to 4 substituents selected from this same list.

[0021] The term "alkenyl" refers to a linear or branched monovalent hydrocarbon radical having at least one site of unsaturation (i.e., a carbon-carbon double bond), where the alkenyl group may be optionally substituted and includes groups having "cis" and "trans" orientations, or, alternatively, "E" and "Z" orientations. In one example, an alkenyl group is an alkyl group having 2 to 18 carbon atoms (C2-C 18 In another embodiment, the alkenyl group is a C-C 12 , C2-C 10, 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, buta-1,3-dienyl, 2-methylbuta-1,3-dienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hex-4-enyl, and hexa-1,3-dienyl. In some embodiments, substituents for "optionally substituted alkenyl" include 1 to 4 of 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, piperazinyl, and pyrimidinyl, where the alkyl, phenyl, and heterocycle portions can be optionally substituted, e.g., with 1 to 4 substituents selected from this same list.

[0022] The term "alkynyl" refers to a linear or branched monovalent hydrocarbon group having at least one site of unsaturation, i.e., a carbon-carbon triple bond, and the alkynyl group may be optionally substituted. In one example, an alkynyl group is an alkyl group having 2 to 18 carbon atoms (C2-C 18 In another embodiment, the alkynyl group is a C-C 12 , C2-C 10, 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, substituents for "optionally substituted alkynyl" include from 1 to 4 occurrences of 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, piperazinyl, and pyrimidinyl, where the alkyl, phenyl, and heterocycle portions may be optionally substituted, e.g., with from 1 to 4 occurrences of a substituent selected from this same list.

[0023] "Alkylene" refers to a saturated, branched or straight chain hydrocarbon group having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkane. In one example, a divalent alkylene group is an alkylene group having 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 group "C0 alkylene" refers to a bond. Examples of alkylene groups include methylene (-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-dimethylethyl-1,2-yl (-C(CH3)2CH2), 1,4-butyl (-CH2CH2CH2CH2), and the like.

[0024] The term "heteroalkyl" refers to a linear or branched monovalent hydrocarbon group consisting of a specified number of carbon atoms, or up to 18 carbon atoms if none are 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 heteroatom can be optionally quaternized. In some embodiments, the heteroatom is selected from O, N, and S, where the nitrogen and sulfur atoms can be optionally oxidized and the nitrogen heteroatom can be optionally quaternized. The heteroatom can be located at any interior position of the heteroalkyl group, including the position at which the alkyl group is attached to the remainder 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. Up to two heteroatoms can be consecutive (e.g., -CH2-NH-OCH3 and -CH2-O-Si(CH3)3). Heteroalkyl groups can be optionally substituted. In some embodiments, substituents for "optionally substituted heteroalkyl" include from 1 to 4 occurrences of 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, piperazinyl, and pyrimidinyl, where the alkyl, phenyl, and heterocycle portions may be optionally substituted, e.g., with from 1 to 4 occurrences of a substituent selected from the same list.

[0025] "Amino" refers to primary (i.e. -NH2), secondary (i.e. -NRH), tertiary (i.e. -NRR) and quaternary (i.e. -N(+)RRR) amines, which may be optionally substituted, where each R is the same or different and is selected from alkyl, cycloalkyl, aryl and heterocyclyl, where the alkyl, cycloalkyl, aryl and heterocyclyl groups are as defined herein. Specific secondary and tertiary amines are alkylamines, dialkylamines, arylamines, diarylamines, aralkylamines and diaralkylamines, where 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 amines are each independently an alkyl group, which may be optionally substituted.

[0026] "Aryl" refers to a carbocyclic aromatic group, whether fused to one or more groups, having the number of carbon atoms indicated, or up to 14 carbon atoms if the number is not indicated. One example includes aryl groups having 6 to 14 carbon atoms. Another example includes aryl groups having 6 to 10 carbon atoms. Examples of aryl groups include phenyl, naphthyl, biphenyl, phenanthrenyl, naphthacenyl, 1,2,3,4-tetrahydronaphthalenyl, 1H-indenyl, 2,3-dihydro-1H-indenyl, and the like (see Lang's Handbook of Chemistry (Dean, JA, ed.) 13th ed. Table 7-2

[1985] ). In particular, aryl is phenyl. Substituted phenyl or substituted aryl means, for example, a phenyl or aryl group substituted by 1, 2, 3, 4 or 5 substituents (e.g., 1-2, 1-3 or 1-4 substituents) selected from the groups specified herein, e.g., F, Cl, Br, I, OH, SH, CN, NH, NHCH, N(CH), NO, N, C(O)CH, COOH, COCH, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methanesulfonylamino, SO, SO, phenyl, piperidinyl, piperazinyl and pyrimidinyl (see the definition of "optionally substituted"), where the alkyl, phenyl and heterocycle portions thereof may be optionally substituted, e.g., by 1-4 of the substituents selected from this same list 1-4 times.Examples of the term "substituted phenyl" include mono- or di(halo)phenyl groups (e.g., 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, and the like), mono- or di(hydroxy)phenyl groups (e.g., 4-hydroxyphenyl, 3-hydroxyphenyl, 2,4-dihydroxyphenyl, protected hydroxy derivatives thereof, and the like), nitrophenyl groups (e.g., 3- or 4-nitrophenyl), cyanophenyl groups (e.g., 4-cyanophenyl), mono- or di(alkyl)phenyl groups (e.g., 4-methylphenyl, 2,4-dimethylphenyl, 2-methylphenyl, 4-(isopropyl)phenyl, 4-ethylphenyl, 3-(n-propyl)phenyl, and the like). ), mono- or di(alkoxy)phenyl groups (e.g., 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 (4-carboxyphenyl, mono- or di(hydroxymethyl)phenyl or (protected hydroxymethyl)phenyl (e.g., 3-(protected hydroxymethyl)phenyl or 3,4-di(hydroxymethyl)phenyl)), mono- or di(aminomethyl)phenyl or (protected aminomethyl)phenyl (e.g., 2-(aminomethyl)phenyl or 2,4-(protected aminomethyl)phenyl), or mono- or di(N-(methylsulfonylamino))phenyl (e.g., 3-(N-methylsulfonylamino))phenyl).The term "substituted phenyl" may also refer to disubstituted phenyl groups with different substituents (e.g., 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.), as well as trisubstituted phenyl groups with different substituents (e.g., 3-methoxy-4-benzyloxy-6-methylsulfonylamino, 3-methoxy-4-benzyloxy-6-phenylsulfonylamino), as well as tetrasubstituted phenyl groups with different substituents (e.g., 3-methoxy-4-benzyloxy-5-methyl-6-phenylsulfonylamino). In some embodiments, the substituent of the aryl (e.g., phenyl) includes an amide. For example, the substituent of the aryl (e.g., phenyl) is -(CH2). 0-4 CONR'R'', where R' and R' are each independently selected from, for example, hydrogen, unsubstituted C1-C6 alkyl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 alkyl substituted with oxo or NR'R'', unsubstituted C1-C6 heteroalkyl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 heteroalkyl substituted with oxo or NR'R'', unsubstituted C6-C6 heteroalkyl, 10 C6-C substituted by aryl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy or NR'R'' 10may be selected from the group including aryl, unsubstituted 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S, or 4-11 membered heterocycloalkyl containing 1-4 heteroatoms selected from O, N and S), and 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S) substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R''; Alternatively, R' and R'' can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring, the ring atoms of which can be optionally substituted with N, O or S, and the ring can be optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR'R''.

[0027] "Cycloalkyl" refers to a non-aromatic, saturated or partially unsaturated hydrocarbon ring, where the cycloalkyl group may be optionally substituted independently with one or more substituents described herein. In one example, the cycloalkyl group is a ring containing 3 to 12 carbon atoms (C3-C4). 12 In other examples, 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 C7-C 12 In another example, the cycloalkyl group, as a spiro ring, is 12Examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, perdeuteriocyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. Exemplary configurations 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 structures. 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 for "optionally substituted cycloalkyl" include 1-4 occurrences of 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, piperazinyl, and pyrimidinyl, where the alkyl, aryl, and heterocyclic portions may be optionally substituted, e.g., with 1-4 occurrences of a substituent selected from this same list. In some embodiments, cycloalkyl substituents include amide. For example, cycloalkyl substituents include -(CH2) 0-4CONR'R'', where R' and R' are each independently selected from, for example, hydrogen, unsubstituted C1-C6 alkyl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 alkyl substituted with oxo or NR'R'', unsubstituted C1-C6 heteroalkyl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 heteroalkyl substituted with oxo or NR'R'', unsubstituted C6-C6 heteroalkyl, 10 C6-C substituted by aryl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy or NR'R'' 10 may be selected from the group including aryl, unsubstituted 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S, or 4-11 membered heterocycloalkyl containing 1-4 heteroatoms selected from O, N and S), and 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S) substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R''; Alternatively, R' and R'' can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring, the ring atoms of which can be optionally substituted with N, O or S, and the ring can be optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR'R''.

[0028] The terms "heterocyclic group", "heterocyclic", "heterocycle", "heterocyclyl" or "heterocyclo" are used interchangeably and refer to a ring system having 3 to 20 ring atoms (e.g., 3 to 10 ring atoms) of mono-, bi-, tricyclic or spiro structure, saturated or unsaturated, aromatic (heteroaryl) or non-aromatic (e.g., heterocycloalkyl) ring atoms, 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 attachment position of the ring system to the rest of the molecule. In one example, a heterocyclyl contains 3 to 11 ring atoms ("members"), including monocyclic, bicyclic, tricyclic and spiro ring systems, 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. In one example, a heterocyclyl contains 1 to 4 heteroatoms. In one example, the heterocyclyl includes 1-3 heteroatoms. In another example, the heterocyclyl includes a 3-7 membered monocyclic ring having 1-2, 1-3, or 1-4 heteroatoms selected from nitrogen, sulfur, or oxygen. In another example, the heterocyclyl includes a 4-6 membered monocyclic ring having 1-2, 1-3, or 1-4 heteroatoms selected from nitrogen, sulfur, or oxygen. In one example, the heterocyclyl includes a 3 membered monocyclic ring. In one example, the heterocyclyl includes a 4 membered monocyclic ring. In one example, the heterocyclyl includes a 5-6 membered monocyclic ring, such as a 5-6 membered heteroaryl. In another example, the heterocyclyl includes a 3-11 membered heterocycloalkyl, such as a 4-11 membered heterocycloalkyl. In some embodiments, the heterocycloalkyl includes at least one nitrogen. In one example, the heterocyclyl group includes 0-3 double bonds. Any nitrogen or sulfur heteroatoms may be optionally oxidized (e.g., NO, SO, SO2), and any nitrogen heteroatoms may be optionally quaternized (e.g., [NR4] + Cl - , [NR4] + OH -Examples of heterocycles are 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, dihydropi pyranyl, tetrahydropyranyl, hexahydrothiopyranyl, hexahydropyrimidinyl, oxadianyl, thiadianyl, thioxanyl, homopiperazinyl, homopiperidinyl, azepanyl, oxepanyl, thiepanyl, oxazepinyl, oxazepanyl, diazepanyl, 1,4-diazepanyl, diazepinylthiazepinyl, thiazepanyl, tetrahydrothiopyranyl, oxazolidinyl, thiazolidinyl, isothiazolidinyl, 1,1-dioxoisothiazolidinonyl, oxazolidin ... xazolidinonyl, imidazolidinonyl, 4,5,6,7-tetrahydro[2H]indazolyl, tetrahydrobenzimidazolyl, 4,5,6,7-tetrahydrobenzo[d]imidazolyl, 1,6-dihydroimidazole[4,5-d]pyrrolo[2,3-b]pyridinyl, thiazinyl, oxazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl, oxathiazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl, imidazolinyl, Dihydropyrimidyl, tetrahydropyrimidinyl, 1-pyrrolidinyl, 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, piperaninedionyl, pyrazolidinylimidazolinyl, 3-azabicyclo[3.1.0]hexanyl, 3,6-diazabicyclo[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 These are azaspiro[3.5]nonanyl, azaspiro[2.5]octanyl, azaspiro[4.5]decanyl, 1-azaspiro[4.5]decan-2-onyl, azaspiro[5.5]undecanyl, tetrahydroindolyl, octahydroindolyl, tetrahydroisoindolyl, tetrahydroindazolyl, and 1,1-dioxohexahydrothiopyranyl. Examples of 5-membered heterocycles containing a sulfur or oxygen atom and 1 to 3 nitrogen atoms are thiazolyl, thiadiazolyl (e.g. 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl), oxazolyl (e.g. oxazol-2-yl), and oxadiazolyl (e.g. 1,3,4-oxadiazol-5-yl and 1,2,4-oxadiazol-5-yl), such as thiazol-2-yl and thiazol-2-yl N-oxide. Examples of 5-membered heterocycles containing 2 to 4 nitrogen atoms are imidazolyl (e.g. imidazol-2-yl), triazolyl (e.g. 1,3,4-triazol-5-yl, 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl), and tetrazolyl (e.g. 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 containing 1 to 3 nitrogen atoms and optionally sulfur or oxygen atoms are, for example, pyridyl (e.g., pyrid-2-yl, pyrid-3-yl and pyrid-4-yl), pyrimidyl (e.g., pyrimid-2-yl and pyrimid-4-yl), triazinyl (e.g., 1,3,4-triazin-2-yl and 1,3,Examples of heterocyclic groups include pyridine N-oxide, pyridazinyl (especially pyridazin-3-yl), pyrazinyl (especially pyridazin-4-yl), pyridazinyl (especially pyridazin-3-yl), and pyrazinyl. Pyridine N-oxide and pyridazine N-oxide and pyridyl, pyrimid-2-yl, pyrimid-4-yl, pyridazinyl, and 1,3,4-triazin-2-yl groups are other examples of heterocyclic groups. The heterocycles may be optionally substituted. For example, substituents for an "optionally substituted heterocycle" include 1-4 of 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, piperazinyl, and pyrimidinyl, where the alkyl, aryl, and heterocycle portions may be optionally substituted, e.g., with 1-4 substituents selected from this same list. In some embodiments, the heterocycle group (e.g., heteroaryl or heterocycloalkyl) substituents include amide. For example, the heterocycle (e.g., heteroaryl or heterocycloalkyl) substituents include -(CH2), 0-4 CONR'R'', where R' and R' are each independently selected from, for example, hydrogen, unsubstituted C1-C6 alkyl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 alkyl substituted with oxo or NR'R'', unsubstituted C1-C6 heteroalkyl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 heteroalkyl substituted with oxo or NR'R'', unsubstituted C6-C6 heteroalkyl, 10 C6-C substituted by aryl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy or NR'R'' 10Aryl, unsubstituted 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S, or 4-11 membered heterocycloalkyl containing 1-4 heteroatoms selected from O, N and S), and 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S) substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR'R''. or R' and R'' may be combined with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring, the ring atoms of which may be optionally substituted with N, O or S, and the ring may be optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR'R''.

[0029] "Heteroaryl" refers to a ring system, either monocyclic, bicyclic, or tricyclic, in which at least one ring is a five- or six-membered, aromatic ring containing one to four heteroatoms selected from nitrogen, oxygen, and sulfur, and in an exemplary embodiment, at least one heteroatom is nitrogen. See, e.g., Lang's Handbook of Chemistry (Dean, JA, ed.) 13 thed. Table 7-2

[1985] . Also included within the definition are any bicyclic groups in which an aryl ring is fused to any of the heteroaryl rings listed above, where either the aryl or heteroaryl ring is attached to the remainder of the molecule. In one embodiment, heteroaryl includes 5-6 membered monocyclic aromatic groups in which one or more ring atoms is nitrogen, sulfur, or oxygen. Examples of 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, imidazole[1,2-a]pyrimidinyl and purinyl, as well as benzo-fused derivatives such as benzoxazolyl, benzofuryl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzimidazolyl and indolyl. Heteroaryl groups can be optionally substituted. In some embodiments, substituents for "optionally substituted heteroaryl" include 1-4 of 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, trifluoromethyl, difluoromethyl, sulfonylamino, methanesulfonylamino, SO, SO2, phenyl, piperidinyl, piperazinyl, and pyrimidinyl, where the alkyl, phenyl, and heterocyclic portions may be optionally substituted, e.g., with 1-4 substituents selected from this same list. In some embodiments, heteroaryl substituents include amide. For example, heteroaryl substituents include -(CH2) 0-4CONR'R'', where R' and R' are each independently selected from, for example, hydrogen, unsubstituted C1-C6 alkyl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 alkyl substituted with oxo or NR'R'', unsubstituted C1-C6 heteroalkyl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 heteroalkyl substituted with oxo or NR'R'', unsubstituted C6-C6 heteroalkyl, 10 C6-C substituted by aryl, halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy or NR'R'' 10 may be selected from the group including aryl, unsubstituted 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S, or 4-11 membered heterocycloalkyl containing 1-4 heteroatoms selected from O, N and S), and 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S) substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R''; Alternatively, R' and R'' can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring, the ring atoms of which can be optionally substituted with N, O or S, and the ring can be optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR'R''.

[0030] In a specific embodiment, the heterocyclyl group is a carbon atom bonded type of heterocyclyl group. For example, the carbon bonded type of heterocyclyl group includes the bonded positions at the 2, 3, 4, 5 or 6 position of the pyridine ring, the 3, 4, 5 or 6 position of the pyridazine ring, the 2, 4, 5 or 6 position of the pyrimidine ring, the 2, 3, 5 or 6 position of the pyrazine ring, the 2, 3, 4 or 5 position of the furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole ring, the 2, 4 or 5 position of the oxazole, imidazole or thiazole ring, the 3, 4 or 5 position of the isoxazole, pyrazole or isothiazole ring, the 2 or 3 position of the aziridine ring, the 2, 3 or 4 position of the azetidine ring, the 2, 3, 4, 5, 6, 7 or 8 position of the quinoline ring, or the 1, 3, 4, 5, 6, 7 or 8 position of the isoquinoline ring.

[0031] In certain embodiments, the heterocyclyl group is N-linked. For example, nitrogen-linked heterocyclyl or heteroaryl groups include aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, 1H-indazole at the 1-position, isoindole or isoindole at the 2-position, morpholine at the 4-position, and carbazole or β-carboline at the 9-position.

[0032] The term "alkoxy" refers to a straight or branched monovalent group represented by -OR, where R is alkyl as defined herein. Alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, mono-di- and tri-fluoromethoxy and cyclopropoxy.

[0033] "Acyl" means a carbonyl-containing substituent represented by the formula -C(O)-R, where R is hydrogen, alkyl, cycloalkyl, aryl, or heterocyclyl, where alkyl, cycloalkyl, aryl, and heterocyclyl are as defined herein. Acyl groups include alkanoyl (e.g., acetyl), aroyl (e.g., benzoyl), and heteroaroyl (e.g., pyridinoyl).

[0034] "Optionally substituted" means that, unless otherwise specified, a group may be unsubstituted or substituted with one or more (e.g., 0, 1, 2, 3, 4, 5 or more, or any range derivable at any time) of the substituents listed for that group, where the substituents may be the same or different. 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.

[0035] Substituents that may be selected for the alkyl group (alone or as part of another substituent, such as alkoxy), and the alkylenyl, alkenyl, alkynyl, heteroalkyl, heterocycloalkyl, and cycloalkyl (also each alone or as part of another substituent) can be a variety of groups, such as those described herein, as well as halogen, oxo, CN, NO, N3, -OR', perfluoroC1-C4 alkoxy, unsubstituted C3-C7 cycloalkyl, and C3-C7 cycloalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R'', and unsubstituted C6-C7 cycloalkyl. 10Aryl (e.g., phenyl), and C-C substituted by halogen, OH, CN, unsubstituted C-C alkyl, unsubstituted C-C alkoxy, oxo, or NR′R″. 10 Aryl, and unsubstituted 3- to 11-membered heterocyclyl (e.g., 5- to 6-membered heteroaryl containing 1-4 heteroatoms selected from O, N, and S, or 4- to 11-membered heterocycloalkyl containing 1-4 heteroatoms selected from O, N, and S), and 3- to 11-membered heterocyclyl (e.g., 5- to 6-membered heteroaryl containing 1-4 heteroatoms selected from O, N, and S) substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R''. aryl, or 4-11 membered heterocycloalkyl containing 1-4 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) 1-4R', R'' and R''' are each independently selected from the group consisting of hydrogen, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R'', unsubstituted C1-C6 heteroalkyl, unsubstituted C1-C6 heteroalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R'', unsubstituted C1-C6 heteroalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R, unsubstituted C6 ... 10 Aryl and C6-C substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy or NR'R. 10 It refers to groups such as aryl, unsubstituted 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S, or 4-11 membered heterocycloalkyl containing 1-4 heteroatoms selected from O, N and S), and 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S) substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR'R''. 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, the ring atoms being optionally substituted with N, O, or S, and the ring being optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R"; for example, -NR'R" is meant to include 1-pyrrolidinyl and 4-morpholinyl.

[0036] Similarly, the substituents that can be selected for the aryl and heteroaryl groups are varied. In some embodiments, the substituents that can be selected for the aryl and heteroaryl groups are halogen, CN, NO, N3, -OR', perfluoroC1-C4 alkoxy, unsubstituted C3-C7 cycloalkyl, and C3-C7 cycloalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R'' and unsubstituted C6-C7 cycloalkyl. 10 Aryl (e.g., phenyl), and C-C substituted by halogen, OH, CN, unsubstituted C-C alkyl, unsubstituted C-C alkoxy, oxo, or NR′R″. 10 Aryl, and unsubstituted 3- to 11-membered heterocyclyl (e.g., 5- to 6-membered heteroaryl containing 1-4 heteroatoms selected from O, N, and S, or 4- to 11-membered heterocycloalkyl containing 1-4 heteroatoms selected from O, N, and S), and 3- to 11-membered heterocyclyl (e.g., 5- to 6-membered heteroaryl containing 1-4 heteroatoms selected from O, N, and S) substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R''. aryl, or 4-11 membered heterocycloalkyl containing 1-4 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 R', R'' and R''' are each independently selected from the group consisting of hydrogen, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R'', unsubstituted C1-C6 heteroalkyl, unsubstituted C1-C6 heteroalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R'', unsubstituted C1-C6 heteroalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R'', unsubstituted C6-C6 heteroalkyl substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R, ... 10 Aryl and C6-C substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy or NR'R. 10 It refers to groups such as aryl, unsubstituted 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S, or 4-11 membered heterocycloalkyl containing 1-4 heteroatoms selected from O, N and S), and 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1-4 heteroatoms selected from O, N and S) substituted by halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo or NR'R''. 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, the ring atoms being optionally substituted with N, O, or S, and the ring being optionally substituted with halogen, OH, CN, unsubstituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, oxo, or NR'R"; for example, -NR'R" is meant to include 1-pyrrolidinyl and 4-morpholinyl.

[0037] The term "oxo" means =O or (=O)2.

[0038] As used herein, a wavy line " " crossing a bond in a chemical structure indicates the location of the bond of the atom where the wavy bond is connected to the remainder of the molecule or to the remainder of a fragment of a molecule in the chemical structure. In some embodiments, when an arrow is used with an asterisk, the wavy line indicates the portion of the bond.

[0039] In certain embodiments, a divalent group is described generally without a specific bond structure. The general description is understood to be meant to include both bond structures unless otherwise specified. For example, the group R 1 -R 2 -R 3 In the formula, the group R 2 When is depicted as -CH2C(O)-, this group represents R 1 -CH2C(O)-R 3 As, and,R 1 -C(O)CH2-R 3 It is understood that they can be combined as above.

[0040] The terms "compound(s) of the invention," "compound(s) of the present invention," and the like, unless otherwise specified, include compounds of formula (I) herein, or referred to as JAK inhibitors, e.g., compounds 1-18, and also include stereoisomers (including atropisomers), geometric isomers, tautomers, solvates, metabolites, isotopes, salts (e.g., pharma- ceutically acceptable salts), and prodrugs thereof. In some embodiments, solvates, metabolites, isotopes, or prodrugs, or combinations thereof, are excluded.

[0041] The phrase "pharmacologically acceptable" refers to molecular substances and compositions that do not produce adverse, allergic, or other side reactions when administered to an animal (e.g., a human, as appropriate).

[0042] The compounds of the present invention may be in the form of a salt, such as a pharma- ceutically acceptable salt. "Pharmaceutically acceptable salt" includes both acid and base addition salts. By "pharmacologically acceptable acid addition salts" is meant salts which retain the biological effectiveness and properties of the free bases and are devoid of biologically or otherwise undesirable properties, and which are formed from inorganic acids (e.g., hydrochloric, hydrobromic, sulfuric, nitric, carbonic, phosphoric, and the like), and from organic acids such as aliphatic, alicyclic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes, e.g., selected from formic, acetic, propionic, glycolic, gluconic, lactic, pyruvic, oxalic, malic, maleic, malonic, succinic, fumaric, tartaric, citric, aspartic, ascorbic, glutamic, anthranilic, benzoic, cinnamic, mandelic, embonic, phenylacetic, methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic, salicylic, and the like.

[0043] "Pharmaceutically acceptable base addition salts" include those derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Specific base addition salts are ammonium, potassium, sodium, calcium, and magnesium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of 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, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Specific organic non-toxic bases include isopropylamine, diethylamine, ethanolamine, tromethamine, dicyclohexylamine, choline, and caffeine.

[0044] In some embodiments, the salt is a hydrochloride, hydrobromide, trifluoroacetate, sulfate, phosphate, acetate, fumarate, maleate, tartrate, butyrate, citrate, pyruvate, succinate, oxaloacetate, methanesulfonate, p-toluenesulfonate, bisulfate, benzenesulfonate, ethanesulfonate, malonate, xinafoate, ascorbate, oleate, nicotinate, saccharate, adipate, formate, glycolate, palmitate, L-lactate, D-lactate, aspartate, malate, L-tartrate, D-tartrate, stearate, furoate (e.g., 2-furoate or 3-furoate), napadisylate (naphthalene-1,5-2 sulfonate or naphthalene-1-(sulfonic acid)-5-sulfonate), edisylate (ethane-1,2-2 sulfonate), or a combination thereof. salt or ethane-1-(sulfonic acid)-2-sulfonate), isethionate (2-hydroxyethylsulfonate), 2-mesitylenesulfonate, 2-naphthalenesulfonate, 2,5-dichlorobenzenesulfonate, D-mandelate, L-mandelate, cinnamate, benzoate, adipate, esylate, malonate, mesitylate (2-mesitylenesulfonate), napsylate (2-naphthalenesulfonate), phthalenesulfonate), camsylate (camphor-10-sulfonate, e.g., (1S)-(+)-10-camphorsulfonate), glutamate, glutarate, hippurate (2-(benzoylamino)acetate), orotate, xylate (p-xylene-2-sulfonate), and pamoate (2,2'-dihydroxy-1,1'-dinaphthylmethane-3,3'-dicarboxylate).

[0045] A "sterile" preparation is aseptic or free of all live microorganisms and their spores.

[0046] "Stereoisomers" refer to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space. Stereoisomers include diastereomers, enantiomers, conformers, and the like.

[0047] "Chiral" refers to a molecule having the property of not being superimposable on its mirror image partner, while "achiral" refers to a molecule whose mirror image partner is superimposable.

[0048] "Diastereomer" refers to a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties (e.g., melting points, boiling points, spectroscopic properties) or biological activity. Mixtures of diastereomers can be separated by high resolution analytical procedures such as, for example, electrophoresis and chromatography (e.g., HPLC).

[0049] "Enantiomers" refers to two stereoisomers of a compound which are non-superimposable mirror images of one another.

[0050] Stereochemical definitions and conventions used herein are generally in accordance with 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. In describing optically active compounds, the prefixes D and L (or R and S) are used to indicate the absolute configuration of the molecule about its chiral center. The prefixes d and l, or (+) and (-) are used to indicate the rotation characteristic of the compound of plane-polarized light, with (-) or l meaning that the compound is levorotatory. Compounds prefixed with (+) or d are dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Some stereoisomers are specifically referred to as enantiomers, and mixtures of such isomers are often referred to as enantiomeric mixtures. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may occur where there has been 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, devoid of optical activity.

[0051] The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton (e.g., keto-enol and imine-enamine isomerizations). Valence tautomers include interconversions by reorganization of some of the bonding electrons.

