HK40010369B - 5-chloro-2-difluoromethoxyphenyl pyrazolopyrimidine compounds which are jak inhibitors - Google Patents

Description

5-Chloro-2-difluoromethoxyphenylpyrazolopyrimidine compounds as JAK inhibitors

This application is a divisional application of PCT application PCT/EP2015/061350, filed on May 22, 2015, entitled “5-Chloro-2-difluoromethoxyphenylpyrazolopyrimidine compounds as JAK inhibitors”. The date on which the PCT application entered the Chinese national phase was November 22, 2016, with application number 201580026680.7.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 62/002,547, filed May 23, 2014, U.S. Provisional Application No. 62/101,234, filed January 8, 2015, U.S. Provisional Application No. 62/130,098, filed March 9, 2015, and International Application No. PCT/CN2015/077176, filed April 22, 2015, each of which is incorporated herein by reference in its entirety.

Technical Field

The field of the invention relates to compounds of formula (00A) and subformulae thereof, which are inhibitors of Janus kinases (e.g., JAK1), and compositions containing these compounds, and methods of use, including but not limited to, diagnosing or treating patients suffering from disorders responsive to inhibition of JAK kinases.

Background Art

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 associated with type I and type II cytokine receptors and regulating cytokine signal transduction. The activation of JAKs associated with the cytokine of the homologous receptor triggers receptors, which results in tyrosine phosphorylation of signal transducers and activators of transcription (STAT) proteins mediated by JAKs, and ultimately leads to transcriptional activation of specific gene sets (Schindler et al., 2007, J.Biol.Chem.282:20059-63). JAK1, JAK2 and TYK2 show a wide range of gene expression patterns, while JAK3 expression is limited to leukocytes. Cytokine receptors typically function as heterodimers, and therefore, more than one type of JAK kinase is typically associated with cytokine receptor complexes. Specific JAKs associated with different cytokine receptor complexes have been determined in many cases by genetic studies and confirmed by other experimental evidence. Exemplary therapeutic benefits of inhibiting JAK enzymes are discussed, for example, in International Application No. WO 2013/014567.

JAK1 was originally identified in a novel kinase screen (Wilks A.F., 1989, Proc. Natl. Acad. Sci. U.S.A. 86: 1603-1607). Genetic and biochemical studies have shown that JAK1 is functionally and physically associated with type I 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 has shown a key role for this kinase in the IFN, IL-10, IL-2/IL-4, and IL-6 pathways. A humanized monoclonal antibody targeting the IL-6 pathway (Tocilizumab) 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).

CD4 T cells play an important role in the pathogenesis of asthma by producing TH2 cytokines including 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 mast cell activation, which exacerbates asthma symptoms (Kearley et al., 2011, Am. J. Resp. Crit. Care Med., 183 (7): 865-875). When combined with the common γ chain or the IL-13Rα1 chain, respectively, the IL-4Rα chain activates JAK1 and binds to IL-4 or IL-13 (Pernis et al., 2002, J. Clin. Invest. 109(10):1279-1283). The common γ chain can also combine with IL-9Rα to bind IL-9, and IL-9Rα also activates JAK1 (Demoulin et al., 1996, Mol. Cell Biol. 16(9):4710-4716). Although the common γ chain activates JAK3, it has been shown that JAK1 is dominant relative to JAK3, and inhibition of JAK1 is sufficient to inactivate signaling through the common γ chain regardless 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 alleviate asthma symptoms in preclinical lung 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).

Biochemical and genetic studies have shown an association between JAK2 and 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 in JAK2 (e.g., JAK2 V617F) are associated with human myeloproliferative diseases.

JAK3 is only associated with the gamma common cytokine receptor chain present in IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21 cytokine receptor complexes. JAK3 is critical for lymphocyte development 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 to immunosuppressive indications (e.g., transplant rejection and rheumatoid arthritis) (Baslund et al., 2005, Arthritis & Rheumatism 52: 2686-2692; Changelian et al., 2003, Science 302: 875-878).

TYK2 is associated with type I interferon (e.g., IFNα), IL-6, IL-10, IL-12, and IL-23 cytokine receptor complexes (Kisseleva et al., 2002, Gene 285: 1-24; Watford, W.T. & O'Shea, J.J., 2006, Immunity 25: 695-697). Consistent with this, primary cells derived from TYK2-deficient humans are defective in type I interferon, IL-6, IL-10, IL-12, and IL-23 signaling. Ustekinumab, a fully human monoclonal antibody targeting the shared p40 subunit of IL-12 and IL-23 cytokines, was recently approved by the European Commission for the treatment of moderate to severe plaque psoriasis (Krueger et al., 2007, N. Engl. J. Med. 356:580-92; Reich et al., 2009, Nat. Rev. Drug Discov. 8:355-356). 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).

There is a need in the art for additional or alternative treatments for disorders mediated by JAK kinases, such as those described above.

Summary of the Invention

Provided herein are 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine compounds that inhibit one or more JAK kinases.

Thus, one aspect of the present invention includes compounds of formula (00A):

and stereoisomers and salts thereof, wherein: R ⁰⁰ is H or CH ₃ ; R ⁰¹ is H or NH ₂ ; R ⁰ is H or NH ₂ ; and ring Q is (i) or (ii):

wherein: ^t1 and ^t2 are each independently 0 or 1; ^XA and ^XB are independently selected from H, _C1 - _C6 alkyl, C2- _C4 alkenyl, ^-NRaRb , _C2 - _C5 ^alkynyl , 3-6- _membered cycloalkyl, 6-10 membered aryl, 3-11 membered heterocycloalkyl, 5-6 membered heterocycloalkenyl and 5-10 membered heteroaryl; wherein when one of ^XA and ^XB is independently _C1 - _C6 alkyl, C2- _C4 alkenyl, _C2 - _C5 alkynyl, 3-6 _- membered cycloalkyl, 6-10 membered aryl, 3-11 membered heterocycloalkyl, 5-6 membered heterocycloalkenyl and 5-10 membered heteroaryl, each ^XA and ^XB is independently optionally substituted by ^Y1 , wherein ^Y1 is selected from:

(a) C ₁ -C ₆ alkyl optionally substituted with T ¹ , wherein T ¹ is selected from OH, halo, CN, imino, 3-6 membered cycloalkyl, 3-11 membered heterocycloalkyl, 3-11 membered heterocycloalkenyl, 5-10 membered heteroaryl, -O-(C ₁ -C ₆ alkyl), C(O)OH, oxetan-3-ylmethyl, -C(O)O-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), -SO ₂ -(C ₁ -C ₆ alkyl), -NR ^a R ^b , -N(+)R ^a R ^b R ^c (wherein R ^c is methyl), -C(O)NR ^a R ^b , -(2-oxoindolin-1-yl), -OC(O)-3-6 membered cycloalkyl and phenyl, wherein T Each alkyl, cycloalkyl, heterocycloalkyl, heterocycloalkenyl, heteroaryl and phenyl group of ¹ is optionally substituted by OH, -C(O)O-(C ₁ -C ₆ alkyl), C ₁ -C ₆ alkyl, halo, CN, oxo, -(C ₁ -C ₆ alkyl)CONR ^a R ^b , -NR ^a R ^b , phenyl or -O-(C ₁ -C ₆ alkyl), which is optionally substituted by OH;

(b) 3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)-3-11-membered heterocycloalkyl, -C(O)-3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)C(O)-3-11-membered heterocycloalkyl, or -OC(O)-4-6-membered heterocycloalkyl; wherein the heterocycloalkyl is optionally substituted with OH, halo, CN, C ₁ -C ₆ alkyl, -(C ₁ -C ₆ alkylene)-CF ₃ , oxo, -C(O)-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , -NR ^a R ^b , -(C ₁ -C ₆ alkylene)- Phenyl or -C(O)-4-6 membered heterocycloalkyl, which is optionally substituted with -NR ^a R ^b ;

(c) N(+)(AA) ₃ , wherein each AA is independently a C ₁ -C ₆ alkyl group optionally substituted with a phenyl group;

(d) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo, NR ^a R ^b or CN;

(e) CN, halo or oxo;

(f) -C(O)-(C ₁ -C ₆ alkyl), -C(O)OH, -C(O)O-(C ₁ -C ₆ alkylene)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , or -C(O)-4-6 membered heterocycloalkyl optionally substituted with -(C ₁ -C ₆ alkyl) or -NR ^a R ^b , or -C(O)O-(C ₁ -C ₆ alkyl) optionally substituted with OH, NR ^a R ^b or 3-11 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with C ₁ -C ₆ alkyl;

(g) OH, -O-phenyl or -O-(C ₁ -C ₆ alkyl), wherein the alkyl group is optionally substituted with: OH or -NR ^a R ^b ;

(h) phenyl optionally substituted with OH, halo, C ₁ -C ₆ alkyl, CF ₃ or CN;

(i) 5-6 membered heteroaryl, which is optionally substituted by OH, halo, C ₁ -C ₆ alkyl, CF ₃ , CN, or 3-11 membered heterocycloalkyl which is optionally substituted by C ₁ -C ₆ alkyl or 3-11 membered heterocycloalkyl;

(j) isoindolin-2-yl optionally substituted by halogen;

(k)-NR ^a R ^b , and

(l) –O-CH ₂ C(O)-3-11-membered heterocycloalkyl;

wherein ^Ra and ^Rb are independently selected from:

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(3-11 membered heterocycloalkyl), -C(O)O-(C ₁ -C ₆ alkyl)-S-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), naphthyl, -NR ^az R ^bz , -C(O)NR ^az R ^bz , oxo, -O-(C ₁ -C ₆ alkyl), 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl, or 3-11 membered heterocycloalkenyl optionally substituted by oxo;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), —S—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo, oxo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) -(C ₁ -C ₆ alkyl) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo, CN or C ₁ -C ₆ alkyl optionally substituted with OH or CN;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

wherein ^Raz and ^Rbz are each independently selected from

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(3-11 membered heterocycloalkyl), -C(O)O-(C ₁ -C ₆ alkyl)-S-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), naphthyl, -oxo, -O-(C ₁ -C ₆ alkyl), 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl, or 3-11 membered heterocycloalkenyl optionally substituted by oxo;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), —S—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo, oxo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) -(C ₁ -C ₆ alkyl) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo, CN or C ₁ -C ₆ alkyl optionally substituted with OH or CN;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

The premise is that when R ⁰ , R ⁰⁰ and R ⁰¹ are each H and ring Q is

wherein t ¹ is 0, then X ^A is not methyl, 2-methylpropan-2-ol, or tetrahydropyranyl; and, in some embodiments, when ring Q is (i) and t ¹ is 0, then X ^A cannot be -NR ^a R ^b .

Furthermore, another aspect of the present invention includes compounds of formula (00A) further defined as compounds of formula (0A):

and stereoisomers and salts thereof, wherein: R ⁰⁰ is H or CH ₃ ; R ⁰¹ is H or NH ₂ ; R ⁰ is H or NH ₂ ; and ring Q is (i) or (ii):

wherein: ^t1 and ^t2 are each independently 0 or 1; ^XA and ^XB are independently selected from H, _C1 - _C6 alkyl, C2- _C4 alkenyl, ^-NRaRb , _C2 - _C5 ^alkynyl , 3-6 _- membered cycloalkyl, 6-10-membered aryl, 3-11-membered heterocycloalkyl, 5-6-membered heterocycloalkenyl and 5-10-membered heteroaryl; wherein when one of ^XA and ^XB is independently _C1 - _C6 alkyl, _C2 - _C5 alkynyl, 3-6-membered cycloalkyl, 6-10-membered aryl, 3-11-membered heterocycloalkyl, 5-6-membered heterocycloalkenyl and 5-10-membered heteroaryl, ^XA and ^XB are each independently optionally substituted by ^Y1 , wherein ^Y1 is selected from:

(a) C ₁ -C ₆ alkyl optionally substituted with T ¹ , wherein T ¹ is selected from OH, halo, CN, imino, 3-6 membered cycloalkyl, 3-11 membered heterocycloalkyl, 3-11 membered heterocycloalkenyl, 5-10 membered heteroaryl, -O-(C ₁ -C ₆ alkyl), C(O)OH, oxetan-3-ylmethyl, -C(O)O-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), -SO ₂ -(C ₁ -C ₆ alkyl), -NR ^a R ^b , -N(+)R ^a R ^b R ^c (wherein R ^c is methyl), -C(O)NR ^a R ^b , -(2-oxoindolin-1-yl), -OC(O)-3-6 membered cycloalkyl and phenyl, wherein T Each alkyl, cycloalkyl, heterocycloalkyl, heterocycloalkenyl, heteroaryl and phenyl group of ¹ is optionally substituted by OH, -C(O)O-(C ₁ -C ₆ alkyl), C ₁ -C ₆ alkyl, halo, CN, oxo, -NR ^a R ^b , phenyl or -O-(C ₁ -C ₆ alkyl), which is optionally substituted by OH;

(b) 3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)-3-11-membered heterocycloalkyl, -C(O)-3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)C(O)-3-11-membered heterocycloalkyl, or -OC(O)-4-6-membered heterocycloalkyl; wherein the heterocycloalkyl is optionally substituted with OH, halo, CN, C ₁ -C ₆ alkyl, -(C ₁ -C ₆ alkylene)-CF ₃ , oxo, -C(O)-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , -NR ^a R ^b , -(C ₁ -C ₆ alkylene)- Phenyl or -C(O)-4-6 membered heterocycloalkyl, which is optionally substituted with -NR ^a R ^b ;

(c) N(+)(AA) ₃ , wherein each AA is independently a C ₁ -C ₆ alkyl group optionally substituted with a phenyl group;

(d) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo or CN;

(e) CN, halo or oxo;

(f) -C(O)-(C ₁ -C ₆ alkyl), -C(O)OH, -C(O)O-(C ₁ -C ₆ alkylene)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , or -C(O)-4-6 membered heterocycloalkyl optionally substituted by -NR ^a R ^b , or -C(O)O-(C ₁ -C ₆ alkyl) optionally substituted by OH, NR ^a R ^b or 3-11 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted by C ₁ -C ₆ alkyl;

(g) OH, -O-phenyl or -O-(C ₁ -C ₆ alkyl), wherein the alkyl group is optionally substituted with: OH or -NR ^a R ^b ;

(h) phenyl optionally substituted with OH, halo, C ₁ -C ₆ alkyl, CF ₃ or CN;

(i) 5-6 membered heteroaryl optionally substituted by OH, halo, C ₁ -C ₆ alkyl, CF ₃ or CN;

(j) isoindolin-2-yl optionally substituted with halo; and

(k)-NR ^a R ^b ，

wherein ^Ra and ^Rb are independently selected from:

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(3-11 membered heterocycloalkyl), -C(O)O-(C ₁ -C ₆ alkyl)-S-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), naphthyl, -NR ^az R ^bz , -C(O)NR ^az R ^bz , oxo, -O-(C ₁ -C ₆ alkyl), 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl, or 3-11 membered heterocycloalkenyl optionally substituted by oxo;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), —S—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo, oxo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) 3-6 membered cycloalkyl, which is optionally substituted by OH, halo, CN or C ₁ -C ₆ alkyl optionally substituted by OH;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

wherein ^Raz and ^Rbz are each independently selected from

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(3-11 membered heterocycloalkyl), -C(O)O-(C ₁ -C ₆ alkyl)-S-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), naphthyl, -oxo, -O-(C ₁ -C ₆ alkyl), 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl, or 3-11 membered heterocycloalkenyl optionally substituted by oxo;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), —S—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo, oxo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) -(C ₁ -C ₆ alkyl) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo, CN or C ₁ -C ₆ alkyl optionally substituted with OH or CN;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

The premise is that when R ⁰ , R ⁰⁰ and R ⁰¹ are each H and ring Q is

wherein t ¹ is 0, then X ^A is not methyl, 2-methylpropan-2-ol, or tetrahydropyranyl; and, in some embodiments, when ring Q is (i) and t ¹ is 0, then X ^A cannot be -NR ^a R ^b .

Another aspect of the present invention includes compounds of formula (00A) further defined as compounds of formula (A),

and stereoisomers and salts thereof, wherein: R ⁰⁰ is H or CH ₃ ; R ⁰¹ is H or NH ₂ ; R ⁰ is H or NH ₂ ; and ring Q is (i) or (ii):

wherein: ^t1 and ^t2 are each independently 0 or 1; ^XA and ^XB are independently selected from H, _C1 - _C6 alkyl, ^-NRaRb , C2- _C5 ^alkynyl , _3-6 -membered cycloalkyl, 6-10-membered aryl, 3-11-membered heterocycloalkyl, 5-6-membered heterocycloalkenyl and 5-10-membered heteroaryl; wherein when one of ^XA and ^XB is independently _C1 - _C6 alkyl, _C2 - _C5 alkynyl, 3-6-membered cycloalkyl, 6-10-membered aryl, 3-11-membered heterocycloalkyl, 5-6-membered heterocycloalkenyl and 5-10-membered heteroaryl, ^XA and ^XB are each independently optionally substituted by ^Y1 , wherein ^Y1 is selected from:

(a) C ₁ -C ₆ alkyl optionally substituted by T ¹ , wherein T ¹ is selected from OH, halo, CN, imino, 3-6 membered cycloalkyl, -O-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl), -SO ₂ -(C ₁ -C ₆ alkyl), -NR ^a R ^b , -N(+) ^Ra R ^b R ^c (wherein R ^c is methyl), -C(O)NR ^a R ^b , -(2-oxoindolin-1-yl), -OC(O)-3-6 membered cycloalkyl and phenyl, wherein each alkyl, cycloalkyl and phenyl of T ¹ is optionally substituted by OH, C ₁ -C ₆ alkyl, halo, CN, oxo, -NR ^a R ^b , phenyl or -O-(C ₁ -C ₆ alkyl) optionally substituted by OH;

(b) 3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)-3-11-membered heterocycloalkyl, -C(O)-3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)C(O)-3-11-membered heterocycloalkyl, or -OC(O)-4-6-membered heterocycloalkyl; wherein the heterocycloalkyl is optionally substituted with OH, halo, CN, C ₁ -C ₆ alkyl, -(C ₁ -C ₆ alkylene)-CF ₃ , oxo, -C(O)-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , -NR ^a R ^b , -(C ₁ -C ₆ alkylene)- Phenyl or -C(O)-4-6 membered heterocycloalkyl, which is optionally substituted with -NR ^a R ^b ;

(c) N(+)(AA) ₃ , wherein each AA is independently a C ₁ -C ₆ alkyl group optionally substituted with a phenyl group;

(d) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo or CN;

(e) CN, halo or oxo;

(f) -C(O)-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl) optionally substituted by OH, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkylene)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , or -C(O)-4-6 membered heterocycloalkyl, which is optionally substituted by -NR ^a R ^b ,

(g) OH, -O-phenyl or -O-(C ₁ -C ₆ alkyl), wherein the alkyl group is optionally substituted with: OH or -NR ^a R ^b ;

(h) phenyl optionally substituted with OH, halo, C ₁ -C ₆ alkyl, CF ₃ or CN;

(i) 5-6 membered heteroaryl optionally substituted by OH, halo, C ₁ -C ₆ alkyl, CF ₃ or CN;

(j) isoindolin-2-yl optionally substituted with halo; and

(k)-NR ^a R ^b ,

wherein ^Ra and ^Rb are independently selected from:

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, naphthyl, -NR ^az R ^bz , -C(O)NR ^az R ^bz , oxo, -O-(C ₁ -C ₆ alkyl), phenyl, a 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) 3-6 membered cycloalkyl, which is optionally substituted by OH, halo, CN or C ₁ -C ₆ alkyl optionally substituted by OH;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

wherein ^Raz and ^Rbz are each independently selected from

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(3-11 membered heterocycloalkyl), -C(O)O-(C ₁ -C ₆ alkyl)-S-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), naphthyl, -oxo, -O-(C ₁ -C ₆ alkyl), 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl, or 3-11 membered heterocycloalkenyl optionally substituted by oxo;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), —S—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo, oxo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) -(C ₁ -C ₆ alkyl) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo, CN or C ₁ -C ₆ alkyl optionally substituted with OH or CN;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

The premise is that when R ⁰ , R ⁰⁰ and R ⁰¹ are each H and ring Q is

wherein t ¹ is 0, then X ^A is not methyl, 2-methylpropan-2-ol, or tetrahydropyranyl; and, in some embodiments, when ring Q is (i) and t ¹ is 0, then X ^A cannot be -NR ^a R ^b .

Also provided are pharmaceutical compositions comprising a 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine as described herein, such as a compound of formula (00A), and a pharmaceutically acceptable carrier, diluent, or excipient.

In some embodiments, the present invention also provides the use of a 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine compound as described herein, such as a compound of formula (00A), in therapy, for example, in the treatment of an inflammatory disease. Also provided is the use of a 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine compound as described herein, such as a compound of formula (00A), in the preparation of a medicament for treating an inflammatory disease. Also provided is a method for preventing, treating, or lessening 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 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine compound, such as a compound of formula (00A).

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing aspects and many of the attendant advantages will become more readily appreciated and understood by reference to the following detailed description when taken in conjunction with the accompanying drawings, in which:

Figure 1 depicts a matched pair analysis of certain compounds of the invention containing OMe (i) or _OCHF2 (ii) groups at the indicated positions.

Detailed Description of the Invention

definition

"Halogen" or "halo" refers to F, Cl, Br or I. Additionally, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl.

The term "alkyl" refers to a saturated, linear or branched, monovalent hydrocarbon radical, wherein the alkyl group may be optionally substituted. In one example, the alkyl group is 1 to 18 carbon atoms (C ₁ -C ₁₈ ). In other examples, the alkyl group is C ₀ -C ₆ , C ₀ -C ₅ , C ₀ -C ₃ , C ₁ -C ₁₂ , C ₁ -C ₁₀ , C ₁ -C ₈ , C ₁ -C ₆ , C ₁ -C ₅ , C ₁ -C ₄ or C ₁ -C ₃ . C ₀ alkyl refers to a bond. Examples of alkyl groups include methyl (Me, -CH ₃ ), ethyl (Et, -CH ₂ CH ₃ ), 1-propyl (n-Pr, n-propyl, -CH ₂ CH ₂ CH ₃ ), 2-propyl (i-Pr, isopropyl, -CH(CH ₃ ) ₂ ), 1-butyl (n-Bu, n-butyl, -CH ₂ CH ₂ CH ₂ CH ₃ ), 2-methyl-1-propyl (i-Bu, isobutyl, -CH ₂ CH(CH ₃ ) ₂ ), 2-butyl (s-Bu, sec-butyl, -CH(CH ₃ )CH ₂ CH ₃ ), 2-methyl-2-propyl (t-Bu, tert-butyl, -C(CH ₃ ) ₃ ), 1-pentyl (n-pentyl, -CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-pentyl (-CH(CH ₃ )CH 2 CH ₂ CH _{3 ),} CH ₃ ), 3-pentyl (-CH(CH ₂ CH ₃ ) ₂ ), 2-methyl-2-butyl (-C(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl (-CH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butyl (-CH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butyl (-CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), 1-hexyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-hexyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₂ CH ₃ ), 3-hexyl (-CH(CH ₂ CH ₃ )(CH ₂ CH ₂ CH ₃ )), 2-methyl-2-pentyl (-C(CH ₃ ) ₂ CH ₂ CH ₂ CH ₃ ), 3-methyl-2-pentyl (—CH(CH ₃ )CH(CH ₃ )CH ₂ CH ₃ ), 4-methyl-2-pentyl (—CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ ), 3-methyl-3-pentyl (—C(CH ₃ )(CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl (—CH(CH ₂ CH ₃ )CH(CH ₃ ) ₂ ), 2,3-dimethyl-2-butyl (—C(CH ₃ ) ₂ CH(CH ₃ ) ₂ ), 3,3-dimethyl-2-butyl (—CH(CH ₃ )C(CH ₃ ) ₃ , 1-heptyl, and 1-octyl. In some embodiments, the substituents of “optionally substituted alkyl” include F, Cl, Br, I, OH, SH, CN, NH ₂ , NHCH ₃ , N(CH ₃ ) ₂ , NO ₂ , N ₃ , C(O)CH ₃ , COOH, CO ₂ CH ₃ , methyl, ethyl, propyl, isopropyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methylsulfonylamino, SO, SO ₂ , phenyl, piperidinyl, piperazinyl and pyrimidinyl, wherein the alkyl, phenyl and heterocyclic moieties may be optionally substituted, for example, with 1 to 4 substituents selected from this same list.

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), wherein the alkenyl group may be optionally substituted and includes groups having "cis" and "trans" orientations, or "E" and "Z" orientations. In one example, the alkenyl group is 2 to 18 carbon atoms ( _C2 - _C18 ). In other examples, the alkenyl group is _C2 - _C12 , _C2 - _C10 , _C2 - _C8 , _C2 - _C6 , or _C2 - _C3 . Examples include, but are not limited to, vinyl (—CH═CH ₂ ), prop-1-enyl (—CH═CHCH ₃ ), prop-2-enyl (—CH ₂ CH═CH ₂ ), 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, buta-1,3-dienyl, 2-methylbut-1,3-diene, 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 examples selected from 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, methylsulfonylamino, SO, _SO2 , phenyl, piperidinyl, piperazinyl, and pyrimidinyl, wherein the alkyl, phenyl, and heterocyclic moieties may be optionally substituted, for example, with 1 to 4 substituents selected from this same list.

The term "alkynyl" refers to a straight or branched monovalent hydrocarbon radical having at least one site of unsaturation (i.e., a carbon-carbon triple bond), wherein the alkynyl radical may be optionally substituted. In one example, the alkynyl radical is 2 to 18 carbon atoms ( _C2 - _C18 ). In other examples, the alkynyl radical is _C2 - _C12 , _C2 - _C10 , _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 of "optionally substituted alkynyl" include 1 to 4 examples selected from 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, methylsulfonylamino, SO, _SO2 , phenyl, piperidinyl, piperazinyl, and pyrimidinyl, wherein the alkyl, phenyl, and heterocyclic moieties can be optionally substituted, for example, with 1 to 4 substituents selected from this same list.

"Alkylene" refers to a saturated, branched, or straight-chain hydrocarbon radical having two monovalent radical centers derived by removing two hydrogen atoms from the same or two different carbon atoms of a parent alkane. In one example, a divalent alkylene radical is 1 to 18 carbon atoms ( _Ci - _Ci8 ). In other examples, a divalent alkylene radical is _C0 - _C6 , _C0 - _C5 , _C0 - _C3 , _C1 - _C12 , _C1 - _C10 , _C1 - _C8 , _C1 - _C6 , _C1 - _C5 , _C1 - _C4 , or _C1 - _C3 . The group _C0 alkylene refers to a bond. Example alkylene groups include methylene (—CH ₂ —), 1,1-ethyl (—CH(CH ₃ )—), (1,2-ethyl (—CH ₂ CH ₂ —), 1,1-propyl (—CH(CH ₂ CH ₃ )—), 2,2-propyl (—C(CH ₃ ) ₂ —), 1,2-propyl (—CH(CH ₃ )CH ₂ —), 1,3-propyl (—CH ₂ CH ₂ CH ₂ —), 1,1-dimethyleth-1,2-yl (—C(CH ₃ ) ₂ CH ₂ —), 1,4-butyl (—CH ₂ CH ₂ CH ₂ CH ₂ —), and the like.

The term "heteroalkyl" refers to a straight or branched monovalent hydrocarbon radical consisting of the specified number of carbon atoms (or, if not specified, up to 18 carbon atoms) and from 1 to 5 heteroatoms selected from O, N, Si, and S, and wherein the nitrogen and sulfur atoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized. In some embodiments, the heteroatom is selected from O, N, and S, wherein the nitrogen and sulfur atoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized. The heteroatom may be located at any interior position of the heteroalkyl group, including the position where 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 may be consecutive, for example, -CH2 _- NH- _OCH3 and _-CH2 -O-Si( _CH3 ) ₃ . Heteroalkyl groups may be optionally substituted. In some embodiments, substituents of "optionally substituted heteroalkyl" include 1 to 4 examples selected from 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, methylsulfonylamino, SO, _SO2 , phenyl, piperidinyl, piperazinyl, and pyrimidinyl, wherein the alkyl, phenyl, and heterocyclic moieties can be optionally substituted, for example, with 1 to 4 substituents selected from this same list.

"Amino" refers to optionally substituted primary amines (i.e., _-NH2 ), secondary amines (i.e., -NRH), tertiary amines (i.e., -NRR), and quaternary amines (i.e., -N(+)RRR), wherein each R is the same or different and is selected from alkyl, cycloalkyl, aryl, and heterocyclyl, wherein alkyl, cycloalkyl, aryl, and heterocyclyl are as defined herein. Particular secondary and tertiary amines are alkylamines, dialkylamines, arylamines, diarylamines, aralkylamines, and diaralkylamines, wherein the alkyl and aryl portions can be optionally substituted. Particular secondary and tertiary amines are methylamine, ethylamine, propylamine, isopropylamine, aniline, benzylamine, dimethylamine, diethylamine, dipropylamine, and diisopropylamine. In some embodiments, each R group of the quaternary amine is independently an optionally substituted alkyl group.

"Aryl" refers to a carbocyclic aromatic group having the specified number of carbon atoms (or, if no number is specified, up to 14 carbon atoms), whether or not fused to one or more groups. One example includes an aryl group having 6-14 carbon atoms. Another example includes an aryl group having 6-10 carbon atoms. Examples of aryl groups include phenyl, naphthyl, biphenyl, phenanthrenyl, naphthacene, 1,2,3,4-tetrahydronaphthyl, 1H-indenyl, 2,3-dihydro-1H-indenyl, and the like (see, for example, Lang's Handbook of Chemistry (Dean, JA, ed.) ^13th ed. Table 7-2 [1985]). A specific aryl group is phenyl. Substituted phenyl or substituted aryl refers to phenyl or aryl substituted with one, two, three, four or five substituents, e.g., 1-2, 1-3 or 1-4 substituents, selected, for example, from the groups indicated herein (see definition of "optionally substituted"), such as F, Cl, Br, I, OH, SH, CN, _NH2 , _NHCH3 , N( _CH3 ) ₂ , _NO2 , _N3 , C(O) _CH3 , COOH, _CO2CH3 , methyl, _ethyl , propyl, isopropyl, butyl, isobutyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methylsulfonylamino, SO, _SO2 , phenyl, piperidinyl, piperazinyl and pyrimidinyl, wherein the alkyl, phenyl and heterocyclic moieties may be optionally substituted, for example, with 1 to 4 substituents selected from this same list. Examples of the term "substituted phenyl" include mono- or di-(halo)phenyl groups such as 2-chlorophenyl, 2-bromophenyl, 4-chlorophenyl, 2,6-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-bromophenyl, 3,4-dibromophenyl, 3-chloro-4-fluorophenyl, 2-fluorophenyl, 2,4-difluorophenyl, and the like; mono- or di-(hydroxy)phenyl groups such as 4-hydroxyphenyl, 3-hydroxyphenyl, 2,4-dihydroxyphenyl, protected-hydroxy derivatives thereof, and the like; nitrophenyl groups such as 3- or 4-nitrophenyl; cyanophenyl groups such as 4-cyanophenyl; mono- or di-(alkyl)phenyl groups such as 4-methylphenyl, 2,4-dimethylphenyl, 2-methylphenyl, 4-(isopropyl)phenyl, 4-ethylphenyl, 3-(n-propyl)phenyl, and the like; mono- or di-(alkoxy)phenyl groups such as phenyl, 3-(protected hydroxymethyl)phenyl, 3,4-dimethoxyphenyl, 3-methoxy-4-benzyloxyphenyl, 3-ethoxyphenyl, 4-(isopropoxy)phenyl, 4-(tert-butoxy)phenyl, 3-ethoxy-4-methoxyphenyl, and the like; 3- or 4-trifluoromethylphenyl; a mono- or dicarboxyphenyl or (protected carboxy)phenyl group, such as 4-carboxyphenyl, a mono- or di-(hydroxymethyl)phenyl or (protected hydroxymethyl)phenyl, such as 3-(protected hydroxymethyl)phenyl or 3,4-di(hydroxymethyl)phenyl; a mono- or di-(aminomethyl)phenyl or (protected aminomethyl)phenyl, such as 2-(aminomethyl)phenyl or 2,4-(protected aminomethyl)phenyl; or a mono- or di-(N-(methylsulfonylamino))phenyl, such as 3-(N-methylsulfonylamino))phenyl. Furthermore, the term "substituted phenyl" refers to disubstituted phenyl groups having different substituents, such as 3-methyl-4-hydroxyphenyl, 3-chloro-4-hydroxyphenyl, 2-methoxy-4-bromophenyl, 4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl, 2-hydroxy-4-chlorophenyl, 2-chloro-5-difluoromethoxy, and the like, as well as trisubstituted phenyl groups having different substituents, such as 3-methoxy-4-benzyloxy-6-methylsulfonylamino, 3-methoxy-4-benzyloxy-6-phenylsulfonylamino, and tetrasubstituted phenyl groups having different substituents, such as 3-methoxy-4-benzyloxy-5-methyl-6-phenylsulfonylamino. In some embodiments, the substituents of the aryl group (e.g., phenyl) comprise amides. For example, an aryl (e.g., phenyl) substituent may be -(CH ₂ ) _0-4 CONR'R", wherein R' and R" each independently refer to a group including, for example, hydrogen; unsubstituted C ₁ -C ₆ alkyl; C ₁ -C ₆ alkyl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C _{1 -C 6} alkoxy, oxo, or NR'R"; unsubstituted C ₁ -C ₆ heteroalkyl; C 1 -C ₆ heteroalkyl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo, or NR'R"; unsubstituted C ₆ -C ₁₀ aryl; C ₆ -C ₁₀ aryl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, or NR'R"; unsubstituted _3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S); and 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S) substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo or NR'R"; or R' and R" can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring, wherein the ring atoms are optionally substituted with N, O or S, and wherein the ring is optionally substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C 1 -C ₆ alkoxy, oxo or NR'R"; ₆ alkoxy, oxo or NR'R" substituted.