[0052] Certain compounds of the present invention can exist in non-solvated form as well as in solvated form, such as hydrated form. "Solvate" refers to an association or complex of one or more solvent molecules with the compound of the present invention. Examples of solvents that form solvates include water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid and ethanolamine. Certain compounds of the present invention can exist in multiple crystalline or amorphous forms. In general, all physical forms are intended to be included within the scope of the present invention. The term "hydrate" refers to a complex where the solvent molecule is water.

[0053] "Metabolite" means a product produced by metabolism in the body of a specified compound or its salt. Such products can result, for example, from the oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, and the like, of the administered compound.

[0054] The metabolites are typically radioisotopes (e.g. 14 C or 3 The compounds of the present invention are identified by preparing isotopes of the compounds of the present invention labeled with 3H, administering them to animals (e.g., rats, mice, guinea pigs, monkeys) or to humans in detectable doses (e.g., greater than about 0.5 mg / kg), allowing them to metabolize for a sufficient period of time (typically about 30 seconds to 30 hours), and isolating the conversion products from urine, blood, or other biological samples. These products are easily isolated because they are labeled (others are isolated using antibodies capable of binding to epitopes remaining in the metabolites). The structures of the metabolites are determined by conventional methods, such as, for example, MS, LC / MS, or NMR analysis. In general, the analysis of the metabolites is performed similarly to conventional drug metabolism studies known to those skilled in the art. The metabolites are useful in diagnostic assays for therapeutic administration of the compounds of the present invention, so long as they are not otherwise detectable in vivo.

[0055] A "subject," "individual," or "patient" is a vertebrate. In certain embodiments, the vertebrate is a mammal. Mammals include, but are not limited to, farm animals (e.g., cows), sport animals, pets (e.g., guinea pigs, cats, dogs, rabbits, and horses), primates, mice, and rats. In certain embodiments, the mammal is a human. In embodiments involving administration of a JAK inhibitor or a pharma- ceutically acceptable salt thereof to a patient as described herein, the patient may be one in need thereof.

[0056] The term "Janus kinase" refers to JAK1, JAK2, JAK3, and TYK2 protein kinases. In some embodiments, a 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.

[0057] The terms "inhibit" and "reduce," or any conjugation of these terms, encompass any significant reduction or complete inhibition to achieve the desired result. For example, for a decrease in activity (e.g., JAK1 activity), the decrease can be up to about or down to about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more compared to a standard, or any numerical range derivable therein.

[0058] "Therapeutically effective amount" refers to the amount of a compound of the present invention or a salt thereof (e.g., a pharma- ceutically acceptable salt thereof), (i) Treat or prevent a particular disease, condition, or disorder; or (ii) reducing, ameliorating or eliminating one or more symptoms of a particular disease, condition or disorder, and optionally, (iii) refers to a content that 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 a content sufficient to reduce or alleviate symptoms of an autoimmune or inflammatory disease (e.g., asthma). In some embodiments, a therapeutically effective amount is a content 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 a drug reduces the number of cancer cells, reduces tumor size, inhibits (i.e., slows down to some extent, and preferably stops) cancer cell invasion into surrounding organs, inhibits (i.e., slows down to some extent, and preferably stops) tumor metastasis, inhibits tumor growth to some extent, or relieves one or more symptoms associated with cancer to some extent. A drug is said to be cytostatic or cytotoxic to the extent that it can prevent growth or kill existing cancer cells. In treating cancer, efficacy can be measured, for example, by assessing the time to disease progression (TTP) or measuring the response rate (RR).

[0059] "Treatment" (and conjugations such as "treat" or "treating") refers to clinical intervention to alter the natural course of an individual or treated cells, which may be performed for prophylactic purposes or during clinical pathology testing. Favorable therapeutic effects include prevention of disease onset or recurrence, alleviation of symptoms, reduction of any direct or indirect pathological consequences of the disease, stabilization (i.e., not worsening) of the disease, reduction in the rate of disease progression, improvement or palliative of the disease state, prolongation of survival compared to expected survival in the absence of treatment, and remission or improved prognosis. In some embodiments, the compounds of the invention or salts thereof (e.g., pharma- ceutically acceptable salts thereof) are used to delay or slow the progression of a disease or disorder. Those in need of treatment include those already with a condition or disorder, as well as those susceptible to a condition or disorder (e.g., due to a genetic mutation) or those in whom a condition or disorder is to be prevented.

[0060] "Inflammatory disease" refers to any disease, disorder, or syndrome in which an excessive or unregulated inflammatory response leads to excessive inflammatory symptoms, host tissue damage, or loss of tissue function. "Inflammatory disease" may also refer to pathological conditions mediated by the influx of leukocytes or neutrophil chemotaxis.

[0061] "Inflammation" refers to the local protective response induced by tissue damage or loss, which functions to destroy, dilute or sequester harmful agents and damaged tissue. Inflammation is particularly associated with the influx of leukocytes or neutrophil chemotaxis. Inflammation can result from infection by pathogenic organisms and viruses, as well as non-infectious means (e.g., trauma or reperfusion after myocardial infarction or stroke), immune responses to external antigens, and autoimmune responses. Thus, the inflammatory diseases that are the subject of treatment with the compound of the present invention or its salt (e.g., its pharmacologic acceptable salt) include disorders associated with reactions in the specific defense system as well as reactions by the non-specific defense system.

[0062] "Specific defense system" refers to the components of the immune system that respond to the presence of specific antigens. Examples of inflammation resulting from the reaction of the specific defense system include classical reactions to foreign antigens, autoimmune diseases, and delayed hypersensitivity reactions mediated by T cells. Chronic inflammatory diseases, rejection of solid transplanted tissues and organs (e.g., kidney and bone marrow transplants, and graft-versus-host disease (GVHD)) are further examples of inflammatory reactions by the specific defense system.

[0063] The term "non-specific defense system" refers to inflammatory diseases mediated by white blood cells (e.g., granulocytes and macrophages) that lack immunological memory. Examples of inflammation resulting at least in part from a reaction by the non-specific 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, acute purulent meningitis or other inflammatory diseases of the central nervous system (e.g., stroke), burns, inflammatory bowel disease, syndromes associated with granulocyte infusion, and cytokine-induced toxicity.

[0064] "Autoimmune disease" refers to any group of disorders in which tissue damage is associated with humoral or cellular responses to the body's own components. Non-limiting examples of autoimmune diseases include rheumatoid arthritis, lupus, and multiple sclerosis.

[0065] As used herein, "allergic disease" refers to any symptoms, tissue damage or loss of tissue function resulting from allergy. As used herein, "arthritic disease" refers to any disease characterized by inflammatory lesions of the joints resulting from a variety of etiologies. As used herein, "dermatitis" refers to any disease of the skin as a large family characterized by inflammation of the skin resulting from a variety of etiologies. As used herein, "graft rejection" refers to any immune response directed against transplanted tissue (e.g., organ or cells, e.g., bone marrow) characterized by loss of function of the transplanted and surrounding tissues, pain, swelling, leukocytosis and thrombocytopenia. The therapeutic methods of the present invention include methods for the treatment of disorders associated with activation of inflammatory cells.

[0066] "Inflammatory cell activation" refers to the induction of a proliferative cellular response by a stimulus (such as, but not limited to, a cytokine, an antigen, or an autoantibody), the production of soluble mediators (such as, but not limited to, a cytokine, an oxygen radical, an enzyme, a prostanoid, or a vasoactive amine), or the expression of new or increased levels of mediators (such as, but not limited to, a major histocompatibility antigen or a cell adhesion molecule) on the cell surface of inflammatory cells (such as, but not limited to, monocytes, macrophages, T lymphocytes, B lymphocytes, granulocytes (i.e., polymorphonuclear leukocytes (e.g., neutrophils, basophils, and eosinophils)), mast cells, dendritic cells, Langerhans cells, and endothelial cells. It will be appreciated by those skilled in the art that activation of one or a combination of these phenotypes of these cells can contribute to the initiation, prolongation, or exacerbation of inflammatory diseases.

[0067] In some embodiments, inflammatory diseases that can be treated according to the methods of the 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 hypersensitivity reactions.

[0068] The terms "cancer" and "cancerous", "neoplasm" and "tumor" and related terms refer to or describe a physiological condition in mammals that is typically characterized by unregulated cell growth. A tumor contains one or more cancer cells. Examples of cancer include carcinoma, blastoma, sarcoma, seminoma, glioblastoma, melanoma, leukemia and bone marrow or lymphatic malignancies. More specific examples of such cancers include squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), and lung cancer, such as small cell lung cancer, non-small cell lung cancer ("NSCLC"), lung adenocarcinoma and lung squamous cell carcinoma. Other cancers include cancers of the skin, keratoacanthoma, follicular carcinoma, hairy cell leukemia, oral cavity, pharynx (oral), lips, tongue, mouth, salivary gland, esophagus, larynx, hepatocellular, stomach, stomach, gastrointestinal system, small intestine, large intestine, pancreas, cervix, ovary, liver, bladder, liver, breast, colon, rectum, colorectal, genitourinary tract, biliary duct, thyroid, nipple, liver, endometrium, uterus, salivary gland, kidney or kidney, prostate, testis, vulva, peritoneum, anus, penis, bone, multiple myeloma, B-cell lymphoma, central nervous system, brain, head and neck, Hodgkin's, and associated metastases. Examples of neoplastic disorders include myeloproliferative disorders such as polycythemia vera, essential thrombocytosis, myelofibrosis such as primary myelofibrosis, and chronic myelogenous leukemia (CML).

[0069] A "chemotherapeutic agent" is an agent that is useful in the treatment of a given disorder (e.g., cancer or inflammatory disease). Examples of chemotherapeutic agents are known in the art, and include, for example, those disclosed in U.S. Patent Application Publication No. 2010 / 0048557, which is incorporated herein by reference. In addition, chemotherapeutic agents include any pharma- ceutically acceptable salt, acid or derivative of the chemotherapeutic agent, as well as combinations of two or more thereof.

[0070] "Package insert" is used to refer to instructions customarily included in the commercial packaging of a therapeutic product, including information regarding instructions, use, dosage forms, administration, contraindications or precautions regarding the use of such therapeutic product.

[0071] Unless otherwise stated, structures depicted herein include compounds which differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine (e.g., 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) can be useful in assaying the distribution of compounds or substrates in tissues. 3 H) and carbon 14 (i.e. 14 C) isotopes can also be useful in terms of their ease of preparation and detectability. 2 Substitution with heavier isotopes such as H may confer certain therapeutic advantages (e.g., longer in vivo half-life or lower dosage requirements) resulting from increased metabolic stability. In some embodiments, one or more hydrogen atoms may be substituted with 2 H or 3 H or one or more carbon atoms are replaced 13 C or 14 C-enhanced carbon is substituted. Positron-emitting isotopes (e.g. 15 O. 13 N, 11 C and 18F) is useful in the analysis of receptor occupancy by substrates in positron emission tomography (PET) studies. Isotopically labeled compounds can generally be prepared by following procedures similar to those disclosed in the schemes herein or in the examples, substituting a non-isotopically labeled reagent for an isotopically labeled reagent.

[0072] It is specifically contemplated that any limitation discussed with respect to one embodiment of the present invention may also apply to any other embodiment of the present invention. Furthermore, any compound or salt thereof (e.g., a pharma- ceutically acceptable salt thereof), or composition of the present invention may be used in any method of the present invention, and any method of the present invention may be used to make or utilize any compound or salt thereof (e.g., a pharma-ceutically acceptable salt thereof), or composition of the present invention.

[0073] Use of the term "or" is used to mean "and / or" unless expressly indicated to refer only to that alternative or that alternative is exclusive, even if the disclosure supports a definition that refers to both the alternative and "and / or."

[0074] Throughout this application, the term "about" is used to indicate that a value includes the standard deviation of error for the device or method being employed to measure the value.

[0075] As used herein, "a" or "an" means one or more, unless otherwise specified. As used herein, "another" means at least a second or more.

[0076] The headings used herein are for editorial purposes only.

[0077] Janus kinase inhibitors In one embodiment, the compound of formula (I): TIFF0007657740000002.tif70170 or a pharma- ceutically acceptable salt thereof is provided, During the ceremony, R 1 is C 1-6 Alkyl, Cyano-C 1-6 Alkyl, C 1-6 Alkoxy-(CO)-, -(CHR a ) m -NR b R c , or -(CHR a ) n -het 1 and R 2 is C 1-6 Alkyl, Hydroxy-C 1-6 Alkyl, halo-C 1-6 Alkyl, C 1-6 Alkoxy-C 1-6 Alkyl, C 3-6 Cycloalkyl, -(CHR a ) p -NR b R c , het 2 , -(CHR a ) q -het 3 , or even unsubstituted R d phenyl, optionally substituted one or two times by R 3 is hydrogen, amino, or C 1-6 is alkyl, R 4 is hydrogen, or C 1-6 is alkyl, R 5 is hydrogen, or C 1-6 is alkyl, R 6 is hydrogen, C 1-6 alkyl or R 2 and R 6 may form a six-membered ring together with the atoms to which they are attached, m is 2 to 3; n is 0 to 2, p is 0 to 2; Each Ra are independently hydrogen or C 1-6 is alkyl, Each R b are independently hydrogen or C 1-6 is alkyl, Each R c are independently hydrogen or C 1-6 is alkyl, het 1 R is tetrahydrofuranyl, azetidinyl, or pyrrolidinyl, each of which may be unsubstituted or e may be substituted once or twice by het 2 is pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, or optionally partially saturated isoquinolinyl, each of which may be unsubstituted or f may be substituted once or twice by het 3 is azetidinyl, pyrrolidinyl, oxetanyl, or piperidinyl, each of which may be unsubstituted or g may be substituted once by Each R d is independently 1-6 Alkyl, Hydroxy, C 1-6 Alkoxy-C 1-6 Alkyl, -(CHR a ) q -NR b R c or phenyl; Each R e is independently 1-6 alkyl, or oxo; Each R f is independently 1-6 Alkyl, Hydroxy-C 1-6 Alkyl, oxo, -(CHR a ) r -NR b R c , -(CHR a ) s -het 4 and Each R g is independently 1-6alkyl, or acetyl; q is 1 to 2; r is 2 to 3; s is 2 to 3; het 4 is azetidin-1-yl, 1-methyl-azetidin-3-yl, quinuclidinyl, 1-methyl-pyrrolidin-2-yl, or 4-methylpiperazin-1-yl.

[0078] In certain embodiments, R 3 is hydrogen.

[0079] In certain embodiments, R 4 is hydrogen.

[0080] In certain embodiments, R 5 is hydrogen.

[0081] In certain embodiments, R 6 is hydrogen.

[0082] In certain embodiments, R 1 is cyanomethyl or methyl.

[0083] In certain embodiments, R 1 is cyanomethyl.

[0084] In certain embodiments, R 1 is methyl.

[0085] In certain embodiments, R 2 is C 1-6 Alkyl, Hydroxy-C 1-6 Alkyl, halo-C 1-6 Alkyl, C 1-6 Alkoxy-C 1-6 Alkyl, C 3-6 Cycloalkyl, -(CHR a ) p -NR b R c , het 2 , -(CHR a )q -het 3 , or even unsubstituted R d phenyl, optionally substituted once by

[0086] In certain embodiments, R 2 is C 1-6 It is an alkyl.

[0087] In certain embodiments, R 2 is hydroxy-C 1-6 It is an alkyl group.

[0088] In certain embodiments, R 2 is Halo-C 1-6 It is an alkyl.

[0089] In certain embodiments, R 2 is C 1-6 Alkoxy-C 1-6 It is an alkyl.

[0090] In certain embodiments, R 2 is C 3-6 It is cycloalkyl.

[0091] In certain embodiments, R 2 is -(CHR a ) p -NR b R c It is.

[0092] In certain embodiments, R 2 het 2 It is.

[0093] In certain embodiments, R 2 is -(CHR a ) q -het 3 It is.

[0094] In certain embodiments, R 2 is unsubstituted or R d is phenyl optionally substituted once by

[0095] In certain embodiments, R 2 is C 1-6 Alkyl, Hydroxy-C 1-6 Alkyl, C 1-6 Alkoxy-C 1-6 Alkyl or halo-C 1-6 There is alkyl.

[0096] In certain embodiments, R 2 is -(CH2)2-NR b R c It is.

[0097] In certain embodiments, R 2 is C 1-6 It is an alkyl.

[0098] In certain embodiments, R 2 is hydroxy-C 1-6 Alkyl or C 1-6 Alkoxy-C 1-6 It is an alkyl.

[0099] In certain embodiments, R 2 is hydroxy-C 1-6 It is an alkyl group.

[0100] In certain embodiments, R 2 is C 1-6 Alkoxy-C 1-6 It is an alkyl.

[0101] In certain embodiments, R 2 is methoxy-C 1-6 It is an alkyl.

[0102] In certain embodiments, R 2 is 2-methoxyethyl.

[0103] In certain embodiments, R 2 is Halo-C 1-6 It is an alkyl.

[0104] In certain embodiments, R2 is 2-hydroxyethyl, difluoromethoxy, 3-hydroxyphenyl, -methoxyphenyl, 2-hydroxypropyl, pyridin-4-yl, methyl, azetidin-yl, 2-hydroxy-1-methyl-ethyl, methylamino, dimethylamino, 1-methyl-pyrazol-4-yl, phenyl, pyrazol-4-yl, 1-methyl-2-oxo-4-pyridyl, or 2-hydroxy-1-methylpropyl.

[0105] In certain embodiments, R 2 is 2-hydroxyethyl, 2-hydroxypropyl, 2-hydroxy-1-methyl-ethyl or 2-hydroxy-1-methylpropyl.

[0106] In certain embodiments, R 2 is 2-hydroxyethyl.

[0107] In certain embodiments, R 2 is 2-hydroxypropyl.

[0108] In certain embodiments, R 2 is 2-hydroxy-1-methyl-ethyl.

[0109] In certain embodiments, R 2 is 2-hydroxy-1-methyl-propyl.

[0110] In certain embodiments, m is 2.

[0111] In certain embodiments, m is 3.

[0112] In certain embodiments, n is 0.

[0113] In certain embodiments, n is 1.

[0114] In certain embodiments, n is 2.

[0115] In certain embodiments, p is 0.

[0116] In certain embodiments, p is 1.

[0117] In certain embodiments, p is 2.

[0118] In certain embodiments, q is 1.

[0119] In certain embodiments, q is 2.

[0120] In certain embodiments, r is 2.

[0121] In certain embodiments, r is 3.

[0122] In certain embodiments, s is 2.

[0123] In certain embodiments, s is 3.

[0124] In certain embodiments, R a is hydrogen.

[0125] In certain embodiments, R a is C 1-6 It is an alkyl.

[0126] In certain embodiments, R b is hydrogen.

[0127] In certain embodiments, R b is C 1-6 It is an alkyl.

[0128] In certain embodiments, R c is hydrogen.

[0129] In certain embodiments, R c is C 1-6 It is an alkyl.

[0130] In certain embodiments, het 1is 1-methylazetidin-4-yl, 2-oxy-tetrahydrofuran-3-yl or pyrrolidinyl.

[0131] In certain embodiments, het 2 is pyridin-4-yl, 1-methyl-pyrazol-4-yl, 1-(2-hydroxypropyl)-pyrazol-4-yl, pyrazol-4-yl, pyridin-3-yl, 6-oxo-1H-pyridin-3-yl, 1-methyl-2-oxo-pyridin-4-yl, 1-quinuclidin-3-ylpyrazol-4-yl, 1-[2-[(1-methylpyrrolidin-2-yl)ethyl]pyrazol-4-yl, 1-[2-(dimethylamino)propyl]pyrazol-4-yl, 1-methyl-azetidin-3-yl)pyrazol-4-yl, 2-methyl-1H-isoquinolin-7-yl, 2-methyl-1H-isoquinolin-6-yl, 1-methyl-imidazol-4-yl or 1H-imidazol-4-yl, pyridin-5-yl.

[0132] In certain embodiments, het 3 is 1-methyl-azetidin-4-yl, azetidin-4-yl, azetidin-1-yl, pyrrolidin-3-yl, oxetan-4-yl, 1-methyl-pyrrolidin-3-yl, 1-acetyl-azetidin-4-yl, piperidin-3-yl or 4-methylpiperazin-1-yl.

[0133] In certain embodiments, the compound of the present invention is of formula (II). TIFF0007657740000003.tif68170, R 1 and R 2 is as defined herein.

[0134] In certain embodiments, the compound of the present invention is of formula (III): TIFF0007657740000004.tif67170, R 2 is as defined herein.

[0135] In certain embodiments, the compound of formula (I) is selected from: TIFF0007657740000005.tif255170TIFF0007657740000006.tif255170TIFF0007657740000007.tif255170TIFF0007657740000008.tif255170TIFF0007657740000009.tif199170, or a pharma- ceutically acceptable stereoisomer or salt thereof.

[0136] In certain embodiments, the compound of the invention is selected from: TIFF0007657740000010.tif214170TIFF0007657740000011.tif211170TIFF0007657740000012.tif255170TIFF0007657740000013.tif206170, or a stereoisomer or a pharma- ceutically acceptable salt thereof.

[0137] In certain embodiments, the compound is selected from the following: TIFF0007657740000014.tif210170TIFF0007657740000015.tif215170TIFF0007657740000016.tif53170, or a stereoisomer or a pharma- ceutically acceptable salt thereof.

[0138] In certain embodiments, the compound of the invention is selected from: structure TIFF0007657740000017.tif157170, or a stereoisomer or a pharma- ceutically acceptable salt thereof.

[0139] Also provided is a pharmaceutical composition comprising a JAK inhibitor described herein, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier, diluent, or excipient.

[0140] Also provided is the use of a JAK inhibitor described herein, or a pharma- ceutically acceptable salt thereof, in therapy, for example, in the treatment of an inflammatory disease (e.g., asthma).Also provided is the use of a JAK inhibitor described herein, or a pharma- ceutically acceptable salt thereof, for the preparation of a medicament for the treatment of 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 described herein, or a pharma- ceutically acceptable salt thereof.

[0141] In one embodiment, the disease or condition for treatment is cancer, polycythemia vera, essential thrombocytosis, myelofibrosis, chronic myelogenous leukemia (CML), rheumatoid arthritis, inflammatory bowel syndrome, Crohn's disease, psoriasis, contact dermatitis, or delayed hypersensitivity reaction.

[0142] In one embodiment, there is provided a use of a JAK inhibitor as described herein, or a pharma- ceutical acceptable salt thereof, for the treatment of cancer, polycythemia vera, basal thrombocytosis, myelofibrosis, chronic myelogenous leukemia (CML), rheumatoid arthritis, inflammatory bowel syndrome, Crohn's disease, psoriasis, contact dermatitis, or delayed hypersensitivity reactions.

[0143] In one embodiment, a composition is provided that is formulated for administration by inhalation.

[0144] In one embodiment, a metered dose inhaler is provided comprising a compound of the invention or a pharma- ceutically acceptable salt thereof.

[0145] In one embodiment, the JAK inhibitors described herein, or pharma- ceutically acceptable salts thereof, are at least 5-fold more potent as inhibitors of JAK1 than as inhibitors of LRRK2.

[0146] In one embodiment, the JAK inhibitors described herein, or pharma- ceutically acceptable salts thereof, are at least 10 times more potent as inhibitors of JAK1 than as inhibitors of LRRK2.

[0147] In one embodiment, there is provided a method of treating hair loss in a mammal, comprising administering to the mammal a JAK inhibitor described herein, or a pharma- ceutically acceptable salt thereof.

[0148] In one embodiment, there is provided a use of a JAK inhibitor, or a pharma- ceutically acceptable salt thereof, as described herein for the treatment of hair loss.

[0149] In one embodiment, there is provided a use of a JAK inhibitor, or a pharma- ceutically acceptable salt thereof, as described herein for the preparation of a medicament for treating hair loss in a mammal.

[0150] The compounds of the present invention may contain one or more asymmetric carbon atoms. Thus, the compounds may exist as diastereomers, enantiomers or mixtures thereof. The synthesis of the compounds may use racemates, diastereomers or enantiomers as starting materials or as intermediates. A mixture of specific diastereomeric compounds may be separated or enriched in one or more specific diastereomers by chromatography or crystallization methods. Similarly, a mixture of enantiomers may be separated or enantiomerically enriched using the same techniques known or other methods. Each asymmetric carbon or nitrogen atom may have an R or S configuration, and both of these configurations are within the scope of the present invention.

[0151] In structures depicted herein, where no specific configuration of a chiral atom is specified, all stereoisomers are intended and encompassed as compounds of the present invention. Where a configuration is specified by a solid wedge or dotted line representing a particular configuration, that stereoisomer is specified and defined. Unless otherwise indicated, when a solid wedge or dotted line is used, the relative configuration is intended.

[0152] Another embodiment includes prodrugs of the compounds described herein that contain known amino-protecting and carboxy-protecting groups that are released, e.g., hydrolyzed, under physiological conditions to yield the compounds of the invention.

[0153] The term "prodrug" refers to a precursor or derivative form of a pharmacologic active substance that is less effective in patients compared to the parent drug, but can be activated or converted to the active parent form by enzymatic reaction or hydrolysis. See Wilman, "Prodrugs in Cancer Chemotherapy" Biochemical Society Transactions, 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., (ed.), 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.

[0154] A particular class of prodrugs are compounds in which the nitrogen atom of an amino, amidino, aminoalkyleneamino, iminoalkyleneamino or guanidino group is replaced by a hydroxy group, an alkylcarbonyl (-CO-R) group, an alkoxycarbonyl (-CO-OR) group, or an acyloxyalkyl-alkoxycarbonyl (-CO-ORO-CO-R) group, where R is a monovalent or divalent group (e.g., alkyl, alkylene or aryl), or a group having the formula -C(O)-O-CP1P2-haloalkyl, where P1 and P2 are the same or different and are hydrogen, alkyl, alkoxy, cyano, halogen, alkyl or aryl. In a particular embodiment, the nitrogen atom is one of the nitrogen atoms of the amidino group. Prodrugs can be prepared by reacting a compound with an activating group (e.g., an acyl group) and binding, for example, a nitrogen atom of the compound to, for example, the carbonyl of the activated acyl group. Examples of activated carbonyl compounds include those containing a leaving group attached to the carbonyl group, such as acyl halides, acyl amines, acyl pyridinium salts, acyl alkoxides, acyl phenoxides (e.g., p-nitrophenoxyacyl), dinitrophenoxyacyl, fluorophenoxyacyl, and difluorophenoxyacyl. The reaction is generally carried out in an inert solvent at low temperatures (e.g., −78° C. to about 50° C.). The reaction may also be carried out in the presence of an inorganic base (e.g., potassium carbonate or sodium bicarbonate) or an organic base (e.g., an amine, such as pyridine, trimethylamine, triethylamine, triethanolamine, etc.).

[0155] Additional types of prodrugs are also included. For example, the free carboxyl groups of the JAK inhibitors described herein can be derivatized as amides or alkyl esters. As another example, in compounds of the invention that contain free hydroxyl groups, the hydroxyl groups can be derivatized as prodrugs by converting them to groups such as, but not limited to, phosphate esters, hemisuccinates, dimethylaminoacetates, or phosphoryloxymethyloxycarbonyl groups, as reviewed 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 are carbonate prodrugs, sulfonate esters, and sulfonate esters of hydroxy groups. The acyl group can be an alkyl ester optionally substituted with groups such as, but not limited to, ether, amine and carbonate functionalities, or, where the acyl group is an amino acid ester as described above, derivatization of the hydroxy group as (acyloxy)methyl and (acyloxy)ethyl ethers is also included. Prodrugs of this type are described in J. Med. Chem., (1996), 39:10.Further specific examples include replacement of the hydrogen atom of an alcohol group with groups such as (C1-C6)alkanoyloxymethyl, 1-((C1-C6)alkanoyloxy)ethyl, 1-methyl-1-((C1-C6)alkanoyloxy)ethyl, (C1-C6)alkoxycarbonyloxymethyl, N-(C1-C6)alkoxycarbonylaminomethyl, succinoyl, (C1-C6)alkanoyl, α-amino(C1-C4)alkanoyl, arylacyl and α-aminoacyl (or α-aminoacyl-α-aminoacyl), where each α-aminoacyl group is independently selected from natural L-amino acids, P(O)(OH)2, -P(O)(O(C1-C6)alkyl)2 or glycosyl (the group resulting from removal of the hydroxyl group of the hemiacetal form of a carbohydrate).

[0156] "Leaving group" refers to a part of an initial reactant in a chemical reaction that leaves the initial reactant in the chemical reaction. Examples of leaving groups include, but are not limited to, halogen atoms, alkoxy, and sulfonyloxy groups. Examples of sulfonyloxy groups include, but are not limited to, 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)).