"Cycloalkyl" refers to a non-aromatic, saturated or partially unsaturated hydrocarbon ring group, wherein the cycloalkyl group may be optionally substituted independently by one or more substituents described herein. In one example, the cycloalkyl group is 3 to 12 carbon atoms ( _C3 - _C12 ). In other examples, the cycloalkyl group is _C3 - _C8 , _C3 - _C10 , or _C5 - _C10 . In other examples, the cycloalkyl group as a monocycle is _C3 - _C8 , _C3 - _C6 , or _C5 - _C6 . In another example, the cycloalkyl group as a bicycle is _C7 - _C12 . In another example, the cycloalkyl group as a spiro system is _C5 - _C12 . Examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, perdeuterated cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. Exemplary arrangements of bicyclic cycloalkyls with 7 to 12 ring atoms include, but are not limited to, [4,4], [4,5], [5,5], [5,6], or [6,6] ring systems. Exemplary bridged bicyclic cycloalkyls include, but are not limited to, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, and bicyclo[3.2.2]nonane. Examples of spirocycloalkyl 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, the substituents of "optionally substituted cycloalkyl" include ₁ to 4 of the following examples: 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, methylsulfonylamino, SO, _SO2 , phenyl, piperidinyl, piperazinyl, and pyrimidinyl, wherein the alkyl, aryl, and heterocyclic moieties may be optionally substituted, for example, with 1 to 4 substituents selected from this same list. In some embodiments, the substituents of the cycloalkyl group include amides. For example, the cycloalkyl substituent can be -(CH ₂ ) _0-4 CONR'R", wherein R' and R" each independently refer to a group including, for example, hydrogen; unsubstituted C ₁ -C ₆ alkyl; C ₁ -C ₆ alkyl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo, or NR'R"; unsubstituted C 1 -C ₆ heteroalkyl; C ₁ -C 6 heteroalkyl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo, or NR'R"; unsubstituted C ₆ -C ₁₀ aryl; C ₆ -C ₁₀ aryl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C _{1 -C 6} alkoxy, or NR'R"; unsubstituted _3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S); and 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S) substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo or NR'R"; or R' and R" can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring, wherein the ring atoms are optionally substituted with N, O or S, and wherein the ring is optionally substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C 1 -C 6 alkoxy, oxo or NR'R _" ; -C ₆ alkoxy, oxo or NR'R" substituted.

"Guandinium" or "guanidinium" refers to the group -NH-C(NH)-NHR, where R is hydrogen, alkyl, cycloalkyl, aryl, or heterocyclyl, wherein alkyl, cycloalkyl, aryl, and heterocyclyl are as defined herein. A specific guanidine is the group -NH-C(NH)-NH ₂ .

"Heterocyclic group,""heterocyclic,""heterocycle," or "heterocyclyl" are used interchangeably and refer to any monocyclic, bicyclic, tricyclic, or spirocyclic, saturated or unsaturated, aromatic (heteroaryl) or non-aromatic (e.g., heterocycloalkyl) ring system having from 3 to 20 ring atoms, wherein the ring atoms are carbon and at least one atom in the ring or ring system is a heteroatom selected from nitrogen, sulfur, or oxygen. A ring system is a heterocycle if any of the ring atoms is a heteroatom, regardless of the point of attachment of the ring system to the rest of the molecule. In one example, a heterocyclyl comprises 3-11 ring atoms ("members") and includes monocyclic, bicyclic, tricyclic, and spirocyclic ring systems wherein the ring atoms are carbon and wherein at least one atom in the ring or ring system is a heteroatom selected from nitrogen, sulfur, or oxygen. In one example, a heterocyclyl contains from 1 to 4 heteroatoms. In one example, a heterocyclyl contains from 1 to 3 heteroatoms. In another example, the heterocyclyl group includes a 3- to 7-membered monocyclic ring having 1-2, 1-3, or 1-4 heteroatoms selected from nitrogen, sulfur, or oxygen. In another example, the heterocyclyl group includes a 4- to 6-membered monocyclic ring having 1-2, 1-3, or 1-4 heteroatoms selected from nitrogen, sulfur, or oxygen. In another example, the heterocyclyl group includes a 3-membered monocyclic ring. In another example, the heterocyclyl group includes a 4-membered monocyclic ring. In another example, the heterocyclyl group includes a 5- to 6-membered monocyclic ring, such as a 5- to 6-membered heteroaryl group. In another example, the heterocyclyl group includes a 3- to 11-membered heterocycloalkyl group, such as a 4- to 11-membered heterocycloalkyl group. In some embodiments, the heterocycloalkyl group includes at least one nitrogen. In one example, the heterocyclyl group includes 0 to 3 double bonds. Any nitrogen or sulfur heteroatom may be optionally oxidized (e.g., NO, SO, SO ₂ ), and any nitrogen heteroatom may be optionally quaternized (e.g., [NR ₄ ] ⁺ Cl ⁻ , [NR ₄ ] ⁺ OH ⁻ ). Example heterocycles are oxiranyl, aziridine, thiiridine, azetidinyl, oxetanyl, thietanyl, 1,2-dithietanyl, 1,3-dithietanyl, pyrrolidinyl, dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydrofuranyl, dihydrothiophenyl, tetrahydrothiophenyl, imidazolidinyl, piperidinyl, piperazinyl, isoquinolinyl, tetrahydroisoquinolinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, dihydro pyranyl, tetrahydropyranyl, hexahydrothiopyranyl, hexahydropyrimidinyl, oxazinyl, thiazinyl, thiaoxanyl, homopiperazinyl, homopiperidinyl, azepanyl, oxepanyl, thiepanyl, oxazepinyl, oxazepanyl, diazepanyl, 1,4-diazepanyl, diazepinyl, thiazepinyl, thiazepanyl, tetrahydrothiopyranyl, oxazolidinyl, thiazolidinyl, isothiazolidinyl, 1,1-dioxoisothiazolidinone, oxazolidinone , imidazolidinone, 4,5,6,7-tetrahydro[2H]indazolyl, tetrahydrobenzimidazolyl, 4,5,6,7-tetrahydrobenzo[d]imidazolyl, 1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridinyl, thiazinyl, oxazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl, oxathiazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl, imidazolinyl, dihydropyrimidinyl, tetrahydropyrimidinyl, 1-pyrrolinyl, 2-pyrrolinyl 1,3-Dioxolanyl, pyrazolinyl, pyrazolidinyl, dithianyl, dithiolanyl, pyrimidinyl, pyrimidinedione, pyrimidine-2,4-dione, piperazinyl, piperazinyl, pyrazolidinyl imidazolinyl, 3-azabicyclo[3.1.0]hexyl, 3,6-diazabicyclo[3.1.1]heptanyl, 6-azabicyclo[3.1.1] heptane, 3-azabicyclo[3.1.1]heptanyl, 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexyl, 2-azabicyclo[3.2.1]octyl, 8-azabicyclo[3.2.1]octyl, 2-azabicyclo[2.2.2]octyl, 8-azabicyclo[2.2.2]octyl, 7-oxabicyclo[2.2.1]heptane, azaspiro[3.5]nonyl, azaspiro[2.5]octyl, azaspiro[4.5]decyl, 1-azaspiro[4.5]decan-2-onyl, azaspiro[5.5]undecyl, tetrahydroindolyl, octahydroindolyl, tetrahydroisoindolyl, tetrahydroindazolyl, 1,1-dioxohexahydrothiopyranyl. Examples of 5-membered heterocycles containing sulfur or oxygen atoms and one to three nitrogen atoms include thiazolyl, including thiazol-2-yl and thiazol-2-yl N-oxide; thiadiazolyl, including 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl; oxazolyl, such as oxazol-2-yl; and oxadiazolyl, such as 1,3,4-oxadiazol-5-yl and 1,2,4-oxadiazol-5-yl. Examples of 5-membered heterocycles containing two to four nitrogen atoms include imidazolyl, such as imidazol-2-yl; triazolyl, such as 1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl; and tetrazolyl, such as 1H-tetrazol-5-yl. Examples of benzo-fused 5-membered heterocycles include benzoxazol-2-yl, benzothiazol-2-yl, and benzimidazol-2-yl. Examples of 6-membered heterocyclic rings contain one to three nitrogen atoms and optionally sulfur or oxygen atoms, such as pyridyl, such as pyridin-2-yl, pyridin-3-yl, and pyridin-4-yl; pyrimidinyl, such as pyrimidin-2-yl and pyrimidin-4-yl; triazinyl, such as 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, particularly pyridazin-3-yl, and pyrazinyl. Pyridine N-oxide and pyridazine N-oxide, and pyridinyl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazinyl, and 1,3,4-triazin-2-yl are other exemplary heterocyclic rings. The heterocyclic ring may be optionally substituted. For example, substituents for "optionally substituted heterocycle" include 1 to 4 of the following examples: 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, methylsulfonylamino, SO, _SO2 , phenyl, piperidinyl, piperazinyl, and pyrimidinyl, wherein the alkyl, aryl, and heterocycle moieties may be optionally substituted with, for example, 1 to 4 substituents selected from this same list. In some embodiments, substituents for heterocycles, such as heteroaryl or heterocycloalkyl, include amides. For example, a heterocycle (e.g., heteroaryl or heterocycloalkyl) substituent can be -(CH ₂ ) _0-4 CONR'R", wherein R' and R" each independently refer to a group including, for example, hydrogen; unsubstituted C ₁ -C ₆ alkyl; C 1 -C 6 alkyl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo, or NR'R"; unsubstituted C ₁ -C ₆ heteroalkyl; C ₁ -C ₆ heteroalkyl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo, or NR'R"; unsubstituted C ₆ -C ₁₀ aryl; C ₆ -C ₁₀ alkyl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C _{1 -C 6} alkoxy, oxo, or NR'R"; ₁₀ aryl; unsubstituted 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S); and 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1 _{to 4} heteroatoms selected from O, N and S or 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S) substituted with halogen, OH, CN, unsubstituted C 1 -C ₆ alkyl, unsubstituted C ₁ -C 6 alkoxy, oxo or NR'R"; or R' and R" can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring, wherein the ring atoms are optionally substituted with N, O or S, and wherein the ring is optionally substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C 1 -C ₆ alkoxy, oxo or NR'R"; ₆ alkoxy, oxo or NR'R" substituted.

"Heteroaryl" refers to any monocyclic, bicyclic, or tricyclic ring system in which at least one ring is a 5- or 6-membered aromatic ring containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur. In an exemplary embodiment, at least one heteroatom is nitrogen. See, for example, Lang's Handbook of Chemistry (Dean, JA, ed.), 13th edition. Table 7-2 [1985]. Included in the definition are any bicyclic groups in which any of the above heteroaryl rings is fused to an aryl ring, wherein the aryl ring or heteroaryl ring is attached to the rest of the molecule. In one embodiment, heteroaryl includes 5-6 membered monocyclic aromatic groups in which one or more of the ring atoms is nitrogen, sulfur, or oxygen. Example heteroaryl groups include thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl, tetrazolo[1,5-b]pyridazinyl, imidazo[1,2-a]pyrimidinyl and purinyl, as well as benzo-fused derivatives such as benzoxazolyl, benzofuranyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzimidazolyl and indolyl. Heteroaryl groups can be optionally substituted. In some embodiments, the substituents of "optionally substituted heteroaryl" include 1 to 4 examples selected from 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, methylsulfonylamino, SO, _SO2 , phenyl, piperidinyl, piperazinyl, and pyrimidinyl, wherein the alkyl, phenyl, and heterocyclic moieties can be optionally substituted with 1 to 4 substituents selected from the same list. In some embodiments, the substituents of heteroaryl include amides. For example, the heteroaryl substituent can be -(CH ₂ ) _{0- 4} CONR'R", wherein R' and R" each independently refer to a group including, for example, hydrogen; unsubstituted C ₁ -C ₆ alkyl; C ₁ -C ₆ alkyl substituted with halogen, OH, CN, unsubstituted C 1 -C 6 alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo, or NR'R"; unsubstituted C ₁ -C ₆ heteroalkyl; C ₁ -C ₆ heteroalkyl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo, or NR'R"; unsubstituted C ₆ -C ₁₀ aryl; C _{6 -C 10} aryl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, or NR'R"; unsubstituted _3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S); and 3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S or 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S) substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo or NR'R"; or R' and R" can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring, wherein the ring atoms are optionally substituted with N, O or S, and wherein the ring is optionally substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C 1 -C ₆ alkoxy, oxo or NR'R"; ₆ alkoxy, oxo or NR'R" substituted.

In a specific embodiment, the heterocyclyl group is attached at a carbon atom of the heterocyclyl group. For example, carbon-bonded heterocyclyl groups include bonding arrangements at the 2, 3, 4, 5, or 6 position of a pyridine ring, the 3, 4, 5, or 6 position of a pyridazine ring, the 2, 4, 5, or 6 position of a pyrimidine ring, the 2, 3, 5, or 6 position of a pyrazine ring, the 2, 3, 4, or 5 position of a furan, tetrahydrofuran, thiophene, thiophene, pyrrole, or tetrahydropyrrole ring, the 2, 4, or 5 position of an oxazole, imidazole, or thiazole ring, the 3, 4, or 5 position of an isoxazole, pyrazole, or isothiazole ring, the 2 or 3 position of an aziridine ring, the 2, 3, or 4 position of an azetidine ring, the 2, 3, 4, 5, 6, 7, or 8 position of a quinoline ring, or the 1, 3, 4, 5, 6, 7, or 8 position of an isoquinoline ring.

In certain embodiments, the heterocyclic group is N-linked. For example, nitrogen-linked heterocyclic or heteroaryl groups include bonding arrangements at the following positions: 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 1, isoindole or isoindoleline 2, morpholine 4, and carbazole or β-carboline 9.

The term "alkoxy" refers to a straight or branched monovalent group represented by the formula -OR, wherein R is an alkyl group as defined herein. Alkoxy includes methoxy, ethoxy, propoxy, isopropoxy, mono-, di- and tri-fluoromethoxy and cyclopropyloxy.

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

Unless otherwise indicated, "optionally substituted" means that the group can be unsubstituted or substituted by one or more (e.g., 0, 1, 2, 3, 4, or 5 or more, or any range derivatizable therein) of the substituents listed for the group, wherein the substituents can be the same or different. In one embodiment, the optionally substituted group has 1 substituent. In another embodiment, the optionally substituted group has 2 substituents. In another embodiment, the optionally substituted group has 3 substituents. In another embodiment, the optionally substituted group has 4 substituents. In another embodiment, the optionally substituted group has 5 substituents.

Optional substituents for alkyl (alone or as part of another substituent such as alkoxy) and alkylene, alkenyl, alkynyl, heteroalkyl, heterocycloalkyl and cycloalkyl (also each alone or as part of another substituent) can be various groups, such as those described herein, and selected from halogen; oxo; CN; NO; N ₃ ; -OR'; perfluoro-C ₁ -C ₄ alkoxy; unsubstituted C ₃ -C ₇ cycloalkyl; C 3 -C ₇ cycloalkyl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo or NR'R"; unsubstituted C ₆ -C ₁₀ aryl (e.g., phenyl); C ₆ -C 10 aryl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C _{1 -C 6} alkoxy or NR'R"; unsubstituted _{3-11 membered heterocyclyl (e.g., 5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S, or 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S); 3-11} membered heterocyclyl (e.g., _5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S, or _4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S) substituted by halogen, OH, CN, unsubstituted C ₁ -C _{6 alkyl, unsubstituted C 1 -C 6} alkoxy, oxo or NR'R";-NR'R";-SR';-SiR'R"R"';-OC(O)R';-C(O)R'; -CO ₂ R';-CONR'R"; -OC -(CH 2 ) 1-4 -OC(O)R'; - _{( CH 2} ) _1-4 -C(O)R'; - ₍ CH _{2 ) 1-4 -CO 2} R _' ; _{and - (} CH _{2 ) 1-4} CONR'R _" or _{a combination thereof ,} The number thereof is from 0 to (2m'+1), wherein m' is the total number of carbon atoms in the group. R', R" and R"' each independently refer to a group including, for example, hydrogen; unsubstituted C ₁ -C ₆ alkyl; C ₁ -C ₆ alkyl substituted by halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo or NR'R"; unsubstituted C 1 -C ₆ heteroalkyl; C ₁ -C ₆ heteroalkyl substituted by halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C 6 alkoxy, oxo or NR'R"; unsubstituted C ₆ -C ₁₀ aryl; C ₆ -C ₁₀ aryl substituted by halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C _{1 -C 6} alkoxy or NR'R"; unsubstituted _3-11 membered heterocyclyl (for example, a 5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S, or a 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S); and a 3-11 membered heterocyclyl (for example, a _5-6 membered heteroaryl containing 1 to ₄ heteroatoms selected from O, N and S, or a 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S) substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C 1 -C 6 alkoxy, oxo or NR'R". When R' and R" are attached to the same nitrogen atom, they can combine with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring, wherein the ring atoms are optionally substituted with N, O, or S, and wherein the ring is optionally substituted with halogen, OH, CN, _C1 - _C6 alkyl, unsubstituted _C1 - _C6 alkoxy, oxo, or NR'R". For example, -NR'R" is intended to include 1-pyrrolidinyl and 4-morpholinyl.

Similarly, the optional substituents for aryl and heteroaryl are varied. In some embodiments, the substituents for aryl and heteroaryl are selected from halogen; CN; NO; N ₃ ; -OR '; perfluoro-C _1- C ₄ alkoxy; unsubstituted C ₃ -C ₇ cycloalkyl; C 3 -C 7 cycloalkyl substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo or NR'R "; unsubstituted C ₆ -C ₁₀ aryl (e.g., phenyl); C ₆ -C _{10 aryl} substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C _{1 -C 6} alkoxy or NR'R "; unsubstituted 3-11 membered heterocyclyl (e.g., containing 1 to 4 members selected from O, N and S substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo or NR'R" (for example, a 5-6 membered heteroaryl group containing 1 to 4 heteroatoms selected from O, N and S, or a 4-11 membered heterocycloalkyl group containing 1 to 4 heteroatoms selected from O, N and S); -NR'R";-SR';-SiR'R"R"';-OC(O)R';-C(O)R'; -CO 2 R';-CONR'R"; -OC(O) _NR'R ";-NR"C(O)R';-NR"'C(O)NR'R";-NR"C(O) ₂ R'; -S(O) ₂ R'; -S(O) _2NR'R ";-NR'S(O) _2R ";-NR"'S(O)_2NR'R";amidino;guanidino;-(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 ) _1-4CONR'R ", or combinations thereof, in a number from 0 to (2m'+1), where m' is the total number of carbon atoms in such a group. R', R" and R"' each independently refer to a group including, for example, hydrogen; unsubstituted C ₁ -C ₆ alkyl; C 1 -C 6 alkyl substituted by halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo or NR'R"; unsubstituted C ₁ -C ₆ heteroalkyl; C ₁ -C ₆ heteroalkyl substituted by halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₁ -C ₆ alkoxy, oxo or NR'R"; unsubstituted C ₆ -C ₁₀ aryl; C ₆ -C ₁₀ aryl substituted by halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C _{1 -C 6} alkoxy or NR'R"; unsubstituted _3-11 membered heterocyclyl (for example, a 5-6 membered heteroaryl containing 1 to 4 heteroatoms selected from O, N and S, or a 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S); and a 3-11 membered heterocyclyl (for example, a _5-6 membered heteroaryl containing 1 to ₄ heteroatoms selected from O, N and S, or a 4-11 membered heterocycloalkyl containing 1 to 4 heteroatoms selected from O, N and S) substituted with halogen, OH, CN, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C 1 -C 6 alkoxy, oxo or NR'R". When R' and R" are attached to the same nitrogen atom, they can combine with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring, wherein the ring atoms are optionally substituted with N, O, or S, and wherein the ring is optionally substituted with halogen, OH, CN, unsubstituted _C1 - _C6 alkyl, unsubstituted _C1 - _C6 alkoxy, oxo, or NR'R". For example, -NR'R" is intended to include 1-pyrrolidinyl and 4-morpholinyl.

The term "oxo" refers to =0 or (=0) ₂ .

As used herein, a wavy line through a bond in a chemical structure indicates the point of attachment of the wavy bond to an atom in the chemical structure that is connected to the rest of the molecule or to the rest of a fragment of a molecule. In some embodiments, an arrow is used in conjunction with an asterisk in a wavy manner to indicate a point of attachment.

In certain embodiments, a divalent group is generally described without a specific bonding configuration. It should be understood that, unless otherwise specified, the general description is intended to include both bonding configurations. For example, in the group R ¹ —R ² —R ³ , if the group R ² is described as —CH ₂ C(O)—, it should be understood that the group can be bonded as R ¹ —CH ₂ C(O)—R ³ and R ¹ —C(O)CH ₂ —R ³ , unless otherwise specified.

The phrase "pharmaceutically acceptable" refers to molecular entities and compositions that do not produce adverse, allergic or other untoward reactions when properly administered to animals, such as humans.

The compounds of the present invention may be in the form of salts, such as pharmaceutically acceptable salts. "Pharmaceutically acceptable salts" include acid and base addition salts. "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic acids and organic acids that retain the biological effectiveness and properties of the free base and are not biologically or otherwise undesirable, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and the like; and organic acids that can be selected from aliphatic, alicyclic, aromatic, aromatic alicyclic, heterocyclic, carboxylic and sulfonic acids, such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, maloneic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, pamoic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

"Pharmaceutically acceptable base addition salts" include those derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Specific base addition salts are ammonium, potassium, sodium, calcium, and magnesium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, tromethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrazine, 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.

In some embodiments, the salt is selected from hydrochloride, hydrobromide, trifluoroacetate, sulfate, phosphate, acetate, fumarate, maleate, tartrate, lactate, citrate, pyruvate, succinate, oxalate, methanesulfonate, p-toluenesulfonate, bisulfate, benzenesulfonate, ethanesulfonate, malonate, xinafoate, ascorbate, oleate, nicotinate, saccharinate, adipate, formate, glycolate, palmitate, L-lactate, D-lactate, aspartate, malate, L-tartrate, D-tartrate, stearate, furoate (e.g., 2-furoate or 3-furoate), naphthalene disulfonate (naphthalene-1,5-disulfonate or naphthalene-1-(sulfonic acid)-5-sulfonate), ethane disulfonate (ethane [0014] Examples of the salts of the present invention include benzoate, adipate, ethanesulfonate, malonate, mesitylenesulfonate (2-mesitylenesulfonate), naphthalenesulfonate (2-naphthalenesulfonate), camphorsulfonate (camphor-10-sulfonate, such as (1S)-(+)-10-camphorsulfonate), glutamate, glutarate, hippurate (2-(benzamido)acetate), orotate, xylenesulfonate (p-xylene-2-sulfonate), and pamoate (2,2'-dihydroxy-1,1'-dinaphthylmethane-3,3'-dicarboxylate).

A "sterile" preparation is sterile or free from all viable microorganisms and their spores.

"Stereoisomers" refer to compounds that 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.

"Chiral" refers to molecules that have the property of non-superimposability of their mirror image partner, while the term "achiral" refers to molecules that are superimposable on their mirror image partner.

"Diastereoisomers" refers to stereoisomers that have two or more chiral centers and whose molecules are not mirror images of one another. Diastereoisomers have different physical properties, such as melting points, boiling points, spectral properties, or biological activities. Mixtures of diastereoisomers can be separated by high-resolution analytical methods such as electrophoresis and chromatography, such as HPLC.

"Enantiomers" refers to two stereoisomers of a compound that are non-superimposable mirror images of one another.

The stereochemical definitions and conventions used herein generally follow those of S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule about its chiral center. The prefixes d and l or (+) and (-) are used to indicate the sign by which the compound rotates plane-polarized light, where (-) or l indicates that the compound is left-handed. Compounds prefixed with (+) or d are right-handed. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers may also be referred to as enantiomers, and mixtures of these isomers are often referred to as enantiomeric mixtures. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur in a chemical reaction or process without stereoselectivity or stereospecificity. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers that is devoid of optical activity.

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, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions via reorganization of some of the bonding electrons.

Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. "Solvate" refers to an association or complex of one or more solvent molecules and a compound of the present invention. Examples of solvents that form solvates include water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. Certain compounds of the present invention may exist in a variety of crystalline or amorphous forms. Generally, all physical forms are within the scope of the present invention. The term "hydrate" refers to a complex in which the solvent molecule is water.

"Metabolite" refers to a product produced by metabolism of a specified compound or a salt thereof in vivo. Such a product may result, for example, from oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, or the like of the administered compound.

Metabolites are typically identified by preparing a radiolabeled (e.g., ¹⁴ C or ³ H) isotope of a compound of the invention, administering it to an animal, such as a rat, mouse, guinea pig, monkey, or human, at a detectable dose (e.g., greater than about 0.5 mg/kg), allowing sufficient time for metabolism to occur (usually about 30 seconds to 30 hours), and isolating its conversion products from urine, blood, or other biological samples. These products are easily isolated because they are labeled (others are isolated by using antibodies that can bind to epitopes that survive in the metabolites). The metabolite structure is determined in a conventional manner, such as by MS, LC/MS, or NMR analysis. Generally, analysis of metabolites is performed in the same manner as conventional drug metabolism studies known to those skilled in the art. Metabolites, as long as they are not otherwise found in vivo, can be used in diagnostic assays for therapeutic doses of the compounds of the invention.

As used herein, "amino-protecting group" refers to a derivative of a group that is typically used to block or protect an amino group while reacting on other functional groups of a compound. Examples of such protecting groups include carbamates, amides, alkyl and aryl groups, as well as imines, and many N-heteroatom derivatives, which can be removed to regenerate the desired amine group. Particular amino-protecting groups are Pmb (p-methoxybenzyl), Boc (tert-butyloxycarbonyl), Fmoc (9-fluorenylmethoxycarbonyl), and Cbz (benzyloxycarbonyl). Other examples of these groups are found in T.W. Greene and P.G.M. Wuts, "Protecting Groups in Organic Synthesis," 3rd edition, John Wiley & Sons, Inc., 1999. The term "protected amino" refers to an amino group substituted with one of the above-mentioned amino-protecting groups.

As used herein, "carboxyl-protecting groups" refer to those groups that are stable to subsequent reaction conditions elsewhere in the molecule and can be removed at an appropriate point without disrupting the remainder of the molecule to yield an unprotected carboxyl group. Examples of carboxyl-protecting groups include ester groups and heterocyclic groups. Ester derivatives of carboxylic acid groups can be used to block or protect the carboxylic acid group while reactions proceed at other functional groups in the compound. Examples of such ester groups include substituted arylalkyl groups, including substituted benzyl groups such as 4-nitrobenzyl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl, 2,4,6-trimethylbenzyl, pentamethylbenzyl, 3,4-methylenedioxybenzyl, benzhydryl, 4,4′-dimethoxybenzhydryl, 2,2′,4,4′-tetramethoxybenzhydryl, alkyl or substituted alkyl esters such as methyl, ethyl, t-butylallyl or t-amyl, triphenylmethyl (trityl), 4-methoxytrityl, 4,4′-dimethoxytrityl, 4,4′,4″-tetramethoxytrityl. Trimethoxytrityl, 2-phenylpropan-2-yl, thioesters such as tert-butylthioester, silyl esters such as trimethylsilyl, tert-butyldimethylsilyl ester, phenacyl, 2,2,2-trichloroethyl, β-(trimethylsilyl)ethyl, β-(di(n-butyl)methylsilyl)ethyl, p-tolylsulfonylethyl, 4-nitrobenzylsulfonylethyl, allyl, cinnamyl, 1-(trimethylsilylmethyl)prop-1-en-3-yl and similar groups. Another example of a carboxyl protecting group is a heterocyclic group such as 1,3-oxazolinyl. Other examples of these groups can be found in T.W. Greene and P.G.M. Wuts, "Protecting Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons, Inc., 1999. The term "protected carboxyl" refers to a carboxyl group substituted with one of the above-mentioned carboxyl protecting groups.

As used herein, "hydroxy-protecting group" refers to a hydroxy derivative that is typically used to block or protect the hydroxy group while reacting on other functional groups of the compound. Examples of such protecting groups include tetrahydropyranyloxy, benzoyl, acetoxy, carbamoyloxy, benzyl, and silyl ether (e.g., TBS, TBDPS) groups. Other examples of these groups are found in TW Greene and PGM Wuts, "Protecting Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons, Inc., 1999. The term "protected hydroxy" refers to a hydroxy group that is substituted by one of the above-mentioned hydroxy protecting groups.

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., cattle), sports 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 comprising administering to a patient a compound of Formula (OOA), (OA), (A), (la), (lb), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, the patient is typically in need thereof.

The term "Janus kinase" refers to JAK1, JAK2, JAK3, and TYK2 protein kinases. In some embodiments, a Janus kinase can be further defined as one of JAK1, JAK2, JAK3, or TYK2. In any embodiment, any one of JAK1, JAK2, JAK3, and TYK2 can 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.

The terms "inhibit" and "reduce" or any variation of these terms include any measurable reduction or complete inhibition that achieves a desired result. For example, an activity (e.g., JAK1 activity) can be reduced by about, up to about, or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more, or any range derivable therefrom, compared to normal.

In some embodiments, a compound of Formula (OOA), (OA), (A), (la), (lb), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, selectively inhibits JAK1 relative to JAK3 and TYK2. In some embodiments, a compound of Formula (OOA), (OA), (A), (la), (lb), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, selectively inhibits JAK1 relative to JAK2, JAK3, or TYK2, or any combination of JAK2, JAK3, or TYK2. In some embodiments, a compound of Formula (OOA), (OA), (A), (la), (lb), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468 selectively inhibits JAK1 and JAK2 relative to JAK3 and TYK2. In some embodiments, a compound of Formula (OOA), (OA), (A), (la), (lb), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468 selectively inhibits JAK1 relative to JAK3. By "selectively inhibit" is meant that the compound is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more, or any range derivable therein, a better inhibitor of the activity of a specific Janus kinase (e.g., JAK1) as compared to the activity of another specific Janus kinase (e.g., JAK1), or at least 2-, 3-, 4-, 5-, 10-, 25-, 50-, 100-, 250-, or 500-fold better inhibitor of the activity of a specific Janus kinase (e.g., JAK1) as compared to the activity of another specific Janus kinase (e.g., JAK1).

"Therapeutically effective amount" refers to an amount of a compound of the invention, e.g., a compound of Formula (OOA), (OA), (A), (la), (lb), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, that (i) treats or prevents a particular disease, disorder, or condition, or (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, disorder, or condition, and optionally (iii) prevents or delays the onset of one or more symptoms of a particular disease, disorder, or condition described herein. In some embodiments, a therapeutically effective amount is an amount sufficient to reduce or alleviate the symptoms of an autoimmune or inflammatory disease (e.g., asthma). In some embodiments, a therapeutically effective amount is an amount of a chemical entity described herein that is sufficient to significantly reduce B-cell activity or number. In the case of cancer, a therapeutically effective amount of a drug can reduce the number of cancer cells; reduce tumor size; inhibit (i.e., slow down to some extent and preferably stop) cancer cell infiltration into peripheral organs; inhibit (i.e., slow down to some extent and preferably stop) tumor metastasis; inhibit tumor growth to some extent; or alleviate one or more symptoms associated with cancer to some extent. To the extent that a drug can prevent growth or kill existing cancer cells, it can be cytostatic or cytotoxic. For cancer treatment, efficacy can be measured, for example, by assessing the time to disease progression (TTP) or determining the response rate (RR).

"Treatment" (and variants such as "treat" or "treating") refers to a clinical intervention that attempts to alter the natural course of the individual or cell being treated, and can be used for prevention or performed during the course of clinical pathology. Desirable effects of treatment include preventing the occurrence or recurrence of the disease, alleviating symptoms, alleviating any direct or indirect pathological consequences of the disease, stabilizing (i.e., not worsening) the disease state, reducing the rate of disease progression, improving or alleviating the disease state, prolonging survival and alleviating or improving prognosis compared to expected survival if not receiving treatment. In some embodiments, a compound of the invention, such as a compound of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, is used to delay the development of a disease or condition or to slow the progression of a disease or condition. Those in need of treatment include those already suffering from the disease or condition as well as those predisposed to having the disease or condition (e.g., through a genetic mutation) or those in which the disease or condition is to be prevented.

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

"Inflammation" refers to a local protective reaction caused by the damage or destruction of tissue, which is used to destroy, dilute or isolate (isolate) the tissue of the damaging agent and the damage. Inflammation is particularly relevant to the influx of leukocytes or neutrophil chemotaxis. Inflammation can be caused by infection and non-infectious modes (such as reperfusion after trauma or myocardial infarction or stroke, immune response to exogenous antigens and autoimmune response) of pathogenic organisms and viruses. Therefore, inflammatory diseases suitable for the treatment of compounds of this invention such as formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX) or (X) compounds or Table 1 or Examples 1-468 compounds include diseases related to the reaction of the specific defense system and the reaction of the nonspecific defense system.

"Specific defense system" refers to the components of the immune system that react to the presence of specific antigens. Examples of inflammation caused by responses of the specific defense system include classical responses to foreign antigens, autoimmune diseases, and delayed-type 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 additional examples of inflammatory responses of the specific defense system.

The term "nonspecific defense system" refers to inflammatory conditions mediated by leukocytes (e.g., granulocytes and macrophages) that are incapable of immunological memory. Examples of inflammation that is at least partially caused by a response of the nonspecific defense system include inflammation associated with, for example, adult (acute) respiratory distress syndrome (ARDS) or multiple organ injury syndrome; reperfusion injury; acute glomerulonephritis; reactive arthritis; skin diseases with an acute inflammatory component; acute purulent meningitis or other central nervous system inflammatory conditions such as stroke; thermal injury; inflammatory bowel disease; granulocyte transfusion-related syndrome; and cytokine-induced toxicity.

"Autoimmune disease" refers to any of a group of diseases in which tissue damage is associated with a humoral or cell-mediated response to the body's own constituents. Non-limiting examples of autoimmune diseases include rheumatoid arthritis, lupus, and multiple sclerosis.

As used herein, "allergic disease" refers to any symptom, tissue damage or loss of tissue function caused by an allergy. As used herein, "arthritic disease" refers to any disease characterized by inflammatory damage to the joints attributable to a variety of causes. As used herein, "dermatitis" refers to any of a large family of skin diseases characterized by inflammation of the skin attributable to a variety of causes. As used herein, "transplant rejection" refers to any immune response to transplanted tissue, such as an organ or cell (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 treating conditions associated with inflammatory cell activation.

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

In some embodiments, inflammatory disorders treatable 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, and delayed hypersensitivity reactions.

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

A "chemotherapeutic agent" is an active agent that can be used to treat a given condition (e.g., cancer or inflammatory conditions). Examples of chemotherapeutic agents are well known in the art and include examples such as those disclosed in U.S. Publ. Appl. No. 2010/0048557, incorporated herein by reference. In addition, chemotherapeutic agents include pharmaceutically acceptable salts, acids, or derivatives of any chemotherapeutic agent, and combinations of two or more thereof.

"Package insert" is used to refer to instructions customarily included in commercial packages of therapeutic products, that contain information about the indications, usage, dosage, administration, contraindications, or warnings concerning the use of such therapeutic products.

Unless otherwise indicated, the terms "compounds of the invention" and "compounds of the invention" and the like include compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, and stereoisomers (including atropisomers), geometric isomers, tautomers, solvates, metabolites, isotopes, salts (e.g., pharmaceutically acceptable salts), and prodrugs thereof. In some embodiments, solvates, metabolites, isotopes, or prodrugs, or any combination thereof, are excluded.