[0157] Synthesis of Janus kinase inhibitor compounds The compounds can be synthesized by the synthetic routes described herein. In certain embodiments, steps known in the chemical arts can be used in addition to or in consideration of the descriptions contained herein. Starting materials are generally commercially available, such as from Aldrich Chemicals (Milwaukee, Wis.), or can be readily prepared using methods known to those skilled in the art (e.g., generally prepared by the methods described in Louis F. Fieser and Mary Fieser, Reagents for Organic Synthesis, v. 1-19, Wiley, NY (1967-1999 ed.), Beilsteins Handbuch der organischen Chemie, 4, Aufl. ed. Springer-Verlag, Berlin (including supplementary materials, and available through the Beilstein online database), or Comprehensive Heterocyclic Chemistry, Editors Katrizky and Rees, Pergamon Press, 1984).

[0158] The compounds may be prepared singly or as a compound library containing at least two (e.g., 5-1,000 compounds or 10-100 compounds). Libraries of compounds may be prepared by combinatorial "split and mix" approaches or multiple parallel synthetic chemical reactions using solution or solid phase, as known to those skilled in the art. Thus, in yet another aspect of the invention, a compound library is provided that contains at least two compounds of the invention.

[0159] For convenience of illustration, the reaction schemes depicted below provide routes for synthesizing the compounds of the present invention as well as key intermediates. For more detailed descriptions of the individual reaction steps, see the Examples section below. Those skilled in the art will appreciate that other synthetic routes may be used. Although some specific starting materials and reagents are depicted in the schemes and discussed below, other starting materials and reagents may be substituted to provide a variety of derivatives or reaction conditions. In addition, many of the compounds prepared by the methods described below may be further modified in light of the present disclosure using conventional chemistry well known to those skilled in the art.

[0160] In the preparation of the compounds of the present invention, protection of remote functional groups (e.g., primary or secondary amines) of intermediates may be necessary. The need for such protection will vary with 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 is readily determined by one skilled in the art. For a general description of protecting groups and their use, see TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

[0161] Other transformations commonly used in the synthesis of compounds of the invention, which can be carried out using a variety of reagents and conditions, include: (1) Reaction of a carboxylic acid with an amine to form an amide. Such a transformation can be carried out using a variety of reagents known to those skilled in the art, but for a comprehensive review see Tetrahedron, 2005, 61, 10827-10852. (2) The reaction of primary or secondary amines with aryl halides or pseudohalides (e.g., triflates), commonly known as "Buchwald-Hartwig cross-coupling", can be carried out using a variety of catalysts, ligands and bases. A review of these methods is given in Comprehensive Organic Name Reactions and Reagents, 2010, 575-581. (3) Palladium-catalyzed cross-coupling reactions between aryl halides and vinyl boronic acids or boronate esters. This transformation is a type of "Suzuki-Miyaura cross-coupling", a reaction type thoroughly reviewed in Chemical Reviews, 1995, 95(7), 2457-2483. (4) Hydrolysis of esters to provide the corresponding carboxylic acids is well known to one of skill in the art and conditions include: for methyl and ethyl esters, use of a strong aqueous base such as lithium, sodium or potassium hydroxide, or a strong aqueous inorganic acid (e.g., HCl); for tert-butyl esters, hydrolysis is carried out using an acid (e.g., HCl in dioxane, or trifluoroacetic acid (TFA) in dichloromethane (DCM)). (5) TIFF0007657740000018.tif137170 Reaction Scheme 1 illustrates the synthesis of compounds of formula I. Nitropyrazole compound 1 can be arylated with 4-bromo-1-(difluoromethoxy)-2-iodobenzene under palladium catalyzed conditions to generate compound 2. The nitro group of compound 2 can be reduced to give aminoaniline 3, for example, by iron and ammonium chloride conditions. Commercially available pyrazolo[1,5-a]pyrimidine-3-carboxylic acid is amide-coupled with an organic base, for example, but not limited to, DIPEA and DMA, in the presence of a coupling reagent, for example, but not limited to, PyAOP, in an organic solvent, for example, but not limited to, DMF, to provide compound 4. The SEM protecting group of compound 4 is removed by using an acid, for example, but not limited to, HCl, in a solvent, for example, but not limited to, 1,4-dioxane, resulting in compound 5. Compound 5 is then reacted with a group R 1 Compound 7 can be prepared by N-alkylation with -X (wherein X is halo, e.g., bromine) to provide compound 6. Compound 7 can be prepared by coupling with an appropriate thiol reagent R under Pd-catalyzed coupling conditions. 2 Compound 6 may also be synthesized by treatment with -SH. Oxidation of the thio group of compound 7 provides compound 8, which is a compound of formula (II) according to the present invention. Alternatively, SEM protected compound 4 can be directly N-alkylated by treatment with a fluoroborate reagent.

[0162] TIFF0007657740000019.tif121170 Reaction Scheme 2 illustrates another route for the preparation of the compounds of the present invention. Nitropyrazylaryl bromide compound 1 is subjected to a thiolation reaction in the presence of a palladium catalyst to give thiolated compound 2, followed by reduction of the nitro group to the corresponding amine to give compound 3. Amide bond coupling with commercially available pyrazolo[1,5-a]pyrimidine-3-carboxylic acid in the presence of a coupling reagent gives compound 4. The SEM protecting group of compound 4 is then removed under acidic conditions to give compound 5, which is then subjected to S-oxidation to give the corresponding sulfone compound 6. N-alkylation of the pyrazole moiety is then performed to provide compound (II) of the present invention.

[0163] In Reaction Scheme 3, aryl bromide compound 1 is treated with a trialkylsilanethiol (e.g., triisopropylsilanethiol) to give thiosilyl compound 2. The thiol is deprotected and alkylated to give sulfide compound 3, which is then S-oxidized to give compound (II) of the present invention.

[0164] In Reaction Scheme 4, aryl bromide compound 1 is treated with trialkylsilanethiol and desilylated in situ under basic conditions to provide thiol compound 2. Compound 2 is S-alkylated to provide sulfide compound 3, which is then oxidized to provide compound II of the present invention.

[0165] TIFF0007657740000022.tif51170 In certain embodiments, trialkylsilylthiol compound 1 can be oxidized directly to provide sulfonic acid compound 2, which can then be reacted with an amine to provide sulfonamide compound 3, as shown in Reaction Scheme 5.

[0166] TIFF0007657740000023.tif77170 In certain embodiments, as shown in Reaction Scheme 6, thiol compound 1 is reacted with bromopyrazole to provide pyrazole thioester compound 2. Compound 2 may be oxidized to provide the corresponding sulfone compound, which may then be N-alkylated to provide compound 4.

[0167] In Reaction Scheme 7, sulfone compound 1 is treated with diisopropyl azodicarboxylate to give isopropyl carboxylate ester compound 2.

[0168] In Reaction Scheme 8, aryl bromide compound 1 is reacted with an N-protected thioalkylamine to give aminoalkylsulfide compound 2. Compound 2 is oxidized to give the corresponding sulfone, which is then deprotected to give aminoalkylsulfone compound 4. Compound 4 can be subjected to reductive animation to give compound 5, or alternatively can be reacted with an acid chloride to give carboxamide compound 6.

[0169] TIFF0007657740000026.tif50170

[0170] In Reaction Scheme 9, a thiol compound 1 is reacted with a substituted epoxide to give a hydroxyalkyl sulfide compound 2. Compound 2 is then S-oxidized to give the corresponding hydroxyalkyl sulfone compound 3.

[0171] It will be appreciated that, in the presence of suitable functional groups, the compounds of the 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 methods include alkylation, arylation, heteroarylation, acylation, sulfonylation, halogenation, nitration, formylation and coupling procedures known in the art.

[0172] In a further example, primary or secondary amine groups can be converted to amide groups (-NHCOR' or -NRCOR') by acylation. Acylation can be carried out by reaction with a suitable acid chloride in the presence of a base (e.g., triethylamine) in a suitable solvent (e.g., dichloromethane) or by reaction with a suitable carboxylic acid in the presence of a suitable coupling reagent such as HATU (O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) in a suitable solvent (e.g., dichloromethane). Similarly, amine groups can be converted to sulfonamide groups (-NHSO2R' or -NR''SO2R') by reaction with a suitable sulfochloride in the presence of a suitable base (e.g., triethylamine) in a suitable solvent (e.g., dichloromethane). Primary or secondary amine groups can be converted to urea groups (-NHCONR'R'' or -NRCONR'R'') by reaction with a suitable isocyanate in the presence of a suitable base, such as triethylamine, in a suitable solvent (e.g., dichloromethane).

[0173] The amine (-NH2) is obtained by reduction of the nitro (-NO2) group, for example by catalytic hydrogenation with hydrogen in the presence of a metal catalyst (e.g. palladium) on a support such as carbon in a solvent (e.g. ethyl acetate or an alcohol such as methanol). Alternatively, the conversion may be carried out by chemical reduction, for example with a metal (e.g. tin or iron) in the presence of an acid (e.g. hydrochloric acid).

[0174] In a further example, amine (-CHNH) groups can be obtained by reduction of nitriles (-CN), for example, by catalytic hydrogenation with hydrogen in the presence of a metal catalyst (e.g., palladium on a carbon support, or Raney nickel) in a solvent such as an ether (e.g., a cyclic ether such as tetrahydrofuran) at a suitable solvent temperature (e.g., from about -78°C to reflux).

[0175] In a further example, amine (-NH2) groups can be obtained by conversion of a carboxylic acid group (-CO2H) to the corresponding acyl azide (-CON3), Curtius rearrangement and hydrolysis of the resulting isocyanate (-N=C=O).

[0176] The aldehyde group (-CHO) can be converted to an amine group (-CHNR'R'') by reductive amination using an amine and a borohydride (e.g., sodium triacetoxyborohydride or sodium cyanoborohydride) in a solvent such as a halogenated hydrocarbon, e.g., dichloromethane, or an alcohol, e.g., ethanol, if necessary, in the presence of an acid (e.g., acetic acid at room temperature).

[0177] In a further example, an aldehyde group may be converted to an alkenyl group (-CH=CHR') using an appropriate phosphorane or phosphonate under standard conditions known to those skilled in the art, such as a Wittig or Wadsworth-Emmons reaction.

[0178] Aldehyde groups can also be obtained by reduction of ester groups (e.g., -CO2Et) or nitriles (-CN) with diisobutylaluminum hydride in a suitable solvent (e.g., toluene).Alternatively, aldehyde groups can also be obtained by oxidation of alcohol groups with any suitable oxidizing agent known to those skilled in the art.

[0179] The ester group (-COR') can be converted to the corresponding acid group (-COH) by acid- or base-catalyzed hydrolysis, depending on the nature of R. When R is t-butyl, for example, acid-catalyzed hydrolysis can be carried out by treatment with an organic acid (e.g. trifluoroacetic acid in an aqueous solvent) or by treatment with an inorganic acid (e.g. hydrochloric acid in an aqueous solvent).

[0180] Carboxylic acid groups (-CO2H) can be converted to amides (CONHR' or -CONR'R'') by reaction with an appropriate amine in the presence of a suitable coupling reagent (eg, HATU) in a suitable solvent (eg, dichloromethane).

[0181] In a further example, a carboxylic acid can be homologated at one carbon (i.e., -CO2H to -CH2CO2H) by conversion to the corresponding acid chloride (-COCl) followed by Arndt-Eistert synthesis.

[0182] In a further example, -OH groups may be generated from the corresponding esters (e.g. -COR') or aldehydes (-CHO) by reduction, for example with a metal hydride complex (e.g. lithium aluminum hydride in diethyl ether or tetrahydrofuran or sodium borohydride in a solvent such as methanol). Alternatively, alcohols may be prepared by reduction of the corresponding acids (-COH) using, for example, lithium aluminum hydride in a solvent such as tetrahydrofuran or using borane in a solvent such as tetrahydrofuran.

[0183] The alcohol group can be converted to a leaving group (e.g., a halogen atom or a sulfonyloxy group, such as an alkylsulfonyloxy (e.g., trifluoromethylsulfonyloxy) or arylsulfonyloxy (e.g., p-toluenesulfonyloxy) group) using conditions known to those skilled in the art. For example, the alcohol is reacted with thioyl chloride in a halogenated hydrocarbon (e.g., dichloromethane) to give the corresponding chloride. A base (e.g., triethylamine) may also be used in the reaction.

[0184] In other examples, alcohol, phenol or amide groups can be alkylated by coupling the phenol or amide with an alcohol in the presence of a phosphine (e.g. triphenylphosphine) and an activator (e.g. diethyl-diisopropyl, or dimethylazodicarboxylate) in a solvent (e.g. tetrahydrofuran). Alternatively, alkylation can be carried out by deprotonation with a suitable base (e.g. sodium hydride) followed by addition of an alkylating agent (e.g. an alkyl halide).

[0185] Aromatic halogen substituents in the compounds may be subjected to halogen-metal displacement by treatment with a base (e.g., a lithium base (e.g., n-butyl or t-butyl lithium)) in a solvent such as tetrahydrofuran, optionally at low temperature (e.g., about −78° C.), followed by quenching with an electrophile to introduce the desired substituent. Thus, for example, a formyl group may be introduced using N,N-dimethylformamide as the electrophile. Alternatively, aromatic halogen substituents may be subjected to a metal (e.g., palladium or copper) catalysis to introduce, for example, an acid, ester, cyano, amide, aryl, heteroaryl, alkenyl, alkynyl, thio or amino substituent. Suitable procedures that may be used include those described in Heck, Suzuki, Stille, Buchwald or Hartwig.

[0186] Aromatic halogen substituents may undergo nucleophilic substitution reactions with suitable nucleophiles, such as amines or alcohols. Suitably, such reactions may be carried out at elevated temperatures in the presence of microwave irradiation.

[0187] Separation method In each exemplary scheme, it may be beneficial to separate the reaction products from each other or from the starting materials. The desired products at each step or successive steps are separated or purified (hereinafter "separated") to the desired degree of homogeneity by techniques common in the art. Typically, such separations include extraction in multiple phases from a solvent or solvent mixture, crystallization or trituration, distillation, sublimation or chromatography. Chromatography can include 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 equipment, small scale analytical, simulated moving bed (SMB) and preparative thin or thick layer chromatography, and small scale thin layer and flash chromatography techniques.

[0188] Another class of separation methods involves treating the mixture with a selected reagent to bind or render separable the desired product, unreacted starting materials, reaction by-products, etc. Such reagents include adsorbents or absorbents (e.g., activated carbon, molecular sieves, ion exchange media, etc.). Alternatively, such reagents can be acids in the case of basic materials, bases in the case of acidic materials, binding reagents (e.g., antibodies), binding proteins, selective chelators (e.g., crown ethers), liquid / liquid ion extraction reagents (LIX), etc.

[0189] The choice of an appropriate separation method depends on the properties of the materials involved: for example, the separation method takes into account boiling point and molecular weight in the case of distillation and sublimation, the presence or absence of polar functional groups in the case of chromatography, the stability of the materials in acidic and basic solvents in the case of multi-phase extraction, etc. The skilled person will apply the technique that allows the most desired separation.

[0190] Diastereomeric mixtures can be separated into their individual diastereoisomers based on their physical chemical differences by methods well known to those skilled in the art, such as chromatography or partial crystallization. Enantiomers can be separated by converting the enantiomeric mixture to a diastereomeric mixture by reaction with a suitable optically active compound, such as a chiral auxiliary (e.g., a chiral alcohol or Mosher's acid chloride), separating the diastereoisomers, and converting the individual diastereoisomers to the corresponding pure enantiomers (e.g., hydrolysis). Some of the compounds of the present invention may also be atropisomers (e.g., substituted biaryls) and are contemplated as part of the present invention. Enantiomers can also be separated using chiral HPLC columns or supercritical fluid chromatography.

[0191] A single stereoisomer (e.g., enantiomer), substantially free of its stereoisomer, can also be obtained by separation of a racemic mixture using methods such as, for example, formation of diastereomers with optically active resolving agents (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)). Racemic mixtures of chiral compounds of the present invention can be separated and isolated by any suitable method, for example, the following: (1) formation of ionic diastereomeric salts with chiral compounds and separation by partial crystallization or other methods; (2) formation of diastereomeric compounds with chiral inducers, separation of diastereomers and conversion to pure stereoisomers; and (3) Separation of substantially pure or enriched stereoisomers under direct chiral conditions. See Drug Stereochemistry, Analytical Methods and Pharmacology, Irving W. Wainer, Ed., Marcel Dekker, Inc., New York (1993).

[0192] Diastereomeric salts can be formed by reaction of enantiomerically pure chiral bases (e.g., brucine, quinine, ephedrine, strychnine, α-methyl-β-phenylethylamine (amphetamine), and the like) with asymmetric compounds bearing acidic functional groups (e.g., carboxylic and sulfonic acids). The diastereomeric salts can be induced to separate by partial crystallization or ionic chromatography. For separation of the optical isomers of amino compounds, addition of chiral carboxylic or sulfonic acids (e.g., camphorsulfonic acid, tartaric acid, mandelic acid, or lactic acid) results in the formation of the diastereomeric salts.

[0193] Alternatively, the substrate to be resolved is reacted with one enantiomer of a chiral compound to form a diastereomeric pair (Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds, John Wiley & Sons, Inc., New York, 1994, p. 322). Diastereomeric compounds are formed by reacting an asymmetric compound with an enantiomerically pure chiral derivatizing agent (e.g., a menthyl derivative), followed by separation and hydrolysis of the diastereomers, thereby yielding pure or enriched enantiomers. Methods for determining optical purity include forming chiral esters (e.g., menthyl esters, such as (-)-menthyl chloroformate, or Mosher esters, such as α-methoxy-α-(trifluoromethyl)phenylacetate) of the racemic mixture in the presence of a base (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 methods for separation of atropisomeric naphthyl-isoquinolines (WO 96 / 15111, incorporated herein by reference). By method (3), a racemic mixture of the 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)). The enriched or purified enantiomer can be characterized by methods used to distinguish other chiral molecules by asymmetric carbon atoms (e.g., optical rotation and circular dichroism). The chiral centers and absolute configurations of the enantiomers can be determined by X-ray crystallography.

[0194] Regioisomers and intermediates for their synthesis may be observed by analytical methods such as, for example, NMR and analytical HPLC. For certain compounds where the energy barrier for interconversion is high enough, E and Z isomers may be separated, for example, by preparative HPLC.

[0195] Pharmaceutical Compositions and Administration The compounds with which the present invention relates are JAK kinase inhibitors (eg JAK1 inhibitors) and are useful in the treatment of a number of diseases, for example inflammatory diseases (eg asthma).

[0196] Thus, other embodiments provide pharmaceutical compositions or medicaments comprising a compound of the invention, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier, diluent or excipient, as well as methods of preparing such compositions and medicaments using the compounds of the invention.

[0197] In one example, the compound of the present invention or a pharma- ceutically acceptable salt thereof may be prepared by mixing at a desired purity at room temperature at an appropriate pH using a physiologically acceptable carrier (i.e., a carrier that is non-toxic to the recipient at dosages and concentrations that can be employed in herbal medicine administration modes). The pH of the formulation depends primarily on the specific use and concentration of the compound, but typically ranges from about 3 to about 8. In one example, the compound of the present invention or a pharma- ceutically acceptable salt thereof is prepared in acetate buffer at pH 5. In other embodiments, the compound of the present invention is sterilized. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation, or as an aqueous solution.

[0198] The compositions are prepared, formulated, and administered in a manner consistent with good medical practice. Considerations in this context include the particular disorder to be treated, the particular mammal to be treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the administration schedule, and other factors known to medical practitioners.

[0199] However, it should be understood that the specific dosage for a particular patient will depend on a variety of factors, including the degree of activity of the specific compound used, the age, weight, overall health, sex, diet, time of administration, method of administration, and rate of excretion, drug combination, and the severity of the specific disease to be treated. As required in the pharmaceutical field, the appropriate level and frequency of administration will be determined in clinical trials. In general, the daily dosage range for oral administration will be in the range of 0.001 mg to about 100 mg per kg human body weight, often 0.01 mg to about 50 mg per kg, for example 0.1 to 10 mg per kg, as a single or divided dose. In general, the daily dosage range for inhalation administration will be in the range of 0.1 μg to about 1 mg per kg human body weight, preferably 0.1 μg to about 50 μg per kg, as a single or divided dose. On the other hand, in some cases, it may be necessary to use dosages outside these limits.

[0200] The compound of the present invention or its pharma- ceutically acceptable salts can be administered by any suitable means, such as oral, topical (including buccal and sublingual), rectal, intravaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal, inhalation, and epidural and intranasal, and, where necessary, by localized treatment, intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. In some embodiments, inhalation administration is employed.

[0201] The compounds of the present invention or pharma- ceutically acceptable salts thereof may be administered in any convenient dosage form, such as tablets, powders, capsules, lozenges, granules, solutions, dispersions, suspensions, syrups, sprays, steam, suppositories, gels, emulsions, patches, etc. Such compositions may contain conventional ingredients known in the art of formulation, such as diluents (e.g., glucose, lactose, or mannitol), carriers, pH modifiers, buffers, sweeteners, fillers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, fragrances, flavorings, other known additives, as well as further active agents.

[0202] Suitable carriers and additives 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 materials such as 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, additives, disintegrants, lubricants, sweeteners, flavorings, dyes, and combinations thereof, as known to those skilled in the art (see Remington's Pharmaceutical Sciences, pp 1289-1329, 1990). Any conventionally known carrier is contemplated for use in therapeutic or pharmaceutical compositions, unless it is incompatible with the active ingredient. Exemplary additives include dicalcium phosphate, mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, or combinations thereof. A pharmaceutical composition may contain various types of carriers or additives, depending on whether it is administered in solid, liquid, or aerosol form, and whether it needs to be sterile for such administration routes.

[0203] For example, tablets and capsules for oral administration may be in unit dosage form and may contain conventional additives such as binders (e.g., syrup, acacia, gelatin, sorbitol, tragacanth gum, or polyvinylpyrrolidone, etc.), fillers (e.g., lactose, sugar, corn starch, calcium phosphate, sorbitol, or glycine), molding lubricants (e.g., magnesium stearate, talc, polyethylene glycol, or silica), disintegrants (e.g., potato starch), or acceptable wetting agents (e.g., sodium lauryl sulfate). Tablets may be coated according to methods known to those skilled in the art. 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 a dry substance for reconstitution with water or other suitable vehicle before use. For example, such liquid preparations may contain conventional additives such as suspending agents (e.g., sorbitol, syrup, methyl cellulose, glucose syrup, hydrogenated edible fats from gelatin), emulsifying agents (e.g., lecithin, sorbitan monooleate or acacia), non-aqueous vehicles (which may include edible oils) (e.g., almond oil, coconut oil, oily esters such as glycerin, propylene glycol or ethyl alcohol), preservatives (e.g., methyl or propyl p-hydroxybenzoates or sorbic acid), and, if desired, conventional flavorings or colorings.

[0204] For topical administration to the skin, the compounds are processed into creams, lotions or ointments. Cream or ointment formulations that can be used for drugs are conventional formulations known to those skilled in the art, for example as described in standard texts of pharmaceutics (e.g. the British Pharmacopoeia).

[0205] The compound of the present invention or its pharma- ceutically acceptable salt can be prepared for inhalation, for example, nasal spray, or inhalation of dry powder or aerosol.For delivery by inhalation, the compound is typically in the form of microparticles, which can be prepared by various techniques, such as spray drying, freeze drying and micronization.The generation of aerosol can be carried out, for example, by using a pressure-driven jet nebulizer or ultrasonic nebulizer, for example, by using a propellant-driven metered-dose aerosol, or by propellant-free administration, for example, by micronized compound from an inhalation capsule or other "dry powder" delivery system.

[0206] For example, the compositions of the present invention can be prepared as suspensions for delivery from nebulizers or as aerosols in liquid propellants, for example, for use in pressurized metered dose inhalers (PMDIs).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).

[0207] In some embodiments, the compositions of the invention are in dry powder form for delivery using a dry powder inhaler (DPI). Many types of DPIs are known.

[0208] Microparticles for administration can be prepared with additives that aid delivery and release.For example, in dry powder formulations, microparticles can be prepared with larger carrier particles to aid the flow from DPI to lungs.Suitable carrier particles are known and include lactose particles, for example, they can have mass median aerodynamic diameter of more than 90 μm.

[0209] In the case of aerosol-based formulations, examples are as follows: Compounds of the Invention * 24mg / canister Lecithin, NF liq. concentration 1.2mg / canister Trichlorofluoromethane, NF 4.025g / canister Dichlorodifluoromethane, NF 12.15g / canister. * or a pharma- ceutically acceptable salt thereof

[0210] Depending on the inhaler system used, the compound of the present invention or its pharma-ceutically acceptable salt can be administered as described.In addition to the compound, the dosage form can further comprise the above-mentioned additives, or for example, propellant (e.g., Frigen in the case of metered dose aerosol), surfactant, emulsifier, stabilizer, preservative, flavoring agent, filler (e.g., lactose in the case of powder inhaler), or other active substance if appropriate.

[0211] For inhalation, many systems are available, which allow the patient to generate and administer aerosols with optimal particle size using the appropriate inhalation technique. For metered dose aerosols, in addition to the use of adapters (spacers, expanders) and pear-shaped containers (e.g., Nebulator®, Volumatic®), and automated devices that generate puffer sprays (Autohaler®), many technical means are available, especially for powder inhalers (e.g., Diskhaler®, Rotadisk®, Turbohaler®, or inhalers described in, for example, U.S. Pat. No. 5,263,475, which is incorporated herein by reference). In addition, the compound of the present invention or its pharma- ceutically acceptable salts may be delivered in a multi-chamber device, which allows the delivery of combination drugs.

[0212] Compound or its pharmaceutically acceptable salt can be administered parenterally in sterile medium.Depending on the vehicle additive and concentration used, compound can be suspended or dissolved in vehicle.Preferably, adjuvants such as local anesthetic, preservative and buffer can be dissolved in vehicle.

[0213] Target-specific inhaled drug delivery The compounds of the present invention can be used for target-specific inhalation delivery. Optimization of drugs for delivery to the lungs by local (inhalation) administration has been recently reviewed (Cooper, AE et al. Curr. Drug Metab. 2012, 13, 457-473).

[0214] Due to the limitations of delivery devices, administration of inhaled drugs may be limited to humans, which requires highly potent molecules with good lung pharmacokinetic properties. For example, high target potency and lung safety concerns associated with high aerosol load (e.g., coughing or discomfort) are important, especially for inhalants, due to factors such as the limited amount of drug that can be delivered in a single puff from an inhaler. For example, in some embodiments, a Ki of about 0.5 nM or less, e.g., in a biochemical assay of JAK1 described herein, and an IC50 of about 20 nM or less, e.g., in a JAK1-dependent cell-based assay described herein, are 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 1 / 2 times less than the predicted human dose of a known compound. Thus, in some embodiments, the compounds described herein (or a pharmaceutically acceptable salt thereof) exhibit such potency values. The following procedure was used to evaluate the potential use of the compounds of the present invention as inhaled drugs.

[0215] IL13 signaling. IL13 signaling is deeply involved in the pathogenesis of asthma. IL13 is a cytokine that requires active JAK1 for signal transduction. Thus, inhibition of JAK1 also inhibits IL13 signaling, which is beneficial for asthma patients. Inhibition of IL13 signaling in animal models (e.g., mouse models) may predict future benefits for human patients suffering from asthma. Thus, it may be beneficial for inhalation of JAK1 inhibitors to show inhibition of IL13 signaling in animal models. Methods for measuring such inhibition are known in the art. For example, as discussed herein and known to those skilled in the art, it is known that JAK1-dependent phosphorylation of STAT6 is a downstream consequence of IL13 stimulation. Thus, in some embodiments, the compounds described herein (or pharma-ceutically acceptable salts thereof) show inhibition of pSTAT6 induction in the lung. To test the pharmacodynamic effect on pSTAT6 levels, compounds of the invention were co-administered intranasally with IL13 to female Balb / c mice. Compounds were prepared in 0.2% (v:v) Tween 80 in saline and mixed 1:1 (v:v) with IL13 immediately before administration. Intranasal administration was performed by preparing target dose levels (3 mg / kg, 1 mg / kg, 0.3 mg / kg, 0.1 mg / kg) in a fixed volume (50 μL) and applying directly to the nostrils by pipetting to lightly anesthetized (isoflurane) mice. 0.25 hours after administration, blood samples (approximately 0.5 mL) were collected by cardiac puncture and plasma was obtained by centrifugation (1500 g, 10 min, +4° C.). Lungs were perfused with chilled phosphate buffered saline (PBS), weighed, and flash frozen in liquid nitrogen. All samples were stored at approximately -80°C until analysis. Thawed lung samples were weighed and homogenized using an Omni-Prep Bead Ruptor at 4°C after the addition of 2 mL of HPLC grade water for each gram of tissue. Plasma and lung samples were extracted by protein precipitation with 3 volumes of acetonitrile containing Tolbutamide (50 ng / mL) and Labetalol (25 ng / mL) as analytical internal standards.After vortexing and centrifugation at 3200g for 30 min at 4°C, the supernatants were 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 analyzed for parent compound by LC-MS / MS against a series of matrix-matched correction and quality control standards. Standards were prepared by spiking test compounds into aliquots of plasma or lung homogenates from control Balb / c mice (2:1 in HPLC-grade water) and extracting as described for experimental samples. Lung:plasma ratios were measured as the ratio of mean lung concentration (μM) to mean plasma concentration (μM) at the sampling time (0.25h).

[0216] To measure pSTAT6 levels, mouse lungs were stored frozen at -80°C until assay and homogenized in 0.6 ml of ice-cold cell lysis buffer (Cell Signalling Technologies, Catalog #9803S) supplemented with 1 mM PMSF and a mix of protease (Sigma Aldrich, Catalog #P8340) and phosphatase (Sigma Aldrich, Catalog #P5726 and P0044) inhibitors. Samples were centrifuged at 16060xg for 4 min at 4°C to remove tissue debris, and protein concentrations of homogenates were measured using the Pierce BCA Protein Assay Kit (Cat #23225). Samples were diluted in ice-cold distilled water to a protein concentration of 5 mg / ml and pSTAT6 levels were assayed by Meso Scale Discovery electrochemiluminescence immunoassay. Briefly, STAT6 capture antibody (R&D Systems, Catalog #MAB 2169) was coated at 150 μg / ml at 5 μl / well onto 96-well Meso Scale Discovery High Binding Plates (Cat #L15XB-3) and air-dried at room temperature for 5 hours. Plates were blocked by adding 30 mg / ml Meso Scale Discovery Blocker A (Cat #R93BA-4) at 150 μl / well and incubating at room temperature for 2 hours on a microplate shaker. Blocked plates were washed 4 times with Meso Scale Discovery TRIS Wash Buffer (Cat #R61TX-1) and then lung homogenate was introduced at 50 μl / well to perform protein loading at 250 μg / well. Assay plates were incubated overnight at 4°C, washed 4 times with TRIS wash buffer, and then sulfo-tagged pSTAT6 detection antibody (BD Pharmingen, Cat#558241) was added at 2.5 μg / ml at 25 μl / well for 2 hours at room temperature on a microplate shaker. Plates were washed 4 times with TRIS wash buffer and 1× Meso Scale Discovery Read Buffer T (Cat#R92TC-1) was added at 150 μl / well.pSTAT6 levels in lung homogenates were quantified by electrochemiluminescence detection on a Meso Scale Discovery SECTOR S 600 instrument.

[0217] JAK and JAK2 inhibition. Compounds that inhibit JAK1 and JAK2 may be useful for the treatment of various 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). Submaximal JAK2 inhibition, however, does not appear to be associated with PAP. Thus, modest JAK1 vs. JAK2 selectivity, or even near-equal 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 with equal potency for JAK1 and JAK2 are desirable. In other embodiments, compounds with 2×-5× higher selectivity for JAK1 over JAK2 may be beneficial as inhaled JAK1 inhibitors. Thus, in some embodiments, the compounds described herein (or pharma- ceutically acceptable salts thereof) exhibit such selectivity. Methods for measuring JAK1 and JAK2 selectivity are known in the art, and information thereon is provided in the Examples herein.

[0218] Kinase profiling. In addition, it may be desirable for an inhaled JAK1 or JAK1 / JAK2 inhibitor to have selectivity over one or more other kinases to reduce the possibility of potential toxicity due to inhibiting off-target kinase pathways. Thus, it may be beneficial for an inhaled JAK1 inhibitor to be selective over a broad panel of non-JAK kinases, for example, in protocols such as the available ThermoFisher Scientific's SelectScreen™ Biochemical Kinase Profiling Service using Adapta™ Screening Protocol Assay Conditions (Revised July 29, 2016), LanthaScreen™ Eu Kinase Binding Assay Screening Protocol and Assay Conditions (Revised June 7, 2016), and / or Z'LYTE™ Screening Protocol and Assay Conditions (Revised September 16, 2016). For example, the compound of the present invention or a pharma- ceutically acceptable salt thereof exhibits at least 50-fold higher selectivity for JAK1 over a panel of non-JAK kinases. Thus, in some embodiments, the compounds described herein (or a pharma- ceutically acceptable salt thereof) exhibit such selectivity.

[0219] Cytotoxicity Assay. Hepatotoxicity, general cytotoxicity, or cytotoxicity by unknown mechanisms are undesirable characteristics for a candidate drug, and inhaled drugs are no exception. It may be beneficial for inhaled JAK1 or JAK1 / JAK2 inhibitors to have inherently low cytotoxicity against various cell types. Typical cell types used to assess cytotoxicity include primary cells (e.g., human hepatocytes) and proliferative cell lines (e.g., Jurkat and HEK-293). Thus, in some embodiments, the compounds described herein (or pharma- ceutically acceptable salts thereof) 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. Cells were seeded at 450 cells / 45μl medium in Greiner 384-well black / clear tissue culture treated plates (Greiner Catalog # 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, compounds were diluted in 100% DMSO (final DMSO concentration on cells=0.5%) to generate a 10-point dose-response curve with a top concentration of 50μM and treated with cells and compounds. The cells were then incubated overnight at 37°C in a CO2 and humidity controlled incubator for 72 hours. After 72 hours of incubation, viability was measured in all wells using CellTiterGlo® (Promega Catalog# G7572). After 20 minutes of incubation at room temperature, plates were read using EnVision™ (Perkin Elmer Life Sciences) in luminescence mode. (b) With human primary hepatocytes: Test compounds were prepared as 10 mM solutions in DMSO. In addition, a positive control, such as Chlorpromazine, was prepared as a 10 mM solution in DMSO. Test compounds were typically evaluated using a 7-point dose-response curve with 2-fold dilutions. The highest concentration tested was typically 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 to 37°C, pelleted and resuspended in InVitroGro™ HT thawing medium (BioreclamationIVT). Hepatocyte viability was assessed by trypan blue exclusion, and cells were seeded in black-walled BioCoat collagen 384-well plates (Conning BD) at a density of 13,000 cells / well in InVitroGro™ CP plating medium (supplemented with 1% Torpedo™ Antibiotic Mix (Bioreclamation IVT) and 5% fetal bovine serum). Cells were incubated overnight for 18 hours (37°C, 5% CO2) before treatment. After 18 hours of incubation, plating medium was removed and hepatocytes were treated with compounds diluted in InVitroGro™ HI medium (serum-free conditions) containing 1% Torpedo™ Antibiotic Mix and 1% DMSO. Hepatocytes were treated with test compound concentrations of, for example, 0.78, 1.56, 3.12, 6.25, 12.5, 25 and 50 μM, in a final volume of 50 μL. Typically, a positive control (e.g., Chlorpromazine) was included in the assay at the same concentration as the test compound. Additional cells were treated with 1% DMSO as a vehicle control. All treatments were timed for 48 hours (37°C, 5% CO2) and each treatment condition was tested in triplicate. After 48 hours of compound treatment, ATP content was measured to determine cell viability using the CellTiter-Glo® Cell Viability Assay (Promega) as an end-point assay. The assay was performed according to the manufacturer's instructions. Luminescence was measured with an EnVision™ Muliplate Reader (PerkinElmer, Waltham, MA, USA).Luminescence data were normalized to vehicle (1% DMSO) control wells. Inhibition curves and estimated IC. 50 Values ​​were obtained by nonlinear regression of log-transformed inhibitor concentrations (7-point dilution series including vehicle) versus normalized response with variable Hill slope, with upper and lower limits set as constants of 100 and 0, respectively (GraphPad Prism™, GraphPad Software, La Jolla, CA, USA).

[0220] hERG inhibition. Inhibition of the hERG (human delayed rectifier potassium ion channel gene) potassium channel can result in long QT syndrome and cardiac arrhythmias. Although the plasma concentration of inhaled JAK1 or JAK1 / JAK2 inhibitors is likely to be low, compounds accumulated in the lungs will exit the lungs into the bloodstream via pulmonary involvement and circulate directly to the heart. Thus, especially immediately after administration, the local concentration of inhaled JAK1 inhibitors in the heart may be transiently higher than the level in total plasma. Thus, it may be beneficial to minimize the hERG suppression of inhaled JAK1 inhibitors. For example, in some embodiments, the IC50 of hERG is preferably more than 30-fold higher than the Cmax of the free drug in plasma. Thus, in some embodiments, the compounds of the present invention (or pharma- ceutically acceptable salts thereof) exhibit minimized hERG inhibition, for example, under the following conditions: (a) hERG 2pt automated patch clamp conditions are used to test the in vitro effects of compounds on hERG expressed in mammalian cells, assessed at room temperature using a QPatch HT® (Sophion Bioscience A / S, Denmark) automated parallel patch clamp system. In some cases, compounds were tested at only one or two concentrations (e.g., 1 or 10 μM). In other cases, a broader concentration-response relationship was established and IC50 estimates were made. For example, test compound concentrations were selected to span a range of approximately 10-90% inhibition in half-log increments. Each test substance concentration was tested in two or more cells (n≧2). Exposure time to each test substance concentration was a minimum of 3 minutes. And / or (b) The ChanTest™ (Charles River Company) protocol of “Effect on Cloned hERG Potassium Channels Expressed in Mammalian Cells” described in the examples of WO2014 / 074775 was modified as follows: cells stably expressing hERG were maintained at −80 mV. The inhibition of onset and steady-state hERG potassium currents by compounds was measured using a pulse pattern with a constant amplitude (preconditioning pulse: +20 mV for 1 s; repolarization test rampoto at −90 mV (−0.5 V / s) repeated at 5 s intervals). Each recording was terminated by a final application of a supramaximal concentration of the reference substance E-4021 (500 nM) (Charles River Company). Residual non-binding currents were digitally subtracted from the data offline, and the potency of the test substance for hERG inhibition was calculated.

[0221] CYP (Cytochrome P450) Inhibition Assay. CYP inhibition is not a desirable feature for inhaled JAK1 or JAK1 / JAK2 inhibitors. For example, a reversible or time-dependent CYP inhibitor may cause an unwanted increase in its own plasma level or that of other co-administered drugs (drug-drug interactions). Furthermore, time-dependent CYP inhibition sometimes causes biotransformation of the parent drug into reactive metabolites. Such reactive metabolites may covalently modify proteins and potentially result in toxicity. Therefore, it may be to the benefit of inhaled JAK1 inhibitors to minimize reversible and time-dependent CYP inhibition. Thus, in some embodiments, the compounds of the present invention (or pharma- ceutically acceptable salts thereof) have minimal or no reversible and / or time-dependent CYP inhibition. Methods for measuring such CYP inhibition are known in the art. Pooled (n=150) human liver microsomes (Corning, Tewksbury, MA) were used to assess CYP inhibition of the compounds described herein using a previously reported method (Halladay et al., Drug Metab. Lett. 2011, 5, 220-230) over a range of compound concentrations from 0.16 to 10 μM. Incubation times and protein concentrations were determined depending on the CYP isotype and probe substrate / metabolite being assessed. For each CYP, the following substrates / metabolites, as well as incubation times and protein concentrations, were used: CYP1A2, phenacetin / acetaminophen, 30 min, 0.03 mg / ml protein; CYP2C9, warfarin / 7-hydroxywarfarin, 30 min, 0.2 mg / ml protein; CYP2C19, mephenytoin / 4-hydroxymephenytoin, 40 min, 0.2 mg / ml protein; CYP2D6, dextromethorphan / dextrorphan, 10 min, 0.03 mg / ml protein; CYP3A4, midazolam / 1-hydroxymidazolam, 10 min, 0.03 mg / ml protein and CYP3A4 testosterone / 6-hydroxytestosterone, 10 min, 0.06 mg / ml protein.These conditions were pre-determined based on linear rates of formation of CYP-specific metabolites. All reactions were initiated with 1 mM NADPH and terminated by the addition of 0.1% formic acid in acetonitrile containing appropriate stably labeled internal standards. Samples were analyzed by LC-MS / MS.

[0222] Binding to Mouse Lung Tissue. A high rate or percentage of binding of a JAK1 / JAK2 inhibitor to lung tissue is undesirable because it reduces the amount of free drug available to inhibit JAK1 or JAK2.

[0223] (a) Tissue binding experiments were performed in triplicate (n=3) using the Single-Use RED Plate according to standard protocols. First, individual drugs were spiked into tissue homogenate (pH 7.4) to a final concentration of 1 μM, and then 300 μL of the drug-tissue homogenate mixture was transferred to the donor well of the RED plate, where 500 μL of phosphate buffered saline (133 mM) was preloaded in the receiver well. The RED plate was sealed with a gas-permeable membrane and placed in a shaking incubator (450 rpm, VWR Symphony™) at 37°C and 5% CO2 for 6 hours. After the incubation was completed, 30 μL sample aliquots were taken from the RED device, and the matrix was equilibrated with an equal volume of tissue homogenate or buffer, and the obtained samples were immediately quenched with ice-cold acetonitrile containing propranolol or labetalol as internal standards (sample:acetonitrile=1:4). 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, after which the supernatant was collected and then diluted with an equal volume of water prior to LC-MS / MS analysis.

[0224] (b) As an alternative procedure, the extent of binding of test compounds to lung tissue of mouse lung homogenates may be measured using equilibrium dialysis in a Pierce RED (rapid equilibrium dialysis) apparatus (Fisher Scientific 89811 & 89809). A 10 mM solution of compound in DMSO was prepared and diluted to 1 mM with DMSO. An aliquot of 1 mM (4 μL) of this was added to lung homogenate (lung tissue:potassium phosphate buffer (0.05 M, pH 7.4) = 1:9 dilution ratio) with solvent to give a final incubation concentration of 5 μM compound, occupying 0.5% (v / v) of the final incubation volume.

[0225] Percentage of bound lung tissue was measured in triplicate for each assay. In triplicate, lung homogenate (200 μL) was loaded into the insert side of the RED device and 350 μL of potassium phosphate buffer was loaded into the opposite side. The RED device was sealed and incubated for 4 hours at approximately 37° C. on an orbital shaker (approximately 150 rpm).

[0226] After incubation, aliquots of lung homogenate (8 μL) and dialysate (72 μL) were matrix matched prior to analysis (72 μL phosphate buffer for lung homogenate and 8 μL lung homogenate for dialysate). 160 μL of acetonitrile containing internal standards was added to precipitate proteins from the samples. For mass balance assessment, the same matrix matching and protein precipitation procedures were performed on a lung homogenate aliquot taken at the beginning of the experiment (t=0 min sample). Quenched samples were centrifuged (4000 rpm, 30 min, 4°C), the resulting supernatant was diluted with water (3:1 (v / v), supernatant:water), and samples were analyzed for parent compound by liquid chromatography-mass spectrometry assay.

[0227] The unbound fraction (fu) of the lung homogenate was determined from the ratio of the dialysate and homogenate peak areas and corrected to account for the lung homogenate diluent (D) using the following formula, allowing estimation of total lung tissue binding: Unbonded fu=(1 / D) / [((1 / apparent fu)-1)+(1 / D)] Corrected binding ratio (%) = (1 - unbound fu) x 100

[0228] Dynamic solubility. Good water solubility of JAK1 / JAK2 inhibitors for inhalation delivery is considered desirable to reduce the content of undissolved powder in the lungs. In one procedure for measuring dynamic solubility, 4 μL of a 10 mM stock solution of the test compound in DMSO is added to 196 μL of pH 7.4 phosphate buffered saline solution in a Millipore Multiscreen® 96-well filter plate, resulting in a test concentration of 200 μM and a residual DMSO concentration of 2%. The filter plate is sealed with aluminum sealing film, shaken at room temperature for 24 hours, and the mixture is vacuum filtered and added to the clear 96-well plate. The filtered sample is diluted 2-fold with pH 7.4 phosphate buffered saline solution, and then the resulting 5 μL solution is analyzed by ultra-high performance liquid chromatography (UHPLC) using chemiluminescence nitrogen detection (CLND) and ultraviolet (UV) detection at a wavelength of 254 nm. Sample concentrations are typically quantified by CLND intensity, which is related to the nitrogen number of the compound. UV detection is primarily used to confirm sample purity, except in rare cases where the test compound does not contain nitrogen. In those cases, a compound-specific calibration curve is compiled based on UV absorption. This curve is then used to determine sample concentration.

[0229] Lipophilicity. Lipophilicity is generally important for the solubility, absorption, tissue penetration, protein binding, distribution, and ADME and PK properties of a candidate drug. Calculated logP (cLogP), the logarithm of the partition coefficient of a compound between n-octanol and water (i.e., log(concentration of compound in n-octanol / concentration of compound in water), can be an important consideration for JAK1 / JAK inhibitors for inhalation delivery.

[0230] Liver microsomal stability. To minimize systemic exposure of inhaled JAK1 / JAK2 inhibitors, it may be beneficial to optimize the rapid metabolism in the liver. Liver microsomal stability assays were performed using a BioCel 1200 liquid handling workstation (Agilent Technologies, Santa Clara, CA). Compounds (1.0 μM) were incubated at 37°C for 5 min in 100 μL of reaction mixture containing 100 mM phosphate buffer (pH 7.4) and 0.5 mg / mL liver microsomes and 1 mM NADPH. At different time intervals (0, 20, 40, and 60 min), aliquots of 20 μL of the reaction mixture were taken and mixed with 4 volumes of acetonitrile (ACN) containing 0.1 μM propranolol as an internal standard to stop the metabolic reaction. Samples were then centrifuged at 3250×g for 40 min to remove precipitated proteins. The supernatant was then transferred to a new 96-well plate, diluted 2-fold with deionized water, and then subjected to LC-MS / MS analysis using an ABI Sciex 5500 QTRAP® mass spectrometer (Applied Biosystems, Foster City, CA) coupled to an Agilent 1260 HPLC (Agilent Technologies, Santa Clara, CA). The percentage of residue was calculated using the peak area ratio of the test compound and the internal standard at different time points compared to the control (T=0 min). See B. Williamson, C. Wilson, G. Dagnell, RJ Riley. Harmonised high throughput microsomal stability assay. J. Pharmacol. Toxicol. Methods. 2017; 84 :31-36.

[0231] Solid state physical properties. For compounds to be delivered via inhalation of dry powder, there is also a requirement to be able to obtain a crystalline form of the compound that can be micronized to a size of 1-5 μm. Particle size is an important factor that determines the accumulation in the lungs of an inhaled compound. Typically, particles with a diameter of less than 5 microns (μm) are defined as respirable. Particles with a diameter greater than 5 μm are considered to accumulate in the oropharynx and therefore are not considered to accumulate in the lungs. Furthermore, particles with a diameter less than 1 μm are considered to be more airborne than larger particles and therefore are considered to be exhaled from the lungs. Therefore, a particle size of 1-5 μm is advantageous for inhaled drugs with the lung as the site of action. 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 particle size measurements below which 10%, 50% or 90% of the sample is respectively. For example, a D50 of 3 μm indicates that 50% of the sample is below 3 μm in size. Mass Mean Aerodynamic Diameter (MMAD). MMAD is the diameter above which 50% of the particles by mass are larger and 50% are smaller. MMAD is a measure of central tendency. Geometric Standard Deviation (GSD). GSD is a measure of the degree of dispersion from the MMAD, or the spread of the aerodynamic particle size distribution.

[0232] A common dosage form for inhaled drugs is a dry powder preparation containing the active pharmaceutical ingredient (API) mixed with a carrier (e.g., lactose, and possibly further excipients, e.g., magnesium stearate). For this and other dosage forms, it may be beneficial for the API itself to have properties that allow it to be machined to a particle size of 1-5 μm suitable for inhalation. Particle agglomeration should be avoided, which can be measured by methods known in the art (e.g., testing D90 values ​​at different pressure conditions). Thus, in some embodiments, the compounds of the invention (or pharma- ceutically acceptable salts thereof) can be prepared in such particle sizes suitable for inhalation with little or no agglomeration.

[0233] For the formulation of some inhaled drugs, it is important to use a specific crystalline form of the API with respect to crystallinity, including lactose blends. Crystallinity and crystalline form can affect many parameters important to inhaled drugs, including but not limited to: Chemical and aerodynamic stability over time, compatibility with inhaled formulation ingredients (e.g. lactose), hygroscopicity, lung retention and lung discomfort. Thus, a stable, reproducible crystalline form may be beneficial for an inhaled drug. Furthermore, the techniques used to machine compounds to the desired particle size are often energy-intensive, converting low-melting crystalline forms to other crystalline forms or rendering them completely or partially amorphous. Crystal forms with melting points below 150°C are considered less compatible with milling, while crystal forms with melting points below 100°C are considered incompatible with milling. Thus, it may be beneficial for an inhalant to have a melting point at least above 100°C, and ideally above 150°C. Thus, in some embodiments, the compounds described herein (or pharma- ceutically acceptable salts thereof) exhibit such properties.

[0234] Furthermore, minimizing molecular weight can help to reduce the effective dose of inhaled JAK1 inhibitor. A low molecular weight will increase the corresponding number of molecules per unit mass of active pharmaceutical ingredient (API). Therefore, it may be beneficial to find the smallest molecular weight inhaled JAK1 inhibitor that retains all other desirable properties of the inhaled drug.

[0235] Finally, the compound must maintain sufficient concentration in the lung to be able to exert a pharmacological effect on the pharmaceutical target for a desired time, and must have low systemic exposure when systemic inhibition of the target is not required.Since the lung has a high inherent permeability to both large molecules (proteins, peptides) and small molecules, as well as a short pulmonary half-life, it may be necessary to reduce the pulmonary absorption rate by modifying one or more properties of the compound (minimizing membrane permeability, optimizing pKa, cLogP, solubility, dissolution rate).Methods for measuring such properties are known in the art.

[0236] Thus, in some embodiments, the compounds of the present invention (or pharma- ceutically acceptable salts thereof) advantageously exhibit one or more of the above-mentioned characteristics. Moreover, in some embodiments, the compounds of the present invention advantageously exhibit one or more of these characteristics compared to known compounds, which may be particularly true for compounds intended for oral, rather than inhaled, drug administration. For example, compounds that have rapid oral absorption are typically not well-maintained when inhaled by lungs.

[0237] Janus kinase inhibitor treatment and methods of use The compound of the present invention or a pharma- ceutically acceptable salt thereof inhibits the activity of Janus kinase (e.g., JAK1 and / or JAK2 kinase). For example, the compound or a pharma- ceutically acceptable salt thereof inhibits phosphorylation of signal transducer and activator of transcription (STAT) by JAK1 kinase and inhibits STAT-mediated cytokine production. The compound of the present invention is useful for inhibiting JAK1 kinase activity of a cell through a cytokine pathway (e.g., IL-6, IL-15, IL-7, IL-2, IL-4, IL-9, IL-10, IL-13, IL-21, G-CSF, IFNα, IFNβ or IFNγ pathway). Thus, in one embodiment, a method is provided for inhibiting Janus kinase activity (e.g., JAK1 activity) of a cell by contacting a compound of the present invention or a pharma- ceutically acceptable salt thereof with the cell.

[0238] The compounds can be used for the treatment of immunological enhancement disorders caused by aberrant 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.

[0239] Thus, one embodiment includes a compound of the invention, or a pharma- ceutically acceptable salt thereof, for use in therapy.

[0240] In some embodiments, the use of the compound of the present invention or its pharmaceutically acceptable salt in the treatment of inflammatory disease is provided.Furthermore, the use of the compound of the present invention or its pharmaceutically acceptable salt for the preparation of a medicament for the treatment of inflammatory disease (e.g., asthma) is provided.The use of the compound of the present invention or its pharmaceutically acceptable salt for the treatment of inflammatory disease (e.g., asthma) is also provided.

[0241] Other embodiments include a method of preventing, treating, or reducing the severity of a disease or condition (e.g., asthma) that responds to inhibition of Janus kinase activity (e.g., JAK1 kinase activity) in a patient. The method can include administering to the patient a therapeutically effective amount of a compound of the invention, or a pharma- ceutically acceptable salt thereof. In one embodiment, the disease or condition that responds to inhibition of Janus kinase (e.g., JAK1 kinase) is asthma.

[0242] In one embodiment, the disease or condition is cancer, stroke, diabetes, hepatomegaly, cardiovascular disease, multiple sclerosis, Alzheimer's disease, cystic fibrosis, a viral disease, an autoimmune disease, atherosclerosis, restenosis, psoriasis, rheumatoid arthritis, inflammatory bowel disease, asthma, an allergic disorder, inflammation, a neurological disorder, a hormone-related disease, a condition associated with organ transplantation (e.g., graft rejection), an immunodeficiency disorder, a destructive bone disorder, a proliferative disorder, an infectious disease, a condition associated with cell death, thrombin-induced platelet aggregation, a liver disease, a pathological immune condition involving T-cell activation, a central nervous system disorder, or a myeloproliferative disease.

[0243] In one embodiment, the inflammatory disease is rheumatoid arthritis, psoriasis, asthma, inflammatory bowel disease, contact dermatitis or delayed hypersensitivity reaction, hi one embodiment, the autoimmune disease is rheumatoid arthritis, lupus or multiple sclerosis.

[0244] In another embodiment, the compounds of the present invention or pharmaceutically acceptable salts thereof may be used to treat pulmonary diseases, such as fibrotic lung diseases or interstitial aquatic lung diseases (e.g., interstitial aquatic pneumonia). In some embodiments, the compounds of the present invention or pharmaceutically acceptable salts thereof may be used to treat idiopathic pulmonary fibrosis (IPF), systemic sclerosis interstitial lung disease (SSc-ILD), non-specific interstitial aquatic pneumonia (NSIP), rheumatoid arthritis-associated interstitial aquatic lung disease (RA-ILD), sarcoidosis, hypersensitivity pneumonitis, or secondary ILDs following connective tissue diseases beyond scleroderma, such as polymyositis, dermatomyositis, rheumatoid arthritis, systemic lupus erythematosus (SLE), or mixed connective tissue disease.

[0245] In one embodiment, the cancer includes cancers of the breast, ovary, cervix, prostate, testis, penis, genitourinary tract, seminoma, esophagus, larynx, stomach, gastrointestinal system, skin, keratoacanthoma, follicular carcinoma, melanoma, lung, small cell lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, squamous cell carcinoma of the lung, colon, pancreas, thyroid, nipple, bladder, liver, biliary duct, kidney, bone, myeloid disorders, lymphatic disorders, hairy cell, oral cavity and pharynx (oral), lips, tongue, mouth, salivary glands, pharynx, small intestine, colon, rectum, anus, kidney, prostate, vulva, thyroid, large intestine, uterus, endometrium, brain, central nervous system, peritoneal cancer, hepatocellular carcinoma, head cancer, neck cancer, Hodgkin's or leukemia, etc.

[0246] In one embodiment, the disease is a myeloproliferative disorder. In one embodiment, the myeloproliferative disorder is polycythemia vera, essential thrombocytosis, myelofibrosis, or chronic myelogenous leukemia (CML).

[0247] Another embodiment includes the use of a compound of the invention or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a disease described herein, such as an inflammatory disease, an immunologically enhanced disorder, or cancer. In one embodiment, the invention provides a method of treating a disease or condition described herein, such as an inflammatory disease, an immunologically enhanced disorder, or cancer, by target-specific inhibition of a JAK kinase, such as JAK1.

[0248] Combination therapy The compounds can be used alone or in combination with other agents for treatment. The second or additional (e.g., third) compounds in a pharmaceutical composition or administration regimen typically exhibit complementary activity to the compounds of the present invention so that they do not adversely affect each other. Such agents are optimally present in a combination of amounts that are effective for the intended purpose. The compounds may be administered together in a pharmaceutical composition in unit form, or may be administered separately, and when administered separately, they may be administered simultaneously or sequentially. Such sequential administration may be close or distant in time.