Unless otherwise indicated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into compounds of the invention, such as compounds of Formula (OOA), (OA), (A), (la), (lb), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as ^2H , ^3H , ^11C , ^13C , ^14C , ^13N , ^15N , ^15O , ^17O , ^18O , ^32P , ^33P , ^35S , ^18F , ^36Cl , ^123I , and ^125I , respectively. Isotopically labeled compounds (e.g., those labeled with ^3H and ^14C ) are useful in compound or substrate tissue distribution assays. Tritiated (i.e. ³ H) and carbon-14 (i.e. ¹⁴ C) isotopes can be useful because of their ease of preparation and detectability. In addition, substitution with heavier isotopes such as deuterium (i.e. ² H) can provide certain therapeutic advantages (e.g., increased in vivo half-life or reduced dosage requirements) resulting from greater metabolic stability. In some embodiments, in a compound of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX) or (X) or a compound of Table 1 or Examples 1-468, one or more hydrogen atoms are replaced by ² H or ³ H, and one or more carbon atoms are replaced by ¹³ C- or ¹⁴ C- enriched carbon. Positron emitting isotopes such as ¹⁵ O, ¹³ N, ¹¹ C and ¹⁸ F can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds can generally be prepared by following procedures analogous to those disclosed in the Schemes or Examples herein, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

It is particularly contemplated that any limitation discussed with respect to one embodiment of the present invention may be applied to any other embodiment of the present invention. In addition, any compound 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 produce or utilize any compound or composition of the present invention.

Use of the term "or" is intended to mean "and/or" unless explicitly indicated that only alternatives are involved or the alternatives are mutually exclusive, although this disclosure supports definitions indicating only alternatives and "and/or."

In this application, the term "about" is used to indicate a value that includes the standard deviation of error for the device or method being employed to determine the value.

As used herein, "a" or "an" refers to one or more, unless expressly stated otherwise. As used herein, "another" refers to at least another or more.

The headings used in this article are for organizational purposes only.

Inhibitors of JANUS kinase

Thus, one aspect of the present invention includes compounds of formula (00A):

and stereoisomers and salts thereof, wherein: R ⁰⁰ is H or CH ₃ ; R ⁰¹ is H or NH ₂ ; R ⁰ is H or NH ₂ ; and ring Q is (i) or (ii):

wherein: ^t1 and ^t2 are each independently 0 or 1; ^XA and ^XB are independently selected from H, _C1 - _C6 alkyl, C2- _C4 alkenyl, ^-NRaRb , _C2 - _C5 ^alkynyl , 3-6- _membered cycloalkyl, 6-10 membered aryl, 3-11 membered heterocycloalkyl, 5-6 membered heterocycloalkenyl and 5-10 membered heteroaryl; wherein when one of ^XA and ^XB is independently _C1 - _C6 alkyl, C2- _C4 alkenyl, _C2 - _C5 alkynyl, 3-6 _- membered cycloalkyl, 6-10 membered aryl, 3-11 membered heterocycloalkyl, 5-6 membered heterocycloalkenyl and 5-10 membered heteroaryl, each ^XA and ^XB is independently optionally substituted by ^Y1 , wherein ^Y1 is selected from:

(a) C ₁ -C ₆ alkyl optionally substituted by T ¹ , wherein T ¹ is selected from OH, halo, CN, imino, 3-6 membered cycloalkyl, 3-11 membered heterocycloalkyl, 3-11 membered heterocycloalkenyl, 5-10 membered heteroaryl, -O-(C ₁ -C ₆ alkyl), C(O)OH, oxetan-3-ylmethyl, -C(O)O-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), -SO ₂ -(C ₁ -C ₆ alkyl), -NR ^a R ^b , -N(+)Ra ^{R b} R ^c wherein R ^c is methyl, -C(O)NR ^a R ^b , -(2-oxoindolin-1-yl), -OC(O)-3-6 membered cycloalkyl and phenyl, wherein each alkyl, cycloalkyl, heterocycloalkyl, heterocycloalkenyl, heteroaryl and phenyl of T ¹ is optionally substituted by OH, -C(O)O-(C ₁ -C ₆ alkyl), C ₁ -C ₆ alkyl, halo, CN, oxo, -(C ₁ -C ₆ alkyl)CONR ^a R ^b , -NR ^a R ^b , phenyl or -O-(C ₁ -C ₆ alkyl), which is optionally substituted with OH;

(b) 3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)-3-11-membered heterocycloalkyl, -C(O)-3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)C(O)-3-11-membered heterocycloalkyl, or -OC(O)-4-6-membered heterocycloalkyl; wherein the heterocycloalkyl is optionally substituted with OH, halo, CN, C ₁ -C ₆ alkyl, -(C ₁ -C ₆ alkylene)-CF ₃ , oxo, -C(O)-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , -NR ^a R ^b , -(C ₁ -C ₆ alkylene)- Phenyl or -C(O)-4-6 membered heterocycloalkyl, which is optionally substituted with -NR ^a R ^b ;

(c) N(+)(AA) ₃ , wherein each AA is independently a C ₁ -C ₆ alkyl group optionally substituted with a phenyl group;

(d) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo, NR ^a R ^b or CN;

(e) CN, halo or oxo;

(f) -C(O)-(C ₁ -C ₆ alkyl), -C(O)OH, -C(O)O-(C ₁ -C ₆ alkylene)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , or -C(O) ^-4-6 membered heterocycloalkyl optionally substituted with -(C ₁ -C ₆ alkyl) or -NR ^a R b, or -C(O)O-(C ₁ -C ₆ alkyl) optionally substituted with OH, NR ^a R ^b , or 3-11 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with C ₁ -C ₆ alkyl;

(g) OH, -O-phenyl or -O-(C ₁ -C ₆ alkyl), wherein the alkyl group is optionally substituted with: OH or -NR ^a R ^b ;

(h) phenyl optionally substituted with OH, halo, C ₁ -C ₆ alkyl, CF ₃ or CN;

(i) 5-6 membered heteroaryl optionally substituted by OH, halo, C ₁ -C ₆ alkyl, CF ₃ , CN, or 3-11 membered heterocycloalkyl optionally substituted by C ₁ -C ₆ alkyl or 3-11 membered heterocycloalkyl;

(j) isoindolin-2-yl optionally substituted by halogen;

(k)-NR ^a R ^b , and

(l) –O-CH ₂ C(O)-3-11-membered heterocycloalkyl;

wherein ^Ra and ^Rb are independently selected from:

(p)H,

(a) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(3-11 membered heterocycloalkyl), -C(O)O-(C ₁ -C ₆ alkyl)-S-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), naphthyl, -NR ^az R ^bz , -C(O)NR ^az R ^bz , oxo, -O-(C ₁ -C ₆ alkyl), 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl, or 3-11 membered heterocycloalkenyl optionally substituted by oxo;

(b) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(c) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O-(C ₁ -C ₆ alkyl), —S-(C ₁ -C ₆ alkyl), or —O-phenyl;

(d) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo, oxo or C ₁ -C ₆ alkyl;

(e) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(f) -(C ₁ -C ₆ alkyl) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo, CN or C ₁ -C ₆ alkyl optionally substituted with OH or CN;

(g) C ₂ -C ₅ alkenyl;

(h) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(i) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(j) phenyl,

(k) -C(O)(C ₁ -C ₆ alkyl),

(1) -C(O)O(C ₁ -C ₆ alkyl),

(m)-C(O)O(3-6 membered cycloalkyl), and

(n)-C(O)-phenyl,

wherein ^Raz and ^Rbz are each independently selected from

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(3-11 membered heterocycloalkyl), -C(O)O-(C ₁ -C ₆ alkyl)-S-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), naphthyl, -oxo, -O-(C ₁ -C ₆ alkyl), 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl, or 3-11 membered heterocycloalkenyl optionally substituted by oxo;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), —S—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo, oxo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) -(C ₁ -C ₆ alkyl) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo, CN, or C ₁ -C ₆ alkyl optionally substituted with OH or CN;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

The premise is that when R ⁰ , R ⁰⁰ and R ⁰¹ are each H and ring Q is

wherein t ¹ is 0, then X ^A is not methyl, 2-methylpropan-2-ol, or tetrahydropyranyl; and, in some embodiments, when ring Q is (i) and t ¹ is 0, then X ^A cannot be -NR ^a R ^b .

Furthermore, another aspect of the present invention includes compounds of formula (00A) further defined as compounds of formula (0A):

and stereoisomers and salts thereof, wherein: R ⁰⁰ is H or CH ₃ ; R ⁰¹ is H or NH ₂ ; R ⁰ is H or NH ₂ ; and ring Q is (i) or (ii):

wherein: ^t1 and ^t2 are each independently 0 or 1; ^XA and ^XB are independently selected from H, _C1 - _C6 alkyl, C2- _C4 alkenyl, ^-NRaRb , _C2 - _C5 ^alkynyl , 3-6 _- membered cycloalkyl, 6-10-membered aryl, 3-11-membered heterocycloalkyl, 5-6-membered heterocycloalkenyl and 5-10-membered heteroaryl; wherein when one of ^XA and ^XB is independently _C1 - _C6 alkyl, _C2 - _C5 alkynyl, 3-6-membered cycloalkyl, 6-10-membered aryl, 3-11-membered heterocycloalkyl, 5-6-membered heterocycloalkenyl and 5-10-membered heteroaryl, ^XA and ^XB are each independently optionally substituted by ^Y1 , wherein ^Y1 is selected from:

(a) C ₁ -C ₆ alkyl optionally substituted with T ¹ , wherein T ¹ is selected from OH, halo, CN, imino, 3-6 membered cycloalkyl, 3-11 membered heterocycloalkyl, 3-11 membered heterocycloalkenyl, 5-10 membered heteroaryl, -O-(C ₁ -C ₆ alkyl), C(O)OH, oxetan-3-ylmethyl, -C(O)O-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), -SO ₂ -(C ₁ -C ₆ alkyl), -NR ^a R ^b , -N(+)R ^a R ^b R ^c (wherein R ^c is methyl), -C(O)NR ^a R ^b , -(2-oxoindolin-1-yl), -OC(O)-3-6 membered cycloalkyl and phenyl, wherein T Each alkyl, cycloalkyl, heterocycloalkyl, heterocycloalkenyl, heteroaryl and phenyl group of ¹ is optionally substituted by OH, -C(O)O-(C ₁ -C ₆ alkyl), C ₁ -C ₆ alkyl, halo, CN, oxo, -NR ^a R ^b , phenyl or -O-(C ₁ -C ₆ alkyl) optionally substituted by OH;

(b) 3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)-3-11-membered heterocycloalkyl, -C(O)-3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)C(O)-3-11-membered heterocycloalkyl, or -OC(O)-4-6-membered heterocycloalkyl; wherein the heterocycloalkyl is optionally substituted with OH, halo, CN, C ₁ -C ₆ alkyl, -(C ₁ -C ₆ alkylene)-CF ₃ , oxo, -C(O)-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , -NR ^a R ^b , -(C ₁ -C ₆ alkylene)- Phenyl or -C(O)-4-6 membered heterocycloalkyl, which is optionally substituted with -NR ^a R ^b ;

(c) N(+)(AA) ₃ , wherein each AA is independently a C ₁ -C ₆ alkyl group optionally substituted with a phenyl group;

(d) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo or CN;

(e) CN, halo or oxo;

(f) -C(O)-(C ₁ -C ₆ alkyl), -C(O)OH, -C(O)O-(C ₁ -C ₆ alkylene)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , or -C(O)-4-6 membered heterocycloalkyl optionally substituted by -NR ^a R ^b , or -C(O)O-(C ₁ -C ₆ alkyl) optionally substituted by OH, NR ^a R ^b , or 3-11 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted by C ₁ -C ₆ alkyl;

(g) OH, -O-phenyl or -O-(C ₁ -C ₆ alkyl), wherein the alkyl group is optionally substituted with: OH or -NR ^a R ^b ;

(h) phenyl optionally substituted with OH, halo, C ₁ -C ₆ alkyl, CF ₃ or CN;

(i) 5-6 membered heteroaryl optionally substituted by OH, halo, C ₁ -C ₆ alkyl, CF ₃ or CN;

(j) isoindolin-2-yl optionally substituted with halo; and

(k)-NR ^a R ^b ,

wherein ^Ra and ^Rb are independently selected from:

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(3-11 membered heterocycloalkyl), -C(O)O-(C ₁ -C ₆ alkyl)-S-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), naphthyl, -NR ^az R ^bz , -C(O)NR ^az R ^bz , oxo, -O-(C ₁ -C ₆ alkyl), 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl, or 3-11 membered heterocycloalkenyl optionally substituted by oxo;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), —S—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo, oxo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) 3-6 membered cycloalkyl, which is optionally substituted by OH, halo, CN or C ₁ -C ₆ alkyl optionally substituted by OH;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

wherein ^Raz and ^Rbz are each independently selected from

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(3-11 membered heterocycloalkyl), -C(O)O-(C ₁ -C ₆ alkyl)-S-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), naphthyl, -oxo, -O-(C ₁ -C ₆ alkyl), 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl, or 3-11 membered heterocycloalkenyl optionally substituted by oxo;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), —S—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo, oxo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) -(C ₁ -C ₆ alkyl) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo, CN, or C ₁ -C ₆ alkyl optionally substituted with OH or CN;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

The premise is that when R ⁰ , R ⁰⁰ and R ⁰¹ are each H and ring Q is

wherein t ¹ is 0, then X ^A is not methyl, 2-methylpropan-2-ol, or tetrahydropyranyl; and, in some embodiments, when ring Q is (i) and t ¹ is 0, then X ^A cannot be -NR ^a R ^b .

In some embodiments of any of the formulae herein, when ring Q is

and ^t1 is 0, the pyrazole is not N-linked to the nitrogen of ^XA .

Another aspect of the present invention provides a compound of formula (00A) further defined as a compound of formula (A),

and stereoisomers and salts thereof, wherein:

R ⁰⁰ is H or CH ₃ ;

R ⁰¹ is H or NH ₂ ;

R ⁰ is H or NH ₂ ; and

Ring Q is (i) or (ii):

in:

^t1 and ^t2 are each independently 0 or 1;

^XA and ^XB are independently selected from H, _C1 - _C6 alkyl, ^-NRaRb , _C2 - _C5 ^alkynyl , 3-6-membered cycloalkyl, 6-10-membered aryl, 3-11-membered heterocycloalkyl, 5-6-membered heterocycloalkenyl, and 5-10-membered heteroaryl;

wherein when one of ^XA and ^XB is independently _C1 - _C6 alkyl, _C2 - _C5 alkynyl, 3-6-membered cycloalkyl, 6-10-membered aryl, 3-11-membered heterocycloalkyl, 5-6-membered heterocycloalkenyl and 5-10-membered heteroaryl, ^XA and ^XB are each independently optionally substituted by ^Y1 , wherein ^Y1 is selected from:

(a) C ₁ -C ₆ alkyl optionally substituted by T ¹ , wherein T ¹ is selected from OH, halo, CN, imino, 3-6 membered cycloalkyl, -O-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl), -SO ₂ -(C ₁ -C ₆ alkyl), -NR ^a R ^b , -N(+)R ^a R ^b R ^c wherein R ^c is methyl, -C(O)NR ^a R ^b , -(2-oxoindolin-1-yl), -OC(O)-3-6 membered cycloalkyl and phenyl,

wherein each alkyl, cycloalkyl and phenyl group of T ¹ is optionally substituted by OH, C ₁ -C ₆ alkyl, halo, CN, oxo, -NR ^a R ^b , phenyl or -O-(C ₁ -C ₆ alkyl), which is optionally substituted by OH;

(b) 3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)-3-11-membered heterocycloalkyl, -C(O)-3-11-membered heterocycloalkyl, -(C ₁ -C ₆ alkylene)C(O)-3-11-membered heterocycloalkyl, or -OC(O)-4-6-membered heterocycloalkyl;

wherein heterocycloalkyl is optionally substituted by OH, halo, CN, C ₁ -C ₆ alkyl, -(C ₁ -C ₆ alkylene)-CF ₃ , oxo, -C(O)-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkylene)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , -NR ^a R ^b , -(C ₁ -C ₆ alkylene)-phenyl, or -C(O)-4-6 membered heterocycloalkyl, which is optionally substituted by -NR ^a R ^b ;

(c) N(+)(AA) ₃ , wherein each AA is independently a C ₁ -C ₆ alkyl group optionally substituted with a phenyl group;

(d) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo or CN;

(e) CN, halo or oxo;

(f) -C(O)-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl) optionally substituted by OH, -C(O)O-(C ₁ -C ₆ alkylene)-phenyl, -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)NR ^a R ^b , or -C(O)-4-6 membered heterocycloalkyl, which is optionally substituted by -NR ^a R ^b ,

(g) OH, -O-phenyl or -O-(C ₁ -C ₆ alkyl), wherein the alkyl group is optionally substituted with OH or -NR ^a R ^b ;

(h) phenyl optionally substituted with OH, halo, C ₁ -C ₆ alkyl, CF ₃ or CN;

(i) 5-6 membered heteroaryl optionally substituted by OH, halo, C ₁ -C ₆ alkyl, CF ₃ or CN;

(j) isoindolin-2-yl optionally substituted with halo; and

(k)-NR ^a R ^b ，

wherein ^Ra and ^Rb are independently selected from:

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, naphthyl, -NR ^az R ^bz , -C(O)NR ^az R ^bz , oxo, -O-(C ₁ -C ₆ alkyl), phenyl, a 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) 3-6 membered cycloalkyl, which is optionally substituted by OH, halo, CN or C ₁ -C ₆ alkyl optionally substituted by OH;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

wherein ^Raz and ^Rbz are each independently selected from

(a)H,

(b) C ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, -C(O)OH, -C(O)O-(C ₁ -C ₆ alkyl), -C(O)O-(3-11 membered heterocycloalkyl), -C(O)O-(C ₁ -C ₆ alkyl)-S-(C ₁ -C ₆ alkyl), -S-(C ₁ -C ₆ alkyl), naphthyl, -oxo, -O-(C ₁ -C ₆ alkyl), 5-6 membered heteroaryl optionally substituted by C ₁ -C ₆ alkyl or halo, or benzo[1,3]dioxol-2-yl, or 3-11 membered heterocycloalkenyl optionally substituted by oxo;

(c) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the alkylene is optionally substituted with OH, halo or CN;

(d) -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkylene group is optionally substituted with halo, and the phenyl group is optionally substituted with OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl), —S—(C ₁ -C ₆ alkyl), or —O-phenyl;

(e) -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with halo, oxo or C ₁ -C ₆ alkyl;

(f) -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with halo, C ₁ -C ₆ alkyl, or -O-phenyl;

(g) -(C ₁ -C ₆ alkyl) 3-6 membered cycloalkyl, which is optionally substituted with OH, halo, CN or C ₁ -C ₆ alkyl optionally substituted with OH or CN;

(h) C ₂ -C ₅ alkenyl;

(i) a 4- to 6-membered heterocycloalkyl group, which is optionally substituted with halogen,

(j) -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl substituted with hydroxymethyl,

(k) phenyl,

(1) -C(O)(C ₁ -C ₆ alkyl),

(m)-C(O)O(C ₁ -C ₆ alkyl),

(n)-C(O)O(3-6 membered cycloalkyl), and

(o)-C(O)-phenyl,

The premise is:

When R ⁰ , R ⁰⁰ and R ⁰¹ are each H and ring Q is

Where ^t1 is 0,

Then ^XA is not methyl, 2-methylpropan-2-ol, or tetrahydropyranyl; and, in some embodiments, when ring Q is (i) and ^t1 is 0, then ^XA cannot be ^{-NRaRb .}

In some embodiments, the compound of formula (00A), (0A), or (A) is further defined as a compound of formula (II):

in:

w is 0 or 1;

R ^0b is H or NH ₂ ;

R ^1b is selected from

a.H,

bC ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, CN, —O—(C ₁ -C ₆ alkyl), naphthyl, 5-6 membered heteroaryl, or —C(O)NR ^v R ^w , wherein R ^v and R ^w are independently H or C ₁ -C ₆ alkyl optionally substituted by halo,

c. -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the cycloalkyl is optionally substituted by CN,

d. -(C ₁ -C ₆ alkylene)-phenyl, wherein the alkyl group is optionally substituted by halo, and wherein the phenyl group is optionally substituted by OH, halo, CF ₃ , C ₁ -C ₆ alkyl, —O—(C ₁ -C ₆ alkyl) or —O-phenyl,

e. -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted by: halo or C ₁ -C ₆ alkyl,

f. -(C ₁ -C ₆ alkylene)-O-phenyl, wherein the phenyl group is optionally substituted with: halo or C ₁ -C ₆ alkyl, or

g. 3-6 membered cycloalkyl;

R ^2b is selected from H, C ₁ -C ₆ alkyl or C ₂ -C ₅ alkenyl optionally substituted by halogen;

or R ^1b and R ^2b together form a 3-11 membered heterocycloalkyl group, which is optionally substituted with OH;

R ^3b is absent or is methyl, wherein when R ^3b is methyl, the nitrogen to which it is attached is N+ and w is 1;

R ^4b is a bond or a C ₁ -C ₆ alkylene group;

R ^5b is selected from H, OH and phenyl; and

Ring F is a 3-7 membered heterocycloalkyl group, wherein p ^1b is 0, 1 or 2, and p ^2b is 0, 1 or 2;

wherein w is equal to 1 only when R ^3b is methyl.

In some embodiments, the compound of formula (00A), (0A) or (A) is further defined as a compound of formula (Ia) or formula (Ib):

in:

R ^0a is H or NH ₂ ;

R ^1a is bonded to the nitrogen atom of ring A in (Ia) or to the carbon atom of ring B in (Ib), and is selected from:

a.H,

bC ₁ -C ₆ alkyl, which is optionally substituted by OH, halo, —O—(C ₁ -C ₆ alkyl), —SO ₂ —(C ₁ -C ₆ alkyl), —(2-oxoindolin-1-yl), —OC(O)-3-6 membered cycloalkyl, —OC(O)-4-6 membered heterocycloalkyl, or phenyl;

c. -(C ₁ -C ₆ alkylene)-O-(C ₁ -C ₆ alkylene)-phenyl, wherein the phenyl group is optionally substituted with halogen;

d. -(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the cycloalkyl is optionally substituted with halo, OH or CN;

e. -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with oxo, C ₁ -C ₆ alkyl, -C(O)-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl), -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)-4-6 membered heterocycloalkyl, or -C(O)N(C ₁ -C ₆ alkyl)(C ₁ -C ₆ alkyl);

f. 4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted by oxo, C ₁ -C ₆ alkyl, -C(O)-(C ₁ -C ₆ alkyl), -C(O)O-(C ₁ -C ₆ alkyl), -SO ₂ -(C ₁ -C ₆ alkyl), -C(O)-4-6 membered heterocycloalkyl, or -C(O)N(C ₁ -C ₆ alkyl)(C ₁ -C ₆ alkyl);

g. -C(O)O-(C ₁ -C ₆ alkylene)-OH; and

h. -C(O)O-(C ₁ -C ₆ alkylene)-phenyl;

i. -NR ^a R ^b , wherein ^Ra and R ^b are independently selected from H and C ₁ -C ₆ alkyl, which is optionally substituted with OH, halo, CN, -C(O)OH, phenyl, and 3-7 membered heterocycloalkyl;

R ^2a is selected from H, C ₁ -C ₆ alkyl optionally substituted with OH, and phenyl;

R ^3a is selected from H, C ₁ -C ₆ alkyl optionally substituted with OH, and phenyl;

Ring A is a 3-7 membered heterocycloalkyl; and

Ring B is a 3-7 membered heterocycloalkyl or 3-7 membered heterocycloalkenyl group, wherein p ^1a is 0, 1 or 2 and p ^2a is 0, 1 or 2,

The premise is that Ring A and Ring B together form a 6-11 membered bicyclic heterocycloalkyl.

In some embodiments, the compound of formula (00A), (0A), or (A) is further defined as a compound of formula (III):

in:

R ^0c is H or NH ₂ ;

^R3 is (i) or (ii):

(i)

in:

R ^1c is selected from H, C ₁ -C ₆ alkyl, -(C ₁ -C ₆ alkyl)-phenyl, -C(O)-(C ₁ -C ₆ alkyl), -C(O)-phenyl and 4-6 membered heterocycloalkyl;

R ^2c is H or C ₁ -C ₆ alkyl; and

R ^3c is a bond or C ₁ -C ₆ alkylene optionally substituted by oxo; or

R ^1c and R ^2c together form a 3-11 membered heterocycloalkyl group, which is optionally substituted by: C ₁ -C ₆ alkyl, oxo or -(C ₁ -C ₆ alkylene)-phenyl; or

R ^1c and R ^3c together form a 3-7 membered heterocycloalkyl group;

(ii) H; C ₁ -C ₆ alkyl, which is optionally substituted by OH, -SO ₂ -(C ₁ -C ₆ alkyl), phenyl, or -O-(C ₁ -C ₆ alkylene)-phenyl; -(C ₁ -C ₆ alkylene)-C(O)O(C ₁ -C ₆ alkyl); or 4-6 membered heterocycloalkyl, which is optionally substituted by -C(O)(C ₁ -C ₆ alkyl); and

Ring G is a 3-7 membered heterocycloalkyl group wherein p ^1c is 0, 1 or 2 and p ^2c is 0, 1 or 2.

In some embodiments, the compound of formula (00A), (0A), or (A) is further defined as a compound of formula (IV):

in:

R ^0d is H or NH ₂ ;

R ^1d is 3-11 membered heterocycloalkyl or -C(O)-3-11 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted by C ₁ -C ₆ alkyl, CF ₃ or fluorine, or R ^1d is -(C ₁ -C ₆ alkylene)-NR ^v R ^w , wherein R ^v and R ^w are independently H or C ₁ -C ₆ alkyl optionally substituted by halo.

In some embodiments, the compound of formula (00A), (0A), or (A) is further defined as a compound of formula (V):

in:

R ^0e is H or NH ₂ ;

R ^1e is selected from

a.H,

bC ₁ -C ₆ alkyl, which is optionally substituted by halo, CN or phenyl,

c.-(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the cycloalkyl is optionally substituted with CN,

d. -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with C ₁ -C ₆ alkyl;

R ^2e is H or C ₁ -C ₆ alkyl;

or R ^1e and R ^2e together form a 3-11 membered heterocycloalkyl group, which is optionally substituted with halo or -NR ^v R ^w , wherein R ^v and R ^w are independently H or C ₁ -C ₆ alkyl optionally substituted with halo;

R ^3e is a bond or C ₁ -C ₆ alkylene, which is optionally substituted with oxo; and

Ring H is a 3-7 membered heterocycloalkyl group, wherein p ^1e is 0, 1 or 2, and p ^2e is 0, 1 or 2.

In some embodiments, the compound of formula (00A), (0A), or (A) is further defined as a compound of formula (VI):

in:

R ^0f is H or NH ₂ ;

R ^1f is selected from C ₁ -C ₆ alkyl, which is optionally substituted with halo, -C(O)OH, oxetan-3-ylmethyl, 3-6 membered cycloalkyl, 3-11 membered heterocycloalkyl, -S-(C ₁ -C ₆ alkyl), 5-10 membered heteroaryl or phenyl, or 3-11 membered heterocycloalkenyl optionally substituted with oxo; and

Ring J is a 6-7 membered heterocycloalkyl group wherein p ^1f is 1 or 2 and p ^2f is 1 or 2.

In some embodiments, the compound of formula (00A), (0A) or (A) is further defined as a compound of formula (VII):

in:

R ^0g is H or NH ₂ ;

R ^1g is selected from C ₁ -C ₆ alkyl;

R ^2g is selected from C ₁ -C ₆ alkyl.

In some embodiments, the compound of formula (00A), (0A), or (A) is further defined as a compound of formula (VIII):

in:

R ^0h is H or NH ₂ ;

R ^1h is selected from H and C ₁ -C ₆ alkyl, which is optionally substituted with CN, 3-6 membered cycloalkyl or 4-6 membered heterocycloalkylene-C(O)O-(C ₁ -C ₆ alkylene)-phenyl;

Ring C is a 3-7 membered cycloalkyl or a 3-7 membered heterocycloalkyl; and

Ring D is a 3-7 membered heterocycloalkyl substituted at the only nitrogen with R ^1h ; and

The prerequisite is that ring C and D together form a 3-11 membered spiroheterocycloalkyl group.

In some embodiments, the compound of formula (00A), (0A) or (A) is further defined as a compound of formula (IX):

in:

R ^0j is H or NH ₂ ;

R ^3j is selected from H and C ₁ -C ₆ alkyl, which is optionally substituted by OH, 3-6 membered cycloalkyl, -O-(C ₁ -C ₆ alkyl), -O-(C ₁ -C ₆ alkyl)-OH, -SO ₂ -(C ₁ -C ₆ alkyl) and phenyl, wherein the phenyl is optionally substituted by CN;

R ^4j is selected from H and C ₁ -C ₆ alkyl optionally substituted with OH;

or R ^3j and R ^4j together form a 4-6-membered heterocycloalkyl group;

R ^5j is selected from H and C ₁ -C ₆ alkyl; and

Ring K is a 6-7 membered heterocycloalkyl group wherein p ^1j is 1 or 2 and p ^2j is 1 or 2.

In some embodiments, the compound of formula (00A), (0A), or (A) is further defined as a compound of formula (X):

in:

R ^0k is H or NH ₂ ;

R ^1k selected from

a.H,

bC ₁ -C ₆ alkyl, which is optionally substituted by halo, CN or phenyl,

c.-(C ₁ -C ₆ alkylene)-3-6 membered cycloalkyl, wherein the cycloalkyl is optionally substituted with CN,

d. -(C ₁ -C ₆ alkylene)-4-6 membered heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted with C ₁ -C ₆ alkyl;

R ^2k is selected from H and C ₁ -C ₆ alkyl;

or R ^1k and R ^2k together form a 3-11 membered heterocycloalkyl group optionally substituted by halo; C ₁ -C ₆ alkyl optionally substituted by OH; or -NR ^v R ^w , wherein R ^v and R ^w are independently H or C ₁ -C ₆ alkyl optionally substituted by halo; and

R ^3k is a bond, a methylene group, or -C(=O)-.

In some embodiments, ^XA and ^XB are independently selected from 3-6-membered cycloalkyl, 6-10 membered aryl, 5-6 membered heterocycloalkenyl, and 5-10 membered heteroaryl, wherein ^XA and ^XB are each independently optionally substituted with Y ¹ .

In some embodiments, one of ^XA or ^XB is 3-11 membered heterocycloalkyl, which is optionally substituted with Y ¹ .

In some embodiments, Ring Q is (i). In some embodiments, Ring Q is (ii).

In some embodiments, ^t1 is 0. In some embodiments, ^t1 is 1.

In some embodiments, R ⁰ , R ⁰⁰ , and R ⁰¹ are each H.

In some embodiments, R ⁰ is NH ₂ .

In some embodiments, Ring Q is (i), ^t1 is 0 or 1, and ^R0 , ^R00 , and ^R01 are each H.

In some embodiments, Ring Q is (ii), ^t2 is 0 or 1, and ^R0 , ^R00 , and ^R01 are each H.

In some embodiments, Ring Q is (i), ^t1 is 1, and ^XA is NR ^a R ^b , wherein ^Ra and R ^b are each independently H, C ₁ -C ₆ alkyl optionally substituted with 5-6 membered heteroaryl, or 3-6 membered cycloalkyl optionally substituted with OH, halo, CN, or C ₁ -C ₆ alkyl optionally substituted with OH.

In some embodiments, Ring Q is (i), ^t1 is 0, and ^XA is _C1 - _C6 alkyl or 3-6 membered cycloalkyl, wherein ^XA is optionally substituted with ^-NRaRb , wherein ^{Ra and Rb} are independently H or _C1 - _C6 alkyl.

In some embodiments, the compound is selected from Table 1 or Examples 1-468.

Also provided are pharmaceutical compositions comprising a 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine compound as described herein, such as a compound of formula (00A), (0A) or (A) or a subformula thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

Also provided are 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine compounds as described herein, for example compounds of formula (OOA), (OA) or (A) or subformulae thereof, for use in therapy.

Also provided is the use of a 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine compound as described herein, such as a compound of formula (00A), (0A) or (A) or a subformula thereof, in treating inflammatory diseases such as asthma.

Also provided is the use of a 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine compound as described herein, such as a compound of formula (00A), (0A) or (A) or a subformula thereof, for the preparation of a medicament for treating inflammatory diseases such as asthma.

Also provided are 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine compounds as described herein, such as compounds of formula (00A), (0A) or (A) or subformulae thereof, for use in treating inflammatory diseases such as asthma.

Also provided are methods for preventing, treating, or lessening the severity of a disease or condition in a patient that is responsive to inhibition of Janus kinase activity, comprising administering to the patient a therapeutically effective amount of a 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine compound, such as a compound of Formula (00A), (0A), or (A), or a subformula thereof, as described herein. In some embodiments, the disease or condition is asthma. In some embodiments, the Janus kinase is JAK1. In some embodiments, the compound is administered by inhalation.

In some embodiments, such as compounds of formula (00A), (0A), or (A), R ⁰⁰ is H or CH ₃ ; R ⁰¹ is H or NH ₂ ; R ⁰ is H or NH ₂ ; and Ring Q is (i) or (ii):

wherein ^t1 and ^t2 are each independently 0 or 1, and ^XA and ^XB are each independently selected from:

In some embodiments, such as compounds of formula (00A), (0A), or (A), R ⁰⁰ is H or CH ₃ ; R ⁰¹ is H or NH ₂ ; R ⁰ is H or NH ₂ ; and Ring Q is (i) or (ii):

wherein ^t1 and ^t2 are each independently 0 or 1, and ^XA and ^XB are each independently selected from:

In any of the embodiments herein, one or more compounds of WO 2011/003065 are excluded.

Also provided are compounds selected from Examples 1-468 or Table 1, or any combination thereof. If there is any discrepancy between a structure and its chemical name, the structure controls.

Table 1: Exemplary compounds of the present invention

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

In the structures shown herein, when the stereochemistry of any particular chiral atom is not specified, all stereoisomers are contemplated and included as compounds of the present invention. When stereochemistry is specified by a solid wedge or dashed line representing a specific configuration, then that stereoisomer is so specified and defined. Unless otherwise indicated, relative stereochemistry is intended when a solid wedge or dashed line is used.

Another aspect includes prodrugs of the compounds of the invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, including known amino protecting groups and carboxyl protecting groups, which are released, e.g., hydrolyzed, to produce the compounds of the invention under physiological conditions.

The term "prodrug" refers to a precursor or derivative form of a pharmaceutically active substance that is less effective in patients than the parent drug and can be enzymatically or hydrolytically activated or converted to a more active parent form. See, for example, 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, phosphorothioate-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.