[0249] For example, other compounds may be combined with a compound of the present invention, or a pharma- ceutically acceptable salt thereof, for the prevention or treatment of inflammatory diseases, such as asthma. Suitable therapeutic agents for combination therapy include, but are not limited to, adenosine A2A receptor antagonists, anti-infectives, nonsteroidal glucocorticoid receptor (GR receptor) agonists, antioxidants, alpha 2 adrenoreceptor 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), cell 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., funebrutinib), spleen tyrosine kinase (SYK) inhibitors, tryptase-β antibodies, ST2 receptor antibodies (e.g., AMG 282), cyclooxygenase inhibitors (COX inhibitors), lipoxygenase inhibitors, leukotriene receptor antagonists, alpha2 adrenoreceptor agonists / M3 receptor antagonist dual agents (MABA compounds), MEK-1 inhibitors, myeloperoxidase inhibitors (MPO inhibitors), muscarinic antagonists, p38 MAPK inhibitors, phosphodiesterase PDE4 inhibitors, phosphatidylinositol 3-kinase inhibitors (PI3-kinase inhibitors), phosphatidylinositol 3-kinase beta inhibitors (PI3-kinase gamma inhibitors), peroxisome proliferator-activated receptor agonists (PPAR gamma agonists), protease inhibitors, retinoic acid receptor modulators (RAR gamma modulators), statins, thromboxane antagonists, TLR7 receptor agonists, or vasodilators.

[0250] In addition, the compounds of the invention, or pharma- ceutically acceptable salts thereof, may be combined with: (1) Corticosteroids: for example, alclomethasone dipropionate, amelomethasone, beclomethasone dipropionate, budesonide, butixocort propionate, bicresonide, clobetasol propionate, desisobutylciclesonide, dexamethasone, etiprednol dicloacetate, fluocinolone acetonide, fluticasone furoate, fluticasone propionate, loteprednol atabate (topical) or mometasone furoate, (2) β2-adrenergic receptor agonists: for example, salbutamol, albuterol, terbutaline, fenoterol, bitolterol, carbuterol, clenbuterol, pirbuterol, rimoterol, terbutaline, tretoquinol, tulobuterol and long-acting β2-adrenergic receptor agonists (for example, metaproterenol, isoproterenol, isoprenaline, salmeterol, indacaterol, formoterol (including formoterol fumarate), arformoterol, carmoterol, abesiterol, vilanterol triphenate or olodaterol); (3) Corticosteroid / long-acting beta-2 agonist combination products: for example, salmeterol / fluticasone propionate (Advair® (also sold as Seretide®)), formoterol / budesonide (Symbicort®), formoterol / fluticasone propionate (Flutiform®), formoterol / ciclesonide, formoterol / mometasone furoate, indacaterol / mometasone furoate, vilanterol triphenate / fluticasone furoate (BREO ELLIPTA) or arformoterol / ciclesonide, (4) Anticholinergics: for example, muscarinic-3 (M3) receptor antagonists (e.g. ipratropium bromide, thiotriptium bromide, acridinium bromide (LAS-34273), glycopyrronium bromide or umeclidinium bromide), (5) M3-anticholinergic / β2-adrenergic receptor agonist combination products: for example, vilanterol / umeclidinium (Anoro® Ellipta®), olodaterol / tiotropium bromide, glycopyrronium bromide / indacaterol (Ultibro® (also sold as Xoterna®)), fenoterol hydrobromide / ipratropium bromide (Berodual®), albuterol sulfate / ipratropium bromide (Combivent®), formoterol fumarate / glycopyrrolate or aclidinium bromide / formoterol, (6) M3-Anticholinergic / β2-Adrenergic Receptor Agonist Dual Agents: e.g., betafenterol succinate, AZD-2115 or LAS-190792; (7) Leukotriene modulators: for example, leukotriene antagonists (e.g., montelukast, zafirlast, or pranlukast), or leukotriene biosynthesis inhibitors (e.g., zileuton), or LTB4 antagonists (e.g., amelvant), or FLAP inhibitors (e.g., fibroflavon, GSK-2190915), (8) Phosphodiesterase-IV (PDE-IV) inhibitors (oral or inhaled): e.g., roflumilast, cilomilast, oglemilast, rolipram, tetomilast, AVE-8112, revamilast, CHF 6001, (9) Antihistamines: for example, selective histamine-1 (H1) receptor antagonists, fexofenadine, cytolysine, loratidine or astemizole or dual H1 / H3 receptor antagonists (e.g. GSK 835726 or GSK 1004723); (10) Antitussives: e.g., codeine or dextramorphan, (11) Mucolytics: e.g., N-acetylcysteine ​​or fodosine, (12) Expectorants / dynamic modulators of mucus: for example, ambroxol, hypertonic solutions (e.g., food brine or mannitol) or surfactants, (13) Peptide mucolytics: for example, recombinant human deoxyribonuclease I (dornase-α and rhDNase) or helicidin, (14) Antibiotics: for example, azithromycin, tobramycin or aztreonam, (15) Non-selective COX-1 / COX-2 inhibitors: e.g. ibuprofen or ketoprofen, (16) COX-2 inhibitors: for example, celecoxib and rofecoxib, (17) VLA-4 antagonists: for example, those described in WO97 / 03094 and WO97 / 02289, which are incorporated herein by reference; (18) TACE inhibitors and TNF-α inhibitors: for example, anti-TNF monoclonal antibodies (e.g., Remicade®) and CDP-870 and TNF receptor immunoglobulin molecules (e.g., Enbrel®); (19) Matrix metalloproteinase inhibitors: for example, MMP-12, (20) Human neutrophil elastase inhibitors: for example, BAY-85-8501, or those described in WO 2005 / 026124, WO 2003 / 053930 and WO2006 / 082412, each of which is incorporated herein by reference; (21) A2b antagonists: for example, those described in WO2002 / 42298, which is incorporated herein by reference; (22) Modulators of chemokine receptor function: for example, antagonists of CCR3 and CCR8; (23) Compounds that modulate the action of other prostanoid receptors: for example, thromboxane A2 antagonists, DP1 antagonists (e.g., laropiprant or asapiprant), CRTH2 antagonists (e.g., OC000459, fevipiprant, ADC 3680 or ARRY 502), (24) PPAR agonists: for example, PPARα agonists (e.g., fenofibrate), PPARδ agonists, PPARγ agonists such as pioglitazone, rosiglitazone and balaglitazone, (25) Methylxanthines: for example, theophylline or aminophylline in combination with methylxanthines / corticosteroids (e.g., theophylline / budesonide, theophylline / fluticasone propionate, theophylline / ciclesonide, theophylline / mometasone furoate and theophylline / beclomethasone propionate); (26) A2a agonists: for example those described in EP1052264 and EP1241176, (27) CXCR2 or IL-8 antagonists: for example, AZD-5069, AZD-4721 or danilixin, (28) IL-R signaling modulators: e.g., Kineret and ACZ885; (29) MCP-1 antagonists: for example, ABN-912, (30) p38 MAPK inhibitors: for example, BCT197, JNJ49095397, losmapimod or PH-797804; (31) TLR7 receptor agonists: for example, AZD8848, (32) PI3-kinase inhibitors: for example, RV1729 or GSK2269557 (nemiralisib), (33) Triple combination products: for example, TRELEGY ELLIPTA (fluticasone furoate, umeclidinium bromide and vilanterol), or (34) Small molecule inhibitors of TRPA1, BTK, or SYK.

[0251] In some embodiments, the compounds of the invention, or pharma- ceutically acceptable salts thereof, can be used in combination with one or more additional drugs, such as antihyperproliferative, anticancer, cytostatic, cytotoxic, anti-inflammatory, or chemotherapeutic agents, including, for example, those described in U.S. Patent Application Publication No. 2010 / 0048557, which is incorporated herein by reference. The compounds of the invention, or pharma-ceutically acceptable salts thereof, can also be used in combination with radiation therapy or surgery, as is known in the art.

[0252] Combinations of any of the above with a compound of the invention, or a pharma- ceutically acceptable salt thereof, are specifically contemplated.

[0253] product Other embodiments include articles of manufacture (e.g., kits) for treating a disease or disorder responsive to inhibition of a Janus kinase (e.g., JAK1 kinase). The kit can include: (a) a first pharmaceutical composition comprising a compound of the invention or a pharma- ceutically acceptable salt thereof; and (b) Instructions for use.

[0254] In other embodiments, the kit further comprises: (c) a second pharmaceutical composition (eg, a pharmaceutical composition comprising a therapeutic agent as described above, such as an agent for the treatment of an inflammatory disease or a chemotherapeutic agent).

[0255] In one embodiment, the instructions describe the simultaneous, sequential or separate administration of said first and second pharmaceutical compositions to a patient in need thereof.

[0256] 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.

[0257] Containers for use include, for example, bottles, vials, syringes, blister packs, and the like. The containers can be made of a variety of materials, such as glass or plastic. The containers contain a compound of the present invention or a pharma- ceutically acceptable salt thereof that is effective for treating a condition, and may have a sterile access port (e.g., the container may be an intravenous solution bag, or a vial with a stopper that can be pierced by a hypodermic needle). The label or package insert indicates that the compound is used to treat the selected condition (e.g., asthma or cancer). In one embodiment, the label or package insert indicates that the compound is used to treat a disorder. In addition, the label or package insert may indicate that the patient to be treated is a patient with a disorder characterized by hypersensitive or abnormal Janus kinase activity (e.g., hypersensitive or abnormal JAK1 activity). The label or package insert may indicate that the compound can be used to treat other disorders.

[0258] Alternatively, or in addition, the kit may further comprise a second (or third) container containing a pharma- ceutically acceptable buffer (e.g., bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, or dextrose solution). It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

[0259] In order to illustrate the present invention, examples are given below. However, it should be understood that these examples are not limiting to the present invention, but merely suggest a method of carrying out the present invention. Those skilled in the art will understand that the described chemical reactions can be easily adapted to prepare other compounds of the present invention, and alternative methods for preparing compounds are also within the scope of the present invention. For example, the synthesis of compounds not exemplified in the present invention can be successfully carried out by modifications obvious to those skilled in the art, such as by appropriately protecting interfering groups, by utilizing other suitable reagents known in the art other than those described, or by making routine changes to reaction conditions. Alternatively, other reactions disclosed herein or known in the prior art are also recognized to have applicability for preparing other compounds of the present invention. EXAMPLES

[0260] Representative compounds shown in Table 1 were prepared using procedures similar to those described in the schemes and examples herein. The absolute stereochemistry of each compound below is omitted: therefore, the structure may exist more than once, each representing a single stereoisomer. Table 1: Exemplary JAK inhibitors of the present invention TIFF0007657740000027.tif253170TIFF0007657740000028.tif244170TIFF000 7657740000029.tif243170TIFF0007657740000030.tif247170TIFF00076577400 00031.tif233170TIFF0007657740000032.tif246170TIFF0007657740000033.t if244170TIFF0007657740000034.tif249170TIFF0007657740000035.tif240170 TIFF0007657740000036.tif241170TIFF0007657740000037.tif244170TIFF000 7657740000038.tif227170TIFF0007657740000039.tif255170TIFF00076577400 00040.tif249170TIFF0007657740000041.tif218170TIFF0007657740000042.t if232170TIFF0007657740000043.tif242170TIFF0007657740000044.tif186170

[0261] LCMS conditions Method A The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Shim-Pack XR-ODS, 50 × 3 mm, 2.2 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000045.tif60170

[0262] Method B The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Shim-Pack XR-ODS, 50 × 3 mm, 2.2 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000046.tif68170

[0263] Method C The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Shim-Pack XR-ODS, 50 × 3 mm, 2.2 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000047.tif59170

[0264] Method D The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Shim-Pack XR-ODS, 50 × 3 mm, 2.2 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000048.tif69170

[0265] Method E The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Shim-Pack XR-ODS, 50 × 3 mm, 2.2 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000049.tif69170

[0266] Method F The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Shim-Pack XR-ODS, 50 × 3 mm, 2.2 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000050.tif70170

[0267] Method G The experiments were carried out using a SHIMADZU 20A HPLC with a C18-reversed phase column (50 x 2.1 mm Ascentis Express C18, 2.7 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid for elution). TIFF0007657740000051.tif60170

[0268] Method H The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Shim-Pack XR-ODS, 50 × 3 mm, 2.2 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000052.tif60170

[0269] Method I Poroshell HPH-C 18 The experiments were carried out using a SHIMADZU 20A HPLC with a column (50 x 3 mm, 2.7 μm particle size) (solvent A: water / 5 mM NH4HCO3; solvent B: acetonitrile for elution). TIFF0007657740000053.tif60170

[0270] Method J C18-reversed phase column (50×3 mm Kinetex XB-C 18 The experiments were carried out using a SHIMADZU LCMS-2020 with a column size of 2.6 μm (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid for elution). TIFF0007657740000054.tif59170

[0271] Method K The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Shim-Pack XR-ODS, 50 × 3 mm, 2.2 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000055.tif60170

[0272] Method L C18-reversed phase column (50×2.1 mm Kinetex XB-C 18 The experiments were carried out using a SHIMADZU LCMS-2020 with a column diameter of 100A, 2.6 μm (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid for elution). TIFF0007657740000056.tif60170

[0273] Method M The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (30×2.1 mm Kinetex C18-100A, 1.7 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid for elution). TIFF0007657740000057.tif61170

[0274] Method N The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (50×3.0 mm Poroshell HPH-C18, 2.7 μm particle size) (solvent A: water+5 mM ammonium bicarbonate; solvent B: acetonitrile was used for elution). TIFF0007657740000058.tif60170

[0275] Method O The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Titank C18, 50×3.0 mm, 3.0 μm particle size) (solvent A: water+5 mM ammonium bicarbonate; solvent B: acetonitrile was used for elution). TIFF0007657740000059.tif61170

[0276] Method P The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Halo C18, 30×2.1 mm, 2.0 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000060.tif60170

[0277] Method Q The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (50 × 3.0 mm YMC-Triart C18, 2.5 μm particle size) (solvent A: water + 0.1% formic acid; solvent B: acetonitrile + 0.1% formic acid was used for elution). TIFF0007657740000061.tif60170

[0278] Method R The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Shim-Pack XR-ODS, 50 × 3 mm, 2.2 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000062.tif59170

[0279] Method S The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (50×3.0 mm Poroshell HPH-C18, 2.7 μm particle size) (solvent A: water+5 mM ammonium bicarbonate; solvent B: acetonitrile was used for elution). TIFF0007657740000063.tif70170

[0280] Method T The experiments were carried out using a SHIMADZU 20A HPLC with a C18-reversed phase column (50 x 2.1 mm Ascentis Express C18, 2.7 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid for elution). TIFF0007657740000064.tif60170

[0281] Method U The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (Shim-Pack XR-ODS, 50 × 3 mm, 2.2 μm particle size) (solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid was used for elution). TIFF0007657740000065.tif71170

[0282] Method V The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (50×3.0 mm Poroshell HPH-C18, 2.7 μm particle size) (solvent A: water+5 mM ammonium bicarbonate; solvent B: acetonitrile was used for elution). TIFF0007657740000066.tif69170

[0283] Method W The experiments were carried out using a SHIMADZU LCMS-2020 with a C18-reversed phase column (50×2.1 mm Water Acquity BEH, 1.7 μm particle size) (solvent A: water + 0.1% formic acid; solvent B: acetonitrile + 0.1% formic acid was used for elution). TIFF0007657740000067.tif61170

[0284] Method X Experiments were performed using an Agilent 1290 UHPLC coupled to an Agilent MSD (6140) mass spectrometer using ESI as the ion source. LC separation was performed using a Phenomenex XB-C18, 1.7um, 50x2.1mm column at a flow rate of 0.4ml / 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 and ended at 98% B over 7 minutes, and was held at 98% B for 1.5 minutes after equilibration for 1.5 minutes. The LC column temperature was 40°C. UV absorption was collected at 220nm and 254nm and full scan of mass spectra was applied for all experiments.

[0285] List of Common Abbreviations ACN Acetonitrile Brine Saturated sodium chloride solution CH3OD Deuterated Methanol CDCl3 Deuterated Chloroform DCM Dichloromethane DIEA or DIPEA Diisopropylethylamine DMA Dimethylacetamide DMAP 4-Dimethylaminopyridine DMF Dimethylformamide DMSO Dimethyl sulfoxide DMSO-d6 Deuterated dimethyl sulfoxide DTAD Di-tert-butyl azodicarboxylate EDC or EDCI 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide ESI electrospray ionization EtOAc Ethyl acetate EtOH Ethanol FA Formic Acid HOAc Acetic acid g grams h time 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 Ethanol L Liter LCMS Liquid Chromatography Mass Spectrometry LiHMDS or LHMDS Lithium hexamethyldisilazide MDAP Mass Spectrometry Automated Purification MeCN Acetonitrile MeOH Methanol μm micrometer min mg milligram mL Milliliters mm millimeters M Molar concentration nm nanometer NMR Nuclear Magnetic Resonance Spectroscopy Pd2(dba)3.CHCl3 Tris(dibenzylideneacetone)dipalladium(0)-chloroform adduct PE Petroleum Ether Prep-HPLC Preparative High Performance Liquid Chromatography PyAOP (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate SCX-2 Strong cation exchange TBAF Tetra-n-butylammonium fluoride THF Tetrahydrofuran TFA Trifluoroacetic acid Xantphos 4,5-bis(diphenylphosphino)-9,9-dimethylxanthine ZnCl2 Zinc chloride

[0286] Intermediate 1 TIFF0007657740000068.tif62170N-(5-(5-bromo-2-(difluoromethoxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of 4-bromo-1-(difluoromethoxy)-2-iodobenzene TIFF0007657740000069.tif29170 To a solution of 4-bromo-2-iodophenol (282 g, 943 mmol) in N,N-dimethylformamide (2000 mL) and water (500 mL) was added sodium 2-chloro-2,2-difluoroacetate (216 g, 1.42 mol) and Cs2CO3 (617 g, 1.89 mol). The reactor 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 (3x1500 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 H 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).

[0287] Step 2: Synthesis of 5-[5-bromo-2-(difluoromethoxy)phenyl]-4-nitro-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazole TIFF0007657740000070.tif49170To a solution of 4-nitro-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazole (100 g, 411 mmol) in anhydrous THF (1000 mL) was added dropwise with stirring at -70 °C under nitrogen to a solution of LiHMDS (490 mL, 1.0 mol / L in THF). The resulting solution was stirred at -50 °C for 1 h and then cooled to -70 °C. ZnCl2 (500 mL, 0.7 mol / L in THF) was added dropwise at -70 °C and the resulting solution was allowed to warm to room temperature and stirred at room temperature for 1 h. To the mixture was added 4-bromo-1-(difluoromethoxy)-2-iodobenzene (150 g, 860 mmol) (Pd(PPh3)4 (24.0 g, 20.8 mmol)). The resulting solution was heated at reflux overnight, cooled to room temperature, and concentrated under reduced pressure. This reaction was repeated once more on this scale, 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). The appropriate fractions were combined and concentrated under reduced pressure. This resulted in 300 g (79%) of 5-[5-bromo-2-(difluoromethoxy)phenyl]-4-nitro-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazole, all as a 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).

[0288] Step 3: Synthesis of 5-(5-bromo-2-(difluoromethoxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-amine TIFF0007657740000071.tif44170To a solution of 5-(5-bromo-2-(difluoromethoxy)phenyl)-4-nitro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (50.1 g, 108 mmol) in ethanol (2000 mL) and water (200 mL), iron powder (60.1 g, 1.07 mol) and NH4Cl (28.0 g, 0.523 mol) were added and the reaction mixture was stirred at reflux temperature under nitrogen for 3 h. The solids were filtered off 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 1x500 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 taken to the next step without further purification. LC / MS (Method G, ESI): [M+H] + = 434.2, R T = 0.93 minutes.

[0289] 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 TIFF0007657740000072.tif60170To a solution of 5-(5-bromo-2-(difluoromethoxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-amine (50.1 g, 115 mmol) in DMA (1500 mL) was 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 (2x). The organic layers were combined, 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). 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 and 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 & 581.1, R T = 1.10 minutes. 1H 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).

[0290] Intermediate 2 TIFF0007657740000073.tif51170N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide 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, 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. This gave 3.80 g of N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. The purity of the intermediate was sufficient to be used in the next step without further purification. LC / MS (Method I, ESI): [M+H] + = 449.0, R T = 1.02 minutes. 1H 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).

[0291] Intermediate 3 TIFF0007657740000074.tif51170N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide To a solution of N-[5-[5-bromo-2-(difluoromethoxy)phenyl]-l-[[2-(trimethylsilyl)ethoxy]methyl]-lH-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 1, 10.1 g, 17.3 mmol) in dichloromethane (200 mL) was added Me3OBF4 (2.81 g, 18.9 mmol) at room temperature. The resulting solution was stirred at room temperature for 2 h. Then, 10 mL of EtOH was added to the reaction mixture and the reaction mixture was stirred for 1 h. To this solution, 5.0 mL of HCl (conc.) was added and stirred for 1 h. The resulting mixture was concentrated under vacuum. The pH value of the solution was adjusted to 8 with sodium bicarbonate (20%). The resulting solution was extracted with 3x300 mL of ethyl acetate and the organic layers were combined, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was loaded onto a silica gel column and eluted 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. 1H 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).

[0292] Intermediate 4 TIFF0007657740000075.tif51170N-(3-(2-(difluoromethoxy)-5-(methylthio)phenyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of 5-[2-(difluoromethoxy)-5-(methylsulfanyl)phenyl]-4-nitro-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazole. TIFF0007657740000076.tif50170Toluene (500 mL), 5-[5-bromo-2-(difluoromethoxy)phenyl]-4-nitro-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazole (60 g, 129 mmol), NaSMe (26 g, 371 mmol), Pd2(dba)3.CHCl3 (6.7 g, 6.47 mmol), and XantPhos (7.5 g, 12.96 mmol) were placed in a 1000 mL round bottom flask purged with nitrogen and maintained at an inert atmosphere. The resulting mixture was stirred at 85° C. overnight. The resulting mixture was concentrated under vacuum. This reaction was repeated three times. The residue was loaded onto a silica gel column and eluted with ethyl acetate / petroleum ether (1:20). The appropriate fractions were combined and concentrated under vacuum. This gave 171 g of 5-[2-(difluoromethoxy)-5-(methylsulfanyl)phenyl]-4-nitro-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazole as a yellow solid. LC / MS (Method F, ESI): [M+H]+ = 432.1, RT = 1.23 min; 1 H NMR (300 MHz, CDC13) δ: (ppm) 8.25 (s, 1H), 7.42 (dd, J = 8.7, 2.4 Hz, 1H), 7.34 (d, J = 2.1 Hz, 1H), 7.23 (d, J = 8.7 Hz, 1H), 6.39 (t, J = 72.9 Hz, 1H), 5.36 - 5.22 (m, 2H), 3.74 - 3.55 (m, 2H), 2.51 (s, 3H), 0.94 - 0.90 (m, 2H), 0.02 (s, 9H).

[0293] Step 2: Synthesis of N-[5-[2-(difluoromethoxy)-5-(methylsulfanyl)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000077.tif451705-[2-(difluoromethoxy)-5-(methylsulfanyl)phenyl]-4-nitro-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazole (171 g, 408 mmol), ethanol (2000 mL), and water (200 mL) were added iron powder (228 g, 4.08 mol) (NH4Cl (120 g, 2.24 mol)). The reaction mixture was stirred at reflux under nitrogen for 3 h and cooled to room temperature. The solid was filtered off. The filtrate was concentrated under vacuum. The residue was dissolved in 3000 mL of ethyl acetate and washed with 1x500 mL of brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under vacuum. This gave 148 g of 5-[2-(difluoromethoxy)-5-(methylsulfanyl)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazol-5-amine as a yellow oil. LC / MS (Method F, ESI): [M+H]+ = 402.1, R T = 0.93 min. A 3000 mL 3-neck round bottom flask was placed with DMA (1500 mL), 5-[2-(difluoromethoxy)-5-(methylsulfanyl)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazol-4-amine (148 g), pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (102 g), HATU (325 g), 4-dimethylaminopyridine (4.5 g), and DIPEA (142 g). The resulting solution was stirred at 60° C. for 3 h, poured into ice water (2000 mL), extracted with 3×2000 mL of ethyl acetate, and the organic layers were combined. The resulting mixture was washed with 1×1000 mL of brine. The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was loaded onto a silica gel column and eluted with ethyl acetate / petroleum ether (4:1) to give 200 g of N-[5-[2-(difluoromethoxy)-5-(methylsulfanyl)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 A, ESI): [M+H]+ = 547.2, RT = 1.10 min;1 H NMR (300 MHz, CDC13) δ: (ppm) 9.63 (s, 1H), 8.77 (dd, J = 7.0, 1.7 Hz, 1H), 8.73 (s, 1H), 8.51 (dd, J = 4.2, 1.8 Hz, 1H), 8.38 (s, 1H), 7.50 (d, J = 2.4 Hz, 1H), 7.39 (dd, J = 8.7, 2.4 Hz, 1H), 7.30 (d, J = 8.7 Hz, 1H), 6.98 (dd, J = 6.9, 4.2 Hz, 1H), 6.39 (t, J = 73.2 Hz, 1H), 5.46 - 5.38 (m, 2H), 3.70 - 3.59 (m, 2H), 2.52 (s, 3H), 0.92-0.85 (m, 2H), 0.03 (s, 9H).

[0294] Step 3: Synthesis of N-(3-(2-(difluoromethoxy)-5-(methylthio)phenyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000078.tif51170 To a solution of N-[5-[2-(difluoromethoxy)-5-(methylsulfanyl)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (60 g) in methanol (600 mL) was added concentrated HCl solution (300 mL). The resulting solution was stirred at 35° C. overnight. The resulting mixture was concentrated under vacuum. The solid was collected by filtration. The solid was suspended in 200 mL of water. The pH value of the solution was adjusted to 8 with saturated sodium bicarbonate. The product was collected by filtration and dried to give 30 g (66%) of N-(3-(2-(difluoromethoxy)-5-(methylthio)phenyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide as a pale yellow solid. LC / MS (Method G, ESI): [M+H]+ = 417.0, R T = 0.80 minutes; 1H NMR (300 MHz, DMSO-d6) δ: (ppm) 13.02 (s, 1H), 9.71 (s, 1H), 9.33 (dd, J = 6.9, 1.5 Hz, 1H), 8.68 (dd, J = 4.1, 1.4 Hz, 1H), 8.66 (s, 1H), 8.24 (s, 1H), 7.47 - 7.36 (m, 3H), 7.27 (dd, J = 6.9, 4.2 Hz, 1H), 7.17 (t, J = 73.8 Hz, 1H), 2.51 (s, 3H).

[0295] Intermediate 5 TIFF0007657740000079.tif61170N-(5-(2-(difluoromethoxy)-5-((triisopropylsilyl)thio)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide 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, 4.00 g, 6.90 mmol) in toluene (20 mL), sodium hydride (415 mg, 10.4 mmol, 60% in mineral oil), Pd2(DBA)3.CHCl3 (735 mg, 0.710 mmol) and XantPhos (800 mg, 1.38 mmol) and tris(propan-2-yl)silanethiol (1.97 g, 10.4 mmol) were added under nitrogen. The resulting solution was stirred at 90° C. for 20 min. The resulting mixture was concentrated under vacuum. The residue was purified by flash chromatography on silica gel, eluting with ethyl acetate / petroleum ether (80 / 20). Appropriate fractions were combined and concentrated in vacuo. This gave 3.30 g (69%) of N-(5-(2-(difluoromethoxy)-5-((triisopropylsilyl)thio)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LC / MS (Method B, ESI): [M+H]+ = 689.4, R T = 1.51 minutes.

[0296] Intermediate 6 TIFF0007657740000080.tif49170N-[3-[2-(difluoromethoxy)-5-[[tris(propan-2-yl)silyl]sulfanyl]phenyl]-1-methyl-lH-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide To a suspension of sodium hydride (260 mg, 10.8 mmol) in toluene (40 mL) was added dropwise tris(propan-2-yl)silanethiol (1.23 g, 6.47 mmol) at room temperature under nitrogen. The mixture was stirred at this temperature for 1 h until it turned clear, and N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 3, 2.50 g, 5.40 mmol), Pd2(DBA)3.CHCl3 (184 mg, 0.178 mmol) and XantPhos (250 mg, 0.432 mmol) were added at room temperature under nitrogen atmosphere. The resulting solution was stirred at 90 °C under N2 for 30 min. The reaction was then quenched by the addition of 50 mL of aqueous NH4Cl. The resulting solution was diluted with 100 mL of EA. The organic layer was separated and washed with 3x50 mL of water and 2x50 mL of brine. The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was loaded onto a silica gel column and eluted with ethyl acetate / petroleum ether (1 / 1) to give 2.35 g (76%) of N-[3-[2-(difluoromethoxy)-5-[[tris(propan-2-yl)silyl]sulfanyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. TLC: PE / EA = 1 / 1, Rf = 0.4.

[0297] Intermediate 7 TIFF0007657740000081.tif511704-(Difluoromethoxy)-3-(1-methyl-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-3-yl)benzene-1-sulfonic acid A 100 mL round bottom flask was placed with N-[3-[2-(difluoromethoxy)-5-[[tris(propan-2-yl)silyl]sulfanyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 6, 360 mg, 0.63 mmol, 1.00 equiv.), dichloromethane (30 mL), and m-CPBA (217 mg, 1.26 mmol, 2.0 equiv.). The resulting solution was stirred at room temperature for 5 h. The resulting mixture was concentrated under vacuum. The residue was loaded onto a silica gel column with dichloromethane / methanol (6:4). This gave 210 mg (72%) of 4-(difluoromethoxy)-3-(1-methyl-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-3-yl)benzene-1-sulfonic acid as a yellow solid. LC / MS (Method I, ESI): [M+H]+ = 465.1, R T = 0.684 minutes.

[0298] Intermediate 8 TIFF0007657740000082.tif52170N-(3-(2-(difluoromethoxy)-5-mercaptophenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide To a solution of N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (intermediate 3, 3000 mg, 6.48 mmol), tris(propan-2-yl)silanethiol (1.48 g, 1.67 mL, 7.77 mmol), XantPhos Pd G2 (575 mg, 0.648 mmol) and XantPhos (375 mg, 0.648 mmol) in toluene (100 mL) cooled to 0° C., NaH (518 mg, 13.0 mmol) was added. The reaction mixture was stirred at 0° C. for 30 min, at room temperature for 1 h, and then at 90° C. for 1 h. The reaction mixture was diluted with dichloromethane, filtered through Celite, eluted with dichloromethane, and the filtrate was concentrated in vacuo. The residue was adsorbed onto silica and purified by flash column chromatography with 10-80% 3:1 MeOHiPrOAc / heptane to give N-(3-(2-(difluoromethoxy)-5-mercaptophenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide as a brown solid (2.32 g, 86%). The product was contaminated with some dimer but was used without further purification. LC / MS (Method W, ESI): [M+H]+ = 417.1, R T = 1.09 minutes.

[0299] Intermediate 9 TIFF0007657740000083.tif50170N-(3-(2-(difluoromethoxy)-5-(methylsulfonyl)phenyl)-1H-pyrazol-4-yl)pyrazolo[1,5]pyrimidine-3-carboxamide To a solution of N-(3-(2-(difluoromethoxy)-5-(methylthio)phenyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 4, 500 mg, 1.20 mmol) in dichloromethane (5.0 mL) was added m-CPBA (623 mg, 3.61 mmol). The resulting solution was stirred at room temperature for 5 min and concentrated in vacuo. The residue was purified by flash chromatography on silica gel, eluting with dichloromethane / methanol (25 / 1). The appropriate fractions were collected and concentrated in vacuo to give 523 mg (97%) of N-[3-[2-(difluoromethoxy)-5-methanesulfonylphenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. LC / MS (Method G, ESI): [M+H]+= 449.2, R T = 0.99 minutes; 1 H NMR (300MHz, DMSO-d6): δ (ppm) 13.18 (s, 1H), 9.68 (s, 1H), 9.33 (dd, J= 6.9, 1.5 Hz, 1H), 8.70 (dd, J= 4.2, 1.5 Hz, 1H), 8.66 (s, 1H), 8.31 (s, 1H), 8.12 - 8.09 (m, 2H), 7.68 (d, J = 8.1 Hz, 1H), 7.47 (t, J = 72.6 Hz, 1H), 7.30 (dd, 7= 6.9, 4.2 Hz, 1H), 3.28 (s, 3H).

[0300] The following examples are numbered according to the examples in Table 1 above.

[0301] Example 28 TIFF0007657740000084.tif49170N-(1-(cyanomethyl)-3-(2-(difluoromethoxy)-5-(methylsulfonyl)phenyl)-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide In a 10 mL vial, N-[3-[2-(difluoromethoxy)-5-methanesulfonylphenyl]-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (Intermediate 9, 100 mg, 0.223 mmol), cesium carbonate (146 mg, 0.447 mmol, 2.00 equiv.), N,N-dimethylformamide (2 mL) and 2-bromoacetonitrile (39.8 mg, 0.332 mmol, 1.49 equiv.) were placed. The resulting solution was stirred at room temperature for 3 h. The reaction was then quenched by the addition of 3 mL of water. The resulting solution was extracted with 3x3 mL of ethyl acetate, and the organic layers were combined and concentrated under vacuum. The crude product (3 mL) was purified by Prep-HPLC (2#-Analytical HPLC-SHIMADZU (HPLC-10)) according to the following conditions: Column: Kinetex EVO C18 Column, 30×150, 5μm; Mobile phase: water (10 mmol / l NHHCO) and ACN (19.0% to 27.0% ACN, 7 min); detector: UV 254 / 220 nm, yielding 40.7 mg (37%) of N-[1-(cyanomethyl)-3-[2-(difluoromethoxy)-5-methanesulfonylphenyl]-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide as a yellow solid. 1 H NMR (300 MHz, DMSO-d6): δ (ppm) 9.76 (s, 1H), 9.36 (dd, J=6.9, 1.5 Hz, 1H), 8.69 - 8.67 (m, 2H), 8.54 (s, 1H), 8.16 (dd, J=8.7, 2.4 Hz, 1H), 8.08 (d, J=1.6 Hz, 1H), 7.74 - 7.26 (m, 3H), 5.64 (s, 2H), 3.32 (s, 3H). LC / MS (Method A, ESI): [M+H]+= 488.2, R T = 1.42 minutes.

[0302] Example 22 TIFF0007657740000085.tif53170N-(3-(5-(cyclopropylsulfonyl)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of N-[5-[5-(cyclopropylsulfanyl)-2-(difluoromethoxy)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide Into a 250 mL round bottom flask purged with nitrogen and maintained at an inert atmosphere was placed N-[5-[5-bromo-2-(difluoromethoxy)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (Intermediate 3, 4 g, 6.90 mmol), Pd2(DBA)3·CHCl3 (1.4 g, 1.35 mmol, 0.20 equiv), XantPhos (1.6 g, 2.77 mmol, 0.401 equiv), potassium carbonate (1.9 g, 13.7 mmol, 2.0 equiv), toluene (100 mL) and cyclopropanethiol (2 g, 27 mmol, 3.9 equiv). The resulting solution was stirred at 100° C. for 20 min. The resulting reaction mixture was cooled to room temperature and concentrated under vacuum. The residue was loaded onto a silica gel column with ethyl acetate / hexane (1 / 1, 4 / 1). The collected fractions were combined and concentrated under vacuum. This gave 3.5 g (89%) of N-[5-[5-(cyclopropylsulfanyl)-2-(difluoromethoxy)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide as a yellow oil. LC / MS (Method J, ESI): [M+H]+= 573.3, R T = 1.34 minutes.

[0303] Step 2: Synthesis of N-[3-[5-(cyclopropylsulfanyl)-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000087.tif52170A 100 mL round bottom flask was placed with N-[5-[5-(cyclopropylsulfanyl)-2-(difluoromethoxy)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (3.87 g, 6.76 mmol), dichloromethane (20 mL) and CF3COOH (10 mL). The resulting solution was stirred at room temperature for 4 h. The resulting mixture was concentrated under vacuum. The resulting solution was diluted with ethyl acetate. The solid was collected by filtration. This gave 3.01 g (crude) of N-[3-[5-(cyclopropylsulfanyl)-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a brown solid. LC / MS (Method H, ESI): [M+H]+= 443.15, R T = 1.24 minutes.

[0304] Step 3: Synthesis of N-[3-[5-(cyclopropylsulfanyl)-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000088.tif51170A 25 mL round bottom flask was placed with N-[3-[5-(cyclopropylsulfanyl)-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (250 mg, 0.565 mmol), N,N-dimethylformamide (4 mL), cesium carbonate (294 mg, 0.902 mmol, 2.00 equiv.), and iodomethane (77 mg, 0.542 mmol, 1.20 equiv.). The resulting solution was stirred at 30° C. for 2 h. The resulting mixture was concentrated under vacuum. The residue was loaded onto a silica gel column with ethyl acetate / petroleum ether (2:1). This gave 150 mg (58%) of N-[3-[5-(cyclopropylsulfanyl)-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. LC / MS (Method H, ESI): [M+H]+= 457.2, R T = 1.24 minutes.

[0305] Step 4: Synthesis of N-[3-[5-(cyclopropanesulfonyl)-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000089.tif52170A 50 mL round bottom flask was placed with N-[3-[5-(cyclopropylsulfanyl)-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (280 mg, 0.613 mmol), dichloromethane (20 mL), and m-CPBA (212 mg, 1.23 mmol, 2.00 equiv). The resulting solution was stirred at room temperature for 5 h. The resulting mixture was concentrated under vacuum. The residue was loaded onto a silica gel column with dichloromethane / methanol (95:5). The crude product was purified by Prep-HPLC (2#-Analytical HPLC-SHIMADZU (HPLC-10)) according to the following conditions: Column: Kinetex EVO C18 Column, 30×150, 5μm; Mobile phase: water (10 mmol / l NH4HCO3) and ACN (25.0%-33.0% ACN, 8 min);Detector: UV 254 / 220 nm. This gave 42.9 mg (14%) of N-[3-[5-(cyclopropanesulfonyl)-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide as a white solid. 1 H NMR (400 MHz, DMSO-d6): δ (ppm) 9.68 (s, 1H), 9.34 (dd, J=6.8, 1.4 Hz, 1H), 8.70 - 8.67 (m, 2H), 8.33 (s, 1H), 8.07 (dd, J=8.8, 2.4 Hz, 1H), 8.02 (d, J=2.4 Hz, 1H), 7.69 - 7.28 (m, 3H), 3.96 (s, 3H), 3.01 - 2.95 (m, 1H), 1.16 - 1.12 (m, 2H), 1.08 - 1.06 (m, 2H). LC / MS (Method A, ESI): [M+H]+= 489.2, R T = 1.47 minutes.

[0306] Example 52 TIFF0007657740000090.tif53170N-(3-(5-(cyclopropylsulfonyl)-2-(difluoromethoxy)phenyl)-1-(1-methylazetidin-3-yl)-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide Step 1: tert-Butyl 3-(3-(5-(cyclopropylsulfonyl)-2-(difluoromethoxy)phenyl)-4-(pyrazolo[l,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-1-yl)azetidine-1-carboxylate To a TIFF0007657740000091.tif53170 vial was added N-[3-[5-cyclopropylsulfonyl-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (51 mg, 0.11 mmol) and cesium carbonate (105 mg, 0.32 mmol), followed by N,N-dimethylformamide (1.1 mL) and tert-butyl-3-iodoazetidine-1-carboxylate (107 mg, 0.065 mL, 0.38 mmol) and the reaction mixture was heated to 60° C. for 16 h. The reaction was quenched by the addition of saturated aqueous ammonium chloride and extracted with EtOAc (3×). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was adsorbed onto silica and purified by flash column chromatography with 10-80% 3:1 MeOH:iPrOAc / heptane to give the desired compound as a yellow oil (35.6 mg, 53%).

[0307] Step 2: N-(1-(azetidin-3-yl)-3-(5-(cyclopropylsulfonyl)-2-(difluoromethoxy)phenyl)-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000092.tif54170 To a solution of tert-butyl 3-[3-[5-cyclopropylsulfonyl-2-(difluoromethoxy)phenyl]-4-(pyrazolo[l,5-a]pyrimidine-3-carbonylamino)pyrazol-1-yl]azetidine-1-carboxylate (35.6 mg, 0.057 mmol) in dichloromethane (1 mL) was added trifluoroacetic acid (0.25 mL) and the reaction was stirred for 2 h at room temperature. The reaction mixture was then concentrated in vacuo and extracted with 2 mL of dichloromethane. MP-carbonate (180 mg, 0.57 mmol) was added and the reaction mixture was stirred for 2 h. The reaction mixture was filtered and the filtrate was concentrated in vacuo to give the desired compound as a white solid. Conversion was deemed complete and so it was taken on crude to the next step. LC / MS (Method W, ESI): [M+H]+= 530.2, R T= 0.83 minutes.

[0308] Step 3: N-(3-(5-(cyclopropylsulfonyl)-2-(difluoromethoxy)phenyl)-1-(1-methylazetidin-3-yl)-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000093.tif54170To a solution of N-[1-(azetidin-3-yl)-3-[5-cyclopropylsulfonyl-2-(difluoromethoxy)phenyl]pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (29.9 mg, 0.057 mmol) in 1,2-dichloroethane (1.88 mL), formaldehyde (1.28 mol / L in water) (6.87 mg, 0.0063 mL, 0.085 mmol) and sodium cyanoborohydride (3.62 mg, 0.0031 mL, 0.058 mmol) were added and the reaction mixture was stirred at room temperature for 1 h. Another portion of formaldehyde (1.28 mol / L in water) (6.87 mg, 0.0063 mL, 0.085 mmol) and sodium cyanoborohydride (3.62 mg, 0.0031 mL, 0.058 mmol) were added and the reaction mixture was stirred overnight. The mixture was diluted with dichloromethane and poured into a separatory funnel containing saturated aqueous sodium bicarbonate. The layers were separated and the aqueous layer was extracted with dichloromethane (2x). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by RP-HPLC to give the title compound (2.1 mg, 7%) as a white solid. LC / MS (Method X, ESI): [M+H]+= 544.2, R T = 3.06 minutes.

[0309] Example 9 TIFF0007657740000094.tif51170N-(3-(2-(difluoromethoxy)-5-((difluoromethyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide Step 1: N-(3-(2-(difluoromethoxy)-5-((difluoromethyl)thio)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000095.tif51170A 50 mL round bottom flask was placed with sodium 2-chloro-2,2-difluoroacetate (617 mg, 4.05 mmol, 1.00 equiv), N,N-dimethylformamide (10 mL, 129 mmol, 1.00 equiv), cesium carbonate (704 mg, 2.16 mmol, 2.00 equiv), and N-[3-[2-(difluoromethoxy)-5-[[tris(propan-2-yl)silyl]sulfanyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (Intermediate 6, 328 mg, 0.59 mmol, 2.00 equiv). The resulting solution was stirred at 100° C. The resulting mixture was concentrated under vacuum. The residue was loaded onto a silica gel column with ethyl acetate / petroleum ether (1:1). This gave 100 mg (5%) of N-[3-[2-(difluoromethoxy)-5-[(difluoromethyl)sulfanyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. LC / MS (Method H, ESI): [M+H]+= 467.2, R T = 1.20 minutes.

[0310] Step 2: N-(3-(2-(difluoromethoxy)-5-((difluoromethyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000096.tif511702 A 5 mL round bottom flask was placed with N-[3-[2-(difluoromethoxy)-5-[(difluoromethyl)sulfanyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (100 mg, 0.22 mmol, 1.00 equiv), dichloromethane (4 mL, 62.9 mmol, 1.00 equiv), m-CPBA (74.4 mg, 0.43 mmol, 2.00 equiv). The resulting solution was stirred at room temperature for 4 h. The resulting mixture was concentrated under vacuum. The residue was loaded onto a silica gel column with dichloromethane / methanol (20:1). This gave 6.2 mg (6%) of N-[3-[5-(difluoromethane)sulfonyl-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. 1 H NMR (300 MHz, DMSO-d6): δ (ppm) 9.66 (s, 1H), 9.35 (dd, J=6.9, 1.5 Hz, 1H), 8.70 - 8.67 (m, 2H), 8.35 (s, 1H), 8.18 (dd, J=8.7, 2.4 Hz, 1H), 8.07 (d, J=2.4 Hz, 1H), 7.81 - 7.22 (m, 4H), 3.97 (s, 1H). LC / MS (Method A, ESI): [M+H]+= 499.2, R T = 1.56 minutes.

[0311] Example 4 TIFF0007657740000097.tif51170N-(3-(5-(azetidin-3-ylsulfonyl)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of tert-butyl 3-((4-(difluoromethoxy)-3-(1-methyl-4-(pyrazolo[l,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-3-yl)phenyl)sulfonyl)azetidine-1-carboxylate TIFF0007657740000098.tif51170In 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[l,5-a]pyrimidine-3-carboxamide (Intermediate 3, 1.00 g, 2.16 mmol), Pd2( DBA)3 (104 mg, 0.114 mmol, 0.053 equiv), tert-butyl 3-sulfanylazetidine-1-carboxylate (500 mg, 2.64 mmol, 1.22 equiv), XantPhos (116 mg, 0.20 mmol, 0.093 equiv), potassium carbonate (552 mg, 3.99 mmol, 1.85 equiv) and toluene (15 mL) were placed. The resulting solution was stirred at 100° C. overnight and then concentrated under vacuum. The residue was loaded onto a silica gel column with dichloromethane / methanol (94 / 6). The collected fractions were combined and concentrated under vacuum. This gave 1.5 g (122%) of tert-butyl 3-[[4-(difluoromethoxy)-3-(1-methyl-4-[pyrazolo[l,5-a]pyrimidin-3-amido]-1H-pyrazol-3-yl)phenyl]sulfanyl]azetidine-1-carboxylate as a red solid. LC / MS (Method A, ESI): [M+H]+= 572.3, R T = 1.28 minutes.

[0312] Step 2: Synthesis of tert-butyl 3-[[4-(difluoromethoxy)-3-(1-methyl-4-[pyrazolo[l,5-a]pyrimidin-3-amido]-1H-pyrazol-3-yl)benzene]sulfonyl]azetidine-1-carboxylate TIFF0007657740000099.tif51170A 50 mL round bottom flask was placed with tert-butyl 3-[[4-(difluoromethoxy)-3-(1-methyl-4-[pyrazolo[l,5-a]pyrimidin-3-amido]-1H-pyrazol-3-yl)phenyl]sulfanyl]azetidine-1-carboxylate (100 mg, 0.175 mmol), dichloromethane (10 mL) and 3-chlorobenzene-1-perbenzoic acid (60 mg, 0.35 mmol, 2.0 equiv). The resulting solution was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum. The residue was loaded onto a silica gel column with dichloromethane / methanol (92 / 8). The collected fractions were combined and concentrated under vacuum. This gave 100 mg (95%) of tert-butyl 3-[[4-(difluoromethoxy)-3-(1-methyl-4-[pyrazolo[l,5-a]pyrimidin-3-amido]-1H-pyrazol-3-yl)benzene]sulfonyl]azetidine-1-carboxylate as a brown solid. LC / MS (Method I, ESI): [M+H]+= 604.3, R T = 1.05 minutes.

[0313] Step 3: Synthesis of N-(3-(5-(azetidin-3-ylsulfonyl)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000100.tif52170A 50 mL round bottom flask was placed with tert-butyl 3-[[4-(difluoromethoxy)-3-(1-methyl-4-[pyrazolo[l,5-a]pyrimidin-3-amido]-1H-pyrazol-3-yl)benzene]sulfonyl]azetidine-1-carboxylate (100 mg, 0.166 mmol, 1.00 equiv.), trifluoroacetic acid (1 mL, 13.5 mmol, 82 equiv.) and dichloromethane (10 mL). The resulting solution was stirred at room temperature for 6 h. The resulting mixture was concentrated under vacuum. The crude product (5 mL) was purified by Flash-Prep-HPLC with the following conditions (IntelFlash-1): Column: silica gel; Mobile phase: increased from HO(NH4HCO3) / CH3CN=90 / 10 to HO(NH4HCO3) / CH3CN=50 / 50 within 10 min; detector: UV 254 nm. This gave 11.6 mg (14%) of N-[3-[5-(azetidine-3-sulfonyl)-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide as an off-white solid. 1 H NMR (400 MHz, DMSO-d6): δ (ppm) 9.67 (s, 1H), 9.35 (dd, J=6.8, 1.6 Hz, 1H), 8.70 - 8.66 (m, 2H), 8.34 (s, 1H), 8.07 (dd, J=4.4, 1.6 Hz, 1H), 8.00 (s, J=2.4 Hz, 1H), 7.70 - 7.29 (m, 3H), 4.67 - 4.59 (m, 1H), 3.97 (s, 3H), 3.80 - 3.77 (m, 2H), 3.60 - 3.56 (m, 2H). LC / MS (Method A, ESI): [M+H]+= 504.2, R T = 1.12 minutes.

[0314] Example 3 TIFF0007657740000101.tif51170N-(3-(2-(difluoromethoxy)-5-((1-methylazetidin-3-yl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide A 50 mL round bottom flask was placed with N-[3-[5-(azetidine-3-sulfonyl)-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (50 mg, 0.099 mmol, 1.00 equiv.), methanol (1 mL), NaBH(AcO)3 (200 mg, 0.94 mmol, 9.5 equiv.) and dichloromethane (10 mL). The resulting solution was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The crude product (5 mL) was purified by Flash-Prep-HPLC with the following conditions (IntelFlash-1): Column: silica gel; Mobile phase: increased from HO(NH4HCO3) / CH3CN=90 / 10 to HO(NH4HCO3) / CH3CN=50 / 50 within 10 min; detector: UV 254 nm. This gave 18.1 mg (35%) of N-[3-[2-(difluoromethoxy)-5-(1-methylazetidine-3-sulfonyl)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide as an off-white solid. 1 H NMR (400 MHz, DMSO-d6): δ (ppm) 9.67 (s, 1H), 9.35 (dd, J=6.8, 1.6 Hz, 1H), 8.70 - 8.66 (m, 2H), 8.34 (s, 1H), 8.07 (dd, J = 4.4, 1.6 Hz, 1H), 8.00 (s, J = 2.4 Hz, 1H), 7.70 - 7.29 (m, 3H), 4.67 - 4.59 (m, 1H), 3.97 (s, 3H), 3.80 - 3.77 (m, 2H), 3.60 - 3.56 (m, 2H). LC / MS (Method A, ESI): [M+H]+= 518.3, R T = 1.14 minutes. TIFF0007657740000102.tif53170

[0315] N-(3-(5-((1-acetylazetidin-3-yl)sulfonyl)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide A 50 mL round bottom flask was placed with N-[3-[5-(azetidine-3-sulfonyl)-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (80 mg, 0.159 mmol), dichloromethane (15 mL), pyridine (60 mg, 0.76 mmol, 4.8 equiv). Acetyl chloride (15 mg, 0.19 mmol, 1.20 equiv) was added to the solution at 0° C. The resulting solution was stirred at room temperature for 30 minutes. The reaction was then quenched by the addition of 10 mL of water. The resulting solution was extracted with 2×20 mL of dichloromethane, and the organic layers were combined, dried, and concentrated under vacuum. The residue was loaded onto a silica gel column with dichloromethane / methanol (93:7). The crude product was purified by Prep-HPLC (2#-Analytical HPLC-SHIMADZU (HPLC-10)) under the following conditions: Column: Kinetex EVO C18 Column, 30×150, 5μm; Mobile phase: water (10 mmol / l NH4HCO3) and ACN (21.0%-25.0% ACN, 8 min);Detector: UV 254 / 220 nm. This gave 5.8 mg (7%) of N-[3-[5-(1-acetylazetidine-3-sulfonyl)-2-(difluoromethoxy)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide as a white solid. 1H NMR (400 MHz, DMSO-d6): δ (ppm) 9.68 (s, 1H), 9.36 (dd, J=6.8, 1.6 Hz, 1H), 8.70 - 8.68 (m, 2H), 8.34 (s, 1H), 8.16 (dd, J=6.8, 2.4 Hz, 1H), 8.07 (d, J=2.4 Hz, 1H), 7.72 - 7.30 (m, 3H), 4.62 - 4.55 (m, 1H), 4.38 - 4.34 (m, 2H), 4.07 - 3.98 (m, 2H), 3.97 (s, 3H), 2.08 (s, 3H). LC / MS (Method A, ESI): [M+H]+= 546.2, R T = 1.32 minutes.

[0316] Example 26 TIFF0007657740000103.tif51170N-[3-[2-(difluoromethoxy)-5-(methylsulfamoyl)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide In a 25 mL, two-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was placed 4-(difluoromethoxy)-3-(1-methyl-4-[pyrazolo[l,5-a]pyrimidin-3-amido]-1H-pyrazol-3-yl)benzene-1-sulfonic acid (Intermediate 7, 150 mg, 0.323 mmol), dichloromethane (10 mL), N,N-dimethylformamide (0.05 mL, 0.646 mmol, 2.00 equiv), oxalyl chloride (0.50 mL, 5.87 mmol, 18.2 equiv), pyridine (100 mg, 1.26 mmol, 3.91 equiv) and methylamine (0.50 mL, 14.4 mmol, 44.7 equiv). The resulting solution was stirred at room temperature for 1 h. The reaction was then quenched by the addition of 10 mL of water. The resulting solution was extracted with 2x20mL of dichloromethane, and the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was loaded onto a silica gel column with dichloromethane / methanol (93:7). The crude product (20mg) was purified by Prep-HPLC (2#-Analytical HPLC-SHIMADZU (HPLC-10)) under the following conditions: Column: Kinetex EVO C18 Column, 30×150, 5μm; Mobile phase: water (10 mmol / l NH4HCO3) and ACN (20%-35% ACN, 7 min);Detector: UV 254 / 220 nm. This gave 8.6 mg (6%) of N-[3-[2-(difluoromethoxy)-5-(methylsulfamoyl)phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide as a white solid. 1 H NMR (400 MHz, DMSO-d6): δ (ppm) 9.70 (s, 1H), 9.35 (dd, J=7.2, 1.6 Hz, 1H), 8.70 - 8.68 (m, 2H), 8.34 (s, 1H), 7.96 - 7.93 (m, 2H), 7.66 - 7.26 (m, 4H), 3.96 (s, 3H), 2.44 (s, 3H). LC / MS (Method A, ESI): [M+H]+= 478.2, R T = 1.39 minutes.

[0317] Example 5 TIFF0007657740000104.tif51170N-(3-(2-(difluoromethoxy)-5-((2-hydroxyethyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of N-(3-(2-(difluoromethoxy)-5-((2-hydroxyethyl)thio)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000105.tif52170To a solution of N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (intermediate 3, 20.0 g, 43.2 mmol), DIEA (11.2 g, 86.5 mmol), Pd2(dba)3·CH3Cl (2.24 g, 2.16 mmol) and XantPhos (2.5 g, 4.32 mmol) in 1,4-dioxane (200 mL) was added 2-mercaptoethanol (3.19 mL, 45.3 mmol) at room temperature under nitrogen. The resulting solution was stirred at 95 °C for 4 h. The resulting mixture was cooled to room temperature and filtered. The filtrate was concentrated under vacuum. The crude residue was purified by flash chromatography on silica gel eluting with MeOH / DCM (3%) to give N-(3-(2-(difluoromethoxy)-5-((2-hydroxyethyl)thio)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide (16.8 g, 36.5 mmol, 84.5% yield) as an off-white solid. LC / MS (Method A, ESI): [M+H]+= 461.1, R T = 1.02 minutes.

[0318] Step 2: Synthesis of N-[3-[2-(difluoromethoxy)-5-(2-hydroxyethylsulfonyl)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000106.tif51170 Potassium peroxomonosulfate (125 g, 204 mmol) was dissolved in water (450 mL) at 25° C. with stirring and then added to a suspension of N-(3-(2-(difluoromethoxy)-5-((2-hydroxyethyl)thio)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide (45 g, 97 mmol) in methanol (900 mL). The resulting suspension was stirred at room temperature for 6 h. Water (1 L) was added and the resulting suspension was concentrated under vacuum to remove methanol. After filtration, the solid was collected and washed with water (100 mL). The solid was dried at 50° C. and slurried with water (500 mL). After filtration, the solid was collected and washed with water (50 mL). The solid was dried at 50° C. overnight to give 44 g. The solid was slurried in EtOH (400 mL) at 80° C. for 3 h and then cooled to room temperature in an oil bath. After filtration, the solid was collected and washed with EtOH (20 mL). The solid was slurried in EtOH (300 mL) at 80° C. for 3 h and then cooled to room temperature in an oil bath with stirring overnight. After filtration, the solid was collected and washed with EtOH (50 mL). The solid was dried at 50° C. overnight to give N-[3-[2-(difluoromethoxy)-5-(2-hydroxyethylsulfonyl)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (43.3 g, 87.9 mmol, 90.4% yield) as an off-white solid. 1 H NMR (400 MHz, DMSO-d6) 9.68 (s, 1H), 9.34 (dd, J = 7.0, 1.6 Hz, 1H), 8.68 (dd, J = 4.2, 1.6 Hz, 1H), 8.66 (s, 1H), 8.33 (s, 1H), 8.11 8.01 (m, 2H), 7.67 - 7.29 (m, 3H), 4.89 (t, J = 5.3 Hz, 1H), 3.96 (s, 3H), 3.71 (m, 2H), 3.53 (t, J = 6.2 Hz, 2H). LC / MS (Method A, ESI): [M+H]+= 493.2, R T= 1.27 minutes.