A specific type of prodrug is a compound in which the nitrogen atom of an amino, amidino, aminoalkyleneamino, iminoalkyleneamino, or guanidino group is substituted with a hydroxyl group, an alkylcarbonyl (-CO-R) group, an alkoxycarbonyl (-CO-OR) group, or an acyloxyalkyl-alkoxycarbonyl (-CO-O-R-O-CO-R) group, wherein R is a monovalent or divalent group, such as an alkyl, alkylene, or aryl group, or a group of the formula -C(O)-O-CP1P2-haloalkyl, wherein P1 and P2 are the same or different and are hydrogen, alkyl, alkoxy, cyano, halogen, alkyl, or aryl. In a specific embodiment, the nitrogen atom is one of the nitrogen atoms of the amidino group of a compound of formula (00A), (0A) or (A) or its subformulae. Prodrugs can be prepared by reacting a compound of the invention, for example, a compound of formula (00A), (0A), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX) or (X) or a compound of Table 1 or Examples 1-468 with an activating group, for example, an acyl group, so that, for example, a nitrogen atom in the compound is bonded to an exemplary carbonyl group of an activated acyl group. Examples of activated carbonyl compounds are those containing a leaving group bonded to a carbonyl group, including, for example, acyl halides, amides, acyl pyridinium salts, acyl alkoxides, acylphenol oxides such as p-nitrophenoxy acyl, dinitrophenoxy acyl, fluorophenoxy acyl and difluorophenoxy acyl. The reaction is typically carried out in an inert solvent at a reduced temperature, such as -78 to about 50°C. The reaction can also be carried out in the presence of an inorganic base, such as potassium carbonate or sodium bicarbonate, or an organic base, such as an amine, including pyridine, trimethylamine, triethylamine, triethanolamine, etc.

Other types of prodrugs are also included. For example, the free carboxyl groups of the compounds of the present invention, such as compounds of formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX) or (X), or compounds of Table 1 or Examples 1-468 containing appropriate free carboxyl groups, can be derivatized into amides or alkyl esters. As another example, compounds of the present invention containing free hydroxyl groups can be derivatized into prodrugs by converting the hydroxyl group into groups such as, but not limited to, phosphate, hemisuccinate, dimethylaminoacetate or phosphoryloxymethoxycarbonyl, as described in Fleisher, D. et al. (1996) Improved oral drug delivery: solubility limitations overcome by the use of prodrugs Advanced Drug Delivery Reviews, 19:115. Also included are carbamate prodrugs of hydroxyl and amino groups, as well as carbonate, sulfonate and sulfate prodrugs of hydroxyl groups. Also contemplated are hydroxyl groups derivatized as (acyloxy)methyl and (acyloxy)ethyl ethers, wherein the acyl group may be an alkyl ester, optionally substituted with groups including, but not limited to, ether, amine, and carboxylic acid functional groups, or wherein the acyl group is an amino acid ester as described above. This type of prodrug is described in J. Med. Chem., (1996), 39:10. More specific examples include replacement of the hydrogen atom of an alcohol group with, for example, a (C ₁ -C ₆ ) alkanoyloxymethyl, 1-((C ₁ -C ₆ ) alkanoyloxy)ethyl, 1-methyl-1-((C ₁ -C ₆ ) alkanoyloxy)ethyl, (C ₁ -C ₆ ) alkoxycarbonyloxymethyl, N-(C ₁ -C ₆ ) alkoxycarbonylaminomethyl, succinyl, ₍ C 1 -C 6 ) alkanoyl, α-amino(C ₁ -C ₄ ) alkanoyl, arylacyl and α-aminoacyl or α-aminoacyl-α-aminoacyl group, wherein each α-aminoacyl group is independently selected from naturally occurring L-amino acids, P(O)(OH) ₂ , -P(O)(O(C _{1 -C 6} ) alkyl) ₂ or a glycosyl group (a group derived from the removal of a hydroxyl group of a carbohydrate in the hemiacetal form).

" leaving group " refers to a part of the first reactant in a chemical reaction, and it is displaced from the first reactant in the chemical reaction. The example of leaving group includes but is not limited to halogen atom, alkoxy and sulfonyloxy. Example sulfonyloxy includes but is not limited to alkylsulfonyloxy (such as methylsulfonyloxy (mesylate group) and trifluoromethylsulfonyloxy (trifluoromethanesulfonate group)) and arylsulfonyloxy (such as p-toluenesulfonyloxy (toluenesulfonate group) and p-nitrosulfonyloxy (nitrosulfonate group (nosylate))).

Synthesis of JANUS kinase inhibitor compounds

The compounds of the present invention can be synthesized by synthetic routes as described herein. In certain embodiments, in addition to the description contained herein or according to the description contained herein, methods well known in the chemical field can be used. Raw materials are generally available from commercial sources such as Aldrich Chemicals (Milwaukee, Wis.), or are easily prepared using methods well known to those skilled in the art (for example, by generally describing in Louis F. Fieser and Mary Fieser, Reagents for Organic Synthesis, v. 1-19, Wiley, N.Y. (1967-1999 ed.), Beilsteins Handbuch der organischen Chemie, 4, Aufl. ed. Springer-Verlag, Berlin, including supplements (also available through the Beilstein online database)), or Comprehensive Heterocyclic Chemistry, Editors Katrizky and Rees, Pergamon Press, prepared by the method in 1984.

Compounds can be prepared individually or as a compound library comprising at least 2, for example 5 to 1,000 compounds or 10 to 100 compounds. Compound libraries can be prepared by a combined "split and mix" method or by using multiple parallel syntheses of solution phase or solid phase chemistry, via methods known to those skilled in the art. Therefore, according to another aspect of the present invention, there is provided a compound library comprising at least 2 compounds of the present invention, for example, compounds of formula (00A), (0A), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX) or (X) compounds or compounds of Table 1 or Examples 1-468.

For illustrative purposes, reaction schemes 1-26 described below provide routes for synthesizing the compounds of this invention and key intermediates. For a more detailed description of each reaction step, see the Examples section below. Those skilled in the art will appreciate that other synthetic routes can be used. Although some specific raw materials and reagents are described in the flow and discussed below, other raw materials and reagents can be substituted to provide multiple derivatives or reaction conditions. In addition, according to the disclosure, conventional chemistry well known to those skilled in the art can be used to further modify many compounds prepared by the following methods.

In the preparation of the compounds of this invention, it may be necessary to protect the remote functional groups (e.g., primary or secondary amines) of the intermediates. The need for such protection will vary according to the properties of the remote functional groups and the conditions of the preparation method. Suitable amino protecting groups include acetyl, trifluoroacetyl, benzyl, phenylsulfonyl, tert-butyloxycarbonyl (BOC), benzyloxycarbonyl (CBz), and 9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protection is readily determined by those skilled in the art. For a general description of protecting groups and their uses, see T.W.Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

Other transformations that are commonly used in the synthesis of compounds of the invention and that can be performed using a variety of reagents and conditions include the following:

(1) Reaction of carboxylic acids with amines to form amides. This transformation can be achieved using various reagents known to those skilled in the art, but a comprehensive review can be found in Tetrahedron, 2005, 61, 10827-10852.

(2) The reaction of primary or secondary amines with aryl halides or pseudohalides such as triflates, often referred to as "Buchwald-Hartwig cross-couplings," can be achieved using a variety of catalysts, ligands, and bases. A review of these methods is provided in Comprehensive Organic Name Reactions and Reagents, 2010, 575–581.

(3) Palladium cross-coupling reaction between aryl halides and vinyl boronic acids or boronic esters. This transformation is a type of "Suzuki-Miyaura cross-coupling", a class of reactions thoroughly reviewed in Chemical Reviews, 1995, 95(7), 2457-2483.

(4) The hydrolysis of esters to the corresponding carboxylic acids is well known to those skilled in the art and includes the following conditions: for methyl and ethyl esters, using aqueous strong bases such as lithium hydroxide, sodium hydroxide, or potassium hydroxide or aqueous strong mineral acids such as HCl; for tert-butyl esters, using acids such as HCl in dioxane or trifluoroacetic acid (TFA) in dichloromethane (DCM) for hydrolysis.

Other exemplary transformations are discussed in the schemes below.

Reaction Scheme 1

Compounds of Formula 6 can be synthesized as shown in Reaction Scheme 1. Trichloroacetonitrile can be reacted with ethyl cyanoacetate to provide compound 34. Compound 1 can be condensed with hydrazine to provide compound 2, which can then be condensed with 1,1,3,3-tetramethoxypropane to provide compound 3. Amine 3 can be doubly Boc-protected to provide compound 4, which can then be hydrolyzed with lithium hydroxide to provide carboxylic acid 5. Carboxylic acid 5 can then be coupled with various amines in the presence of PyAOP, DIEA, and DMAP to provide compounds of Formula 6.

Reaction Scheme 2

Reaction Scheme 2 illustrates a method for synthesizing compounds of Formula 5. α-Bromoketones can be generated from Compound 1 using reagents such as bromine. Alkylation of di-tert-butyl iminodicarbonate with sodium hydride and various α-bromoketones 2 produces Compound 3. Compound 3 can be heated with DMFDMA to yield Compound 4. Compound 4 is cyclized with hydrazine in ethanol to provide the pyrazole compound 5. Compound 5 is reacted with pyrazolo[1,5-a]pyrimidine-3-carboxylic acid in the presence of PyAOP, DIEA, and DMAP to yield the compound of Formula 6.

Reaction Scheme 3

An alternative method for synthesizing compounds of Formula 6 is described in Reaction Scheme 3. Alkylation of potassium phthalimide with α-bromoketone 1 produces compound 2. Condensation with DMFDMA provides compound 3. Compound 3 can be cyclized with hydrazine to provide compound 4, which can then be coupled with pyrazolo[1,5-a]pyrimidine-3-carboxylic acid using HATU under amide-forming conditions to provide compounds of Formula 5. Alkylation of compound 5 with an alkyl halide in the presence of cesium carbonate provides compounds of Formula 6.

Reaction Scheme 4

Reaction Scheme 4 illustrates an alternative synthesis of compounds of Formula 5. Nitro-SEM pyrazole compound 1, prepared as described in Reaction Scheme 5, can be regioselectively deprotonated with lithium hexamethyldisilazide at low temperature and quenched with iodine to afford 2. The nitro group of compound 2 can be reduced in the presence of iron and ammonium chloride, followed by Boc protection to yield compound 3. Compound 3 can be coupled with an aryl boronic acid or aryl boronic ester under Suzuki conditions to afford compound 4. Cleavage of the Boc group with tin tetrachloride affords compounds of Formula 5.

Reaction Scheme 5

An alternative method for synthesizing compounds of Formula 6a and 6b is shown in Reaction Scheme 5. Commercially available 4-nitro-1H-pyrazoles can be protected with [β-(trimethylsilyl)ethoxy]methyl (SEM) by treatment with sodium hydride and (2-(chloromethoxy)ethyl)trimethylsilane. The resulting compound 1 can be arylated with an aryl bromide or iodide under palladium catalysis to produce 4-nitro-5-aryl-pyrazoles of Formula 2. The nitro group of compound 2 can be reduced in the presence of iron and ammonium chloride to produce aminopyrazole 3. Amide bond coupling with commercially available pyrazolo[1,5-a]pyrimidine-3-carboxylic acid in the presence of PyAOP, DIEA, and DMAP provides compound 4. Removal of the SEM protecting group with aqueous HCl in ethanol produces compound 5, which can be alkylated with an alkyl halide or with a Michael acceptor in the presence of a suitable base such as cesium carbonate to provide compounds of Formula 6a and 6b.

Reaction Scheme 6

An alternative method for synthesizing compounds of Formula 4a and 4b is shown in Reaction Scheme 6. Commercially available 4-nitro-1H-pyrazole can be reacted with an alkyl bromide in the presence of cesium carbonate at 55°C for 12 hours to provide compound 1. Compound 1 can be reacted with an aryl bromide in N,N-dimethylacetamide in the presence of palladium(II) acetate, di(1-adamantyl)-n-butylphosphine, potassium carbonate, and trimethylacetic acid to provide compounds 2a and 2b. The ratio of product 2a:2b varies depending on the substituent R1, but the reaction generally favors the formation of product 2b. Compounds 32a and 32b can be reduced to compounds 3a and 3b in ethanol and water in the presence of iron and ammonium chloride. Coupling of compounds 3a and 3b with pyrazolo[1,5-a]pyrimidine-3-carboxylic acid in the presence of PyAOP, DIEA, and DMAP can provide compounds of Formula 4a and 4b.

Reaction Scheme 7

Amide bond coupling of 1 with 5-chloro-pyrazolo[1,5-a]pyrimidine-3-carbonyl chloride (prepared according to Journal of Medicinal Chemistry, 55(22), 10090-10107; 2012) in the presence of triethylamine provides compounds of Formula 2. Treatment of 2 with aqueous ammonia produces compounds of Formula 3. Removal of the SEM protecting group by aqueous HCl in methanol produces compounds of Formula 4, which can be alkylated with an alkyl halide or with a Michael acceptor in the presence of a suitable base such as cesium carbonate to give compounds of Formula 5a and 5b.

Reaction Scheme 8

The method for synthesizing compounds of Formula 4a and 4b is shown in Reaction Scheme 8. Amide bond coupling of compound 1 with commercially available 6-methyl-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid in the presence of HATU and DIPEA provides compound 2. Removal of the SEM protecting group by aqueous HCl in ethanol produces compound 3, which can be alkylated with an alkyl halide or with a Michael acceptor in the presence of a suitable base such as cesium carbonate to provide compounds of Formula 4a and 4b.

Reaction Scheme 9

Compounds of Formula 4 can be synthesized as shown in Reaction Scheme 9. Pyrazole compound 1 (prepared as described herein) can be alkylated with tert-butyl bromoacetate in the presence of cesium carbonate to provide intermediate 2. 2 can be treated with trifluoroacetic acid to provide acid 3, which can then react with a primary or secondary amine in the presence of a coupling reagent such as N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (HATU) to provide compounds of Formula 4.

Reaction Scheme 10

According to Scheme 10, compound 1 can be reacted with an α-haloamide in the presence of a base such as cesium carbonate, followed by deprotection under acidic aqueous conditions to provide compounds of type 2. Compounds of type 2 can be reacted with primary or secondary amines using a reducing agent such as sodium triacetoxyborohydride under reducing conditions to provide compounds of types 4 and 3, respectively. Intermediate 3 can be further modified by reacting with an aldehyde in the presence of an appropriate reducing agent such as sodium triacetoxyborohydride to provide compound 4.

Reaction Scheme 11

Pyrazole compounds of formula 1 can be modified by copper-catalyzed coupling with secondary or tertiary amine side chains, for example, containing phenyl iodide, to provide compounds 2.

Reaction Scheme 12

Compound 3 is treated with thiourea in a suitable solvent such as ethanol under heating to give thiazole compound 4. Aminothiazole 4 can be converted to bromide 5 by treatment with tBuONO, CuBr ₂ in a suitable solvent such as acetonitrile. Compound 5 is then hydrolyzed using a base such as aqueous potassium hydroxide in a compatible solvent such as ethanol to give acid (6). Compound 7 can be prepared by treating compound 6 with diphenylphosphoryl azide (DPPA) in tert-butanol, followed by deprotection under acidic conditions. Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid is activated with thionyl chloride in the presence of a base such as DIPEA in a solvent such as THF and then reacted with 7 to give compound 8. Aryl methyl ether 8 is demethylated with, for example, BBr ₃ in DCM, followed by alkylation with sodium chlorodifluoroacetate and a suitable base and solvent combination such as cesium carbonate in DMF to give compound 10.

Reaction Scheme 13

Alternatively, compounds of Formula 6 can be synthesized as shown in Reaction Scheme 13. For example, compound 1 can be prepared by treating a commercially available substituted acetophenone with diethyl carbonate and then brominating it using, for example, bromine in dioxane. Compound 1 is treated with an appropriately substituted thioamide or thiourea to give thiazole compound 2. Compound 2 can be hydrolyzed using a base such as aqueous potassium hydroxide in a compatible solvent such as THF to give acid compound 35. Compound 4 can be prepared by reacting compound 3 with diphenylphosphoryl azide (DPPA) in tert-butanol. Compound 4 is deprotected under acidic conditions to give amino compound 5. Compounds of Formula 6 can be prepared by treating compound 5 with pyrazolo[1,5-a]pyrimidine-3-carbonyl chloride in pyridine.

Reaction Scheme 14

Compounds of type 1 can be reacted with suitable nucleophiles such as appropriately monosubstituted cyclic diamines, acetals containing cyclic amines, and cyclic amines containing a substituted second amino group, which can be achieved in a solvent such as DMA with heating to give compounds 7, 2, and 6, respectively. Compounds of type 2 can be deprotected under acidic aqueous conditions to provide structures of type 3. Compounds 3 can be reductively aminated with monosubstituted or disubstituted amines, and for monosubstituted amino products, they can be further modified by reductive alkylation with appropriate aldehydes in the presence of a reducing agent such as sodium cyanoborohydride to give compounds of type 5.

Reaction Scheme 15

Compound 1 can undergo a Sonogashria reaction with an N-substituted propargylamine or propargyl alcohol in the presence of a palladium source, a copper(i) halide, and a base in a suitable solvent with heating to provide compounds 2 and 3, respectively. For example, this can be a combination _of Pd( _PPh3 ) _2Cl2 , Cu(I)I, and _Et3N in THF. Alcohol 3 can be converted to bromide 4 using _CBr4 and _PPh3 in a suitable solvent such as DCM. The bromide can then be displaced by primary and secondary amines to provide structures of the type 5.

Reaction Scheme 16

As shown in Scheme 16, compounds of type 1 can react with 1,2-dibromoethane to give alkyl bromide 2. Reaction of 2 with an amine nucleophile can produce structures of type 4.

Reaction Scheme 17

As shown in Scheme 17, compounds of type 1 can be alkylated with amine-containing electrophiles to produce compounds 2. Manipulation of functional groups on the pendant amine can produce structures of types 3 and 4.

Reaction Scheme 18

As shown in Scheme 18, compounds of type 1 can be reacted with allyl dihalides to produce compounds 2. Further reaction of the pendant allyl halide with an amine nucleophile can produce structures of type 4.

Reaction Scheme 19

As shown in Scheme 19, compounds of type 1 can be reacted with substituted piperazines and a suitable dehydrating agent (e.g., HATU) to produce structures of type 4. Alternatively, compounds of type 1 can be coupled with a suitable monoprotected piperazine to produce compound 2. Compound 2 can then be deprotected and further reacted with a suitable electrophile to produce structures of type 4.

Reaction Scheme 20

As shown in Scheme 20, compounds of type 1 can be reacted with epoxides in the presence of a Lewis acid such as Yb(OTf) ₃ to produce structures such as 2 and 3. Deprotection can then produce structures of type 4 and 5.

Reaction Scheme 21

As shown in Scheme 21, compounds of type 1 can be reacted with epoxides in the presence of a Lewis acid such as Yb(OTf) ₃ to produce structures such as 2. The resulting alcohol can then be oxidized to produce structures of type 3.

Reaction Scheme 22

As shown in Scheme 22, compounds of type 1 are reacted with a suitable electrophile in the presence of a base, followed by deprotection to produce structures such as 2. Further treatment with a reactive amine can then produce structures of type 3.

Reaction Scheme 23

As shown in Scheme 23, the reaction of compounds of type 1 with propargyl bromide or propargyl chloride in the presence of a base can produce compounds of type 2. In the presence of copper halide and an organic base, 2 reacts with an azide such as 3, followed by deprotection, to afford triazole compounds of type 4. Further treatment with a reactive amine can then yield structures of type 5.

Reaction Scheme 24

As shown in Scheme 24, the reaction of compounds of type 1 with 2-(halomethyl)oxirane can produce compounds of type 2. The reaction of compounds of type 2 with nucleophilic amines can give compounds of type 3. Oxidation of 3 gives compounds of type 4.

Reaction Scheme 25

As shown in Scheme 25, reaction of compounds of type 1 with an appropriate allylic alkylating agent in the presence of a base yields compounds of type 2. Deprotection of the amino protecting group under acidic conditions followed by reductive amination or alkylation of the reactive amine affords compounds of type 3.

Reaction Scheme 26

As shown in Scheme 26, reaction of compounds of type 1 with a suitable allylic alkylating agent produces compounds of type 2. Removal of the ketal protecting group under acidic conditions followed by reaction of the liberated ketone with an amine under reductive amination conditions yields compounds of type 3. Subsequent reductive amination or alkylation steps produce structures of type 3.

It will be appreciated that, where suitable functional groups are present, the compounds of the formulae or any intermediates used in their preparation may be further derivatized by one or more standard synthetic methods using condensation, substitution, oxidation, reduction, or cleavage reactions. Specific substitution methods include conventional alkylation, arylation, heteroarylation, acylation, sulfonylation, halogenation, nitration, formylation, and coupling methods.

In another example, a primary or secondary amine group can be converted to an amide group (-NHCOR' or -NRCOR') by acylation. Acylation can be performed by reaction with an appropriate acyl chloride in a suitable solvent such as dichloromethane in the presence of a base such as triethylamine, or by reaction with an appropriate carboxylic acid in a suitable solvent such as dichloromethane in the presence of a suitable coupling agent such as HATU (O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate). Similarly, an amine group can be converted to a sulfonamide group (-NHSO2R' or -NR" _SO2R ') by reaction with an appropriate sulfonyl chloride in the presence of a suitable base such as triethylamine in a suitable solvent such as dichloromethane. A primary or secondary amine group can be converted to a urea group ( _-NHCONR'R " or -NRCONR'R") by reaction with an appropriate isocyanate in the presence of a suitable base such as triethylamine in a suitable solvent such as dichloromethane.

For example, amines ( _-NH2 ) can be obtained by reducing the nitro ( _-NO2 ) group using, for example, hydrogen in a solvent such as ethyl acetate or an alcohol such as methanol in the presence of a metal catalyst such as palladium on a support such as carbon. Alternatively, the conversion can be carried out by chemical reduction using, for example, a metal such as tin or iron in the presence of an acid such as hydrochloric acid.

In another example, an amine ( _-CH2NH2 ) group can be obtained, for example, by catalytic hydrogenation reduction of a nitrile (-CN) using, for example, hydrogen in the presence of a metal catalyst such as palladium or Raney nickel _on a support such as carbon in a solvent such as an ether such as a cyclic ether such as tetrahydrofuran at a suitable temperature such as about -78°C to the reflux temperature of the solvent.

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

Aldehyde groups (-CHO) can be converted to amine groups (-CH2NR'R") by reductive amination using an amine and a borohydride such as sodium triacetoxyborohydride or sodium cyanoborohydride in a solvent such as a halogenated hydrocarbon such as dichloromethane or an alcohol such as ethanol, optionally in the presence of _an acid such as acetic acid at ambient temperature.

In another example, an aldehyde group can be converted to an alkenyl group (-CH=CHR') by using a Wittig or Wadsworth-Emmons reaction using an appropriate phosphorane or phosphonate under standard conditions known to those skilled in the art.

Aldehyde groups can be obtained by reducing ester groups (eg _-CO2Et ) or nitrile (-CN) using diisobutylaluminum hydride in a suitable solvent such as toluene. Alternatively, aldehyde groups can be obtained by oxidizing alcohol groups using any suitable oxidizing agent known to those skilled in the art.

The ester group ( _-CO2R ') can be converted into the corresponding acid group ( _-CO2H ) by acid- or base-catalyzed hydrolysis, depending on the nature of R. If R is tert-butyl, acid-catalyzed hydrolysis can be achieved, for example, by treatment with an organic acid such as trifluoroacetic acid in an aqueous solvent, or by treatment with an inorganic acid such as hydrochloric acid in an aqueous solvent.

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

In another example, carboxylic acids can be synthesized by conversion to the corresponding acid chloride (-COCl) followed by one _carbon homologation (ie, _-CO2H to _-CH2CO2H ) via the Arndt-Eistert synthesis.

In another example, the -OH group can be generated from the corresponding ester (e.g., -CO ₂ R') or aldehyde (-CHO) by reduction using, for example, a complex metal hydride such as lithium aluminum hydride in diethyl ether or tetrahydrofuran, or sodium borohydride in a solvent such as methanol. Alternatively, the alcohol can be prepared by reducing the corresponding acid (-CO ₂ H) using, for example, lithium aluminum hydride in a solvent such as tetrahydrofuran, or by using borane in a solvent such as tetrahydrofuran.

Using conditions well known to those skilled in the art, alcohol group can be converted into leaving group, for example halogen atom or sulfonyloxy such as alkylsulfonyloxy, for example trifluoromethylsulfonyloxy or arylsulfonyloxy such as p-tolylsulfonyloxy. For example, alcohol can react with sulfuryl chloride in halogenated hydrocarbon (for example dichloromethane) to obtain corresponding chloride. Also can use base (for example triethylamine) in the reaction.

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

The aromatic halogen substituent in the compound can be by using a base such as a lithium base such as n-butyllithium or tert-butyllithium, optionally at low temperatures such as about -78 ° C, in a solvent such as tetrahydrofuran, treated, halogen-metal exchange, and then quenched with an electrophilic reagent to introduce the desired substituent. Therefore, for example, formyl can be introduced as an electrophilic reagent by using N,N-dimethylformamide. The aromatic halogen substituent can alternatively be subjected to a metal (such as palladium or copper) catalyzed reaction to introduce, for example, an acid, ester, cyano, amide, aryl, heteroaryl, alkenyl, alkynyl, sulfenyl or amino substituent. Suitable methods that can be adopted include those described by Heck, Suzuki, Stille, Buchwald or Hartwig.

Aromatic halogen substituents can also undergo nucleophilic displacement following reaction with a suitable nucleophile such as an amine or alcohol. Advantageously, this reaction can be carried out at elevated temperature in the presence of microwave irradiation.

Separation method

In each exemplary process, it may be advantageous to separate the reaction products from each other or from the starting materials. The desired product of each step or series of steps is separated or purified (hereinafter referred to as separation) to the desired uniformity by techniques commonly used in the art. Typically, such separations include multiphase extraction, crystallization or grinding from a solvent or solvent mixture, distillation, sublimation, or chromatography. Chromatography can include any number of methods, including, for example: reverse phase and normal phase; size exclusion; ion exchange; supercritical fluid; high, medium, and low pressure liquid chromatography methods and apparatus; small-scale analysis; simulated moving bed (SMB) and preparative thin or thick layer chromatography, as well as small-scale thin layer and flash chromatography techniques.

Another type of separation method comprises treating the mixture with a selected reagent to bind the desired product, unreacted starting material, reaction by-products, etc. or to otherwise render them separable. These reagents include adsorbents or absorbents such as activated carbon, molecular sieves, ion exchange media, etc. Alternatively, the reagent can be an acid (in the case of a basic material), a base (in the case of an acidic material), a binding agent such as an antibody, a binding protein, a selective chelating agent such as a crown ether, a liquid/liquid ion extraction reagent (LIX), etc.

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

Diastereomeric mixtures can be separated into their respective diastereomers based on their physicochemical differences by methods well known to those skilled in the art, such as by chromatography or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. In addition, some compounds of the present invention may be atropisomers (e.g., substituted biaryls) and are considered part of the present invention. Enantiomers can also be separated by using chiral HPLC columns or supercritical fluid chromatography.

A single stereoisomer, such as an enantiomer, substantially free of its stereoisomers can be obtained by resolving the racemic mixture using, for example, methods for forming diastereomers using 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 the chiral compounds of the present invention can be separated and isolated by any suitable method, including: (1) forming diastereomeric salts with chiral compounds and separating by fractional crystallization or other methods, (2) forming diastereomeric compounds with chiral derivatizing agents, separating the diastereomers, and converting them to pure stereoisomers, and (3) directly separating the substantially pure or enriched stereoisomers under chiral conditions. See: Drug Stereochemistry, Analytical Methods and Pharmacology, Irving W. Wainer, Ed., Marcel Dekker, Inc., New York (1993).

Diastereomeric salts can be formed by reacting enantiomerically pure chiral bases such as strychnine, quinine, ephedrine, strychnine, α-methyl-β-phenylethylamine (amphetamine), and the like with asymmetric compounds having acidic functional groups such as carboxylic acids and sulfonic acids. The separation of diastereomeric salts can be induced by fractional crystallization or ion chromatography. To separate the optical isomers of amino compounds, the addition of chiral carboxylic or sulfonic acids such as camphorsulfonic acid, tartaric acid, mandelic acid, or lactic acid can result in the formation of diastereomeric salts.

Alternatively, the substrate to be resolved is reacted with one enantiomer of a chiral compound to form a pair of diastereoisomers (Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds, John Wiley & Sons, Inc., New York, 1994, p. 322). Diastereomeric compounds can be formed by reacting an asymmetric compound with an enantiomerically pure chiral derivatizing reagent (e.g., a menthyl derivative), followed by separation of the diastereomers and hydrolysis to yield the pure or enriched enantiomer. Methods for determining optical purity include preparing a chiral ester of the racemic mixture, such as a menthyl ester, e.g., (-) menthyl chloroformate or Mosher's ester, α-methoxy-α-(trifluoromethyl)phenyl acetate (Jacob, J. Org. Chem. 47: 4165 (1982)), in the presence of a base and analyzing the NMR spectrum for the presence of the two atropisomeric enantiomers or diastereomers. According to the method for separating atropisomeric naphthyl-isoquinolines (WO 96/15111, incorporated herein by reference), the stable diastereomers of atropisomeric compounds can be separated and isolated by normal and reverse phase chromatography. For method (3), a racemic mixture of two enantiomers can be separated by chromatography using a chiral stationary phase (Chiral Liquid Chromatography W.J. Lough, Ed., Chapman and Hall, New York, (1989); Okamoto, J. of Chromatogr. 513:375-378 (1990)). The enriched or purified enantiomers can be distinguished by methods used to distinguish other chiral molecules with asymmetric carbon atoms, such as optical rotation and circular dichroism. The absolute stereochemistry of the chiral center and the enantiomers can be determined by x-ray crystallography.

Positional isomers, such as the E and Z forms, of compounds of Formula (00A), (0A), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X) and the intermediates used to synthesize them can be observed by identification methods such as NMR and analytical HPLC. For certain compounds where the interconversion barrier is sufficiently high, the E and Z isomers can be separated, for example, by preparative HPLC.

Pharmaceutical compositions and administration

The compounds to which the present invention relates are JAK kinase inhibitors, such as JAK1 inhibitors, and are useful in the treatment of several diseases, such as inflammatory diseases, such as asthma.

Thus, another embodiment provides pharmaceutical compositions or medicaments containing a compound of the invention, such as a compound of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, and a pharmaceutically acceptable carrier, diluent, or excipient, as well as methods of preparing such compositions and medicaments using the compounds of the invention.

In one example, a compound of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, can be formulated into a galenic administration form by mixing with a physiologically acceptable carrier (i.e., a carrier that is non-toxic to the recipient at the dosage and concentration employed) at an appropriate pH and the desired purity at ambient temperature. The pH of the formulation will depend primarily on the specific use and concentration of the compound, but will generally be in the range of about 3 to about 8. In one example, a compound of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, is formulated in acetate buffer at pH 5. In another embodiment, the compound of the invention, e.g., a compound of Formula (OOA), (OA), (A), (la), (lb), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, is sterile. The compound can be stored, for example, as a solid or amorphous composition, as a lyophilized formulation, or as an aqueous solution.

The compositions are formulated, dosed, and administered in a manner consistent with good medical practice. Factors for consideration herein include the particular disease being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disease, the site of active agent delivery, the method of administration, the schedule of administration, and other factors known to physicians.

Should be understood that the specific dosage level of any particular patient will depend on multiple factors, including the activity, age, body weight, general health, sex, diet, application time, route of administration, excretion rate, drug combination and the severity of the particular disease being treated of the specific compound used.Optimal dose level and administration frequency will be determined by clinical trials, as needed in the pharmaceutical field.Usually, the daily dosage range for oral administration will be in the following range: about 0.001mg to about 100mg/kg human body weight, generally 0.01mg to about 50mg per kg body weight, for example 0.1 to 10mg per kg body weight, carried out in single dose or divided dose.In general, the daily dosage range for inhalation administration will be in the following range: about 0.1 μ g to about 1mg/kg human body weight, preferably 0.1 μ g to 50 μ g/kg, carried out in single dose or divided dose.On the other hand, it may be necessary to use a dosage that exceeds these limits in some cases.

The compounds of the present invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, can be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal, inhalation, and epidural and intranasal, and if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In some embodiments, administration is by inhalation.

The compounds of the present invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, can be administered in any convenient administration form, such as tablets, powders, capsules, lozenges, granules, solutions, dispersions, suspensions, syrups, sprays, vapors, suppositories, gels, emulsions, patches, etc. Such compositions may contain conventional components of pharmaceutical formulations, such as diluents (e.g., glucose, lactose, or mannitol), carriers, pH adjusters, buffers, sweeteners, fillers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, fragrances, flavorings, other known additives, and other active agents.

Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, for example, Ansel, Howard C., et al., Ansel's Drug 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 solvents, dispersion media, coating materials, surfactants, antioxidants, preservatives (e.g., antibacterials, antifungals), isotonic agents, absorption delaying agents, salts, preservatives, drugs, drug stabilizers, gels, adhesives, excipients, disintegrants, lubricants, sweeteners, flavorings, dyes, similar materials and combinations thereof, as known to those of ordinary skill in the art (see, e.g., Remington's Pharmaceutical Sciences, pp 1289-1329, 1990). Unless any conventional carrier is incompatible with the active ingredient, its use in treatment or pharmaceutical compositions is considered. Exemplary excipients include dicalcium phosphate, mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, or a combination thereof. Pharmaceutical compositions can include different types of carriers or excipients, depending on whether they are administered in solid, liquid, or aerosol form, and whether they need to be sterile for these routes of administration.

For example, tablets and capsules for oral administration can be in unit dose presentation form and can contain conventional excipients such as binders, for example, syrup, gum arabic, gelatin, sorbitol, gum tragacanth, or polyvinyl pyrrolidone; fillers, for example, lactose, sugar, corn starch, calcium phosphate, sorbitol, or glycine; tableting lubricants, for example, magnesium stearate, talc, polyethylene glycol, or silicon dioxide; disintegrants, for example, potato starch, or acceptable wetting agents such as sodium lauryl sulfate. Tablets can be coated according to methods well known in normal pharmaceutical practice. Oral liquid preparations can be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups, or elixirs, or can be provided as a dry product for reconstitution with water or other suitable solvents before use. Such liquid preparations may contain conventional additives, for example, suspending agents, for example, sorbitol, syrup, methylcellulose, glucose syrup, gelatin, hydrogenated edible fats; emulsifiers, for example, lecithin, sorbitan monooleate or acacia; non-aqueous vehicles (which may include edible oils), for example, almond oil, fractionated coconut oil, oily esters such as glycerol, propylene glycol or ethanol; preservatives, for example, methyl or propyl paraben or sorbic acid, and, if desired, conventional flavoring or coloring agents.

For topical application to the skin, the compounds may be formulated into creams, lotions or ointments. Pharmaceutically acceptable cream or ointment formulations are conventional formulations well known in the art, for example as described in standard pharmaceutical textbooks such as the British Pharmacopoeia.

Compounds of the invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, can also be formulated for inhalation, e.g., as nasal sprays or dry powders or aerosol inhalers. For delivery by inhalation, the compound is typically in the form of microparticles, which can be prepared by a variety of techniques including spray drying, freeze drying, and micronization. Aerosol generation can be achieved using, for example, a pressure-driven jet nebulizer or ultrasonic nebulizer, e.g., by using a propellant-driven metered aerosol or propellant-free administration of the micronized compound, e.g., from an inhalation capsule or other "dry powder" delivery system.

As an example, the compositions of the present invention can be prepared as a suspension for delivery from a nebulizer or as an aerosol in a liquid propellant, for example for use in a pressurized metered dose inhaler (PMDI). Suitable propellants for PMDI are known to those skilled in the art and include CFC-12, HFA-134a, HFA-227, HCFC-22 (CCl ₂ F ₂ ) and HFA-152 (CH ₄ F ₂ and isobutane).

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

Microparticles for delivery by administration can be formulated with excipients that aid delivery and release. For example, in a dry powder formulation, the microparticles can be formulated with large carrier particles that aid flow from a DPI into the lungs. Suitable carrier particles are known and include lactose particles; they can have, for example, a mass median aerodynamic diameter greater than 90 μm.

In the case of aerosol-based formulations, an example is:

*For example, formula (00A), (0A), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI),

(VII), (VIII), (IX) or (X) or the compound of Table 1 or Examples 1-468.

Depending on the inhalation system used, compounds such as compounds of the formula (00A), (0A), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX) or (X) or compounds of Table 1 or Examples 1 to 468 can be administered as described. In addition to the compounds, the administration forms may further contain excipients as described above, or for example propellants (e.g. Frigen in the case of metered aerosols), surfactants, emulsifiers, stabilizers, preservatives, flavorings, fillers (e.g. lactose in the case of powder inhalers) or, if appropriate, other active compounds.

In order to improve the effect of the present invention, the invention provides a kind of inhalation device of the present invention.For the purpose of sucking, a large amount of systems can be used, use the inhalation technique that is suitable for the patient and can be produced by it and use the aerosol of optimum particle size.Except using adapter (spacer, expander) and pear-shaped container (for example,) and the automatic device of emission puff spray for metered aerosol, particularly in the situation of powder inhaler, multiple technical scheme is available (for example or as in U.S. Patent No. 5,263,475, the inhaler described in it is incorporated herein by reference).In addition, the compounds of this invention for example formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX) or (X) compound or table 1 or embodiment 1-468 compound can be sent in multi-chamber device, thereby allow to send combination active agent.

Compounds such as Formula (00A), (0A), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X) or compounds of Table 1 or Examples 1-468 can also be administered parenterally in a sterile medium. Depending on the solvent and concentration used, the compound can be suspended or dissolved in the solvent. Advantageously, adjuvants such as local anesthetics, preservatives, or buffers can be dissolved in the solvent.

Targeted inhaled drug delivery

The optimization of drugs delivered to the lungs by topical (inhalation) administration has recently been reviewed (Cooper, A.E. et al. Curr. Drug Metab. 2012, 13, 457-473). Due to the limitations of the delivery device, the dose of inhaled drugs may be low in humans (approximately <1 mg/day), which requires highly effective molecules. For compounds intended to be delivered by dry powder inhalation, it is also necessary to be able to produce a crystalline form of the compound that can be micronized to a size of 1-5 μm. In addition, the compound needs to maintain a sufficient concentration in the lungs within a given time period so that the pharmacological effect of the desired duration can be exerted, and systemic inhibition is an undesirable pharmacological target with low systemic exposure. The lungs have an inherent high permeability to macromolecules (proteins, peptides) and small molecules with a short lung half-life, so it is necessary to reduce the lung absorption rate by modifying one or more characteristics of the compound: minimizing membrane permeability, reducing the dissolution rate, or introducing a certain degree of alkalinity into the compound to enhance binding to phospholipid-rich lung tissue or by trapping in acidic subcellular compartments such as lysosomes (pH 5). Thus, in some embodiments, compounds of the present invention exhibit one or more of these characteristics.

Matched pair analysis

Figure 1 depicts a matched pair analysis of certain compounds of the invention containing OMe (i) or _OCHF2 (ii) groups at the indicated positions. Any two points connected by a line represent two compounds having identical _R1 and _R2 groups and differing only in substitution by OMe or _OCHF2 at the indicated position. Figure 1 demonstrates that the _OCHF2- substituted analogs (ii) are consistently more potent than the corresponding OMe analogs (i) in a JAK1 biochemical assay (described below).

Methods of treatment using JANUS kinase inhibitors and uses of JANUS kinase inhibitors

Compounds of the present invention, such as compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, inhibit the activity of Janus kinases, such as JAK1 kinase. For example, compounds of the present invention, such as compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, inhibit the phosphorylation of signal transducers and activators of transcription (STATs) by JAK1 kinase and STAT-mediated cytokine production. Compounds of the present invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, can be used to inhibit JAK1 kinase activity in cells via cytokine pathways, 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 pathways. Thus, in one embodiment, a method is provided for contacting a cell with a compound of the present invention, e.g., a compound of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, to inhibit Janus kinase activity (e.g., JAK1 activity) in the cell.

Compounds of the invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, can be used to treat immunological disorders driven 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.

Thus, one embodiment includes a compound of the invention, e.g., a compound of Formula (OOA), (OA), (A), (la), (lb), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, for use in therapy.

In some embodiments, provided are uses of compounds of the invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, for treating inflammatory diseases. Also provided are uses of compounds of the invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, for preparing a medicament for treating an inflammatory disease, such as asthma. Also provided are compounds of the invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, for treating an inflammatory disease, such as asthma.

Another embodiment includes a method for preventing, treating, or reducing the severity of a disease or condition (e.g., asthma) in a patient that is responsive to inhibition of Janus kinase activity (e.g., JAK1 kinase activity). The method can include administering to the patient a therapeutically effective amount of a compound of the invention, e.g., a compound of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468. In one embodiment, the disease or condition responsive to inhibition of a Janus kinase, e.g., JAK1 kinase, is asthma.

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

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

In one embodiment, the cancer is breast cancer, ovarian cancer, cervical cancer, prostate cancer, testicular cancer, penile cancer, genitourinary tract cancer, seminoma, esophageal cancer, laryngeal cancer, stomach cancer, gastric cancer, gastrointestinal cancer, skin cancer, keratoacanthoma, follicular cancer, melanoma, lung cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, lung squamous cell carcinoma, colon cancer, pancreatic cancer, thyroid cancer, papillary cancer, bladder cancer, liver cancer, biliary tract cancer, kidney cancer, bone cancer, bone marrow disease, lymphoma, hairy cell cancer, buccal and pharyngeal (oral cavity) cancer, lip cancer, tongue cancer, mouth cancer, salivary gland cancer, pharyngeal cancer, small intestine cancer, colon cancer, rectum cancer, anal cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, large intestine cancer, endometrial cancer, uterine cancer, brain cancer, central nervous system cancer, peritoneal cancer, hepatocellular carcinoma, head cancer, neck cancer, Hodgkin's cancer, or leukemia.

In one embodiment, the disease is a myeloproliferative disorder. In one embodiment, the myeloproliferative disorder is polycythemia vera, essential thrombocythemia, myelofibrosis, and chronic myeloid leukemia (CML).

Another embodiment includes the use of a compound of the invention, e.g., a compound of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, in the preparation of a medicament for treating a disease described herein (e.g., an inflammatory disorder, an immune disorder, or cancer). In one embodiment, the invention provides a method for treating a disease or condition described herein (e.g., an inflammatory disorder, an immune disorder, or cancer) by targeted inhibition of a JAK kinase, e.g., JAK1.

Combination therapy

The compounds of the present invention, such as compounds of formula (00A), (0A), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, can be used alone or in combination with other active agents for treatment. The second compound of the pharmaceutical composition or dosage regimen typically has an activity that is complementary to the compounds of the present invention so that they do not adversely affect each other. These active agents are suitably present in combination in an amount effective for the intended purpose. The compounds can be administered together or separately in a single pharmaceutical composition, and when administered separately, this can occur simultaneously or sequentially. This sequential administration can be close or distant in time.

For example, other compounds can be used in combination with the compounds of interest of the present invention for the prevention or treatment of inflammatory diseases, such as asthma. Therefore, the present invention also relates to pharmaceutical compositions comprising a therapeutically effective amount of a compound of the present invention and one or more other therapeutic agents. Suitable therapeutic agents for combined treatment with the compounds of the present invention include, but are not limited to: adenosine A2A receptor antagonists; anti-infectives; nonsteroidal glucocorticoid receptor (GR receptor) agonists; antioxidants; β2 adrenergic receptor agonists; CCR1 antagonists; chemokine antagonists (not CCR1); corticosteroids; CRTh2 antagonists; DP1 antagonists; formyl peptide receptor antagonists; histone deacetylase activators; chloride channel hCLCA1 blockers; epithelial sodium channel blockers (ENAC blockers; intercellular adhesion molecule 1 blockers (ICAM blockers); IKK2 inhibitors; JNK inhibitors; cyclooxygenase inhibitors (COX inhibitors); lipoxygenase inhibitors; leukotriene receptor antagonists; dual beta2 adrenergic receptor agonists/M3 receptor antagonists (MABA compounds); MEK-1 inhibitors; myeloperoxidase inhibitors (MPO inhibitors); muscarinic antagonists; p38 MAPK inhibitors; phosphodiesterase (PDE4) inhibitors; phosphatidylinositol 3-kinase delta inhibitors (PI3-kinase delta inhibitors); phosphatidylinositol 3-kinase gamma 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.

In addition, the compounds of the present invention, such as compounds of Formula (OOA), (OA), (A), (la), (lb), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, can be combined with the following active agents: (1) corticosteroids, such as alclometasone dipropionate, amelometasone, beclomethasone dipropionate, budesonide, buticotol propionate, biclesonide, blobetasol propionate, desisobutyrylciclesonide, dexamethasone, dtiprednol dicloacetate, fluocinolone acetonide, fluticasone furoate, fluticasone propionate, loteprednol carbonate ethyl (topical), or mometasone furoate; (2) β2- Adrenergic receptor agonists such as salbutamol, albuterol, terbutaline, fenoterol, bitolterol, carbuterol, clenbuterol, pirbuterol, rimoterol, terbutaline, tritoquinol, tulobuterol and long-acting β2-adrenergic receptor agonists such as metaproterenol, isoproterenol, isoprenaline, salmeterol, indacaterol, formoterol (including formoterol fumarate), arformoterol, carmoterol, abediterol, vilanterol trifenate, olodaterol; (3) corticosteroids/ Long-acting beta2 agonist combination products such as salmeterol/fluticasone propionate (also marketed as salmeterol/budesonide), formoterol/fluticasone propionate, formoterol/ciclesonide, formoterol/mometasone furoate, indacaterol/mometasone furoate, vilanterol trifenate/fluticasone furoate, or arformoterol/ciclesonide; (4) anticholinergics, for example, muscarinic-3 (M3) receptor antagonists such as ipratropium bromide, tiotropium bromide, aclidinium (LAS-34273), glycopyrrolate, and umeclidinium bromide; (5) M3-anticholinergic/β2-adrenergic receptor agonist combination products, such as vilanterol/umeclidinium olodaterol/tiotropium, glycopyrrolate/indacaterol (also sold as ), fenoterol hydrobromide/ipratropium bromide, salbutamol sulfate/ipratropium bromide, formoterol fumarate/glycopyrrolate, or aclidinium bromide/formoterol; (6) dual pharmacology M3-anticholinergic/β2-adrenergic receptor agonists such as batefenterol succinate, AZD-2115, or LAS-190792; (7) Leukotriene modifiers, for example, leukotriene antagonists such as montelukast, zafirulast or pranlukast or leukotriene biosynthesis inhibitors such as zileuton or LTB4 antagonists such as amelubant, or FLAP inhibitors such as fiboflapon, GSK-2190915; (8) phosphodiesterase-IV (PDE-IV) inhibitors (oral or inhaled), for example, roflumilast, cilomilast, oglemilast, rolipram, tetomilast, AVE-8112, revamilast, CHF 6001; (9) antihistamines, for example, selective histamine-1 (H1) receptor antagonists such as fexofenadine, citirizine, loratadine or astemizole or dual H1/H3 receptor antagonists such as GSK 835726 or GSK 1004723; (10) antitussives, such as codeine or dextramorphan; (11) mucolytics, such as N-acetylcysteine or fudostein; (12) expectorants/viscoelastic modifiers, such as ambroxol, hypertonic solutions (such as saline or mannitol), or surfactants; (13) mucolytic peptides, such as recombinant human deoxyribonuclease I (DNAse-α and rhDNase) or spirocidin; (14) antibiotics, such as azithromycin, tobramycin, or aztreonam; (15) non-selective COX-1/COX-2 inhibitors, such as ibuprofen or ketoprofen; (16) COX-2 inhibitors, such as celecoxib and rofecoxib; (17) VLA-4 antagonists, such as those described in WO97/03094 and WO97/02289, each of which is incorporated herein by reference; (18) TACE inhibitors and TNF-α inhibitors, such as anti-TNF monoclonal antibodies such as and CDP-870 and TNF receptor immunoglobulin molecules, such as (19) matrix metalloproteinase inhibitors, such as MMP-12; (20) human neutrophil elastase inhibitors, such as BAY-85-8501 or those described in WO2005/026124, WO2003/053930 and WO06/082412, each of which is incorporated herein by reference; (21) A2b antagonists, such as those described in WO2002/42298, each of which is incorporated herein by reference; (22) modulators of chemokine receptor function, such as antagonists of CCR3 and CCR8; (23) compounds that modulate the action of other prostaglandin receptors, such as thromboxane _A2 antagonists; DP1 antagonists, such as laropiprant or asapiprant CRTH2 antagonists such as OC000459, fevipiprant, ADC 3680 or ARRY 502; (24) PPAR agonists, including PPARα agonists (e.g., fenofibrate), PPARδ agonists, PPARγ agonists such as pioglitazone, rosiglitazone, and balaglitazone; (25) methylxanthines such as theophylline or aminophylline and methylxanthine/corticosteroid combinations such as theophylline/budesonide, theophylline/fluticasone propionate, theophylline/ciclesonide, theophylline/mometasone furoate, and theophylline/beclomethasone propionate; (26) A2a agonists such as those described in EP1052264 and EP1241176; (27) CXCR2 or IL-8 antagonists such as AZD-5069, AZD-4721, danirixin; (28) IL-R signaling modulators such as kineret and ACZ 885; (29) MCP-1 antagonists, such as ABN-912; (30) p38 MAPK inhibitors, such as BCT197, JNJ49095397, losmapimod or PH-797804; (31) TLR7 receptor agonists such as AZD 8848; (32) PI3-kinase inhibitors, such as RV1729 or GSK2269557.

In some embodiments, the compounds of the present invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, can be used in combination with one or more additional drugs, such as anti-hyperproliferative agents, anticancer agents, cytostatic agents, cytotoxic agents, anti-inflammatory agents, or chemotherapeutic agents, such as those disclosed in U.S. Patent Application Publication No. 2010/0048557, incorporated herein by reference. The compounds of the present invention, e.g., compounds of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or compounds of Table 1 or Examples 1-468, can also be used in combination with radiation therapy or surgery, as is known in the art.

Products

Another embodiment includes an article of manufacture (e.g., a kit) for treating a disease or condition responsive to inhibition of a Janus kinase, such as JAK1 kinase. The kit may comprise:

(a) a first pharmaceutical composition comprising a compound of the present invention, e.g., a compound of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468; and

(b) Instructions for use.

In another embodiment, the kit further comprises:

(c) a second pharmaceutical composition, eg, a pharmaceutical composition comprising an active agent as described above for use in therapy, eg, an active agent for treating an inflammatory disorder or a chemotherapeutic agent.

In one embodiment, the instructions describe administering the first and second pharmaceutical compositions simultaneously, sequentially, or separately to a patient in need thereof.

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.

Containers for use include, for example, bottles, vials, syringes, blister packs, and the like. The container can be formed from a variety of materials, such as glass or plastic. The container contains a compound of the invention, such as a compound of Formula (OOA), (OA), (A), (Ia), (Ib), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X), or a compound of Table 1 or Examples 1-468, or a combination thereof, which is effective for treating a condition and can have a sterile access port (e.g., the container can be an intravenous solution bag or a vial with a stopper pierceable by a hypodermic needle). The label or package insert indicates that the compound or composition is used to treat a selected condition, such as asthma or cancer. In one embodiment, the label or package insert indicates that the compound or composition can be used to treat a condition. In addition, the label or package insert can indicate that the patient to be treated is a patient with a condition characterized by overactive or irregular Janus kinase activity (e.g., overactive or irregular JAK1 activity). The label or package insert can also indicate that the compound or composition can be used to treat other conditions.

Alternatively or additionally, the kit may further comprise a second (or third) container comprising a pharmaceutically acceptable buffer (e.g., bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, or dextrose solution). It may also include other materials desirable from a commercial and user perspective, including other buffers, diluents, filters, needles, and syringes.

The following examples are included to illustrate the present invention. However, it should be understood that these examples do not limit the present invention and are intended only to provide methods for implementing the present invention. Those skilled in the art will recognize that the chemical reactions described can be easily adapted to prepare other compounds of the present invention, and alternative methods for preparing the compounds are within the scope of the present invention. For example, the synthesis of non-exemplary compounds of the present invention can be successfully performed by modifications apparent to those skilled in the art, such as by appropriately protecting interfering groups, by utilizing other suitable reagents known in the art in addition to those described, or by making conventional modifications to the reaction conditions. Alternatively, other reactions disclosed herein or known in the art will be considered suitable for preparing other compounds of the present invention.

DETAILED DESCRIPTION

Example

Although the present invention has been described and illustrated with a certain degree of particularity, it should be understood that this disclosure is made by way of example only and that many variations in the combination and arrangement of parts may be ascertained by those skilled in the art without departing from the spirit and scope of the invention as defined by the claims.

General experimental details:

Unless otherwise stated, all solvents and commercial reagents were used as received. When the product was purified by silica gel chromatography, a glass column or SPE SiII column manually packed with silica gel (Kieselgel 60, 220-440 mesh, 35-75 μm) was used. 'Isolute SPE Si column' refers to a pre-packed polypropylene column containing unbonded activated silica with irregular particles having an average size of 50 μm and a nominal porosity. When an SCX-2 column was used, "SCX-2 column" refers to a pre-packed polypropylene column containing a non-end-capped propylsulfonic acid functionalized silica strong cation exchange adsorbent.

Preparative HPLC conditions

HPLC System 1: C18 reversed-phase column (250 x 21.2 mm Gemini column, 5 μm particle size), gradient elution: A: water + 0.1% formic acid; B: acetonitrile + 0.1% formic acid, flow rate typically 20 mL/min, with increasing gradient of B. UV detection at 254 nm. The compound was obtained as the formate salt as described above.

HPLC System 2: Phenylhexyl reversed-phase column (250 x 21.2 mm Gemini column, 5 μm particle size), gradient elution: A: water + 0.1% formic acid; B: acetonitrile + 0.1% formic acid, flow rate typically 20 mL/min, with B increasing in a 1%/min gradient. UV detection at 254 nm. The compound was obtained as the formate salt as described above.

HPLC system 3: C18 reverse phase capped column (250 x 21.2 mm Gemini column, 5 μm particle size), gradient elution: A: water + 0.1% ammonia; B: acetonitrile + 0.1% ammonia, flow rate typically 20 mL/min with increasing gradient of B. UV detection at 254 nm. The compound was obtained as the free base.

NMR analysis method

^1H NMR spectra were recorded at ambient temperature using one of the following:

i. Varian Unity Inova (400 MHz) spectrometer with 400 4NUC 5 mm probe.

ii. Bruker Avance DRX400 (400 MHz) spectrometer with PABBO 5 mm probe.

iii. Varian Unity Inova (400 MHz) spectrometer with 5 mm reverse detection triple resonance probe.

iv. Bruker Avance DRX (400 MHz) spectrometer with 5 mm reverse-detected triple-resonance TXI probe.

Chemical shifts are expressed in ppm relative to tetramethylsilane.

LC-MS analysis method

LC-MS information is provided in Table 2.

LC-MS Method 1: Waters Platform LC with a C18-reverse phase column (30×4.6 mm Phenomenex Luna 3 μm particle size), eluted with A: water + 0.1% formic acid; B: acetonitrile + 0.1% formic acid. Gradient:

Detection - MS, ELS, UV (100 μl split to MS with online UV detector)

MS Ionization Methods - Electrospray (Positive and Negative)

LC-MS method 2: Waters Micromass ZMD with a C18-reverse phase column (30×4.6 mm Phenomenex Luna 3 μm particle size), eluted with A: water + 0.1% formic acid; B: acetonitrile + 0.1% formic acid. Gradient:

Detection - MS, ELS, UV (100 μl split to MS with online UV detector)

MS Ionization Methods - Electrospray (Positive and Negative)

LC-MS method 3: VGPlatform II quadrupole spectrometer with a C18-reverse phase column (30×4.6 mm Phenomenex Luna 3 μm particle size), eluted with A: water + 0.1% formic acid; B: acetonitrile + 0.1% formic acid. Gradient:

Detection - MS, ELS, UV (200 μl/min split to ESI source with online HP1050 DAD detection)

MS Ionization Methods - Electrospray (Positive and Negative)

LC-MS method 4: Finnigan AQA with a C18-reverse phase column (30×4.6 mm Phenomenex Luna 3 μm particle size), eluting with A: water + 0.1% formic acid; B: acetonitrile + 0.1% formic acid. Gradient:

Detection-MS, ELS, UV

MS Ionization Method - Electrospray (Positive Ion)

LC-MS method 5: Waters Micromass ZQ2000 quadrupole mass spectrometer with a C18-reversed phase column (100×2.1 mm Acquity BEH C18 1.7μ, Acquity BEHShield RP18 1.7μ, or Acquity HSST3 1.8μ), maintained at 40°C, eluted with A: water + 0.1% formic acid; B: acetonitrile + 0.1% formic acid. Gradient:

Detection – MS, UV PDA

MS Ionization Method - Electrospray (Positive Ion)

LC-MS Method 6: Waters Acquity UPLC with a C18-reverse phase column (100 x 2.1 mm Acquity BEH C18, 1.7 μm particle size), eluted with solvent A: water + 0.1% formic acid; solvent B: acetonitrile + 0.1% formic acid at 40°C. Gradient:

Detection-UV (220nm)

MS ionization method - ESI ⁺

LC-MS method 7: SHIMADZU20A HPLC with a C18-reverse phase column (30 x 2.1 mm Xtimate™-C18, 3 μm particle size), eluted with solvent A: water + 0.038% trifluoroacetic acid; solvent B: acetonitrile + 0.02% trifluoroacetic acid. Gradient:

Detection-UV (220nm)

MS ionization method - ESI ⁺

LC-MS method 8: SHIMADZU20A HPLC with a C18-reverse phase column (30 x 2.1 mm Xtimate™-C18, 3 μm particle size), eluted with solvent A: water + 0.038% trifluoroacetic acid; solvent B: acetonitrile + 0.02% trifluoroacetic acid. Gradient:

Detection-UV (220nm)

MS ionization method - ESI ⁺

LC-MS method 9: SHIMADZU20A HPLC with a C18-reverse phase column (30 x 2.1 mm Xtimate™-C18, 3 μm particle size), eluted with solvent A: water + 0.038% trifluoroacetic acid; solvent B: acetonitrile + 0.02% trifluoroacetic acid. Gradient:

Detection-UV (220nm)

MS ionization method - ESI ⁺

LC-MS method 10: SHIMADZU 20A HPLC with a C18-reverse phase column (30 x 2.1 mm Xtimate TM-C18, 3 μm particle size), eluted with solvent A: water + 0.038% trifluoroacetic acid; solvent B: acetonitrile + 0.02% trifluoroacetic acid. Gradient:

Detection-UV (220nm)

MS ionization method - ESI ⁺

LC-MS method 11: Agilent 1200 HPLC with Xtimate C18 column (3um, 30x2.1mm), eluting with A: water + 0.038% trifluoroacetic acid; B: acetonitrile + 0.02% trifluoroacetic acid.

Detection – MS, UV (PDA detector)

MS Ionization Method - Electrospray (Positive Ion)

LC-MS method 12: Agilent 1200 HPLC with an Xtimate C18 column (3 μm, 30×2.1 mm), eluting with A: water + 0.038% trifluoroacetic acid; B: acetonitrile + 0.02% trifluoroacetic acid.

Detection – MS, UV (PDA detector)

MS Ionization Method - Electrospray (Positive Ion)

LC-MS method 13: Agilent 1200 HPLC with an Xtimate C18 column (3 μm, 30×2.1 mm), eluting with A: water + 0.038% trifluoroacetic acid; B: acetonitrile + 0.02% trifluoroacetic acid.

Detection – MS, UV (PDA detector)

MS Ionization Method - Electrospray (Positive Ion)

LC-MS method 14: SHIMADZU 20A HPLC with a C18-reverse phase column (50 x 2.0 mm Shim-pack XR-ODS, 1.6 μm particle size), eluted with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient:

Detection-UV (254nm)

MS ionization method - ESI ⁺

LC-MS method 15: SHIMADZU 20A HPLC with a C18-reverse phase column (50 x 3.0 mm Shim-pack XR-ODS, 1.6 μm particle size), eluted with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient:

Detection-UV (254nm)

MS ionization method - ESI ⁺

LC-MS method 16: SHIMADZU 20A HPLC with a C18-reverse phase column (50 x 3.0 mm Shim-pack XR-ODS, 2.2 μm particle size), eluted with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient:

Detection-UV (254nm)

MS ionization method - ESI ⁺

LC-MS method 17: SHIMADZU 20A HPLC with a C18-reverse phase column (50 x 3.0 mm Shim-pack XR-ODS, 2.2 μm particle size), eluted with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient:

Detection-UV (254nm)

MS ionization method - ESI ⁺

LC-MS method 18: SHIMADZU LCMS-2020 HPLC column (150x4.6 mm Venusil XBP Silica, 5.0 μm particle size), eluted with solvent A: hexane; solvent B: ethanol.

Detection-UV (254nm)

MS ionization method - ESI ⁺

LC-MS method 19: SHIMADZU 20A HPLC with a C18-reverse phase column (50x3 mm Xtimate TM-C18, 2.2 μm particle size), eluted with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 20: SHIMADZU 20A HPLC with a C18-reverse phase column (50x3 mm Xtimate TM-C18, 2.2 μm particle size), eluted with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 21: SHIMADZU 20A HPLC with a C18-reverse phase column (50x3 mm Xtimate TM-C18, 2.2 μm particle size), eluted with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 22: SHIMADZU 20A HPLC with a C18-reverse phase column (50x3 mm Xtimate TM-C18, 2.2 μm particle size), eluted with solvent A: water + 0.05% formic acid; solvent B: acetonitrile + 0.05% formic acid. Gradient:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 23: SHIMADZU 20A HPLC with a C18-reverse phase column (50x3 mm Xtimate TM-C18, 2.2 μm particle size), eluted with solvent A: water + 0.1% formic acid; solvent B: acetonitrile + 0.05% formic acid. Gradient:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 24: SHIMADZU 20A HPLC with a C18-reverse phase column (50x3 mm Xtimate TM-C18, 2.2 μm particle size), eluted with solvent A: water + 0.04% ammonium hydroxide; solvent B: acetonitrile. Gradient:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 25: SHIMADZU 20A HPLC with a C18-reverse phase column (30 x 2.1 mm Xtimate™-C18, 3 μm particle size), eluted with solvent A: water + 0.05% trifluoroacetic acid; solvent B: acetonitrile + 0.05% trifluoroacetic acid. Gradient:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 26: SHIMADZU 20A HPLC with a C18-reverse phase column (50 x 3.0 mm Xtimate™-C18, 2.6 μm particle size), eluted with solvent A: water/0.1% formic acid; solvent B: acetonitrile/0.1% formic acid. Gradient:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 27: SHIMADZU 20A HPLC with a C18-reverse phase column (50 x 2.1 mm Xtimate™-C18, 2.7 μm particle size), eluted with solvent A: water/0.05% formic acid; solvent B: acetonitrile/0.1% formic acid:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 28: SHIMADZU 20A HPLC with a C18-reverse phase column (50 x 3.0 mm Xtimate TM-C18, 2.5 μm particle size), eluted with solvent A: water/5 mM NH ₄ HCO ₃ ; solvent B: acetonitrile:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 29: SHIMADZU LCMS-2020 C18 reverse phase column (50x3.0 mm, Shim-pack XR-ODS, 2.5 μm particle size), eluted with solvent A: water/0.05% TFA ₃ ; solvent B: acetonitrile/0.05% TFA:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 30: SHIMADZU LCMS-2020 C18 reverse phase column (50x2.1 mm, Shiseido CAPCELLCORE C18, 2.7 μm particle size), eluted with solvent A: water/0.05% TFA ₃ ; solvent B: acetonitrile/0.05% TFA:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 31: SHIMADZU LCMS-2020 C18 reverse phase column (50x2.1 mm, Waters BEH C18, 1.7 μm particle size), eluted with solvent A: water/0.1% TFA ₃ ; solvent B: acetonitrile/0.1% TFA:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 32: SHIMADZU LCMS-2020 C18 reverse phase column (50x3.0 mm, Gemini-NX 3μ C18110A, 3.0 μm particle size), eluted with solvent A: water/6.5 mM NH ₄ HCO ₃ pH 10; solvent B: acetonitrile

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 33: SHIMADZU LCMS-2020 C18 reverse phase column (50x2.1 mm, Waters BEH C18, 1.7 μm particle size), eluted with solvent A: water/0.05% TFA ₃ ; solvent B: acetonitrile/0.05% TFA:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 34: SHIMADZU LCMS-2020 C18 reverse phase column (50x2.1 mm, Waters BEH C18, 1.7 μm particle size), eluted with solvent A: water/0.05% TFA ₃ ; solvent B: acetonitrile/0.05% TFA:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 35: SHIMADZU LCMS-2020 C18 reverse phase column (50x3.0 mm, Shim-pack XR-ODS, 2.2 μm particle size), eluted with solvent A: water/0.05% TFA ₃ ; solvent B: acetonitrile/0.05% TFA:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC-MS method 36: SHIMADZU 20A HPLC with a C18-reverse phase column (50 x 3.0 mm, Gemini-NX 3μ C18 110A, 3.0 μm particle size), eluted with solvent A: water/5 mM NH ₄ HCO ₃ ; solvent B: acetonitrile:

Detection - UV (220 and 254 nm)

MS Ionization Method – Electrospray (Positive Ion)

LC/MS Method 37: Agilent 10-min LCMS method:

Experiments were performed on an Agilent 1100 HPLC coupled to an Agilent MSD mass spectrometer using ESI as the ionization source. LC separations were performed using a Phenomenex XB-C18, 1.7 μm, 50 × 2.1 mm column at a flow rate of 0.4 ml/min. Solvent A was water with 0.1% FA, and solvent B was acetonitrile with 0.1% FA. The gradient consisted of 2% to 98% solvent B over 7 minutes, followed by a 1.5-minute equilibration and a 1.5-minute hold at 97% B. The LC column temperature was 40°C. UV absorbance was collected at 220 nm and 254 nm, and full-scan mass spectra were used for all experiments.

LC/MS Method 38: Waters 10-min LCMS method:

Experiments were performed on a Waters Acquity UPLC equipped with a Waters LCT Premier XE mass spectrometer using ESI as the ionization source. LC separations were performed using an Acquity UPLC BEH C18, 1.7 μm, 2.1 × 50 mm column, with a flow rate of 0.6 ml/min. Solvent A was water with 0.05% TFA, and solvent B was acetonitrile with 0.05% TFA. The gradient consisted of 2–98% solvent B over 7 minutes, followed by a 1.5-minute equilibration and a 1.5-minute hold at 98% B. The LC column temperature was 40°C. UV absorbance was collected at 220 nm and 254 nm, and full-scan mass spectra were used for all experiments.

LC/MS Method 39: Shimadzu 5-min LCMS method:

Experiments were performed on a Shimadzu LC with an LC-30AD solvent pump, an SPD-M30A UV detector, and a 2020MS using ESI and APCI as ionization sources. LC separations were performed using a Waters UPLC BEH C18, 1.7 mm, 2.1 × 50 mm column with a flow rate of 0.7 ml/min. Solvent A was water with 0.1% FA, and solvent B was acetonitrile with 0.1% FA. The gradient consisted of 2–98% solvent B over 4.5 minutes, followed by a 0.5-minute equilibration hold at 98% B for 0.5 minutes. The LC column temperature was 40°C. UV absorbance was collected at 254 nm, and full-scan mass spectra were used for all experiments.

Preparative quality-oriented automated purification conditions

MDAP Method 1: Agilent 1260 Infinite Purification System. Agilent 6100 Series Single Quadrupole LC/MS. XSEELECT CSH Prep C18 5μm OBD, 30x150mm, RT. Elution was performed at 60ml/min using solvent A: 0.1% formic acid in water; solvent B: 0.1% formic acid in acetonitrile, 10% to 95% over 22 minutes, with a focused gradient. Injection of 20-60mg/ml DMSO solutions (with optional formic acid and water)

MDAP Method 2: Agilent 1260 Infinite Purification System. Agilent 6100 Series Single Quadrupole LC/MS. XBridge Prep C18 5μm OBD, 30x150mm, RT. Elution was performed at 60ml/min using solvent A: 0.1% ammonia; solvent B: 0.1% ammonia in acetonitrile, 10% to 95% over 22 minutes, with a focused gradient. Injection of 20-60mg/ml DMSO solutions (with optional formic acid and water).

abbreviation

CH ₃ OD deuterated methanol

CDCl ₃ -deuterated chloroform

DCM dichloromethane

DIPEA Diisopropylethylamine

DMAP 4-dimethylaminopyridine

DMF dimethylformamide

DMSO dimethyl sulfoxide

DMSO-d6 deuterated dimethyl sulfoxide

EtOAc

EtOH

HOAc acetic acid

g grams

HATU (O-(7-Azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate)

HCl

IMS Industrial methylated spirits

L liter

MDAP mass-directed automated purification

MeOH methanol

mg milligrams

mL milliliters

SCX-2 Strong Cation Exchange

THF Tetrahydrofuran

TFA trifluoroacetic acid

Example A

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxyphenyl)-1H-pyrazol-4-yl]amide

To a solution of 2-bromo-4-chlorophenol (4.98 g, 24.0 mmol) in DMF (25 mL) was added sodium chlorodifluoroacetate (8.42 g, 55.2 mmol), cesium carbonate (10.97 g, 33.67 mmol) and water (2.5 mL). The reaction was stirred at 100° C. for 16 hours. The reaction mixture was partitioned between ethyl acetate and water, and the organic portion was washed with brine, dried (MgSO ₄ ), and evaporated. The crude product was purified by flash chromatography on silica gel, eluting with 0-20% EtOAc in heptane, to afford 2-bromo-4-chloro-1-(difluoromethoxy)benzene as a clear, colorless oil (2.98 g, 48%). LCMS (ESI) showed no m/z signal; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 7.90 (d, 1H), 7.54 (dd, 1H), 7.38 (d, 1H), 7.28 (t, 1H).