[0319] Example 34 TIFF0007657740000107.tif51170Isopropyl 3-(2-(difluoromethoxy)-5-(methylsulfonyl)phenyl)-4-(pyrazolo[l,5-a]pyrimidine-3-carboxamide)-1H-pyrazole-1-carboxylate A 100 mL three-neck round bottom flask was placed with N-[3-[2-(difluoromethoxy)-5-methanesulfonylphenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.223 mmol), triphenylphosphine (76 mg, 0.29 mmol, 1.3 equiv), DIAD (58.6 mg, 0.290 mmol, 1.30 equiv) and toluene (6 mL). The resulting solution was stirred at 95° C. overnight. The resulting mixture was concentrated under vacuum. The residue was loaded onto a silica gel column with ethyl acetate / petroleum ether (4:1). This gave 20 mg (17%) of isopropyl 3-(2-(difluoromethoxy)-5-(methylsulfonyl)phenyl)-4-(pyrazolo[l,5-a]pyrimidine-3-carboxamido)-1H-pyrazole-1-carboxylate as a white solid. 1 H NMR (300 MHz, DMSO-d6) 9.87 (s, 1H), 9.37 (dd, J = 7.2, 1.5 Hz, 1H), 8.78 (s, 1H), 8.71 (s, 1H), 8.64 (dd, J = 4.2, 1.8 Hz, 1H), 8.25 (dd, J = 8.9, 2.7 Hz, 1H), 8.16 (d, J = 2.4 Hz, 1H), 7.80 - 7.26 (m, 3H), 5.26 - 5.18 (m, 1H), 3.33 (s, 1H), 1.42 (d, J = 6.3 Hz, 1H). LC / MS (Method A, ESI): [M+H]+= 535.2, R T = 1.68 minutes.

[0320] Example 12 TIFF0007657740000108.tif50170N-(3-(2-(difluoromethoxy)-5-((3-methoxyphenyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of N-[3-[2-(difluoromethoxy)-5-[(3-methoxyphenyl)sulfanyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide TIFF0007657740000109.tif51170In a 30 mL capped test tube purged and maintained with an inert atmosphere of nitrogen, N-[3-[2-(difluoromethoxy)-5-[[tris(propan-2-yl)silyl]sulfanyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (Intermediate 6, 1.10 g, 1 A mixture of 1.00 g, 4.30 mmol, 2.24 equiv., 1-bromo-3-methoxybenzene (809 mg, 4.33 mmol, 2.25 equiv.), Pd2(dba)3.CHCl3 (117 mg, 0.113 mmol, 0.059 equiv.), cesium carbonate (1.40 g, 4.30 mmol, 2.24 equiv.) was placed in 10 mL of 1-bromo-3-methoxybenzene (809 mg, 4.33 mmol, 2.25 equiv.), Pd2(dba)3.CHCl3 (117 mg, 0.113 mmol, 0.059 equiv.), and cesium carbonate (1.40 g, 4.30 mmol, 2.24 equiv.). The resulting solution was stirred at 90° C. for 6 h. The resulting mixture was concentrated under vacuum. The residue was loaded onto a silica gel column with dichloromethane / methanol (95 / 5). The collected fractions were combined and concentrated under vacuum. This gave 1.2 g (120%) of N-[3-[2-(difluoromethoxy)-5-[(3-methoxyphenyl)sulfanyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a red solid.

[0321] Step 2: Synthesis of N-(3-(2-(difluoromethoxy)-5-((3-methoxyphenyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide. TIFF0007657740000110.tif51170A 50 mL round-bottom flask was placed with N-[3-[2-(difluoromethoxy)-5-[(3-methoxyphenyl)sulfanyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (800 mg, 1.53 mmol), dichloromethane (10 mL), and 3-chlorobenzene-1-perbenzoic acid (753 mg, 4.36 mmol, 1.00 equiv.). The resulting solution was stirred at room temperature for 2 h. The mixture was concentrated and the crude was purified by column chromatography to give N-(3-(2-(difluoromethoxy)-5-((3-methoxyphenyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide. 1 H NMR (300 MHz, DMSO-d6): δ (ppm) 9.67 (s, 1H), 9.32 (dd, J=7.2, 1.5 Hz, 1H), 8.65 - 8.60 (m, 2H), 8.31 (s, 1H), 8.20 (dd, J=8.7, 2.4 Hz, 1H), 8.08 - 7.20 (m, 7H), 3.95 (s, 3H), 3.81 (s, 3H). LC / MS (Method A, ESI): [M+H]+= 555.3, R T = 1.65 minutes.

[0322] Example 11 TIFF0007657740000111.tif52170N-(3-(2-(difluoromethoxy)-5-((3-hydroxyphenyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide In a 50 mL round bottom flask purged and maintained with an inert atmosphere of nitrogen was placed N-[3-[2-(difluoromethoxy)-5-[(3-methoxybenzene)sulfonyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (410 mg, 0.739 mmol), dichloromethane (10 mL) and tribromoborane (1.5 mL, 12.0 mmol, 16.2 equiv). The resulting solution was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum and purified by column chromatography. This afforded 76.5 mg (19%) of N-[3-[2-(difluoromethoxy)-5-[(3-hydroxybenzene)sulfonyl]phenyl]-1-methyl-1H-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide as an off-white solid. 1 H NMR (400 MHz, DMSO-d6): δ (ppm) 10.26 (s, 1H), 9.67 (s, 1H), 9.32 (dd, J=7.2, 1.5 Hz, 1H), 8.66 - 8.63 (m, 2H), 8.33 (s, 1H), 8.12 (dd, LC / MS (Method A, ESI): [M+H]+= 541.3, R T = 1.51 minutes.

[0323] Example 35 TIFF0007657740000112.tif51170N-(3-(2-(difluoromethoxy)-5-((1-methyl-1H-pyrazol-4-yl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of N-[3-[2-(difluoromethoxy)-5-(1-methylpyrazol-4-yl)sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide To a TIFF0007657740000113.tif52170 vial was added N-[3-[2-(difluoromethoxy)-5-sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[l,5-a]pyrimidine-3-carboxamide (Intermediate 8, 1.05 g, 2.52 mmol) and XantPhos Pd G3 (201 mg, 0.202 mmol). N,N-Dimethylformamide (16.8 mL) was added, followed by 4-bromo-1-methyl-1H-pyrazole (837 mg, 0.537 mL, 5.04 mmol) and N,N-diisopropylethylamine (978 mg, 1.32 mL, 7.56 mmol) and nitrogen was bubbled through the reaction mixture for 3 min. The reaction mixture was heated to 180° C. in a microwave reactor for 30 min. The reaction mixture was quenched by the addition of saturated aqueous ammonium chloride and extracted with ethyl acetate (3×). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was adsorbed onto silica and purified by flash column chromatography with 10-80% 3:1 MeOH:iPrOAc / heptane to give N-[3-[2-(difluoromethoxy)-5-(1-methylpyrazol-4-yl)sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow foam (555 mg, 44%). LC / MS (Method W, ESI): [M+H]+= 497.2 R T = 1.09 minutes.

[0324] Step 2: Synthesis of N-(3-(2-(difluoromethoxy)-5-((1-methyl-1H-pyrazol-4-yl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000114.tif50170To a solution of N-[3-[2-(difluoromethoxy)-5-(1-methylpyrazol-4-yl)sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (651 mg, 1.31 mmol) in dichloromethane (16.4 mL) was added 3-chloro-perbenzoic acid (1.18 g, 5.25 mmol) and the reaction was stirred at room temperature for 30 min. The reaction mixture was concentrated in vacuo and the residue was dissolved in ethyl acetate and washed with saturated aqueous ammonium chloride. The aqueous layer was extracted with EtOAc (2x) and the combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The reaction was repeated to give N-[3-[2-(difluoromethoxy)-5-(1-methylpyrazol-4-yl)sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (555 mg) and the two portions were combined for recrystallization. The combined lots were completely dissolved in 410 mL of boiling EtOH. Heating was stopped and the solution was allowed to cool slowly to room temperature with stirring, then cooled in an ice bath. The precipitate was filtered, collected, and dried under vacuum to give N-(3-(2-(difluoromethoxy)-5-((1-methyl-1H-pyrazol-4-yl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (372 mg, 29%). 1H NMR (400 MHz, DMSO-d6) δ 9.64 (s, 1H), 9.33 (dd, J = 7.0, 1.7 Hz, 1H), 8.67 - 8.61 (m, 2H), 8.51 (s, 1H), 8.32 (s, 1H), 8.10 (dd, J = 8.8, 2.5 Hz, 1H), 8.03 (d, J = 2.5 Hz, 1H), 7.97 (d, J = 0.9 Hz, 1H), 7.64 (d, J = 8.8 Hz, 1H), 7.42 (t, J = 72.5 Hz, 1H), 7.28 (dd, J = 7.0, 4.3 Hz, 1H), 3.95 (s, 3H), 3.83 (s, 3H). LC / MS (Method X, ESI): [M+H]+= 529.1 R T = 3.96 minutes.

[0325] Example 41 TIFF0007657740000115.tif50170N-(3-(5-((1H-pyrazol-4-yl)sulfonyl)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of N-[3-[2-(difluoromethoxy)-5-(1H-pyrazol-4-ylsulfanyl)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide To a TIFF0007657740000116.tif50170 vial was added N-[3-[2-(difluoromethoxy)-5-sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 8, 1000 mg, 2.40 mmol), 4-bromo-1H-pyrazole (706 mg, 4.80 mmol) and XantPhos Pd G3 (240 mg, 0.240 mmol). N,N-Dimethylformamide (16.0 mL) was added followed by N,N-diisopropylethylamine (931 mg, 1.26 mL, 7.20 mmol) and nitrogen was bubbled through the reaction mixture for 3 min. The reaction mixture was heated to 180° C. in a microwave reactor for 45 min. The reaction mixture was quenched by the addition of saturated aqueous ammonium chloride and extracted with ETOAc (3x). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was adsorbed onto silica and purified by flash column chromatography with 10-80% 3:1 MeOH:iPrOAc / heptane to give N-[3-[2-(difluoromethoxy)-5-(1H-pyrazol-4-ylsulfanyl)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an orange solid (517 mg, 45%).

[0326] Step 2: Synthesis of N-(3-(5-((1H-pyrazol-4-yl)sulfonyl)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000117.tif50170To a solution of N-[3-[2-(difluoromethoxy)-5-(1H-pyrazol-4-ylsulfanyl)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (517 mg, 1.07 mmol) in dichloromethane (10 mL), 3-chloroperbenzoic acid (961 mg, 4.29 mmol) was added and the reaction was stirred at room temperature for 30 minutes. The reaction mixture was diluted with dichloromethane and poured into a separatory funnel containing saturated aqueous sodium bicarbonate. The layers were separated and the aqueous layer was extracted with dichloromethane (2x). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was adsorbed onto silica and purified by flash column chromatography using 10-80% 3:1 MeOH:iPrOAc / heptane to give N-(3-(5-((1H-pyrazol-4-yl)sulfonyl)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide as a brown solid (383 mg, 69%). 1 H NMR (400 MHz, DMSO-d6) δ 13.78 (s, 1H), 9.64 (s, 1H), 9.32 (dd, J = 7.0, 1.7 Hz, 1H), 8.65 (s, 1H), 8.63 (dd, J = 4.3, 1.7 Hz, 1H), 8.40 - 8.18 (m, 3H), 8.11 (dd, J = 8.8, 2.5 Hz, 1H), 8.05 (d, J = 2.5 Hz, 1H), 7.63 (d, J = 8.7 Hz, 1H), 7.41 (t, J = 72.5 Hz, 1H), 7.27 (dd, J = 7.0, 4.2 Hz, 1H), 3.95 (s, 3H). LC / MS (Method X, ESI): [M+H]+=515.1 R T = 3.71 minutes.

[0327] Example 53 TIFF0007657740000118.tif50170N-(3-(5-((1-(2-(azetidin-1-yl)ethyl)-1H-pyrazol-4-yl)sulfonyl)-2-(difluoromethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide To a solution of N-[3-[2-(difluoromethoxy)-5-(1H-pyrazol-4-ylsulfonyl)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (50 mg, 0.097 mmol), PS-triphenylphosphine (85.0 mg, 0.20 mmol), di-tert-butyl azodicarboxylate (49.2 mg, 0.21 mmol) in tetrahydrofuran (1.5 mL) was added 2-(azetidin-1-yl)ethanol (20.3 mg, 0.019 mL, 0.19 mmol) and the reaction mixture was heated to 60° C. for 90 min. The reaction mixture was filtered and the filtrate was concentrated in vacuo. The crude mixture was extracted with dichloromethane (3 mL) and HCl (2.0 mol / L in diethyl ether, 0.73 mL, 1.46 mmol) and the reaction mixture was stirred for 1 h at room temperature. The reaction mixture was concentrated in vacuo and extracted with dichloromethane. MP-carbonate (464 mg, 1.458 mmol) was added and the reaction mixture was stirred overnight. The mixture was filtered, concentrated in vacuo and purified by reverse phase HPLC to give the title compound (25.4 mg, 44%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 9.64 (s, 1H), 9.33 (dd, J = 7.0, 1.6 Hz, 1H), 8.66 - 8.63 (m, 2H), 8.52 (s, 1H), 8.31 (s, 1H), 8.24 (s, 2H), 8.11 (dd, J = 8.7, 2.5 Hz, 1H), 8.03 (d, J = 2.5 Hz, 1H), 7.99 - 7.94 (m, 1H), 7.64 (d, J = 8.7 Hz, 1H), 7.42 (t, J = 72.5 Hz, 1H), 7.28 (dd, J = 7.0, 4.3 Hz, 1H), 4.02 (t, J = 6.0 Hz, 2H), 3.95 (s, 3H), 2.97 (t, J = 7.0 Hz, 4H), 1.87 - 1.76 (m, 2H). LC / MS (Method X, ESI): [M+H]+=598.2 R T = 3.07 minutes.

[0328] Example 40 TIFF0007657740000119.tif51170N-(3-(2-(difluoromethoxy)-5-(phenylsulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of N-(3-(2-(difluoromethoxy)-5-(phenylthio)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide To a solution of N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 3, 200 mg, 0.43 mmol), thiophenol (57 mg, 0.053 mL, 0.52 mmol), XantPhos Pd G2 (38.3 mg, 0.0432 mmol) and XantPhos (25 mg, 0.0432 mmol) in toluene (11 mL) cooled to 1700 °C, NaH (34.5 mg, 0.86 mmol) was added and the reaction mixture was stirred at 0 °C for 30 min, at room temperature for 1 h and then heated to 90 °C for 2 h. The reaction mixture was diluted with dichloromethane, filtered through Celite, eluted with dichloromethane, and the filtrate was concentrated in vacuo. The crude N-(3-(2-(difluoromethoxy)-5-(phenylthio)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide was used without further purification. LC / MS (Method W, ESI): [M+H]+=593.1 R T = 1.34 minutes.

[0329] Step 2: Synthesis of N-(3-(2-(difluoromethoxy)-5-(phenylsulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000121.tif51170To a solution of N-[3-[2-(difluoromethoxy)-5-phenylsulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (213 mg, 0.432 mmol) in dichloromethane (4.0 mL), 3-chloroperbenzoic acid (203 mg, 0.91 mmol) was added and the reaction was stirred at room temperature for 2 days. 3-chloroperbenzoic acid (68 mg, 0.302 mmol) was added and after 1 h a second portion was added (25 mg). After 1 h the reaction mixture was diluted with dichloromethane and poured into a separatory funnel containing saturated aqueous sodium bicarbonate. The layers were separated and the aqueous layer was extracted with dichloromethane (2x). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by reverse phase HPLC to give the title compound (107.2 mg, 47%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.61 (s, 1H), 9.32 (dd, J = 7.0, 1.5 Hz, 1H), 8.64 (s, 1H), 8.62 (dd, J = 4.3, 1.7 Hz, 1H), 8.31 (s, 1H), 8.15 (dd, J = 8.8, 2.5 Hz, 1H), 8.06 (d, J = 2.5 Hz, 1H), 8.03 - 7.93 (m, 2H), 7.71 - 7.22 (m, 6H), 3.95 (s, 3H). LC / MS (Method [M+H]+=525.1 R T = 4.58 minutes.

[0330] Example 44 TIFF0007657740000122.tif50170N-(3-(2-(difluoromethoxy)-5-((1-methyl-2-oxo-1,2-dihydropyridin-4-yl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of N-[3-[2-(difluoromethoxy)-5-[(1-methyl-2-oxo-4-pyridyl)sulfanyl]phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000123.tif51170To a vial was added N-[3-[2-(difluoromethoxy)-5-sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 3, 50 mg, 0.12 mmol) and XantPhos Pd G3 (9.60 mg, 0.0096 mmol). N,N-Dimethylformamide (1.5 mL) was added, followed by 4-Bromo-1-methylpyridin-2(1H)-one (47.5 mg, 0.240 mmol) and N,N-Diisopropylethylamine (46.6 mg, 0.0628 mL, 0.360 mmol), and nitrogen was bubbled through the reaction mixture for 3 min. The reaction mixture was heated to 180° C. in a microwave reactor for 30 min. The reaction mixture was quenched by the addition of saturated aqueous ammonium chloride and extracted with EtOAc (3×). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo to give N-[3-[2-(difluoromethoxy)-5-[(1-methyl-2-oxo-4-pyridyl)sulfanyl]phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide.

[0331] Step 2: Synthesis of N-(3-(2-(difluoromethoxy)-5-((1-methyl-2-oxo-1,2-dihydropyridin-4-yl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000124.tif51170To a solution of N-[3-[2-(difluoromethoxy)-5-[(1-methyl-2-oxo-4-pyridyl)sulfanyl]phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (63 mg, 0.120 mmol) in dichloromethane (2.0 mL), 3-chloroperbenzoic acid (108 mg, 0.48 mmol) was added and the reaction was stirred at room temperature for 15 minutes. The reaction mixture was diluted with dichloromethane and poured into a separatory funnel containing saturated aqueous sodium bicarbonate. The layers were separated and the aqueous layer was extracted with dichloromethane (2x). The combined organic layers were dried over magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by RP-HPLC to afford the title compound (29.2 mg, 44%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.63 (s, 1H), 9.33 (dd, J = 6.9, 1.6 Hz, 1H), 8.66 - 8.63 (m, 2H), 8.32 (s, 1H), 8.22 (dd, J = 8.8, 2.5 Hz, 1H), 8.09 (d, J = 2.5 Hz, 1H), 7.90 (d, J = 7.1 Hz, 1H), 7.68 (d, J = 8.7 Hz, 1H), 7.48 (t, J = 72.3 Hz, 1H), 7.28 (dd, J = 7.0, 4.3 Hz, 1H), 6.94 (d, J = 2.2 Hz, 1H), 6.60 (dd, J = 7.1, 2.1 Hz, 1H), 3.96 (s, 3H), 3.40 (s, 3H). LC / MS (Method X, ESI): [M+H]+=556.1 R T = 3.18 minutes.

[0332] Example 15 TIFF0007657740000125.tif51170(S)-N-(3-(2-(difluoromethoxy)-5-((2-hydroxypropyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of (S)-N-(3-(2-(difluoromethoxy)-5-((2-hydroxypropyl)thio)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000126.tif51170 To a solution of N-[3-[2-(difluoromethoxy)-5-sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate 8, 950 mg, 2.28 mmol) in methanol (10 mL) was added (S)-(-)-propylene oxide (250 mg, 4.3 mmol) and DIEA (900 mg, 6.98 mmol) and the mixture was stirred for 3 h at 80° C. After cooling to room temperature, the reaction mixture was concentrated under vacuum. The residue was purified by reverse phase HPLC [mobile phase A: water (0.1% NH4HCO3), mobile phase B: acetonitrile; gradient: 50 min, 10%B to 40%B] to give N-[3-[2-(difluoromethoxy)-5-[(2S)-2-hydroxypropyl]sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (730 mg, 1.54 mmol, 67% yield) as a pale yellow solid. LC / MS (Method O, ESI): [M+H]+=475.2 R T = 1.43 minutes.

[0333] Step 2: Synthesis of (S)-N-(3-(2-(difluoromethoxy)-5-((2-hydroxypropyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000127.tif50170To a solution of N-[3-[2-(difluoromethoxy)-5-[(2S)-2-hydroxypropyl]sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (730 mg, 1.54 mmol) in methanol (10 mL), a solution of oxone (2.0 g, 3.25 mmol) in water (5.0 mL) was added and the mixture was stirred at room temperature for 3 h. The organic solvent was removed under vacuum and 10 mL of water was added. The solid was collected by filtration. The filter cake was washed with water (5.0 mL x 5) and dried under vacuum. The residue was purified by preparative HPLC (column: XBridge Prep OBD C18 Column (30 × 150 mm, 5 μm); mobile phase A: water (10 mmol / l NH4HCO3) (mobile phase B): ACN; flow rate: 60 mL / min; gradient: 16% B to 37% B in 8 min; 254 nm; R T The product was purified by HPLC using a 1:7.45 min. reaction. The product was stirred in 3.0 mL of ethanol at room temperature overnight to give N-[3-[2-(difluoromethoxy)-5-[(2S)-2-hydroxypropyl]sulfonyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (783 mg, 1.52 mmol) as an off-white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.69 (s, 1H), 9.35 (dd, J = 6.8, 1.6 Hz, 1H), 8.70 - 8.68 (m, 2H), 8.34 (s, 1H), 8.09 - 8.04 (m, 2H), 7.68 - 7.29 (m, 3H), 4.90 (d, J = 5.6 Hz, 1H), 4.10 - 4.04 (m, 1H), 3.97 (s, 3H), 3.50 - 3.40 (m, 2H), 1.12 (d, J = 6.0 Hz, 3H). LC / MS (Method O, ESI): [M+H]+=507.2 R T = 1.26 minutes.

[0334] Example 25 TIFF0007657740000128.tif51170(R)-N-(3-(2-(difluoromethoxy)-5-((2-hydroxypropyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of (R)-N-(3-(2-(difluoromethoxy)-5-((2-hydroxypropyl)thio)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000129.tif50170 To a solution of N-[3-[2-(difluoromethoxy)-5-sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (1.10 g, 2.64 mmol) in methanol (10 mL) was added (R)-(+)-propylene oxide (300 mg, 5.17 mmol) and DIEA (1.00 g, 7.75 mmol) and the reaction mixture was stirred for 3 h at 80° C. After cooling to room temperature, the solvent was removed under vacuum. The residue was purified by reverse phase HPLC [mobile phase A: water (0.1% NH4HCO3), mobile phase B: acetonitrile; gradient: 50 min, 10%B to 40%B] to give N-[3-[2-(difluoromethoxy)-5-[(2S)-2-hydroxypropyl]sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (780 mg, 1.64 mmol, 62% yield) as a pale yellow solid. LC / MS (Method O, ESI): [M+H]+=475.2 R T = 1.43 minutes.

[0335] Step 2: Synthesis of (R)-N-(3-(2-(difluoromethoxy)-5-((2-hydroxypropyl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000130.tif50170To a solution of N-[3-[2-(difluoromethoxy)-5-[(2R)-2-hydroxypropyl]sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (780 mg, 1.64 mmol) in methanol (10 mL), a solution of oxone (2.0 g, 3.26 mmol) in water (5.0 mL) was added and the mixture was stirred at room temperature for 3 h. The organic solvent was removed under vacuum. Water (10 mL) was added and the solid was collected by filtration. The filter cake was washed with water (5.0 mL x 5) and then dried under vacuum. The residue was purified by preparative HPLC (column: XBridge Prep OBD C18 Column (30 × 150 mm, 5 μm); mobile phase A: water (10 mmol / l NH4HCO3) (mobile phase B): ACN; flow rate: 60 mL / min; gradient: 16% B to 37% B in 8 min; 254 nm; R T The product was purified by HPLC using a 1:7.45 min. chromatography. The product was stirred in 3.0 mL of ethanol at room temperature overnight to give N-[3-[2-(difluoromethoxy)-5-[(2S)-2-hydroxypropyl]sulfonyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (771 mg, 1.51 mmol 92% yield) as an off-white solid. LC / MS (Method O, ESI): [M+H]+=507.2 R T = 1.26 minutes. 1 H NMR (400 MHz, DMSO-d6) δ 9.69 (s, 1H), 9.35 (dd, J = 6.8, 1.6 Hz, 1H), 8.70 - 8.68 (m, 2H), 8.34 (s, 1H), 8.10 - 8.04 (m, 2H), 7.68 - 7.29 (m, 3H), 4.90 (d, J = 5.6 Hz, 1H), 4.10 - 4.04 (m, 1H), 3.97 (s, 3H), 3.50 - 3.40 (m, 2H), 1.12 (d, J = 6.0 Hz, 3H).

[0336] Examples 23 and 24 TIFF0007657740000131.tif51170(R)-N-(3-(2-(difluoromethoxy)-5-((1-hydroxypropan-2-yl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide and (S)-N-(3-(2-(difluoromethoxy)-5-((1-hydroxypropan-2-yl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide Step 1: Synthesis of N-(3-(2-(difluoromethoxy)-5-((1-hydroxypropan-2-yl)thio)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000132.tif51170To a solution of N-[3-[5-bromo-2-(difluoromethoxy)phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (2.00 g, 4.32 mmol) in 1,4-dioxane (100 mL) was added Pd2(dba)3 (396 mg, 0.431 mmol), Xantphos (500 mg, 0.864 mmol) and DIPEA (1.67 g, 13.0 mmol) at room temperature under nitrogen. The resulting solution was stirred at RT for 15 min and heated to 90 °C. 2-Sulfanylpropan-1-ol (580 mg, 6.29 mmol) was added to the reaction mixture and the resulting mixture was stirred at 90 °C for 10 h. The reaction was cooled to room temperature and the suspension was filtered through Celite. The filtrate was concentrated under vacuum. The residue was purified by flash chromatography on silica gel eluting with DCM / EA (0%-100%) to give N-[3-[2-(difluoromethoxy)-5-(2-hydroxy-1-methyl-ethyl)sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (1.74 g, 3.67 mmol, 85% yield) as an off-white solid. LC / MS (Method I, ESI): [M+H]+=507.2 R T = 0.97 minutes.

[0337] Step 2: Synthesis of N-(3-(2-(difluoromethoxy)-5-((1-hydroxypropan-2-yl)sulfonyl)phenyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide TIFF0007657740000133.tif51170 A solution of N-[3-[2-(difluoromethoxy)-5-(2-hydroxy-1-methyl-ethyl)sulfanyl-phenyl]-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (850 mg, 1.79 mmol) in methanol (8.0 mL) was stirred at 0° C. for 5 min. Then, a solution of oxone (2.20 g, 3.58 mmol) in water (8.0 mL) was added at 0° C. The reaction solution was stirred at room temperature for 1 h. The reaction mixture was concentrated and filtered. The filter cake was mixed with DCM (10 mL) and filtered. The filtrate was concentrated under vacuum to give the product (779 mg, 1.54 mmol, 86% yield). This reaction was repeated on the same scale and combined to give 1.10 g (61% yield of racemic compound). The combined products were purified by chiral SFC according to the following conditions: Column: Lux 5um Cellulose-4.5×25cm, 5μm; Mobile phase A: CO2; Mobile phase B: EtOH:ACN=2:1 (2 mM NH3-MeOH); Flow rate: 150mL / min; Gradient: 40%B; 220nm; R T 1:9.03 minutes; RT2: 10.79 min. Fractions containing the two enantiomers were combined, concentrated, and both were recrystallized by stirring in 2 mL of isopropanol at room temperature for 2 days. The product was isolated by filtration to give the title compound, which has been arbitrarily assigned as to configuration.

[0338] Peak 1: (R)-(N-[3[2-(difluoromethoxy)-5-(2-hydroxy-1-methyl-ethyl)sulfonyl-phenyl)-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (341 mg) was obtained as an off-white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.67 (s, 1H), 9.34 (dd, J = 7.0, 1.6 Hz, 1H), 8.79 - 8.55 (m, 1H), 8.66 (s, 1H), 8.32 (s, 1H), 8.17 - 7.85 (m, 2H), 7.79 - 7.16 (m, 3H), 4.96 (t, J = 5.6 Hz, 1H), 3.96 (s, 3H), 3.70 (m, 1H), 3.60 - 3.27 (m, 2H), 1.22 (d, J = 6.7 Hz, 3H). LC / MS (Method X, ESI): [M+H]+=507.1 R T = 3.59 minutes.

[0339] Peak 2: (S)-(N-[3[2-(difluoromethoxy)-5-(2-hydroxy-1-methyl-ethyl)sulfonyl-phenyl)-1-methyl-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (411 mg) was obtained as off-white. 1 H NMR (400 MHz, DMSO-d6) δ 9.67 (s, 1H), 9.34 (dd, J = 7.0, 1.6 Hz, 1H), 8.79 - 8.45 (m, 2H), 8.32 (s, 1H), 8.17 - 7.86 (m, 2H), 7.86 - 7.16 (m, 3H), 4.96 (t, J = 5.5 Hz, 1H), 3.96 (s, 3H), 3.70 (m, 1H), 3.60 - 3.28 (m, 2H), 1.22 (d, J = 6.7 Hz, 3H). LC / MS (Method X, ESI): [M+H]+=507.1 R T = 3.56 minutes.

[0340] Assay Test drug Test drug samples were provided as solutions at a concentration of 10 mM in dimethylsulfoxide (DMSO) and stored in the dark at room temperature prior to use.

[0341] Biochemical assays for JAK1 and JAK2 In vitro biochemical assays use a LabChip® EZ Reader II microfluidic mobility shift device (PerkinElmer; Waltham, MA) for detection and quantification of JAK-catalyzed phosphorylation of synthetic peptides. Substrate peptide Y-1B has the sequence 5-FAM-VALVDGYFRLTT-NH2. Y-1B is fluorescently labeled at the N-terminus with 5-FAM (5-carboxyfluorescein) and contains a single tyrosine residue (Y) that can be phosphorylated by JAK activity. Substrate peptide stocks are prepared in DMSO at 5 mM. Purified recombinant human JAK1 kinase domain protein (residues 854-1154) was expressed in insect cells and obtained from Proteros Biostructures GmbH (Martinsried, Germany). Recombinant human JAK2 kinase domain protein (residues 812-1132) was expressed in insect cells and purified at Genentech, Inc. (South San Francisco, Calif.).

[0342] 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 M Y-1B peptide substrate, 25 μM adenosine triphosphate (ATP), 1 nM total JAK1 or 0.2 nM total JAK2, and test compound up to 1000 nM at a final concentration of 2% (volume to volume [v / v]) in DMSO. In each titration experiment, test compounds were tested in replicates at 12 concentrations each. Blank reactions contained ATP, peptide, and DMSO but no JAK or test compound, and uninhibited control reactions contained ATP, peptide, JAK, and DMSO but no test compound.

[0343] The peptide+ATP mixture (24 μL) was added to 1 μL of test compound in DMSO (or DMSO alone). The reaction was initiated by adding 25 μL of JAK enzyme to the inhibitor / peptide / ATP mixture, and the resulting solution was then mixed thoroughly. The reactions were incubated at room temperature (22° C.-23° C.) in a final volume of 50 μL per well in a 384-well plate. After 30 min of 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.

[0344] In each reaction, the residual Y-1B substrate and phosphorylated peptide products generated were separated using an EZ Reader II instrument. Electrophoretic separation of substrate and product molecules was performed using downstream and upstream voltages of 500 and 2600 V, respectively, at a driving pressure of 1.3 psi. The 5-FAM groups present in the substrate and product peptides were excited at 488 nm, fluorescence was detected at 530 nm, and peak heights were recorded.

[0345] 4. Data Analysis The extent (or %) of conversion of substrate to product was calculated from the corresponding peak heights in the electropherogram using HTS Well Analyzer software, Version 5.2 (PerkinElmer) and the following equation (Equation 1): formula 1 Conversion % = [P÷(S+P)] × 100 where S and P represent the peak heights of the substrate and product, respectively. After subtracting the baseline signal of blank wells containing no JAK from the signals of all test wells, the % conversion data were converted to fractional activity as shown in Equation 2, where vi and vo represent the % conversion in the presence and absence of test compound, respectively. The % conversion observed in the uninhibited control reaction containing JAK and DMSO vehicle but no test compound was defined as a fractional activity of 1 (no inhibitor, vi=vo), while blank wells without JAK were defined as a fractional activity=0. The fractional activity was plotted against test compound concentration and the apparent inhibition constant of tight binding (K) was calculated using XLfit software (IDBS; Guildford, UK). i app ) The data were fitted to a quadratic equation (see Equation 2) (Williams JW, Morrison JF. The kinetics of reversible tight-binding inhibition. Methods Enzymol 1979;63:43767), which was used to calculate the fractional activity and K i app was used to calculate

[0346] formula 2 TIFF0007657740000134.tif19170, [E] T and [i] T are the active enzyme (initial estimates are 0.15 nM for JAK1 and 0.048 nM for JAK2) and the total concentration of inhibitor (the parameter that is varied), respectively. Finally, by applying the competitive inhibition relationship (Equation 3), K i ,K i app Calculated from.

[0347] formula 3 K i =K i app / (1+[ATP] / K m app ) where [ATP] is the concentration of ATP = 25 μM and K m app The apparent ATP Michaelis constant for JAK1 is 32.1 μM, whereas for JAK2, K m app = 11.7 μM. By applying the tight binding equation 2, which describes the inhibitor reduction, and the competitive inhibition relationship, equation 3, the sensitivity of the assay can be extended to at least a calculated K of 0.008 nM for JAK1 and 0.0015 nM for JAK2.

[0348] Kinase selectivity In vitro kinase selectivity of test agents was assessed at a concentration of 1 μM in a panel of recombinant human kinase activity and binding experiments, including cytoplasmic and receptor tyrosine kinases, serine / threonine kinases, and lipid kinases (SelectScreen Kinase Profiling Services, ThermoFisher Scientific, Madison, WI). Kinase activity assays measure peptide phosphorylation (Z'-LYTE®) or ADP production (Adapta®), while binding experiments monitor displacement of an ATP site binding probe (LanthaScreen®). The ATP concentration used in the activity assays is typically determined based on the experimentally determined apparent Michaelis constant (K m app ) value, whereas the competitive binding tracer concentration used in binding experiments is usually within two-fold of the experimentally determined dissociation constant (K d) value. Inhibitors are tested in replicates for each kinase, and the average % inhibition value is recorded. For kinases that are inhibited near or above 50% at the initial 1 μM test concentration, a 10-point inhibitor titration is performed using the same assay to determine the 50% inhibition (IC 50 The inhibitor concentration at which an IC50 value of 37.5 nM (half the total enzyme concentration) was determined. The total JAK1 concentration used in this assay panel was 75 nM. If 100% of the 75 nM JAK1 protein is catalytically active, the limit of JAK1 inhibitor sensitivity from the vendor JAK1 assay is theoretically an IC50 value of 37.5 nM (half the total enzyme concentration). 50 However, the SelectScreen® JAK1 assay yielded JAK1 IC values ​​significantly lower than 37.5 nM for some inhibitors. 50 values ​​that are consistent with our internal results. Thus, the active JAK1 enzyme concentration in the SelectScreen® assay must be much lower than the 75 nM total apparent JAK1 protein concentration used in the assay, and the observed sensitivity of this assay is within a theoretical IC of 37.5 nM. 50 This is much better than the sensitivity limit.

[0349] 4. Data Analysis To fit the data to concentration-kinase inhibition plots, SelectScreen® Kinase Profiling Services employed XLfit Software (IDBS) Model No. 205 (sigmoidal concentration-response model), which is a 4-parameter logistic fit model described in Equation 4. formula 4 y=A+{(B A )÷[1+(C ÷x) D ]} where x is the inhibitor concentration, y is the % inhibition observed, A is the minimum y value, B is the maximum y value, and C is the IC 50where σ is the Hill slope and D is the slope of the Hill curve. In some cases, a 3-parameter logistic fit was used. For example, if the plateau of the curve for infinitely low inhibitor concentration did not fit between -20% and 20% inhibition, the lower plateau was set to 0% inhibition, whereas if the plateau of the curve for infinitely high inhibitor concentration did not fit between 70% and 130% inhibition, the upper plateau was set to 100% inhibition.

[0350] Phosphorylated STAT JAK1 and JAK2 pathway selectivity assay with TF-1 cell line TF-1 human erythroleukemia cells (ATCC; Manassas, VA; Catalog No. 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, cultures were transferred to Opti-MEM™, 1× NEAA, 1 mM sodium pyruvate and 0.5% charcoal-stripped FBS (poor medium). Inhibitor stock solutions (5 mM in DMSO) were serially diluted 1:2 in DMSO to give a 10-point concentration titration (500× test concentration), which was further diluted 50-fold in assay medium (RPMI with 1× NEAA and 1 mM sodium pyruvate) to give a 10-point concentration titration (2% in DMSO). Cells (300,000 cells / well in 35 μL of assay medium) were seeded into 384-well Greiner plates. Ten concentrations of diluted inhibitor (5 μL) were added to the cells and the plates were incubated at 37° C. for 30 min in a humidified incubator. Cells were incubated at 100° C. for 30 min at each EC 90Cells were stimulated with human recombinant cytokines at concentrations of 0.1 mM. 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 min. For the P-STAT5 TF-1 erythropoietin (EPO) assay, 10 μL of 110 IU / mL EPO (Gibco Life Technologies, Catalog No. PHC2054) was added to the cells, which were then incubated at 37°C for 30 min. For both assays, after incubation, 5 μL of ice-cold 10× cell lysis buffer (Cell Signaling Technologies; Danvers, MA; Catalog No. 9803S) containing 1 mM phenylmethylsulfonyl fluoride (PMSF) was added to the cells. Assay plates were frozen at -80°C for a minimum of 1 hour. For the IL-13 assay, P-STAT6 was measured by coating goat anti-rabbit (GAR) plates (Meso Scale Discovery [MSD]; Rockville, MD; Catalog No. MSD L21RA-1) with rabbit anti-human whole STAT6 antibody (Cell Signaling Technologies; Catalog No. 9362S), incubating cell lysates on the coated plates overnight at 4°C, and detecting with mouse anti-P-STAT6 (Tyr641) Clone16E12 antibody (MilliporeSigma; Burlington, MA; Catalog No. 05-590 (custom labeled by MSD with a SULFO-tag) using standard MSD plate processing, washing, and detection protocols. For the EPO assay, P-STAT5 was detected using the phospho-STAT5a,b Whole Cell Lysate Kit (MSD; Catalog No. K150IGD-1). Electrochemiluminescence (ECL) signals of the wells were measured using a MESO SECTOR ELISA Kit. Measurements were taken using an S600 (MSD) reader.

[0351] 4. Data Analysis The mean ECL value of the negative control (cells stimulated with cytokines and treated with 20 μM of control inhibitor) was subtracted from the ECL values ​​of all wells, and the percent of the ECL value of the test compound wells relative to the mean ECL value of the positive control (cells stimulated with cytokines and treated with DMSO) was determined, and the IC of the test compound was calculated using a 4-parameter logistic fit model shown in Equation 4. 50 Data analysis was performed by determining the following:

[0352] P-STAT6 BEAS-2B+IL-13 cell assay To test the effects of JAK1 inhibitors in cell lines important to the cell biology of human asthma, an IL-13 stimulated STAT6 phosphorylation assay with the human lung bronchial epithelial BEAS-2B cell line was developed.

[0353] BEAS-2B cells (ATCC® CRL-9609™) were grown in Bronchial Epithelial Growth Medium (BEGM) (Lonza Catalog No. CC-3170; Walkersville, MD; or PromoCell Catalog No. C-21060; Heidelberg, Germany). Test compound stock solutions (0.5 mM in DMSO) were serially diluted 1:2 in DMSO to obtain a 10-point concentration titration (500× test concentration), which was further diluted 50-fold in BEGM to obtain a 10× concentration curve (2% in DMSO). Cells were seeded at 100,000 cells / well in 200 μL of BEGM in a 96-well plate and incubated at 37° C. for 48 hours in a humidified incubator. Media was aspirated from the cells and replaced with 70 μL of fresh BEGM. Diluted test compounds (10 μL, or 2% DMSO in assay medium) were added to the cells and the plates were incubated for 1 hour at 37° C. in a humidified incubator. 20 μL of 250 ng / mL human recombinant IL-13 (Bio Techne Cat. No. 213-ILB) was then added to the cells and incubated for 15 minutes at 37° C. Media was aspirated from the cells and 60 μL of ice-cold 1× cell lysis buffer (Cell Signaling Technologies; Cat. No. 9803S) containing 1 mM PMSF was added to the cells. Assay plates were incubated at −80° C. for a minimum of 1 hour. P-STAT6 was measured by coating GAR plates (MSD, Cat. No. L45RA-1) with rabbit anti-human total STAT6 antibody (Cell Signaling Technologies; Cat. No. 9362S), incubating cell lysates on the coated plates overnight at 4°C, and detecting with mouse anti-phospho-STAT6 (Tyr641) Clone16E12 antibody (Millipore; Cat. No. 05-590 (custom labeled by MSD with SULFO-tag) using standard MSD plate processing, washing and detection protocols. Plates were read on a MESO SECTOR S600.

[0354] 4. Data Analysis Data analysis was performed by subtracting the negative control values ​​from all wells and using the positive control values ​​to determine the percent relative to control, IC 50 was determined with a 4-parameter logistic fit model as shown in Equation 4.

[0355] Cytotoxicity assay A549 (ATCC® CCL-185™), Jurkat clone E6-1 (ATCC® TIB-152™) and HEK-293T (ATCC® CRL-1573™) cells were used, maintained at less than confluent density in T175 flasks. Logarithmic growth phase cells were seeded at 450 cells / 45 μL of medium in Greiner 384-well black / clear tissue culture treated plates (Greiner Catalog # 781091). After dispensing the cells, the plates were equilibrated at room temperature for 30 minutes, after which the cell plates were placed in a CO2 and humidity controlled incubator at 37° C. overnight. The next day, cells were treated with test agents diluted in 100% DMSO in a 10-point titration to a maximum concentration of 50 μM (final 0.5% DMSO concentration on cells). Cells and compounds were incubated for 72 hours in a 37°C, CO2- and humidity-controlled incubator, after which cell viability was measured by adding CellTiter-GloR (Promega G7572) reagent to all wells. Plates were incubated for 20 minutes at room temperature, and then the luminescence of the wells was measured on an EnVision plate reader (Perkin Elmer Life Sciences).

[0356] Data from the above JAK1 and JAK2 assays with the compounds of Table 1 are shown in Table 2 below. TIFF0007657740000135.tif255170TIFF0007657740000136.tif255170TIFF0007657740000137.tif178170

[0357] As can be seen from Table 2, all of the compounds of the present invention are highly active in biochemical JAK1 and JAK2 assays, and in most cases also in one or more of the JAK1 and JAK2 cell-based assays. Some of the compounds are substantially equipotent at JAK1 and JAK2.

[0358] Table 3 below provides mouse lung tissue binding (LTB) data (% bound) and cytotoxicity data for selected compounds from Table 1. TIFF0007657740000138.tif253170TIFF0007657740000139.tif105170

[0359] From the data in Table 3, it can be seen that the 1-methylpyrazole compounds (i.e., R 1 is methyl) to the corresponding desmethyl (R 1 The methyl vs. desmethyl analogs (R 1 = Methyl vs R 1 = hydrogen) for Examples 6, 9 and 17 in Tables 1 and 2. TIFF0007657740000140.tif232170TIFF0007657740000141.tif194170

[0360] From the data in Table 3, it was also found that the sulfone compounds of the present invention surprisingly exhibited improved lung tissue binding compared to the corresponding sulfide analogs. Table 5 below shows the results of the sulfone vs. sulfide analog (R 1 = Methyl vs R 1 = hydrogen) for Examples 13, 28 and 35 in Tables 1 and 2. TIFF0007657740000142.tif247170TIFF0007657740000143.tif110170

[0361] The sulfone compounds of the present invention also exhibit lower LogD and greater kinetic solubility compared to the corresponding sulfide analogs.

[0362] From the data in Tables 2 and 3, R 1 is methyl and R 2 It has also been found that compounds of the invention in which is hydroxyalkyl or alkoxyalkyl exhibit good balanced affinity for JAK1 and JAK2, and further exhibit relatively low lung tissue binding and cytotoxicity.

[0363] Animal models House dust mite (HDM) model Female C57BL / 6J mice, 7-8 weeks of age, were purchased from Jackson West. House Dust Mite (HDM (D. Pteronyssinus)) was purchased from Greer Laboratory, mixed with 2 mg of alum (Thermo Scientific) diluted in sterile PBS and normalized to 0.918 μg DerP1 content per mouse, and immunized intraperitoneally on days 0 and 14. Mice were sensitized on days 21 and 24 with HDM in PBS (again normalized to 0.918 μg DerP1 content) administered by intratracheal inhalation. Prior to inhalation sensitization of each HDM (and in a subset of groups on days 22 and 23), animals receive test compound via nose-only inhalation (using an Electro-Medical Measurement Systems (EMMS) dry powder inhalation device equipped with a Wright dust feeder and a 4-tier / 24-port or 2-tier / 12-port, directional-flow, nose-only inhalation tower) 1 hour prior to sensitization. Control animals receive nose-only inhalation with air only. 24 hours after the final treatment, mice are bled from the orbital plexus for plasma PK and then euthanized by CO2 inhalation. After euthanasia, BAL fluid is collected for total (by FACS using a known number of spike-in reference beads) and differential (Wright Giemsa stain, cytospin) cell counts. Lungs and spleens are harvested for PK, weighed, and frozen. Five or six animals were used per group.

[0364] In addition, to confirm the dose delivered to the lung, three PK satellite groups of untreated animals are administered test compound via inhalation only through nose for one or four consecutive days.Immediately after the last inhalation administration, PK satellite animals are bled from the orbital plexus of mice for plasma PK, and then euthanized by CO2 inhalation.Lungs and spleens are harvested, weighed, and frozen for PK analysis.

[0365] Rat OVA model Six-week-old male Brown Norway rats from Charles River-Kingston. Rats are immunized intraperitoneally on day 0 with 150 μg OVA (Sigma) mixed with 40 mg alum (Thermo Scientific) diluted in sterile PBS. 28 days after sensitization, rats are sensitized with 2% OVA in PBS aerosolized via a nebulizer for 30 minutes on three consecutive days. Prior to each OVA sensitization, animals receive JAK1 / JAK2 test compounds via nose-only inhalation (using a Wright dust feeder and / or Electro-Medical Measurement Systems (EMMS) dry powder inhalation device with a 4-tier / 24-port, directional-flow, nose-only inhalation tower) one hour prior to sensitization. Control animals receive MCT buffer orally or air only nose-only inhalation. 24 hours after the final treatment, rats are euthanized by CO2 inhalation. They are bled via the abdominal aorta for plasma PK and whole blood FACS analysis. After euthanasia, BAL fluid is collected for total (by FACS using a known number of spike-in reference beads) and differential (Wright Giemsa stain, cytospin) cell counts. Lungs are harvested for PK, weighed and frozen. 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 % natural killer cells (CD161a positive). Six animals are used per group, except for five animals in the untreated control group.

[0366] In addition, to confirm the dose delivered to the lung, three PK satellite groups of untreated animals are administered JAK1 / JAK2 test compound via nose-only inhalation for 1 or 3 days, respectively.Immediately after the final inhalation administration, the PK satellite animals are euthanized by CO2 inhalation.They are bled from the abdominal aorta for plasma PK.Lungs and spleens are harvested, weighed, and frozen for PK analysis.

[0367] The plasma and lung levels of test compounds and their ratios are measured by the following method. BALB / c mice from Charles River Laboratories are used in the assay. Test compounds are individually prepared in 0.2% Tween 80 solution in saline, and the solution is administered to the trachea of ​​the mice by oral aspiration. At each time point (typically 0.167, 2, 6, 24 hours after administration), blood samples are taken via cardiac puncture, and whole lungs are excised from the mice. Blood samples are centrifuged (Eppendorf centrifuge, 5804R) at 4°C and approximately 12,000 rpm for 4 minutes, and plasma is collected. Lungs are padded dry, weighed, and homogenized in a 1:3 dilution with sterile water. Plasma and lung concentrations of test compounds are measured by LC-MS analysis using analytical standards prepared as standard curves in test matrix. Lung-to-plasma ratios were determined as the ratio of lung AUC (μg hr / g) to plasma AUC (μg hr / mL), where AUC is commonly defined as the area under the curve of test compound concentration versus time.

[0368] Mouse plasma and lung pharmacokinetics The pharmacokinetics of the compounds are measured in female Balb / c mice following administration by a single intranasal (IN) bolus solution / suspension administration of a target dose of 0.3 mg / kg prepared in 0.2% Tween 80 solution in saline. Female Balb / c mice aged 7-8 weeks may be purchased from Charles River. Mice are housed in specific pathogen-free conditions until use in the study.

[0369] Animals are not fasted prior to dosing. Blood samples are taken by cardiac puncture with EDTA-coated microtainers under anesthesia (intraperitoneal injection of pentobarbitone) from three animals each at 0.083, 2, 7 and 24 hours post-dosing. Blood samples are centrifuged (1500 g, 4° C., 10 min) to separate plasma. Plasma samples are frozen at approximately −80° C. After intranasal dosing, spleens are harvested, weighed and rapidly frozen prior to pulmonary perfusion. After confirmation of death, lungs of dosed animals are perfused with chilled PBS to remove residual blood from the pulmonary vasculature. Lungs are then excised and weighed (all weights are recorded). All tissue samples are frozen by immersion in liquid nitrogen. Tissue samples are kept frozen (at approximately −80° C.) until analysis.

[0370] Prior to PK analysis, thawed tissue samples (spleen and lung) are weighed and homogenized, and 4 mL of HPLC grade water is added for each gram of tissue using an Omni-Prep Bead Ruptor (Omni Inc., Kennesaw, GA) at 4°C. Plasma and tissue homogenate samples are extracted by protein precipitation with 4 volumes of acetonitrile containing tolbutamide (200 ng / mL) or labellorol (100 ng / mL) as an internal standard. Samples are mixed and centrifuged at 3200g for 30 min at 4°C to remove precipitated proteins, and the supernatants are diluted appropriately (e.g., 1:1, v / v) with HPLC grade water in 96-well plates. Representative aliquots of plasma, spleen and lung samples are analyzed 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. Standards are prepared by spiking compounds into aliquots of control plasma, spleen and lung tissue homogenates and extracting as described for the experimental samples. In all matrices, the assay detection limits are between 0.168 mg / mL and 4000 ng / mL.

[0371] Concentrations below the lower limit of quantification (LLOQ) are treated as zero in the calculation of the mean and SD. The mean concentrations measured in the samples are used to construct semi-logarithmic concentration-time curve profiles. Pharmacokinetic (PK) analysis is performed using non-compartmental methods in Biobook (E-Workbook IDBS).

[0372] A mouse model of Alternaria alternata-induced pulmonary eosinophilic inflammation Airway eosinophilia is a hallmark of human asthma. Alternaria alternata, a fungal aeroallergen that can exacerbate asthma in humans, induces pulmonary eosinophilic inflammation in mice (Havaux et al. Clin Exp Immunol. 2005, 139(2):179-88). In mice, Alternaria has been shown to indirectly activate tissue-resident type 2 lymphoid cells in the lungs, which respond (e.g., to IL-2 and IL-7) to release JAK-dependent cytokines (e.g., IL-5 and IL-13) and modulate eosinophilic inflammation (Bartemes et al. J Immunol. 2012, 188(3):1503-13).

[0373] Taconic 7-9 week old male C57 mice are used for the study. On the day of the study, animals are lightly anesthetized by isoflurane and administered vehicle or test compound via oropharyngeal aspiration. After administration, animals are placed in lateral recumbency and monitored for full recovery from anesthesia before being returned to their own cage. One hour later, animals are lightly anesthetized again and sensitized with vehicle or alternaria extract via oropharyngeal aspiration, then monitored for recovery from anesthesia before being returned to their own cage. 48 hours after alternaria administration, bronchoalveolar lavage fluid (BALF) is collected and eosinophils in BALF are counted using an Advia 120 Hematology system (Siemens).

[0374] The activity of compound in the model is shown by the reduction in the eosinophil levels present in the BALF of animals 48 hours after treatment, compared with vehicle-treated control animals with alternaria lesions.Data is expressed as % inhibition of the eosinophil response of the BALF of vehicle-treated with alternaria lesions.To calculate % inhibition, the number of BALF eosinophils in each condition is converted to the average eosinophils in the BALF of vehicle-treated with alternaria lesions, and subtracted from 100%.

Claims

1. Formula (III) (In the formula, R ２ is hydroxy-C １－６ is alkyl) or a stereoisomer or a pharma- ceutically acceptable salt thereof.

2. R ２ 2. The compound of claim 1, or a stereoisomer or pharma- ceutically acceptable salt thereof, wherein is hydroxyethyl or hydroxypropyl.

3. R ２ 2. The compound of claim 1, or a stereoisomer or pharma- ceutically acceptable salt thereof, wherein is 2-hydroxyethyl, 2-hydroxypropyl, 2-hydroxy-1-methyl-ethyl, or 2-hydroxy-1-methylpropyl.

4. R ２ The compound according to claim 1 or 3, or a stereoisomer or a pharma- ceutically acceptable salt thereof, wherein is 2-hydroxyethyl.

5. R ２ The compound according to claim 1 or 3, or a stereoisomer or a pharma- ceutically acceptable salt thereof, wherein is 2-hydroxypropyl.

6. R ２ The compound according to claim 1 or 3, or a stereoisomer or a pharma- ceutically acceptable salt thereof, wherein is 2-hydroxy-1-methyl-ethyl.

7. R ２ The compound according to claim 1 or 3, or a stereoisomer or a pharma- ceutically acceptable salt thereof, wherein is 2-hydroxy-1-methylpropyl.

8. A compound of the formula:

2. The compound of claim 1, selected from:

9. A compound of the formula:

9. The compound of claim 1 or 8, wherein:

10. A compound of the formula:

9. The compound of claim 1 or 8, wherein:

11. A compound of the formula:

9. The compound of claim 1 or 8, wherein:

12. A compound of the formula:

9. The compound of claim 1 or 8, wherein:

13. A compound of the formula:

9. The compound of claim 1 or 8, wherein:

14. A compound of the formula:

9. The compound of claim 1 or 8, wherein:

15. A pharmaceutical composition for preventing, treating or reducing the severity of 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 disorders, inflammation, neurological disorders, hormone-related diseases, conditions associated with organ transplantation (e.g., graft rejection), immunodeficiency disorders, destructive bone disorders, proliferative disorders, infectious diseases, conditions associated with cell death, thrombin-induced platelet aggregation, liver diseases, pathological immune conditions involving T-cell activation, central nervous system disorders or myeloproliferative disorders, comprising a compound according to any one of claims 1 to 14 or a stereoisomer or pharma-ceutically acceptable salt thereof.

16. 15. A pharmaceutical composition comprising microparticles of a compound according to any one of claims 1 to 14, or a stereoisomer or pharma- ceutically acceptable salt thereof.

17. 17. The pharmaceutical composition of claim 16, wherein the microparticles are prepared by spray drying, freeze drying or micronization.

18. 18. The pharmaceutical composition of claim 16 or 17, further comprising an additive.

19. A pharmaceutical for treating an inflammatory disease comprising a compound according to any one of claims 1 to 14, or a stereoisomer or a pharma- ceutically acceptable salt thereof.

20. The pharmaceutical composition described in claim 19, wherein the inflammatory disease is asthma.

21. 15. Use of a compound according to any one of claims 1 to 14, or a stereoisomer or a pharma- ceutically acceptable salt thereof, for the preparation of a medicament for the treatment of an inflammatory disease.

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

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