To a solution of 4-nitro-1-(2-trimethylsilylethoxymethyl)-1H-pyrazole (prepared as described in WO2011003065) (46.5 g, 191 mmol) in DMA (350 mL) was added 2-bromo-4-chloro-1-difluoromethoxybenzene (64.0 g, 248 mmol), palladium(II) acetate (2.15 g, 9.6 mmol), di-(adamantyl)-n-butylphosphine (5.0 g, 13.4 mmol), potassium carbonate (79.2 g, 573 mmol), and trimethylacetic acid (5.27 g, 51.6 mmol). The mixture was degassed with nitrogen for 10 minutes and then heated at 130°C for 8 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, washed with water and brine, dried (MgSO ₄ ), filtered, and evaporated. The resulting crude material was purified by flash chromatography on silica gel using 0-10% EtOAc in cyclohexane to afford 5-(5-chloro-2-difluoromethoxyphenyl)-4-nitro-1-(2-trimethylsilylethoxymethyl)-1H-pyrazole (62.4 g, 78%). ¹ H NMR (400 MHz, CDCl ₃ ) δ: (ppm) 8.24 (s, 1H), 7.52-7.53 (m, 2H), 6.39 (t, 1H), 5.29-5.30 (m, 2H), 3.63-3.64 (m, 2H), 0.90 (s, 9H).

To a solution of 5-(5-chloro-2-difluoromethoxyphenyl)-4-nitro-1-(2-trimethylsilylethoxymethyl)-1H-pyrazole (62 g, 148 mmol) in ethanol (600 mL) was added water (200 mL), ammonium chloride (32 g, 590 mmol), and iron powder (41 g, 740 mmol). The mixture was heated at 80°C for 2 hours and then cooled to room temperature. The remaining solid was removed by filtration. The filtrate was evaporated under reduced pressure, diluted with water, and extracted twice with DCM. The combined organic extracts were washed with water and brine, dried ( _MgSO4 ), and evaporated to give a black oil. The oil was purified by flash chromatography on silica gel, eluting with 0-25% EtOAc in DCM. The appropriate fractions were collected and the solvent removed in vacuo to afford 5-(5-chloro-2-difluoromethoxyphenyl)-1-(2-trimethylsilyl-ethoxymethyl)-1H-pyrazol-4-ylamine as a brown oil (30.8 g, 54%). ¹ H NMR (400 MHz, CDCl ₃ ) δ: (ppm) 7.56 (d, 1H), 7.44 (dd, 1H), 7.34 (s, 1H), 7.30-7.25 (m, 1H), 6.37 (t, 1H), 5.29 (s, 2H), 3.56 (t, 2H), 0.88 (dd, 2H), 0.00 (s, 9H).

A solution of 5-(5-chloro-2-difluoromethoxyphenyl)-1-(2-trimethylsilyl-ethoxymethyl)-1H-pyrazol-4-ylamine (60.0 g, 154 mmol) in THF (100 mL) was added dropwise over 30 minutes to an ice/water-cooled mixture of pyrazolo[1,5-a]pyrimidine-3-carbonyl chloride (27.8 g, 153 mmol) and DIPEA (49.5 g, 383 mmol) in THF (300 mL). After the addition was complete, the mixture was stirred at room temperature for 1 hour. The solvent was evaporated and the residue was diluted with 0.5N aqueous HCl and extracted with ethyl acetate. The combined organic extracts were purified by removing the residual solid and the filtrate was washed with 1M _{aqueous K₂CO₃} , water, and brine, dried ( _Na₂SO₄ ), and evaporated to give a red solid. The solid was triturated with 10% diethyl ether in _cyclohexane . The solid was collected by filtration, washed with a 1:1 solution of diethyl ether in cyclohexane, and air-dried to afford pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [5-(5-chloro-2-difluoromethoxyphenyl)-1-(2-trimethylsilylethoxymethyl)-1H-pyrazol-4-yl]amide as an off-white solid (59.2 g, 73%). ¹ H NMR (300MHz, CDCl ₃ ): δ (ppm) 9.61 (s, 1H), 8.77-8.78 (m, 1H), 8.51 (dd, 1H), 8.36 (s, 1H), 7.65 (d, 1H), 7.52 (dd, 1H), 7.36 (d,1H),7.29(s,1H),7.01(dd,1H),6.42(t,1H),5.39-5.41(m,2H),3.60-3.64(m,2H),0.87-0.89(m,2H),0.09(s,9H).

A suspension of pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [5-(5-chloro-2-difluoromethoxyphenyl)-1-(2-trimethylsilylethoxymethyl)-1H-pyrazol-4-yl]amide (59.0 g, 110 mmol) in methanol (420 mL) was treated with 6N HCl (80 mL), and the mixture was heated at 60°C for 4 hours. The solvent was evaporated and the residue was triturated with water. The solid was collected by filtration, washed with water, and air-dried. The solid was triturated with a minimum volume of acetonitrile, collected by filtration, washed with diethyl ether, and dried under high vacuum at 60°C to afford the title compound as a yellow solid (42.9 g, 96%). ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.71 (s, 1H), 9.34 (dd, 1H), 8.68-8.69 (m, 1H), 8.66 (s, 1H), 8.25 (s, 1H), 7.62 (dd, 2H), 7.43-7.46 (m, 1H),7.29(dd,1H),7.23(d,1H).

Example B

tert-Butyl (3-((3-(5-chloro-2-(difluoromethoxy)phenyl)-1H-pyrazol-4-yl)carbamoyl)pyrazolo[1,5-a]pyrimidin-2-yl)carbamate

To a solution of 2-tert-butoxycarbonylamino-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (2.78 g, 0.01 mol) in DMF (40 mL) were added DIPEA (3.9 g, 0.03 mol), HATU (3.8 g, 0.01 mol), and 5-(5-chloro-2-difluoromethoxyphenyl)-1H-pyrazol-4-ylamine (2.6 g, 0.01 mmol), and the reaction mixture was stirred at room temperature overnight. The reaction mixture was poured into water and the precipitate was collected to obtain the title compound as a solid (4 g, 77%). LCMS (Method 7) [M+Na] ⁺ =541.9, R _T =1.20min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 13.13 (s, 1H), 9.58 (s, 2H), 9.2 (dd, 1H, J = 7.2, 1.6 Hz), 8.58 (d, 1H, J = 2), 8.27 (s, 1H), 7.60 (m, 2H), 7.23-7.04 (m, 3H), 1.48 (s, 9H).

Example C

{3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid

A solution of pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxyphenyl)-1H-pyrazol-4-yl]amide (42.0 g, 104 mmol) in DMF (400 mL) was treated with cesium carbonate (37.2 g, 114 mmol) and tert-butyl bromoacetate (22.3 g, 114 mmol) and allowed to stir at room temperature for 18 hours. The reaction mixture was diluted with water and extracted twice with ethyl acetate. The combined organic extracts were washed with water and brine, dried ( _Na2SO4 ), filtered, and evaporated to afford an orange solid. The resulting solid was triturated with diethyl ether and then recrystallized from acetone to afford a white solid (33.2 g, 62%). _The mother liquors were combined and evaporated to afford an orange oil. The residual oil was purified by flash chromatography on silica gel using 0-50% EtOAc in DCM. Appropriate fractions were combined and evaporated to give a white solid (12.3 g, 23%). Combined, the resulting products yielded tert-butyl {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetate (45.2 g, 85%). LCMS (Method 3) [M+H] ⁺ = 519.1, _RT = 3.72 min. ¹ H NMR (400 MHz, CDCl ₃ ) δ: (ppm) 9.86 (s, 1H), 8.75-8.76 (m, 2H), 8.56 (dd, 1H), 8.40 (s, 1H), 7.70 (d, 1H), 7.42 (dd, 1H), 6.99 (dd, 1H), 6.47 (t, 1H), 4.86 (s, 2H), 1.49 (s, 9H).

To a solution of tert-butyl {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetate (45.0 g, 86.7 mmol) in dichloromethane (350 mL) was added TFA (100 mL), and the mixture was stirred at room temperature for 18 hours. The solvent was evaporated, and the resulting residue was triturated with diethyl ether. The resulting solid was collected by filtration, washed with diethyl ether, and air-dried to afford the title compound as a white solid (40.0 g, 99%). LCMS (Method 3) [M+H] ⁺ = 463.1, _RT = 2.89 min. ¹ H NMR (400 MHz, CDCl ₃ ) δ: (ppm) 9.86 (s, 1H), 8.78 (dd, 1H), 8.71 (s, 1H), 8.56 (dd, 1H), 8.40 (s, 1H), 7.70 (d, 1H), 7.42 (dd, 1H), 7.30 (s, 1H), 6.99 (dd, 1H), 6.47 (t, 1H), 4.86 (s, 2H), 1.49 (s, 9H).

Example D

[4-[(2-Amino-pyrazolo[1,5-a]pyrimidine-3-carbonyl)-amino]-3-(5-chloro-2-difluoromethoxy-phenyl)-pyrazol-1-yl]-acetic acid

A solution of tert-butyl (3-((3-(5-chloro-2-(difluoromethoxy)phenyl)-1H-pyrazol-4-yl)carbamoyl)pyrazolo[1,5-a]pyrimidin-2-yl)carbamate (1.0 g, 1.92 mmol) in DMF (10 mL) was treated with cesium carbonate (0.69 g, 2.12 mmol) and tert-butyl bromoacetate (0.43 g, 2.12 mmol) and stirred at room temperature for 2.5 hours. The reaction mixture was diluted with water, and the resulting precipitate was collected by filtration. The resulting solid was purified by flash chromatography on silica gel, eluting with 0-70% EtOAc in cyclohexane. Appropriate fractions were combined and evaporated to give [4-[(2-tert-butoxycarbonylamino-pyrazolo[1,5-a]pyrimidine-3-carbonyl)-amino]-3-(5-chloro-2-difluoromethoxy-phenyl)-pyrazol-1-yl]-acetic acid tert-butyl ester as a white solid (0.91 g, 74%). LCMS (Method 3) [M+H] ⁺ = 633.9, _RT = 4.46 min. ¹ H NMR (400 MHz, CDCl ₃ ) δ: (ppm) 9.78 (s, 1H), 9.70 (s, 1H), 8.76 (dd, 1H, J = 1.8, 6.8 Hz), 8.47 (dd, 1H, J = 1.8, 6.8 Hz). 4.3Hz),8.34(s,1H),7.69(d,1H,J＝2.5Hz),7.41(dd,1H,J＝2.7,8.7Hz),7.28(d, 1H, J＝8.8Hz), 6.92 (dd, 1H, J＝4.4, 6.9Hz), 6.47 (t, 1H, J＝74.0Hz), 4.85 (s, 2H), 1.56 (s, 9H), 1.49 (s, 9H).

To a solution of [4-[(2-tert-butoxycarbonylamino-pyrazolo[1,5-a]pyrimidine-3-carbonyl)-amino]-3-(5-chloro-2-difluoromethoxy-phenyl)-pyrazol-1-yl]-acetic acid tert-butyl ester (0.91 g, 1.43 mmol) in dichloromethane (7 mL) was added TFA (15 mL), and the mixture was stirred at room temperature for 6 hours. The solvent was evaporated and the resulting residue was azeotroped with dichloromethane/methanol to afford the title compound as a pale yellow solid (0.66 g, 97%). LCMS (Method 3) [M+H] ⁺ = 477.8, _RT = 2.95 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 13.15 (br s, 1H), 9.56 (s, 1H), 8.93 (dd, 1H, J = 1.5, 6.8 Hz), 8.37-8.34 (m, 1H), 8.35 (s, 1H), 7.63 (dd, 1H, J = 2.8, 8.8 Hz), 7.56 (d, 1H, J = 2.8 Hz), 7.46 (d, 1H, J = 8.9Hz), 7.26 (t, 1H, J = 73.4Hz), 7.00 (dd, 1H, J = 4.5, 6.8Hz), 6.58 (br s, 2H), 5.05 (s, 2H).

Example E

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [2-bromo-4-(5-chloro-2-difluoromethoxy-phenyl)-thiazol-5-yl]-amide

Sodium hydride (60% dispersion in mineral oil, 2.17 g, 54.2 mmol) was added portionwise to a stirred solution of 1-(5-chloro-2-methoxy-phenyl)-ethanone (10.0 g, 54.2 mmol) in THF (100 mL) at 0°C. The mixture was then stirred for 10 minutes before diethyl carbonate (7.68 g, 65.0 mmol) was added and stirred for an additional hour. The mixture was allowed to warm to room temperature for 2 hours and then heated to 65°C for 2 hours. Diethyl ether was added, and the organics were washed with water and brine before being evaporated to dryness. The resulting residue was purified by silica gel flash chromatography (50 to 100% dichloromethane in cyclohexane) to yield 3.41 g of ethyl 3-(5-chloro-2-methoxyphenyl)-3-oxopropanoate. LCMS (Method 1) [M+H] ⁺ = 257.2, _RT = 3.55 min. ¹ H NMR (400 MHz, CDCl ₃ ): δ (ppm) 7.59 (d, 1H), 7.38 (dd, 1H), 6.89 (d, 1H), 4.18 (q, 2H), 3.95 (s, 2H), 3.88 (s, 3H), 1.24 (t, 3H).

Bromine (0.70 mL, 13.6 mmol) was added to a solution of ethyl 3-(5-chloro-2-methoxyphenyl)-3-oxopropanoate (3.39 g, 13.2 mmol) in dioxane (25 mL) and stirred for 1 hour. The reaction was poured into ice water and extracted with ethyl acetate. The organics were washed with water and brine and evaporated to dryness to give ethyl 2-bromo-3-(5-chloro-2-methoxyphenyl)-3-oxopropanoate. LCMS (Method 1) [M+H] ⁺ = 337.2, _RT = 3.84 min.

A mixture of ethyl 2-bromo-3-(5-chloro-2-methoxyphenyl)-3-oxopropanoate (assumed to be 13.2 mmol) and thiourea (1.01 g, 13.3 mmol) in ethanol (25 mL) was heated at reflux for 3 hours and then cooled to room temperature for 18 hours. The resulting solid was removed by filtration and the filtrate was evaporated under vacuum. DCM was added to the residue, and the organic matter was washed with sodium bicarbonate (saturated aqueous solution), water, and brine and evaporated to dryness. The residue was triturated (DCM) to give ethyl 2-amino-4-(5-chloro-2-methoxyphenyl)thiazole-5-carboxylate (1.30 g, 31%) as a yellow solid. LCMS (Method 1) [M+H] ⁺ = 313.2, _RT = 3.15 min. 1H NMR (400 MHz, DMSO-d ₆ ): δ (ppm) 7.77 (s, br, 2H), 7.39 (dd, 1H), 7.22 (d, 1H), 7.05 (d, 1H), 4.00 (q, 2H), 3.70 (s, 3H), 1.04 (t, 3H).

A mixture of copper bromide (1.07 g, 4.79 mmol) in acetonitrile (20 mL) was degassed with nitrogen and cooled to 0 ° C. Then, tert-butyronitrile (0.80 mL, 6.00 mmol) was added, followed by a suspension of ethyl 2-amino-4-(5-chloro-2-methoxyphenyl)thiazole-5-carboxylate (1.25 g, 3.99 mmol) in acetonitrile (20 mL) and stirred at room temperature for 18 hours. The reaction was concentrated under vacuum, ethyl acetate was added, and the organics were washed with sodium bicarbonate (saturated aqueous solution) and brine, then evaporated to dryness to provide ethyl 2-bromo-4-(5-chloro-2-methoxyphenyl)thiazole-5-carboxylate (1.40 g, 93%). LCMS (Method 1) [M+H] ⁺ = 378.1, _RT = 4.26 min. ^1H NMR (400 MHz, DMSO-d ₆ ): δ (ppm) 7.50 (dd, 1H), 7.42 (d, 1H), 7.14 (d, 1H), 4.16 (q, 2H), 3.73 (s, 3H), 1.12 (t, 3H).

A mixture of ethyl 2-bromo-4-(5-chloro-2-methoxyphenyl)thiazole-5-carboxylate (1.40 g, 3.72 mmol), potassium hydroxide (278 mg) in THF (40 mL), and water (10 mL) was stirred at ambient temperature for 20 hours. The mixture was treated with 1 M HCl aq. (ca. 8 mL, 2 eq.), DCM was added, and the organics were separated and evaporated to dryness to afford 2-bromo-4-(5-chloro-2-methoxyphenyl)thiazole-5-carboxylic acid (1.23 g, 95%) as a yellow solid. LCMS (Method 2) [M+H] ⁺ = 350.1, _RT = 3.28 min. 1H NMR (400 MHz, DMSO-d ₆ ): δ (ppm) 7.47 (dd, 1H), 7.39 (d, 1H), 7.13 (d, 1H), 3.73 (s, 3H).

A solution of 2-bromo-4-(5-chloro-2-methoxyphenyl)thiazole-5-carboxylic acid (1.22 g, 3.50 mmol), diphenylphosphoryl azide (963 mg, 3.50 mmol) and triethylamine (354 mg, 3.50 mmol) in tert-butanol (30 mL) was stirred at 85° C. for 4 hours. After cooling, the reaction was partitioned between ethyl acetate and water, and the organics were separated, then washed with brine and evaporated to dryness. The resulting residue was purified by silica gel flash chromatography (50 to 100% dichloromethane in cyclohexane) to provide tert-butyl 2-bromo-4-(5-chloro-2-methoxyphenyl)thiazol-5-ylcarbamate (970 mg, 66%). LCMS (Method 1) [M+H- ^t Bu] ⁺ = 364.8, _RT = 4.61 min. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ (ppm) 7.43 (dd, 1H), 7.30 (d, 1H), 7.11 (d, 1H), 3.77 (s, 3H), 1.45 (s, 9H).

TFA (4.0 mL) was added to a solution of tert-butyl 2-bromo-4-(5-chloro-2-methoxyphenyl)thiazol-5-ylcarbamate (360 mg, 0.86 mmol) in DCM (10 mL) and water (3 drops). The reaction mixture was stirred at room temperature for 1.5 hours and then evaporated to dryness. The residue was taken up in DCM and washed with sodium bicarbonate (saturated aqueous solution), water, and brine, and concentrated under vacuum to afford 2-bromo-4-(5-chloro-2-methoxyphenyl)thiazol-5-amine as an orange residue. LCMS (Method 1) [M+H] ⁺ = 321.3, _RT = 3.63 min. 1H NMR (400 MHz, DMSO-d ₆ ): δ (ppm) 7.36 (d, 1H), 7.34-7.32 (m, 1H), 7.10 (d, 1H), 3.83 (s, 3H).

The title compound was prepared according to the synthetic method described for pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxyphenyl)-1H-pyrazol-4-yl]amide using 2-bromo-4-(5-chloro-2-methoxyphenyl)thiazol-5-amine and pyrazolo[1,5-a]pyrimidine-3-carbonyl chloride and further purified by silica gel flash chromatography (0 to 40% ethyl acetate in DCM) to give N-(2-bromo-4-(5-chloro-2-methoxyphenyl)thiazol-5-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 1) [M+H] ⁺ =465.8, R _T =4.01min.1H NMR (400MHz, DMSO-d ₆ ): δ10.68(s,br,1H),9.41(dd,1H),8.78(s,1H),8.76(dd,1H), 7.57(dd,1H),7.50(d,1H),7.37-7.34(m,2H),3.81(s,3H).

To a solution of pyrazolo[1,5- _a ]pyrimidine-3-carboxylic acid [2-bromo-4-(5-chloro-2-methoxy-phenyl)-thiazol-5-yl]-amide (4.3 g, 9.3 mmol) in DCM (90 mL) was added boron tribromide (1 M in DCM, 45 mL, 45 mmol) dropwise at -78°C under an N atmosphere. The mixture was stirred at -78°C for 1 hour, then slowly warmed to room temperature and stirred for an additional 16 hours. The mixture was carefully poured into aqueous sodium bicarbonate solution, stirred for 15 minutes, filtered, and the solid collected and dried to afford pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [2-bromo-4-(5-chloro-2-hydroxy-phenyl)-thiazol-5-yl]-amide as a light brown solid (6 g, >100%). LCMS (Method 3) [M+H] ⁺ =450.1, R _T =3.71min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: 15.96 (s, 1H), 9.18(dd,1H,J＝1.6,6.9Hz),8.70(dd,1H,J＝1.8,4.0Hz),8.59(s,1H),7.80(d,1H ,J＝2.8Hz),7.15(dd,1H,J＝4.0,7.1Hz),7.05(dd,1H,J＝2.8,8.6Hz),6.77(d,1H, J＝8.6Hz).

To a solution of pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [2-bromo-4-(5-chloro-2-hydroxy-phenyl)-thiazol-5-yl]-amide (1.1 g, 2.4 mmol) in DMF (20 mL) and water (2 mL) was added cesium carbonate (1.1 g, 3.4 mmol) followed by sodium chlorodifluoroacetate (839 mg, 5.5 mmol). The mixture was stirred at 100°C for 16 hours, after which additional cesium carbonate (2.2 g, 6.8 mmol) and sodium chlorodifluoroacetate (1.7 g, 11 mmol) were added and stirring continued at 100°C for ₆ hours. The mixture was cooled to room temperature and partitioned between ethyl acetate and water, and the phases were separated. The organic layer was washed with brine. Once the combined aqueous layers were extracted with ethyl acetate, the combined organic layers were dried ( _Na2SO4 ), filtered, and evaporated. The resulting residue was purified by flash silica gel chromatography, eluting with 50% ethyl acetate in cyclohexane, to provide the title compound as a yellow solid (710 mg, 59%). LCMS (Method 3) [M+H] ⁺ = 500.1, _RT = 3.97 min. ^1H NMR (400 MHz, DMSO-d ₆ ) δ: 10.84 (s, 1H), 9.42 (dd, 1H, J = 1.6, 6.9 Hz), 8.79 (s, 1H), 8.66 (dd, 1H, J = 1.6, 4.3 Hz), 7.76 (d, 1H, J = 2.8 Hz), 7.51 (d, 1H, J = 8.8 Hz), 7.36 (dd, 1H, J = 4.2, 7.0 Hz), 7.22 (t, 1H, J = 73.1 Hz).

Example 1

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxyphenyl)-1-piperidin-4-yl-1H-pyrazol-4-yl]amide hydrochloride

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxyphenyl)-1H-pyrazol-4-yl]amide (200 mg, 0.49 mmol) was dissolved in DMF (5 mL), tert-butyl 4-(tolyl-4-sulfonyloxy)piperidine-1-carboxylate (263 mg, 0.74 mmol) and cesium carbonate (240 mg, 0.74 mmol) were added, and the mixture was heated at 90°C for 2 hours. The mixture was cooled to room temperature, diluted with water and extracted with DCM (x3). The combined organic extracts were washed with brine, dried ( _Na2SO4 ), filtered and evaporated. The resulting yellow oil was purified by flash chromatography _on silica gel, eluting with 0-2% MeOH in DCM. The appropriate fractions were collected and evaporated to yield tert-butyl 4-{3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}piperidine-1-carboxylate as a yellow oil. The crude product was carried forward to the next step without further purification. LCMS (Method 4) [M+Na] ⁺ = 610.0; _RT = 4.30 min.

Tert-butyl 4-{3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}piperidine-1-carboxylate (600 mg, 0.97 mmol) was dissolved in DCM (3 mL) and TFA (3 mL) was added. The mixture was stirred at room temperature for 30 minutes. The solvent was evaporated and the residue was dissolved in MeOH and loaded onto an SCX-2 column, which had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH. Evaporation gave a glass that was purified by HPLC (Method 3). The appropriate fractions were combined and evaporated to give an off-white solid. The solid was dissolved in MeOH and 1.25M HCl in methanol (1 mL) was added. The solvent was evaporated, azeotroped three times with methanol, and triturated with ethyl acetate to give the title compound as an off-white solid (144 mg, 28%). LCMS (Method 5) [M+H] ⁺ =488.0, R _T =2.93min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.76 (s, 1H), 9.35 (dd, 1H, J = 1.6, 7.0Hz), 8.98(d,1H,J＝9.9Hz),8.69(dd,1H,J＝1.7,4.3Hz),8.67(s,1H),8.38(s,1H), 7.65-7.62(m,2H),7.30(dd,1H,J＝4.2,7.2Hz),7.26(t,1H,J＝73.6Hz),4.66- 4.57(m,1H),3.47-3.42(m,2H),3.17-3.03(m,2H),2.32-2.18(m,4H).

Example 2

2-Amino-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxy-phenyl)-1-piperidin-4-yl-1H-pyrazol-4-yl]-amide

The title compound was prepared from tert-butyl (3-((3-(5-chloro-2-(difluoromethoxy)phenyl)-1H-pyrazol-4-yl)carbamoyl)pyrazolo[1,5-a]pyrimidin-2-yl)carbamate according to the method described in Example 1 to give an amber solid. LCMS (Method 3) [M+H] ⁺ =503.3, R _T =2.32min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.55 (s, 1H), 8.93 (dd, 1H, J = 1.5, 6.8Hz), 8.37 (dd, 1H, J = 1.5, 4.5Hz),8.29(s,1H),7.64-7.58(m,2H),7.44(d,1H,J＝8.7Hz),7.25(t,1H,J＝ 73.5Hz),7.00(dd,1H,J＝4.5,6.7Hz),6.57(s,2H),4.33-4.22(m,1H),3.07(d,2H, J＝12.6Hz),2.66-2.57(m,2H),2.06-1.96(m,2H),1.89-1.75(m,2H).

Example 3

cis-Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxyphenyl)1-[2-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide

A solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (20.0 g, 43.2 mmol) in DMF (150 mL) was treated with DIPEA (6.7 g, 5.2 mmol), cis-2-methyloctahydropyrrolo[3,4-c]pyrrole (6.27 g, 49.7 mmol), and HATU (18.9 g, 49.7 mmol), and the mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with 0.5 M _{aqueous K₂CO₃} and extracted with ethyl acetate (x2). The combined organic extracts were washed with water (x2) and brine, dried ( _Na₂SO₄ ), and evaporated to give a solid. The resulting solid was purified by flash chromatography _on silica gel, eluting with 0-10% 2M _NH₃ /MeOH in DCM. Collection of the appropriate fractions followed by evaporation gave the title compound as a cream solid (17.1 g, 69%). LCMS (Method 5) [M+H] ⁺ = 570.9, _RT = 2.82. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 9.70 (s, 1H), 9.30 (dd, 1H, J = 7.0, 1.6 Hz), 8.62 - 8.63 (m, 2H), 8.27 (s, 1H), 7.59 (dd, 1H, J = 8.8, 2.7 Hz), 7.52 (d, 1H, J = 2.7 Hz), 7.42 (d, 1H, J = 8.8 Hz), 7.25 (dd, 1H, J = 7.0, 4.2 Hz), 7.11 (t, 1H, J = 73.3 Hz), 5.07 (m, 2H). (d,2H,J＝5.9Hz),3.69(dd,1H,J＝10.8,8.6Hz),3.56(dd,1H,J＝12.2,8.8Hz), 3.36(dd,1H,J＝10.8,4.4Hz),3.22(dd,1H,J＝12.3,4.5Hz),2.45-2.46(m,3H), 2.37-2.39(m,3H),2.17(s,3H).

Example 4

cis-Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxyphenyl)-1-[2-(hexahydropyrrolo[3,4-c]pyrrol-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide

{3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (3.35 g, 7.25 mmol) was dissolved in DMF (30 mL), and HATU (2.85 g, 9.42 mmol) was added. cis-tert-butyl hexahydropyrrolo[3,4-c]pyrrole-2-carboxylate (2.0 g, 9.42 mmol) and DIPEA (1.63 mL, 9.42 mmol) were added, and the mixture was stirred at room temperature for 17 hours. The volatiles were evaporated, and the resulting residue was azeotroped with toluene. The solid was partitioned between water and DCM. The organic layer was separated, dried (Na ₂ SO ₄ ), and evaporated. The residue was dissolved in DCM (10 mL), and TFA (10 mL) was added. The solution was stirred at room temperature for 4 hours, then evaporated. The residue was dissolved in MeOH and loaded onto an SCX-2 column which had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH. The solvent was evaporated and the product was crystallized from MeOH/ _Et2O . The desired product was obtained as an off-white solid (3.7 g, 90%). LCMS (Method 5) [M+H] ⁺ = 556.9, _RT = 2.83 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.35 (dd, 1H, J = 1.6, 7.0 Hz), 9.38-9.22 (m, 2H), 8.69 (dd, 2H, J = 1.7, 4.2 Hz), 8.67 (s, 1H), 8.31 (s, 1H), 7.64 (dd, 1H, J = 2.7, 8.8 Hz), 7.56 (d, 1H, J = 2.6 Hz), 7.47 (d, 1H, J = 8.8 Hz), 7.30 (dd, 2H, J = 4.3, 7.0 Hz), 7.27(t,1H,J=73.4Hz),5.15(s,2H),3.80-3.67(m,2H),3.65-3.53(m,2H),3.49- 3.32(m,3H),3.16-2.94(m,4H).

Example 5

cis-Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxyphenyl)-1-[2-(5-ethyl-hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide hydrochloride

To a solution of cis-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxyphenyl)-1-[2-(hexahydropyrrolo[3,4-c]pyrrol-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide (100 mg, 0.18 mmol) in acetonitrile (4 mL) was added potassium carbonate (37 mg, 0.27 mmol) and bromoethane (20 μL, 0.27 mmol). The reaction was heated at 50°C for 2 hours and then cooled to room temperature. The mixture was applied directly to an SCX-2 column. Elution with 2M ammonia in MeOH gave the desired product. The product was purified by HPLC (Method 1) and the pure fractions were combined and passed through an SCX-2 column. The free base was obtained after evaporation of the solvent by elution with 2M ammonia in MeOH. The solid was dissolved in 1.25M methanolic HCl and the volatiles were evaporated. The HCl salt was crystallized from MeOH/Et ₂ O to give a white solid (32 mg, 40%). LCMS (Method 5) [M+H] ⁺ = 585.0, _RT = 2.88 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 10.44 (d, 1H, J = 35.0 Hz), 9.79-9.74 (m, 1H), 9.35 (dd, 1H, J = 1.3, 7.0 Hz), 8.69 (dd, 1H, J = 1.5, 4.2 Hz), 8.67(s,1H),8.31(d,1H,J＝3.8Hz),7.64(dd,1H,J＝2.5,8.8Hz),7.57(d,1H,J＝ 2.6Hz),7.47(d,1H,J＝8.9Hz),7.30(dd,1H,J＝4.3,7.0Hz),7.27(t,1H,J＝73.6 Hz),5.22-5.10(m,2H),3.43(s,12H),3.84-2.77(m,12H),1.29-1.19(m,3H).

Example 6

cis-Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxyphenyl)-1-[2-(5-cyclopropylmethylhexahydropyrrolo[3,4-c]pyrrol-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide

To a solution of cis-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxyphenyl)-1-[2-(hexahydropyrrolo[3,4-c]pyrrol-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide (100 mg, 0.18 mmol) in 2,2,2-trifluoroethanol (3 mL) was added cyclopropanecarbaldehyde (67 μL, 0.90 mmol). After stirring at room temperature for 15 minutes, sodium borohydride (21 mg, 0.54 mmol) was added, and the reaction was heated at 90°C for 2 hours. The reaction was quenched by the addition of MeOH, and the mixture was loaded onto an SCX-2 column that had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH. Evaporation gave the crude product, which was purified by HPLC (Method 1). Pure fractions were loaded onto an SCX-2 column that had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH. The pure product was evaporated to give the title compound as the free base, which was crystallized from MeOH/ _Et2O and obtained as a yellow solid (55 mg, 50%). LCMS (Method 5) [M+H] ⁺ =611.1, R _T =3.02min. ¹ HNMR (400MHz, DMSO-d ₆ ) δ: (ppm) 10.94- 10.88(m,1H),9.57-9.55(m,1H),8.95-8.92(m,1H),8.38-8.35(m,1H),8.29- 8.28(m,1H),7.65-7.61(m,1H),7.56-7.54(m,1H),7.47(d,1H,J=8.9Hz),7.45 -7.07(m,1H),7.08(d,1H,J＝1.9Hz),7.02-6.99(m,1H),5.24-5.07(m,2H), 3.86-3.52 (m, 5H), 3.52-3.22 (m, 2H), 3.17-3.11 (m, 1H), 3.07-3.00 (m, 2H), 3.00-2.79 (m, 1H), 1.27-1.01 (m, 1H), 0.63-0.56 (m, 2H), 0.42-0.34 (m, 2H). (Atropisomers exist)

Example 7

5-Amino-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[2-((3aR,6aS)-5-methyl-hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-2-oxo-ethyl]-1H-pyrazol-4-yl}-amide hydrochloride

A mixture of 5-chloro-pyrazolo[1,5-a]pyrimidine-3-carbonyl chloride (500 mg, 2.3 mmol) (prepared according to the method in Journal of Medicinal Chemistry, 55(22), 10090-10107; 2012) and 5-(5-chloro-2-difluoromethoxyphenyl)-1-(2-trimethylsilyl-ethoxymethyl)-1H-pyrazol-4-ylamine (868 mg, 2.3 mmol) was suspended in triethylamine (0.35 mL, 2.5 mmol) and DCM (10 mL), and the mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with dichloromethane, and the resulting organic layer was washed with saturated aqueous sodium bicarbonate solution and brine, dried (Na ₂ SO ₄ ) and evaporated to give an oil. The resulting oil was purified by flash chromatography on silica gel, eluting with 0-2% MeOH in DCM. The appropriate fractions were collected and evaporated to give 5-chloro-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [5-(5-chloro-2-difluoromethoxy-phenyl)-1-(2-trimethylsilanyl-ethoxymethyl)-1H-pyrazol-4-yl]-amide as a brown oil (1.19 g, 91%). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 9.07 (s, 1H), 8.72 (s, 1H), 8.67 (d, 1H, J=7.3 Hz), 8.38 (s, 1H), 7.60 (d, 1H, J=2.3 Hz), 7.51 (dd, 1H, J=2.6, 8.8 Hz), 7.38(d,1H,J＝8.9Hz),6.95(d,1H,J＝7.2Hz),6.43(t,1H,J＝72.8Hz),5.41(d, 1H, J＝10.9Hz), 5.31 (d, 1H, J＝11.0Hz), 3.76-3.49 (m, 2H), 1.27 (s, 1H), 0.94- 0.84 (m, 2H), 0.00 (s, 9H).

5-Chloro-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [5-(5-chloro-2-difluoromethoxy-phenyl)-1-(2-trimethylsilanyl-ethoxymethyl)-1H-pyrazol-4-yl]-amide (756 mg, 1.32 mmol) was dissolved in THF (10 mL) and concentrated aqueous ammonia solution (10 mL) was added. The mixture was heated at 50°C for 3 hours and then cooled to ambient temperature. The solvent was evaporated and the residue was azeotroped with methanol (x2) to give 5-amino-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [5-(5-chloro-2-difluoromethoxy-phenyl)-1-(2-trimethylsilanyl-ethoxymethyl)-1H-pyrazol-4-yl]-amide as a white solid (724 mg, 99%). LCMS (Method 4) [M+H] ⁺ = 549.9, _RT = 4.11 min.

A suspension of 5-amino-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [5-(5-chloro-2-difluoromethoxy-phenyl)-1-(2-trimethylsilanyl-ethoxymethyl)-1H-pyrazol-4-yl]-amide (720 mg, 1.31 mmol) in methanol (20 mL) was treated with concentrated aqueous HCl (3 mL) and the mixture was heated at 80°C for 1 hour. The solvent was evaporated and the residue was azeotroped with methanol (x3). The resulting solid was triturated with ethyl acetate, collected by filtration, washed with ethyl acetate and diethyl ether, and air-dried. The residual solid was purified by flash chromatography on silica gel, eluting with 0-12% 2M _NH3 /MeOH in DCM. The appropriate fractions were collected and evaporated to give 5-amino-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxy-phenyl)-1H-pyrazol-4-yl]-amide as a brown solid (378 mg, 69%). LCMS (Method 4) [M+H] ⁺ =419.9, _RT =2.93

5-Amino-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxy-phenyl)-1H-pyrazol-4-yl]-amide was converted to the title compound according to the procedures described in Example C and Example 1 to give a white solid. LCMS (Method 5) [M+H] ⁺ =586.0, R _T =2.58min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.76 (s, 1H), 9.35 (dd, 1H, J = 1.6, 7.1Hz), 9.38-9.21 (m, 2H), 8.69(dd,1H,J＝1.7,4.3Hz),8.67(s,1H),8.31(s,1H),7.64(dd,1H,J＝2.7,8.8 Hz),7.56(d,1H,J＝2.6Hz),7.48(d,1H,J＝8.6Hz),7.30(dd,1H,J＝4.3,6.9Hz), 7.27(t,1H,J=73.3Hz),5.15(s,2H),3.93-3.67(m,3H),3.65-3.32(m,6H),3.19- 2.94(m,4H).

Example 8

6-Methyl-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[2-((3aR,6aS)-5-methyl-hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-2-oxo-ethyl]-1H-pyrazol-4-yl}-amide hydrochloride

The title compound was prepared from commercially available 6-methyl-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid according to the procedure described for Example B and Example 1 to give a white solid. LCMS (Method 5) [M+H] ⁺ = 585.0, _RT = 3.03 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 10.74-10.49 (m, 1H), 9.70 (s, 1H), 9.21 (dd, 1H, J = 1.1, 2.0 Hz), 8.60 (d, 1H, J = 2.0 Hz), 8.58 (s, 1H), 8.31 (s, 1H), 7.65 (dd, 1H, J = 2.7, 8.8 Hz), 7.55 (d, 1H, J = 2.7 Hz), 7.49 (d, 1H, J = 8.8 Hz), 7.28 (m, 1H), 7.70 (s, 1H), 7.70 (s, 1H), 7.91 (s, 1H), 7.97 (s, 1H), 7.99 (s, 1H), 7.10 (s, 1H), 7. (t,1H,J＝73.3Hz),5.19(d,1H,J＝16.8Hz),5.13(d,1H,J＝16.8Hz),3.94-2.88 (m,10H),2.82(s,3H),2.39(s,3H).

Example 63

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxy-phenyl)-1-(2-{4-[(2-cyano-ethyl)-methyl-amino]-piperidin-1-yl}-2-oxo-ethyl)-1H-pyrazol-4-yl]-amide

A solution of pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxy-phenyl)-1H-pyrazol-4-yl]-amide (32.0 g, 79.1 mmol) in DMF (400 mL) was treated with 2-chloro-1-(1,4-dioxa-8-aza-spiro[4.5]dec-8-yl)-ethanone (20.8 g, 94.9 mmol) and Cs ₂ CO ₃ (51.5 g, 158.1 mmol) and the mixture was stirred at room temperature for 19 hours. The reaction was diluted with water (~1.6 L) and the resulting precipitate was collected by filtration. The solid was washed with water and dried under reduced pressure to give pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[2-(1,4-dioxa-8-aza-spiro[4.5]dec-8-yl)-2-oxo-ethyl]-1H-pyrazol-4-yl}-amide as a light brown solid (33.3 g, 72%). LCMS (Method 3) [M+H] ⁺ = 588.2, _RT = 3.02 min. ¹ H NMR (400 MHz, CDCl ₃ ) δ: (ppm) 9.85 (s, 1H), 8.78 (dd, 1H, J = 1.6, 6.9Hz),8.71(s,1H),8.56(dd,1H,J＝1.8,4.0Hz),8.42(s,1H),7.69(d,1H,J＝2.5 Hz),7.41(dd,1H,J＝2.7,8.7Hz),7.30-7.26(m,1H),7.00(dd,1H,J＝4.2,6.9Hz), 6.48(t,1H,J＝74.0Hz),5.06(s,2H),4.00-3.95(m,4H),3.74(dd,2H,J＝5.8,5.8Hz),3.61(dd,2H,J＝5.8,5.8Hz),1.74-1.64(m,4H).

A suspension of pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[2-(1,4-dioxa-8-aza-spiro[4.5]dec-8-yl)-2-oxo-ethyl]-1H-pyrazol-4-yl}-amide (46.7 g, 79.5 mmol) in dioxane (280 mL) was cooled in an ice bath and then treated with concentrated hydrochloric acid (210 mL) at a rate that maintained the internal temperature below 22° _C . Once the addition was complete, the reaction was allowed to warm to room temperature and stirred for 4 hours. The reaction was cooled in an ice bath, diluted with ethyl acetate and water, and the pH of the aqueous phase was adjusted to ~8 by portionwise addition of solid _Na2CO3 . The mixture was extracted with ethyl acetate (x4) and _the combined organic extracts were dried ( _Na2CO3 ) and evaporated to give pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[2-oxo-2-(4-oxo-piperidin-1-yl)-ethyl]-1H-pyrazol-4-yl}-amide in dioxane as a light brown solid (48.8 g, 97%). LCMS (Method 3) [M+H] ⁺ = 544.2, _RT = 2.85 min. ^1H NMR (400 MHz, CDCl3 ₎ δ: (ppm) 9.87 (s, 1H), 8.79 (dd, 1H, J = 1.8, 7.0 Hz), 8.70 (s, 1H), 8.57(dd,1H,J＝1.8,4.0Hz),8.47(s,1H),7.67(d,1H,J＝2.8Hz),7.43(dd,1H,J＝ 2.7,8.7Hz),7.29(d,1H,J＝8.8Hz),7.01(dd,1H,J＝4.3,7.0Hz),6.48(t,1H,J＝ 74.0Hz), 5.14 (s, 2H), 3.93 (t, 2H, J = 6.2Hz), 3.85 (t, 2H, J = 6.2Hz), 3.70 (s, 8H), 2.50 (t, 2H, J = 6.2Hz), 2.42 (t, 2H, J = 6.1Hz).

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[2-oxo-2-(4-oxo-piperidin-1-yl)-ethyl]-1H-pyrazol-4-yl}-amide. A solution of dioxane (36 g, 57.0 mmol) in DCM (500 mL) was treated with 3-amino-propionitrile (5.0 mL, 68.4 mmol) and acetic acid (50 mL). The mixture was cooled in an ice bath, then sodium triacetoxyborohydride (18.1 g, 85.4 mmol) was added portionwise. The reaction was warmed to room temperature and stirred for 1.5 hours. The mixture was diluted with methanol and loaded onto an SCX-2 pad that had been treated with MeOH. After rinsing with MeOH, the product was eluted with a 2M ammonia solution in MeOH. The basic fractions were combined and evaporated to give pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (3-(5-chloro-2-difluoromethoxy-phenyl)-1-{2-[4-(2-cyano-ethylamino)-piperidin-1-yl]-2-oxo-ethyl}-1H-pyrazol-4-yl)-amide as a light brown solid (31.1 g, 91%). LCMS (Method 3) [M+H] ⁺ =598.2, _RT =2.25 min. ^{1 H} NMR (400 MHz, CDCl ₃ ) δ: (ppm) 9.84 (s, 1H), 8.78 (dd, 1H, J=1.6, 6.9 Hz), 8.69 (s, 1H),8.58-8.55(m,1H),8.40(s,1H),7.68(d,1H,J＝2.5Hz),7.41(dd,1H,J＝2.7, 8.7Hz),7.28(d,1H,J＝8.7Hz),7.28(s,1H),7.00(dd,1H,J＝4.2,6.9Hz),6.50(t, 1H,J＝74.1Hz),5.08(d,1H,J＝15.4Hz),5.02(d,1H,J＝15.5Hz),4.37(d,1H,J ＝13.4Hz),3.88(d,1H,J＝13.4Hz),3.22-3.12(m,1H),3.02(t,1H,J＝6.4Hz), 2.98-2.86(m,2H),2.82-2.72(m,1H),2.52-2.46(m,2H),1.96-1.83(m,2H), 1.36-1.20(m,2H).

A solution of pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (3-(5-chloro-2-difluoromethoxy-phenyl)-1-{2-[4-(2-cyano-ethylamino)-piperidin-1-yl]-2-oxo-ethyl}-1H-pyrazol-4-yl)-amide (31.1 g, 52.0 mmol) in DCM (500 mL) was treated with 37% aqueous formaldehyde (21.3 mL, 286.2 mmol). Once the addition was complete, the reaction was cooled in an ice bath before sodium triacetoxyborohydride (44.1 g, 208.2 mmol) was added portionwise. The reaction mixture was allowed to warm to room temperature and stirred for 1.5 hours, after which the reaction was filtered and the filtrate diluted with methanol. The mixture was diluted with methanol and loaded onto an SCX-2 pad, which had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH. The basic fractions were combined and evaporated. The resultant residue was purified by flash silica chromatography eluting with 0-10% 2M _NH3 /MeOH in DCM. Appropriate fractions were combined and evaporated. The resulting residue was recrystallized from ethyl acetate to give the title compound as a light brown solid (30.7 g, 96%). LCMS (Method 5) [M+H] ⁺ = 612.2, _RT = 2.84 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 7.0, 1.6 Hz), 8.70-8.67 (m, 1H), 8.68 (s, 1H), 8.32 (s, 1H), 7.63 (dd, 1H, J = 8.8, 2.7 Hz), 7.56 (d, 1H, J = 2.7 Hz), 7.47 (d, 1H, J = 8.8 Hz), 7.29 (dd, 1H, J = 7.0, 4.2 Hz),7.26(t,1H,J＝73.4Hz),5.22-5.24(m,2H),4.38(d,1H,J＝12.9Hz),3.96 (d,1H,J＝13.5Hz),3.06(t,1H,J＝12.7Hz),2.62-2.65(m,6H),2.22(s,3H), 1.72(d,2H,J＝11.9Hz),1.34-1.40(m,2H).

Example 142

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [1-[1-(3-acetylaminopropyl)piperidin-4-yl]-3-(5-chloro-2-difluoromethoxyphenyl)-1H-pyrazol-4-yl]amide hydrochloride

A mixture of pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxyphenyl)-1-piperidin-4-yl-1H-pyrazol-4-yl]amide (100 mg, 0.20 mmol), (3-bromo-propyl)-carbamic acid tert-butyl ester (71 mg, 0.3 mmol) and potassium carbonate (45 mg, 0.32 mmol) in DMF (2 mL) was heated at 60° C. for 1.5 hours. The reaction mixture was cooled to ambient temperature, diluted with water and extracted with dichloromethane. The organic layer was separated, washed with brine, dried (Na ₂ SO ₄ ) and evaporated. The resulting residue was chromatographed on silica gel, eluting with a 2 M ammonia in methanol gradient in dichloromethane (0-6%) to provide tert-butyl [3-(4-{3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}piperidin-1-yl)propyl]carbamate as a yellow oil (131 mg, 100%). LCMS (Method 4) [M+H] ⁺ = 675.0, _RT = 2.86 min.

Tert-butyl [3-(4-{3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}piperidin-1-yl)propyl]carbamate (129 mg, 0.20 mmol) was dissolved in DCM (2 mL) and TFA (2 mL) was added. The reaction was stirred at room temperature for 1 hour and then evaporated to dryness. The residue was dissolved in MeOH and loaded onto an SCX-2 column, which had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH. The basic fractions were evaporated to give a residue, which was dissolved in pyridine (2 mL). Acetyl chloride (21 μL, 0.30 mmol) was added and the solution was allowed to stand at room temperature for 5 days. The volatiles were evaporated and azeotroped with toluene. The residue was dissolved in MeOH and loaded onto an SCX-2 column, which had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH. Evaporation gave a glassy solid which was purified by HPLC (Method 3). The pure amine was dissolved in MeOH and 1.25M methanolic HCl was added. The volatiles were evaporated and the solid product was triturated with EtOAc/Et ₂ O to give the title compound as a white solid (40 mg, 32%). LCMS (Method 5) [M+H] ⁺ = 587.0, _RT = 2.91 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 9.89 (s, 1H), 9.76 (s, 1H), 9.35 (dd, 1H, J = 1.5, 7.0 Hz), 8.69 (dd, 1H, J = 1.6, 4.2 Hz), 8.67 (s, 1H), 8.39 (s, 1H), 8.04 (t, 1H, J = 5.8 Hz), 7.67-7.61 (m, 2H), 7.46 (d, 2H, J = 8.4 Hz), 7.31 (dd, 1H, J = 4.2, 6.9Hz), 7.26 (t, 1H, J = 73.4Hz), 4.64-4.54 (m, 1H), 3.64 (d, 2H, J = 12.2Hz), 3.21 -3.01 (m, 6H), 2.40-2.29 (m, 4H), 1.84 (m, 5H).

Example 302

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[4-(4-dimethylamino-piperidine-1-carbonyl)-phenyl]-1H-pyrazol-4-yl}-amide hydrochloride

A solution of 4-iodo-benzoyl chloride (4.72 g, 17.71 mmol) in DCM (70 mL) was treated with dimethyl-piperidin-4-yl-amine (2.27 g, 17.71 mmol) at 0°C. The resulting suspension was warmed to room temperature and stirred for 2.5 hours. The reaction mixture was diluted with 1M _{aqueous Na₂CO₃} solution, and the phases were separated. The aqueous phase was extracted twice with DCM, and the combined organic phases _were dried ( _Na₂SO₄ ) and evaporated to afford (4-dimethylamino-piperidin-1-yl)-(4-iodo-phenyl)-methanone as a white solid (6.2 g, 98%). LCMS (Method 3) [M+H] ^⁺ = 359.2, _RT = 1.82 min.

A microwave tube was charged with pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxy-phenyl)-1H-pyrazol-4-yl]-amide (100 mg, 0.25 mmol), (4-dimethylamino-piperidin-1-yl)-(4-iodo-phenyl)-methanone (107 mg, 0.30 mmol), copper(I) iodide (15 mg, 0.08 mmol), potassium carbonate (73 mg, 0.53 mmol), and trans-N,N'-dimethyl-1,2-cyclohexanediamine (24 μL, 0.15 mmol). The tube was sealed and flushed with argon, followed by the addition of toluene (1.0 mL). The reaction mixture was stirred at 110°C for 18 hours. The reaction was cooled to room temperature, diluted with water, and the resulting precipitate was collected by filtration, washed with ethyl acetate, and dried under reduced pressure. The resulting solid was purified by MDAP (Method 1) by taking up the solid in a MeOH/DCM mixture and loading it onto an SCX-2 column treated with MeOH. The column was washed with MeOH and then eluted with 2M _NH3 /MeOH. The basic fractions were combined, evaporated, and the resulting solid was suspended in MeOH and treated with 1.25M HCl in MeOH. The suspension was evaporated, and the solid was triturated with ethyl acetate and dried under reduced pressure to give the title compound as a yellow solid (67 mg, 40%). LCMS (Method 5) [M+H] ⁺ = 635.2, _RT = 3.27 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 9.88 (s, 1H), 9.38 (dd, 1H, J = 7.1, 1.8 Hz), 9.04 (s, 1H), 8.71 (s, 1H), 8.70 (dd, 1H, J = 4.3, 1.7 Hz), 8.04-7.99 (m, 2H), 7.79 (d, 1H, J = 2.7 Hz), 7.72 (dd, 1H, J = 8.8,2.6Hz),7.63-7.57(m,2H),7.53(d,1H,J＝8.8Hz),7.33(dd,1H,J＝7.0,4.3Hz ),7.31(t,1H,J=73.1Hz),4.65(brs,1H),2.98(s,2H),2.67(s,6H),2.14-1.82(m, 2H),1.69-1.54(m,2H).

Example 188

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(4-dimethylamino-piperidin-1-yl)-thiazol-5-yl]-amide

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [2-bromo-4-(5-chloro-2-difluoromethoxy-phenyl)-thiazol-5-yl]-amide (100 mg, 0.2 mmol) and 4-dimethylaminopiperidine (128 mg, 1 mmol) were dissolved in DMA (1 ml) and heated in a microwave at 160 ° C for 1 hour. The residue was dissolved in MeOH and loaded onto an SCX-2 column, which had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH. The resulting yellow glass was purified by MDAP (Method 1). The appropriate fractions were combined and evaporated to give a yellow solid. The residue was dissolved in MeOH and loaded onto an SCX-2 column, which had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH to give the title compound as an orange-yellow solid (43 mg, 39%). LCMS (Method 5) [M+H] ⁺ =548.1, R _T =3.20min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm)10.23(s,1H),9.36(dd,1H,J＝1.6,7.0Hz),8.68(s,1H),8.63(dd,1H,J＝1.6,4.2Hz),7.66(d,1H,J＝2.7Hz), 7.59(dd,1H,J＝2.6,8.8Hz),7.41(d,1H,J＝9.0Hz),7.31(dd,1H,J＝4.2,6.7Hz), 7.18(t,1H,J＝73.5Hz), 3.93(d,2H,J＝12.7Hz), 3.02(dd,2H,J＝10.3,12.3Hz), 3.05-2.95(m,1H),2.40-2.26(m,6H),1.92-1.89(m,2H),1.58-1.47(m,2H).

Example 189

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (4-(5-chloro-2-difluoromethoxy-phenyl)-2-{4-[(2-cyano-ethyl)-methyl-amino]-piperidin-1-yl}-thiazol-5-yl)-amide

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [2-bromo-4-(5-chloro-2-difluoromethoxy-phenyl)-thiazol-5-yl]-amide (820 mg, 1.64 mmol) and 1,4-dioxa-8-azaspiro[4.5]decane (1.05 ml, 8.2 mmol) were dissolved in DMA (10 ml) and heated in a microwave at 165° C. for 1 hour. The reaction mixture was partitioned between ethyl acetate and water and the phases were separated. The organic layer was washed with brine. The combined aqueous layers were extracted once with ethyl acetate and the combined organic layers were dried (Na ₂ SO ₄ ), filtered and the solvent removed. The crude product was chromatographed on silica gel, eluting with 50-60% ethyl acetate in cyclohexane. The appropriate fractions were combined and evaporated to give pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(1,4-dioxa-8-aza-spiro[4.5]dec-8-yl)-thiazol-5-yl]-amide as a yellow solid (541 mg, 59%). LCMS (Method 3) [M+H] ⁺ =563.3, _RT =3.68 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 10.26 (s, 1H), 9.37 (dd, 1H, J=1.6, 6.9 Hz), 8.69 (s, 1H), 8.64 (dd, 1H, J=1.5, 4.4 Hz), 7.68 (s, 1H). (d,1H,J＝2.5Hz),7.59(dd,1H,J＝2.7,8.7Hz),7.42(d,1H,J＝8.5Hz),7.31(dd, 1H, J＝4.3, 7.0Hz), 7.18 (t, 1H, J＝73.8Hz), 3.94 (s, 4H), 3.52 (dd, 4H, J＝5.7, 5.7 Hz), 1.75 (dd, 4H, J＝5.7, 5.7Hz).

[4-(5-Chloro-2-difluoromethoxy-phenyl)-2-(1,4-dioxa-8-aza-spiro[4.5]dec-8-yl)-thiazol-5-yl]-amide (536 mg, 0.95 mmol) was dissolved in dioxane (10 ml) and concentrated hydrochloric acid (10 ml) and stirred at room temperature for 3 hours. The mixture was cooled to 0°C and basified to approximately pH 13 with 50% aqueous sodium hydroxide solution, then partitioned between ethyl acetate and water, and the phases were separated. The organic layer was washed with brine. Once the combined aqueous layers were extracted with ethyl acetate, the combined organic layers were dried (Na ₂ SO ₄ ), filtered and the solvent removed to give pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(4-oxo-piperidin-1-yl)-thiazol-5-yl]-amide as an orange solid (465 mg, 94%). LCMS (Method 3) [M+H] ⁺ = 519.3, _RT = 3.39 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 10.30 (s, 1H), 9.38 (dd, 1H, J = 1.5, 7.1 Hz), 8.70 (s, 1H), 8.65 (dd, 1H, J = 1.4, 4.3 Hz), 7.70 (d, 1H, J = 2.5Hz),7.61(dd,1H,J＝2.8,8.8Hz),7.43(d,1H,J＝9.2Hz),7.32(dd,1H,J＝4.2 ,7.1Hz),7.20(t,1H,J＝74.0Hz),3.81(dd,4H,J＝6.2,6.2Hz),2.54(d,4H,J＝6.1 Hz).

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(4-oxo-piperidin-1-yl)-thiazol-5-yl]-amide (90 mg, 0.17 mmol) was dissolved in DCM (2 ml) and N-methyl-β-alanine nitrile (20 μl, 0.21 mmol), acetic acid (200 μl) and macroporous polymer-supported cyanoborohydride (166 mg, 0.36 mmol) were added successively and stirred at room temperature for 16 hours. The mixture was diluted with MeOH and loaded onto an SCX-2 column that had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH. The resulting yellow glass was purified by MDAP (Method 1). Appropriate fractions were combined and evaporated to give a yellow glass. The residue was dissolved in MeOH and loaded onto an SCX-2 column that had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2 M ammonia in MeOH to give the title compound as a yellow solid (46 mg, 46%). LCMS (Method 5) [M+H] ⁺ = 587.2, _RT = 3.19 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 10.23 (s, 1H), 9.36 (dd, 1H, J = 1.6, 7.0 Hz), 8.68 (s, 1H), 8.64 (dd, 1H, J = 1.6, 4.3 Hz), 7.67 (d, 1H, J = 2.6 Hz), 7.59 (dd, 1H, J = 2.7, 8.9 Hz), 7.41 (d, 1H, J = 8.6 Hz), 7.31 (dd, 1H, J = 4.3, 7.1 Hz),7.18(t,1H,J＝73.9Hz),3.94(d,2H,J＝12.8Hz),3.04-2.96(m,2H),2.72- 2.60(m,5H),2.24(s,3H),1.79(d,2H,J=10.9Hz),1.59-1.47(m,2H).

Example 205

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(4-methyl-piperazin-1-yl)-thiazol-5-yl]-amide

The title compound was prepared in analogy to the method of pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(4-dimethylamino-piperidin-1-yl)-thiazol-5-yl]-amide to give the title compound as an orange solid. LCMS (Method 5) [M+H] ⁺ =520.2, R _T =3.02min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 10.25 (s, 1H), 9.37 (dd, 1H, J = 1.6, 7.0Hz), 8.69 (s, 1H), 8.64 (dd,1H,J＝1.6,4.3Hz),7.67(d,1H,J＝2.6Hz),7.59(dd,1H,J＝2.7,8.8Hz),7.42 (d,1H,J＝9.2Hz),7.31(dd,1H,J＝4.2,7.3Hz),7.18(t,1H,J＝73.7Hz),3.41(dd, 4H,J＝4.9,4.9Hz),2.48-2.42(m,4H),2.24(s,3H).

Example 209

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(3-dimethylamino-prop-1-ynyl)-thiazol-5-yl]-amide

Under nitrogen, to a solution of pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [2-bromo-4-(5-chloro-2-difluoromethoxy-phenyl)-thiazol-5-yl]-amide (150 mg, 0.3 mmol), bis(triphenylphosphine)palladium(II) dichloride (11 mg, 0.015 mmol), and copper(I) iodide (5 mg, 0.024 mmol) in THF (1 mL) was added propargyl alcohol (35 μL, 0.6 mmol) followed by triethylamine (1 mL). The resulting mixture was stirred at 50°C for 3 hours and then cooled to room temperature. The mixture was partitioned between ethyl acetate and water, the phases separated, and the organic phase washed with brine, dried (Na ₂ SO ₄ ), and evaporated. The resulting residue was purified by flash column chromatography on silica gel, eluting with 80% ethyl acetate in cyclohexane, to afford pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(3-hydroxy-prop-1-ynyl)-thiazol-5-yl]-amide as a yellow solid (112 mg, 78%). LCMS (Method 3) [M+H] ⁺ = 476.2, _RT = 3.19 min.

To a suspension of pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(3-hydroxy-prop-1-ynyl)-thiazol-5-yl]-amide (110 mg, 0.23 mmol) and triphenylphosphine (105 mg, 0.4 mmol) in DCM (3 mL) was added carbon tetrabromide (132 mg, 0.4 mmol) in portions. The reaction was stirred at room temperature, then a 2M solution of methylamine in THF (1 mL) was added. The resulting mixture was stirred at room temperature for 3 hours. The mixture was evaporated, and the residue was taken up in MeOH and loaded onto an SCX-2 column, which had been treated with MeOH. The column was washed with MeOH, then eluted with a 2M solution of NH ₃ in MeOH. The basic fractions were combined and evaporated. The residue was purified by MDAP (Method 1) and after evaporation the material was taken up in MeCN and loaded onto an SCX-2 column which had been treated with MeCN. The column was washed with MeCN and then eluted with 2M _NH3 in MeOH. The basic fractions were combined and evaporated to give the title compound as an orange solid (18 mg, 15%). LCMS (Method 5) [M+H] ⁺ = 503.0, _RT = 3.16 min. ^1H NMR (400 MHz, DMSO- _d6 ) δ: (ppm) 10.85 (s, 1H), 9.41 (dd, 1H, J = 7.1, 1.3 Hz), 8.79 (s, 1H), 8.63 (dd, 1H, J = 4.4, 1.5 Hz), 7.76 (d, 1H, J = 2.6 Hz), 7.72 (dd, 1H, J = 8.8, 2.6Hz), 7.52 (d, 1H, J = 8.8Hz), 7.36 (dd, 1H, J = 7.0, 4.3Hz), 7.19 (t, 1H, J = 73.2Hz), 3.61 (s, 2H), 2.29 (s, 6H).

Example 211

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxyphenyl)-1-[2-(6-methyl-2,6-diazaspiro[3.4]octan-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[2-(hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide was prepared using a method similar to that used to synthesize pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[2-(hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide. LCMS (Method 5) [M+H] ⁺ = 556.9, _RT = 2.84 min.

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[2-(2,6-diaza-spiro[3.4]octan-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide (70 mg, 0.13 mmol) and paraformaldehyde (19 mg, 0.63 mmol) were stirred in 2,2,2-trifluoroethanol for 15 minutes. Sodium borohydride (14 mg, 0.38 mmol) was added and the reaction mixture was heated at 90°C for 2 hours. The reaction mixture was cooled to room temperature and quenched with MeOH. The mixture was loaded onto an SCX-2 column, which had been treated with MeOH. After rinsing with MeOH, the product was eluted with a 2M ammonia solution in MeOH. Evaporation gave the crude product, which was purified by HPLC (Method 1). Pure fractions were loaded onto an SCX-2 column, which had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2M ammonia in MeOH. Evaporation gave the free base which was crystallized from MeOH/ _Et2O . The title compound was obtained as a white solid (28 mg, 39%). LCMS (Method 5) [M+H] ⁺ = 570.9, _RT = 2.86 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.5, 7.0 Hz), 8.69-8.66 (m, 2H), 8.34 (s, 1H), 8.15 (s, 1H), 7.64 (dd, 1H, J = 2.7, 8.9 Hz), 7.57 (d, 1H, J = 2.7 Hz), 7.47 (d, 1H, J = 8.8 Hz), 7.29 (dd, 2H, J = 4.3, 7.1 Hz), 7.25 (t, 1H, J = 73.5Hz), 4.94 (s, 2H), 4.04 (q, 2H, J = 8.8Hz), 3.90-3.81 (m, 2H), 2.69 (s, 2H), 2.27 (s, 3H), 2.03 (dd, 2H, J = 6.8, 6.8Hz).

Example 227

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxyphenyl)-1-[2-(4-methylpiperazin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide

The title compound was prepared in a manner analogous to cis-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxyphenyl)1-[2-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide using {3-(5-chloro-2-difluoromethoxy-phenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid and methylpiperazine to give the title compound as a light yellow solid. LCMS (Method 5) [M+H] ⁺ =545.1, R _T =2.89min. ¹ H NMR (400MHz, CDCl ₃ ) δ: (ppm) 9.85 (s, 1H),8.78(dd,1H,J＝1.6,7.0Hz),8.71(s,1H),8.56(dd,1H,J＝1.7,4.1Hz),8.41 (s,1H),7.69(d,1H,J＝2.6Hz),7.41(dd,1H,J＝2.6,8.7Hz),7.28(d,1H,J＝9.1 Hz),7.00(dd,1H,J＝4.2,7.0Hz),6.47(t,1H,J＝74.1Hz),5.04(s,2H),3.72-3.65 (m,2H),3.57(dd,2H,J＝4.8,4.8Hz),2.47-2.39(m,4H),2.31(s,3H).

Example 233

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxy-phenyl)-1-[2-(4-ethyl-piperazin-1-yl)-2-oxo-ethyl]-1H-pyrazol-4-yl}-amide

The title compound was prepared in a manner analogous to cis-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {3-(5-chloro-2-difluoromethoxyphenyl)1-[2-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2-yl)-2-oxoethyl]-1H-pyrazol-4-yl}amide using {3-(5-chloro-2-difluoromethoxy-phenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid and ethylpiperazine to give the title compound as a white solid. LCMS (Method 5) [M+H] ⁺ =559.1, R _T =2.89min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s,1H),9.34(dd,1H,J＝1.6,7.0Hz),8.69-8.67(m,2H),8.31(s,1H),7.63(dd,1H, J＝2.7,8.8Hz),7.56(d,1H,J＝2.6Hz),7.46(d,1H,J＝9.0Hz),7.29(dd,1H,J＝ 4.2,7.1Hz),7.25(t,1H,J=73.5Hz),5.23(s,2H),3.50-3.49(m,4H),2.41-2.32 (m, 6H), 1.02 (t, 3H, J = 7.1 Hz).

Example 253

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid {4-(5-chloro-2-difluoromethoxy-phenyl)-2-[4-(4-dimethylamino-piperidine-1-carbonyl)-phenyl]-thiazol-5-yl}-amide hydrochloride

Under _N2 atmosphere, pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [2-bromo-4-(5-chloro-2-difluoromethoxy-phenyl)-thiazol-5-yl]-amide (100 mg, 0.2 mmol), 4-carboxyphenylboronic acid (40 mg, 0.24 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (16 mg, 0.02 mmol), and potassium carbonate (110 mg, 0.8 mmol) were dissolved in dioxane (3.2 ml) and water (0.8 ml) and heated in a microwave at 120°C for 30 mins. The mixture was partitioned between ethyl acetate and water, and the phases were separated. The aqueous layer was acidified with 1M HCl, and the resulting precipitate was filtered, and the solid was collected and dried.

The resulting residue was dissolved in DMF (2 ml) and DIPEA (52 μl, 0.3 mmol), HATU (91 mg, 0.24 mmol) was added, and stirred at room temperature for 5 mins, followed by the addition of 4-dimethylaminopiperidine (31 mg, 0.24 mmol). The resulting mixture was stirred at room temperature for a further 16 hours. The mixture was diluted with MeOH and loaded onto an SCX-2 column that had been treated with MeOH. After rinsing with MeOH, the product was eluted with a 2M ammonia solution in MeOH. The resulting yellow glass was purified by HPLC (MDAP, Method 1). The appropriate fractions were combined and evaporated to give an off-white solid. The residue was dissolved in MeOH and loaded onto an SCX-2 column that had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2 M ammonia in MeOH to give the title compound as an off-white solid (27 mg, 21%). LCMS (Method 5) [M+H] ⁺ = 652.3, _RT = 3.35 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 10.82 (1H, s), 9.42 (1H, dd, J = 1.4, 6.9 Hz), 8.80 (1H, s), 8.65 (1H, dd, J = 1.4, 4.3 Hz), 8.03 (2H, d, J = 8.2 Hz), 7.85 (1H, d, J = 2.6 Hz), 7.73 (1H, dd, J = 2.7, 8.8 Hz), 7.56-7.51 (3H, m), 7.37 (1H,dd,J＝4.3,6.9Hz),7.25(1H,t,J＝73.5Hz),4.71-4.39(m,1H),3.73-3.65(1H, m),3.52-3.41(1H,m),3.13-3.11(1H,m),2.89-2.88(1H,m),2.51(s,6H),2.06- 1.74(2H,m),1.49(2H,dd,J＝7.6,14.7Hz).

Example 260

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(4-dimethylamino-piperidin-1-ylmethyl)-thiazol-5-yl]-amide

Under _N2 atmosphere, pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [2-bromo-4-(5-chloro-2-difluoromethoxy-phenyl)-thiazol-5-yl]-amide (600 mg, 1.2 mmol), potassium vinyl trifluoroborate (177 mg, 1.32 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (48 mg, 0.06 mmol), and DIPEA (627 μl, 3.6 mmol) were dissolved in 2-propanol (9 ml) and water (4.5 ml) and heated in a microwave at 100°C for 40 mins. The mixture was partitioned between ethyl acetate and water, and the phases were separated. The organic layer was washed with brine, dried ( _Na2SO4 ), filtered, and _the solvent removed.

The resulting residue was suspended in acetone (12 ml) and water (1.2 ml), followed by the addition of potassium osmate dihydrate (25 mg, 0.06 mmol) and N-methylmorpholine N-oxide (280 mg, 2.4 mmol), and the mixture was stirred vigorously at room temperature for 16 hours. The reaction was quenched by the addition of solid sodium metabisulfite (2 g), then partitioned between ethyl acetate and water, and the phases separated. The organic layer was washed with brine. The combined aqueous layers were extracted once with ethyl acetate, and the combined organic layers were dried (Na ₂ SO ₄ ), filtered, and the solvent removed.

The resulting residue was dissolved in THF (15 ml) and water (15 ml), sodium periodate (514 mg, 2.4 mmol) was added, and the mixture was vigorously stirred at room temperature for 1.5 hours. The reaction mixture was partitioned between ethyl acetate and water, and the phases were separated. The organic layer was washed with brine. The combined aqueous layers were extracted once with ethyl acetate, and the combined organic layers _were dried ( _Na2SO4 ), filtered, and the solvent removed to afford the title compound as an orange solid (509 mg, 94%).

A portion of the resulting solid (100 mg, 0.22 mmol) was dissolved in DCM (2 ml) and 4-dimethylaminopiperidine (33 mg, 0.26 mmol), acetic acid (200 μl) and macroporous polymer-supported cyanoborohydride (216 mg, 0.44 mmol) were added successively and stirred at room temperature for 16 hours. The mixture was diluted with MeOH and loaded onto an SCX-2 column that had been treated with MeOH. After rinsing with MeOH, the product was eluted with a 2M ammonia solution in MeOH. The resulting yellow glass was purified by HPLC (MDAP, Method 1). Appropriate fractions were combined and evaporated to give a yellow glass. The residue was dissolved in MeOH and loaded onto an SCX-2 column that had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2 M ammonia in MeOH to give the title compound as a pale yellow solid (52 mg, 42%). LCMS (Method 5) [M+H] ⁺ = 562.1, _RT = 2.56 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 10.59 (s, 1H), 9.39 (dd, 1H, J = 1.6, 7.0 Hz), 8.75 (s, 1H), 8.62 (dd, 1H, J = 1.6, 4.3 Hz), 7.48 (d, 1H, J = 8.8 Hz), 7.34 (dd, 1H, J = 4.2, 7.0 Hz), 7.17 (t, 1H, J = 73.4 Hz), 3.78 (s, 2H), 2.98 (d, 2H, J＝11.5Hz), 2.19 (s, 6H), 2.17-2.08 (m, 3H), 1.79-1.72 (m, 2H), 1.50-1.38 (m, 2H).

Example 264

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [4-(5-chloro-2-difluoromethoxy-phenyl)-2-(5-methyl-hexahydro-pyrrolo[3,4-c]pyrrole-2-carbonyl)-thiazol-5-yl]-amide

Pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [2-bromo-4-(5-chloro-2-difluoromethoxy-phenyl)-thiazol-5-yl]-amide (200 mg, 0.4 mmol), 2-methyl-octahydro-pyrrolo[3,4-c]pyrrole (70 mg, 0.56 mmol), Herrmann's catalyst (4.5 mg, 0.0048 mmol), tri-tert-butylphosphine hydrofluoroborate (3.5 mg, 0.011 mmol), hexacarbonyl molybdenum (53 mg, 0.2 mmol) and DBU (40 μl, 0.27 mmol) were dissolved in THF (2 ml) and heated in a microwave at 125° C. for 15 mins. The residue was dissolved in MeOH and loaded onto an SCX-2 column that had been treated with MeOH. After rinsing with MeOH, the product was eluted with a 2M ammonia solution in MeOH. The resulting yellow glass was purified by HPLC (MDAP, Method 1). Appropriate fractions were combined and evaporated to give a yellow solid. The residue was dissolved in MeOH and loaded onto an SCX-2 column, which had been treated with MeOH. After rinsing with MeOH, the product was eluted with 2 M ammonia in MeOH to give the title compound as a pale yellow solid (18 mg, 8%). LCMS (Method 5) [M+H] ⁺ = 574.1, _RT = 3.06 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 10.88 (s, 1H), 9.42 (dd, 1H, J = 1.6, 7.0 Hz), 8.81 (s, 1H), 8.61 (dd, 1H, J = 1.5, 4.3 Hz), 7.84 (d, 1H, J = 2.6 Hz), 7.73 (dd, 1H, J = 2.6, 8.8 Hz), 7.54 (d, 1H, J = 8.8 Hz), 7.36 (dd, 1H, J = 4.3, 7.0 Hz), 7.22 (t, 1H, J = 73.1Hz), 4.24 (dd, 1H, J = 8.6, 12.3Hz), 4.09 (dd, 1H, J = 3.9 ,12.3Hz),3.80(dd,1H,J＝8.9,12.6Hz),3.59-3.47(m,1H),3.02-2.92(m,1H), 2.88-2.80(m,1H),2.65-2.48(m,4H),2.29(s,3H).

Example 297

2-Amino-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [1-(3-amino-cyclobutyl)-3-(5-chloro-2-difluoromethoxy-phenyl)-1H-pyrazol-4-yl]-amide

The title compound was prepared in a manner analogous to 2-amino-pyrazolo[1,5-a]pyrimidine-3-carboxylic acid [3-(5-chloro-2-difluoromethoxy-phenyl)-1-piperidin-4-yl-1H-pyrazol-4-yl]-amide using tert-butyl (3-((3-(5-chloro-2-(difluoromethoxy)phenyl)-1H-pyrazol-4-yl)carbamoyl)pyrazolo[1,5-a]pyrimidin-2-yl)carbamate and trans-tolyl-4-sulfonic acid 3-tert-butoxycarbonylamino-cyclobutyl ester to give the title compound as a white solid. LCMS (Method 5) [M+H] ⁺ =489.1, R _T =2.87min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.53 (s, 1H), 8.93 (dd, 1H, J = 1.6, 6.8Hz), 8.37-8.34 (m, 2H), 7.63-7.60(m,2H),7.45(d,1H,J＝7.2Hz),7.25(t,1H,J＝72.6Hz),7.00(dd,1H, J＝4.5,6.5Hz),6.56(s,2H),4.51-4.41(m,1H),3.18-3.07(m,1H),2.75-2.67(m,2H),2.24-2.14(m,2H).

Example 304

N-[1-(2-carbamimidoylethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine

-3-Formamide

A mixture of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (500 mg, 1.24 mmol), 3-bromopropionitrile (412 mg, 3.08 mmol), and Cs ₂ CO ₃ (1.21 g, 3.70 mol) in N,N-dimethylformamide (15 mL) was flushed with a gentle stream of nitrogen. The resulting mixture was stirred in a sealed tube at 65° C. for 16 h and then poured into water (200 mL). The crude product was collected by filtration and then purified by flash chromatography on silica gel using ethyl acetate/petroleum ether (1:2) as the eluent. The appropriate fractions were collected and evaporated to give N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-cyanoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid (320 mg, 57%). LCMS (Method 17) [M+H] ⁺ =458.1, _RT =1.67 min.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-cyanoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (200 mg, 0.44 mmol) and methanol (280 mg, 8.74 mmol) in toluene (10 mL) was added acetyl chloride (341 mg, 4.34 mmol) dropwise and stirred at 0°C. The resulting solution was stirred at room temperature for 2 hours. A solution of concentrated ammonium hydroxide (459 mg) in methanol (3 mL) was added dropwise with stirring at 0°C. The resulting solution was stirred at room temperature overnight and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel using dichloromethane/methanol (5/1) as the eluent. Appropriate fractions were collected and concentrated to give a white solid (130 mg), which was further purified by high pH Prep-HPLC to give N-[1-(2-carbamimidoylethyl)-3-[5-chloro-2-(difluoromethoxy)-phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid (36.8 mg). LCMS (Method 17) [M+H] ⁺ =475.1 (Note: under Method 17, product decomposition was observed), LCMS (Method 18) _RT =6.52 min. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: (ppm) 9.72 (s, 1H), 9.36 (dd, J=1.8, 7.2 Hz, 1H), 8.68-8.66 (m, 2H), 8.35 (s, 1H), 7.64-7.60(m,2H),7.48-7.24(m,3H),6.39(s,2H),4.44(t,J＝7.2Hz,2H),2.67(t,J＝7.2Hz,2H).

Example 306

Ethyl 2-[(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)amino]acetate

A mixture of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (3.00 g, 7.41 mmol), Cs ₂ CO ₃ (9.90 g, 30.38 mmol), tetrahydrofuran (90 mL) and 1,2-dibromoethane (7.00 g, 37.26 mmol) was stirred in an oil bath at 70° C. for 3 hours. The mixture was cooled to ambient temperature and then concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel using ethyl acetate/petroleum ether (1:1) as eluent to afford N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]-pyrimidine-3-carboxamide as an off-white solid (2.2 g, 58%). TLC: R _f = 0.6; ethyl acetate/petroleum ether = 1:1; LCMS (Method 14) [M+H] ⁺ = 513.2, _RT = 1.03 min.

A mixture of N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)-phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (200 mg, 0.39 mmol), triethylamine (394 mg, 3.89 mmol) and ethyl 2-aminoacetate hydrochloride (271 mg, 1.94 mmol) in ethanol (20 mL) was stirred at 80 ° C. for 24 hours and then concentrated under reduced pressure. The crude product was purified by high pH Prep-HPLC to give the title compound as an off-white solid (31.9 mg, 15%). LCMS (Method 14) [M+H] ⁺ = 534.2, _RT = 1.13 min. ¹ H NMR (300 MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.11 (dd, J = 1.5, 7.0 Hz, 1H), 8.66-6.65 (m, 2H), 8.36 (s, 1H), 7.68 (d, J = 2.4 Hz, 1H), 7.58 (dd, J = 2.7, 8.7 Hz, 1H), 7.42 (d, J = 8.7 Hz, 1H), 7.39 (dd, J = 4.5, 7.2 Hz, 1H), 6.55 (t, J = 73.5 Hz, 1H), 4.35 (t, J = 73.5 Hz, 1H). 5.7Hz, 2H), 4.17 (q, J = 7.2Hz, 2H), 3.41 (s, 2H), 3.15 (t, J = 6.0Hz, 2H), 1.25 (t, J = 7.2Hz, 3H).

Example 310

2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]acetic acid

A solution of potassium hydroxide (200 mg, 3.56 mmol) in water (2 mL) was added to a solution of ethyl 2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]acetate (200 mg, 0.32 mmol) in ethanol (20 mL). The resulting solution was stirred at room temperature for 2 hours and neutralized with 1M aqueous HCl to pH ~7. The resulting solution was concentrated in vacuo. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): XBridge Prep C18 OBD column, 5 μm, 19 x 150 mm; mobile phase: water containing 10 mmol of NH4HCO3 and MeCN (30.0% to 60.0% in 10 minutes, reaching 95.0% in 1 minute, maintaining 95.0% for 1 minute, and decreasing to 30.0% in 2 minutes); detector: UV 254/220 nm. This yielded 20.6 mg (11%) of 2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]acetic acid as a white solid. LCMS (Method 26) [M+H] ⁺ =589.2, R _T =0.82min. ¹ H NMR (400MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.10 (dd, 1H, J = 1.6, 7.2Hz), 8.66 (s, 1H), 8.65(d,1H,J＝2.8Hz),8.38(s,1H),7.69(d,1H,J＝2.4Hz),7.57(dd,1H,J＝2.4, 8.8Hz),7.42(d,1H,J＝8.8Hz),7.21(J＝4.0,6.8Hz),6.63(t,1H,J＝73.6Hz), 5.30(s,2H),3.93-3.86(m,4H),3.50(s,2H),3.32-3.13(m,4H).

Example 311

2-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)piperidin-4-yl]amino]acetic acid

A mixture of tert-butyl N-(piperidin-4-yl)carbamate (627 mg, 3.13 mmol) and N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]-pyrimidine-3-carboxamide (400 mg, 0.78 mmol) in DMF (5 mL) was stirred at 100 ° C for 5 h and cooled to room temperature. Water (50 mL) was added. The precipitate was collected by filtration and dried. This gave 400 mg (81%) of tert-butyl N-[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]ethyl)piperidin-4-yl]carbamate as a yellow solid. LCMS (Method 21) [M+H] ⁺ = 631.1, _RT = 1.21 min.

Saturated HCl in dioxane (15 mL) was added to tert-butyl N-[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)piperidin-4-yl]carbamate (400 mg, 0.63 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The pH of the remaining solution was adjusted to 8-9 with saturated _{aqueous Na₂CO₃} . The resulting mixture was concentrated under vacuum. The residue was purified by flash chromatography on silica gel using MeOH/DCM (1:1) as the eluent to afford 330 mg (98%) of N-[1-[2-(4-aminopiperidin-1-yl)ethyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. LCMS (Method 28) [M+H] ⁺ = 531.1, _RT = 0.49 min.

Potassium carbonate (98 mg, 0.71 mmol) was added to a solution of N-[1-[2-(4-aminopiperidin-1-yl)ethyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (290 mg, 0.55 mmol) and tert-butyl 2-bromoacetate (96 mg, 0.49 mmol) in DMF (5 mL). The resulting mixture was stirred at 50° C. overnight. The resulting mixture was concentrated under vacuum. The residue was purified by flash chromatography on silica gel using 15% MeOH in DCM as eluent to afford 150 mg (43%) of tert-butyl 2-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)piperidin-4-yl]amino]-acetate as a yellow solid. LCMS (Method 24) [M+H] ⁺ = 645.2, _RT = 1.39 min.

A solution of the tert-butyl ester (150 mg, 0.23 mmol) from the previous step in DCM (2 mL) and TFA (2 mL) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19 x 150 mm; Mobile phase, water containing 0.05% FA and MeCN (20.0% to 27.0% in 10 min, to 95.0% in 1 min, hold at 95.0% for 1 min, decrease to 20.0% in 2 min); Detector, UV 254/220 nm. This gave 63.5 mg (43%) of the formate salt of 2-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)piperidin-4-yl]amino]acetic acid as an off-white solid. LCMS (Method 25) [M+H] ⁺ =589.1, R _T =0.85min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.73 (s, 1H), 9.34 (d, 1H, J = 8.0Hz), 8.79-8.65 (m, 2H),8.46-8.35(m,1H),7.64-6.98(m,5H),4.30(s,2H),3.10-2.82(m,5H),2.54-2.53(m,2H),2.17-1.93(m,4H),1.58-1.45(m,2H).

Example 312

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[2-(methylsulfanyl)ethyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

2-(Methylsulfanyl)ethan-1-amine (0.5 mL, 5.34 mmol) was added to a solution of N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (120 mg, 0.23 mmol) in CH ₃ CN (3 mL). The resulting solution was stirred at 80° C. for 2 h and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing ₁₀ mmol of _NH₄HCO₃ and MeCN (40.0% to 57.0% in 10 min, to 95.0% in 1 min, hold at 95.0% for 1 min, decrease to 40.0% in 2 min); Detector, UV 254/220 nm. This yielded 58.2 mg (48%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[2-(methylsulfanyl)ethyl]-amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. LCMS (Method 20) [M+H] ⁺ =522.2, R _T =2.50min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.73 (s, 1H), 9.34 (dd, 1H, J＝ 1.6,6.8Hz),8.67(dd,1H,J＝1.6,4.4Hz),8.67(s,1H),8.36(s,1H),7.68-7.61(m, 2H),7.46-7.44(m,1H),7.28(dd,1H,J＝4.4,7.2Hz),7.06(t,1H,J＝73.2Hz),4.23 (t,2H,J＝6.4Hz),2.98(t,2H,J＝6.0Hz),2.71(t,2H,J＝6.8Hz),2.53(t,2H,J＝6.8Hz),2.03(s,3H).

Example 313

N-[1-[2-(Benzylamino)ethyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

The title compound was prepared from N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide and benzylamine using a synthetic method similar to that of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[2-(methylsulfanyl)ethyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 20) [M+H] ⁺ =538.2, R _T =2.70 min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.84 (s, 1H), 9.34 (dd, 1H, J = 1.6, 6.8 Hz),8.67(dd,1H,J＝1.6,4.4Hz),8.67(s,1H),8.37(s,1H),7.64-7.59(m,2H), 7.46-7.42(m,1H),7.32-7.19(m,6H),7.06(t,1H,J＝73.2Hz),4.25(t,2H,J＝6.4Hz),3.73(s,2H),2.93(t,2H,J＝6.4Hz).

Example 314

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[(pyridin-2-ylmethyl)amino]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

The title compound was prepared from N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide and pyridin-2-ylmethanamine using a synthetic method similar to that of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[2-(methylsulfanyl)ethyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 20) [M+H] ⁺ =539.2, R _T =2.53min. ¹ HNMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J＝ 1.6,6.8Hz),8.68(dd,1H,J＝1.6,4.0Hz),8.67(s,1H),8.49(d,1H,J＝4.0Hz), 8.37(s,1H),7.72(dd,1H,J＝1.6,7.6Hz),7.70-7.61(m,2H),7.46-7.06(m,5H), 4.27(t,2H,J＝6.0Hz),3.83(s,2H),2.98(t,2H,J＝6.0Hz).

Example 315

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[(pyridin-4-ylmethyl)amino]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

The title compound was prepared from N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[2-(methylsulfanyl)ethyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide and pyridin-4-ylmethanamine using a synthetic method similar to that of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[2-(methylsulfanyl)ethyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 25) [M+H] ⁺ =539.1, _RT =1.45 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 9.74 (s, 1H), 9.34 (dd, 1H, J ＝1.6,6.8Hz),8.68(dd,1H,J＝1.6,4.4Hz),8.45(d,2H,J＝6.0Hz),8.38(s,1H), 7.64-7.59(m,2H),7.44(d,1H,J＝8.8Hz),7.32-7.28(m,3H),7.07(t,1H,J＝73.2 Hz),4.26(t,2H,J＝6.0Hz),3.76(s,2H),2.93(t,2H,J＝6.0Hz)

Example 316

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-[4-(1-phenylethyl)piperazin-1-yl]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of tert-butyl piperazine-1-carboxylate (1.00 g, 5.37 mmol) and 1-phenylethan-1-one (774 mg, 6.44 mmol) in methanol (30 mL) was added NaBH ₃ CN (511 mg, 8.13 mmol). The resulting solution was stirred at 50° C. overnight. Water (50 mL) was added. The methanol was removed under vacuum. The remaining solution was extracted with ethyl acetate (×2). The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel column chromatography, eluting with ethyl acetate/petroleum ether (1:1). This gave 750 mg (48%) of tert-butyl 4-(1-phenylethyl)piperazine-1-carboxylate as a yellow oil. LCMS (Method 20) [M+H] ⁺ = 291.1, _RT = 1.14 min.

A solution of tert-butyl 4-(1-phenylethyl)piperazine-1-carboxylate (750 mg, 2.58 mmol) and saturated HCl in dioxane (20 mL) was stirred at room temperature for 3 h. The solid was collected by filtration and dried. This gave 320 mg (55%) of 1-(1-phenylethyl)piperazine hydrochloride as a white solid.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) in DMF (4 mL) was added 1-(1-phenylethyl)piperazine hydrochloride (47 mg, 0.21 mmol), DIEA (67 mg, 0.52 mmol), and HATU (79.2 mg, 0.21 mmol). The resulting mixture was stirred at room temperature overnight and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing ₁₀ mmol of _NH₄HCO₃ and MeCN (40.0% to 65.0% in 10 min, to 95.0% in 1 min, hold at 95.0% for 1 min, decrease to 40.0% in 2 min); Detector, UV 254/220 nm. This yielded 31.3 mg (28%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-[4-(1-phenylethyl)piperazin-1-yl]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 21) [M+H] ⁺ =635.0, R _T =1.56min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.74 (s, 1H), 9.34 (dd, 1H, J = 1.6, 7.2Hz), 8.67 (d, 1H,J＝4.8Hz),8.28(s,1H),7.62(dd,1H,J＝2.8,8.8Hz),7.53(d,1H,J＝2.8Hz), 7.45(d,1H,J＝8.8Hz),7.35-7.26(m,6H),7.07(t,1H,J＝73.2Hz),5.19(s,2H), 3.49-3.46(m,5H),2.49-2.33(m,4H),1.31(d,3H,J＝6.4Hz).

Example 317

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(2-methylpropyl)piperazin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

The title compound was prepared from {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid and 1-(2-methylpropyl)piperazine using a synthetic method similar to that of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-[4-(1-phenylethyl)piperazin-1-yl]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 25) [M+H] ⁺ =587.1, R _T =1.80min. ¹ HNMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.33 (dd, 1H, J＝ 1.8,6.9Hz),8.68(dd,1H,J＝1.5,3.9Hz),8.67(s,1H),8.31(s,1H),7.62(dd,1H,J ＝3.0,8.7Hz),7.55(d,1H,J＝2.7Hz),7.45(d,1H,J＝9.0Hz),7.28(dd,1H,J＝4.2, 7.2Hz),7.02(t,1H,J＝73.2Hz),5.23(s,2H),3.57-3.42(m,4H),2.40-2.21(m,4H), 2.05(d,2H,J＝7.2Hz),1.81-1.76(m,1H),0.86(d,6H,J＝6.3Hz).

Example 318

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(3-methyloxetan-3-yl)methyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of 3-methyloxetane-3-carbaldehyde (17.2 mg, 0.17 mmol) and N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(piperazin-1-yl)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (70 mg, 0.13 mmol) in methanol (5 mL) was added NaBH ₃ CN (12.5 mg, 0.20 mmol). The resulting solution was stirred at room temperature overnight. The reaction was then quenched by the addition of 5 mL of water. The resulting mixture was concentrated under vacuum. The residue was dissolved in ethyl acetate (30 mL), washed with brine, dried, and concentrated. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): column, XBridge Prep C18 OBD column, 5um, 19*150mm; mobile phase, water containing _{10mmol NH4HCO3} and MeCN (55.0% to 65.0% in 10min, to 95.0% in 1min, keep 95.0% for 1min, decrease to 55.0% in 2min); detector, UV 254/220nm. This gave 39.8 mg (49%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(3-methyloxetan-3-yl)methyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 25) [M+H] ⁺ =615.2, _RT =0.93 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: (ppm) 9.92 (s, 1H), 9.36-9.27 (m, 1H), 8.91-8.68 (m, 2H), 8.31 (s, 1H), 7.73-7.46 (m, 3H), 7.37-7.06 (m, 2H),5.23(s,2H),4.52-4.20(m,4H),3.60-3.11(m,6H),2.43-2.20(m,4H),1.46(s, 3H).

Example 319

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(4-propylpiperazin-1-yl)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of 1-propylpiperazine dihydrobromide (60.4 mg, 0.21 mmol) and {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) in DMF (2 mL) was added DIEA (112 mg, 0.866 mmol) and HATU (79.2 mg, 0.21 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridgePrep C18 OBD column, 5 μm, 19×150 mm; Mobile phase: Water containing 10 mmol of NH₄HCO₃ and MeCN (50.0% to 61.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 50.0% in 2 minutes); Detector: UV 254/220 nm. This yielded 69.1 mg (70%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(4-propylpiperazin-1-yl)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 20) [M+H] ⁺ =573.2, R _T =2.86min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J = 1.6, 6.8Hz), 8.68 (dd, 1H, J = 1.6, 7.6Hz),8.67(s,1H),8.31(s,1H),7.62(dd,1H,J＝2.4,8.8Hz),7.55(d,1H,J＝2.4 Hz),7.45(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.08(t,1H,J＝73.6Hz), 5.23(s,2H),3.48(t,4H,J＝6.0Hz),2.40-2.34(m,4H),2.24(t,2H,J＝7.2Hz), 1.48-1.41(m,2H),0.85(t,3H,J＝7.2Hz).

Example 320

N-[1-[2-(4-Benzylpiperazin-1-yl)-2-oxoethyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of 2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]acetic acid (100 mg, 0.17 mmol) in DMF (4 mL) and 1-benzylpiperazine (33.6 mg, 0.19 mmol) was added DIEA (44.8 mg, 0.35 mmol) and HATU (79.2 mg, 0.21 mmol). The resulting solution was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase: Water containing ₁₀ mmol of _NH₄HCO₃ and MeCN (37.0% MeCN to 50.0% in 10 minutes, to 95.0% in 1 minute, maintained at 95.0% for 1 minute, and decreased to 37.0% in 2 minutes); Detector: UV 254/220 nm. This yielded 53.8 mg (50%) of N-[1-[2-(4-benzylpiperazin-1-yl)-2-oxoethyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 21) [M+H] ⁺ =621.0, R _T =1.53min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.6, 7.2Hz), 8.69-8.68(m,2H),8.30(s,1H),7.62(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J＝2.8 Hz), 7.45 (d, 1H, J = 8.8Hz), 7.65-7.24 (m, 6H), 7.08 (t, 1H, J = 73.2Hz), 5.23 (m, 2H), 3.52-3.46 (m, 6H), 2.49-2.33 (m, 4H).

Example 321

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[2-(morpholin-4-yl)ethyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) and 4-[2-(piperazin-1-yl)ethyl]morpholine (41 mg, 0.21 mmol) in DMF (5 mL) was added DIEA (45 mg, 0.35 mmol) and HATU (79 mg, 0.21 mmol). The resulting solution was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19*150 mm; Mobile phase, water containing ₁₀ mmol _NH4HCO3 and MeCN (37.0% MeCN to 52.0% in 10 minutes, to 95.0% in 1 minute, maintain 95.0% for 1 minute, decrease to 37.0% in 2 minutes); Detector, UV 254/220 nm. This gave 62.0 mg (56%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[2-(morpholin-4-yl)ethyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 20) [M+H] ⁺ = 644.2, R _T = 2.34min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J = 1.6, 7.2Hz),8.68-8.67(m,2H),8.31(s,1H),7.63(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J ＝2.8Hz),7.46(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.09(t,1H,J＝ 73.6Hz), 5.23 (s, 2H), 3.54 (t, 4H, J = 4.4Hz), 3.50-3.46 (m, 4H), 2.49-2.39 (m, 12H).

Example 322

Methyl 2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]acetate

To a solution of tert-butyl piperazine-1-carboxylate (5 g, 26.85 mmol) in DMF (50 mL) were added Cs ₂ CO ₃ (11 g, 33.76 mmol) and methyl 2-bromoacetate (3.4 g, 22.2 mmol). The resulting mixture was stirred at room temperature for 6 h. Water (50 mL) and DCM (100 mL) were added. The phases were separated. The aqueous phase was extracted with DCM (100 mL). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. This gave 7.1 g (crude) of tert-butyl 4-(2-methoxy-2-oxoethyl)piperazine-1-carboxylate as a yellow oil. LCMS (Method 27) [M+H] ⁺ = 259.2, _RT = 1.08 min.

A solution of tert-butyl 4-(2-methoxy-2-oxoethyl)piperazine-1-carboxylate (1.50 g, 5.81 mmol) and saturated HCl in dioxane (20 mL) was stirred at room temperature for 3 h. The solid was collected by filtration and dried. This gave 950 mg (84%) of methyl 2-(piperazin-1-yl)acetate hydrochloride as a white solid.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) in DMF (2 mL) was added methyl 2-(piperazin-1-yl)acetate hydrochloride (41 mg, 0.21 mmol), DIEA (67.2 mg, 0.52 mmol), and HATU (79.2 mg, 0.21 mmol). The resulting solution was stirred at room temperature for 2 h and concentrated under vacuum. The residue was passed through a short pad of silica gel and eluted with a 10% MeOH solution in DCM. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19*150 mm; Mobile phase, water containing ₁₀ mmol of _NH₄HCO₃ and MeCN (40.0% to 59.0% in 10 min, to 95.0% in 1 min, maintained at 95.0% for 1 min, and decreased to 40.0% in 2 min); Detector, UV 254/220 nm. This gave 49.2 mg (47%) of methyl 2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]acetate as a white solid. LCMS (Method 20) [M+H] ⁺ =603.2, R _T =2.83min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76(s,1H),9.34(dd,1H,J＝1.6,7.2Hz),8.69-8.67(m,2H),8.31(s,1H),7.62(dd, 1H,J＝2.8,8.8Hz),7.56(d,1H,J＝2.4Hz),7.45(d,1H,J＝8.8Hz),7.28(dd,1H,J ＝4.4,7.2Hz),7.08(t,1H,J＝73.2Hz),5.24(s,2H),3.63(s,3H),3.52-3.48(m,4H), 3.30(s,2H),2.67-2.51(m,4H).

Example 323

N-[3-[5-chloro-2-(difluoromethoxy)pyridin-3-yl]-1-[(5-oxooxolan-2-yl)methyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

An 8-mL microwave tube was charged with a mixture of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (150 mg, 0.37 mmol), 5-(chloromethyl)oxolan-2-one (74.6 mg, 0.55 mmol), and Cs ₂ CO ₃ (242 mg, 0.74 mmol) in DMF (2 mL). The tube was evacuated and refilled with nitrogen three times. The final reaction mixture was microwaved at 120° C. for 30 min. The resulting mixture was concentrated under vacuum. The residue was passed through a short pad of silica gel and eluted with ethyl acetate. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19*150 mm; Mobile phase, water containing 0.05% FA and MeCN (39.0% to 53.0% in 10 minutes, to 95.0% in 1 minute, maintain 95.0% for 1 minute, decrease to 39.0% in 2 minutes); Detector, UV 254/220 nm. This yielded 35.8 mg (19%) of N-[3-[5-chloro-2-(difluoromethoxy)pyridin-3-yl]-1-[(5-oxooxolan-2-yl)methyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 20) [M+H] ⁺ =503.1, R _T =1.65min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J = 1.5, 6.9Hz), 8.68-8.66(m,2H),8.38(s,1H),7.63(dd,1H,J＝2.7,8.7Hz),7.61(d,1H,J＝2.7 Hz),7.45(d,1H,J＝8.7Hz),7.28(dd,1H,J＝4.2,6.9Hz),7.00(t,1H,J＝73.2Hz), 4.98-4.94(m,1H),4.48(d,2H,J＝5.1Hz),2.46-2.23(m,3H),2.07-1.98(m,1H).

Example 324

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(piperazin-1-yl)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (200 mg) in DMF (5 mL) was added tert-butyl piperazine-1-carboxylate (77.5 mg, 0.42 mmol), DIEA (89.6 mg, 0.69 mmol), and HATU (158.3 mg, 0.42 mmol). The resulting solution was stirred at room temperature for 3 h and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 3% MeOH in DCM, to afford 279 mg of tert-butyl 4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]acetyl)piperazine-1-carboxylate as a yellow solid. LCMS (Method 22) [M+H] ⁺ = 631.4, _RT = 1.49 min.

A solution of tert-butyl 4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazine-1-carboxylate (279 mg, 0.44 mmol) and saturated HCl in dioxane (10 mL) was stirred at room temperature overnight. The reaction mixture was concentrated under vacuum. The residue was redissolved in methanol and neutralized with DIEA. The neutralized solution was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19*150 mm; Mobile phase, water containing ₁₀ mmol _NH4HCO3 and MeCN (35.0% MeCN to 48.0% in 10 minutes, to 95.0% in 1 minute, maintain 95.0% for 1 minute, decrease to 35.0% in 2 minutes); Detector, UV 254/220 nm. This yielded 95.6 mg (41%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(piperazin-1-yl)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 20) [M+H] ⁺ =531.2, R _T =2.67min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J = 1.6, 6.8Hz), 8.69-8.67 (m,2H),8.30(s,1H),7.62(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J＝2.8Hz),7.45(d, 1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.08(t,1H,J＝73.2Hz),5.22(s,2H), 3.47-3.41(m,4H),2.73-2.67(m,4H).

Example 325

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(dimethylcarbamoyl)methyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of tert-butyl piperazine-1-carboxylate (1 g, 5.37 mmol) in DMF (15 mL) were added Cs ₂ CO ₃ (3.5 g, 10.74 mmol) and 2-bromo-N,N-dimethylacetamide (1.80 g, 10.84 mmol). The resulting mixture was stirred at room temperature overnight. Water (40 mL) and EtOAc (100 mL) were added. The phases were separated. The aqueous phase was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 3% MeOH in DCM. This gave 880 mg (60%) of tert-butyl 4-[(dimethylcarbamoyl)methyl]piperazine-1-carboxylate as a yellow oil. TLC: R _f = 0.3; dichloromethane/methanol = 1/10.

A solution of tert-butyl 4-[(dimethylcarbamoyl)methyl]piperazine-1-carboxylate (880 mg, 3.24 mmol) and saturated HCl in dioxane (20 mL) was stirred at room temperature overnight. The solid was collected by filtration. This yielded 450 mg (67%) of N,N-dimethyl-2-(piperazin-1-yl)acetamide hydrochloride as a white solid.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) in DMF (4 mL) was added N,N-dimethyl-2-(piperazin-1-yl)acetamide hydrochloride (72 mg, 0.35 mmol), DIEA (67.2 mg, 0.52 mmol), and HATU (79.2 mg, 0.21 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing 0.05% FA and MeCN (27.0% to 36.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 27.0% in 2 minutes); Detector, UV 254/220 nm. This yielded 73.4 mg (64%) of the formate salt of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(dimethylcarbamoyl)methyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 20) [M+H] ⁺ = 616.3, R _T =2.41min. ¹ H NMR (400MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.09 (dd, 1H, J = 1.6, 7.2Hz),8.66-8.64(m,2H),8.37(s,1H),8.11(s,1H),7.68(d,1H,J＝2.8Hz),7.57 (dd,1H,J＝2.8,8.8Hz),7.42(d,1H,J＝8.8Hz),7.21(dd,1H,J＝4.4,7.2Hz),6.63 (t,1H,J＝73.6Hz),5.29(s,2H),3.84-3.77(m,4H),3.75(s,2H),3.08(s,3H), 3.06-2.96(m,7H).

Example 326

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(2-hydroxyphenyl)methyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

_NaBH3CN (112 mg, 1.78 mmol) was added portionwise to a solution of 2-hydroxybenzaldehyde (55 mg, 0.45 mmol) and N-[1-(2-aminoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (200 mg, 0.45 mmol) in methanol (10 mL). The resulting solution was stirred at room temperature overnight. The reaction was then quenched by the addition of 0.5 mL of water. The resulting mixture was concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with 3% MeOH in DCM. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep _C18 OBD column, 5 μm, 19*150 mm; Mobile phase, water containing 10 mmol _NH4HCO3 and MeCN ( _38.0 % to 45.0% in 10 minutes, to 95.0% in 1 minute, maintain 95.0% for 1 minute, decrease to 38.0% in 2 minutes); Detector, UV 254/220 nm. This gave 28.8 mg (12%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(2-hydroxyphenyl)methyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 20) [M+H] ⁺ =554.2, R _T =2.79min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.74 (s, 1H), 9.34 (dd, 1H, J = 1.6, 7.2Hz), 8.69-8.67(m,2H),8.37(s,1H),7.63(dd,1H,J＝2.4,8.0Hz),7.61(d,1H,J＝2.8 Hz),7.44(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,7.2Hz),7.09-7.04(m,2H),7.06 (t,1H,J＝73.6Hz),6.73-6.69(m,2H),4.29(t,2H,J＝6.0Hz),3.82(s,2H),2.98(t,2H,J＝6.0Hz).

Example 327

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(3-hydroxyphenyl)methyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

_NaBH3CN (28 mg, 0.45 mmol) was added to a solution of N-[1-(2-aminoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.22 mmol) in MeOH (10 mL) and 3-hydroxybenzaldehyde (55 mg, 0.45 mmol). The resulting solution was stirred at room temperature overnight. The reaction was then quenched by the addition of 0.2 mL of water. The resulting mixture was concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with 2.5% MeOH in DCM. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing 0.05% FA and MeCN (32.0% to 36.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 32.0% in 2 minutes); Detector, UV 254/220 nm. This yielded 25.7 mg (19%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(3-hydroxyphenyl)-methyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide formate as a white solid. LCMS (Method 20) [M+H] ⁺ =554.2, R _T =1.58min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.37 (s, 1H), 9.34 (dd, 1H, J = 1.6, 7.2Hz), 8.68-8.67(m,2H),8.37(s,1H),7.61(dd,1H,J＝2.4,8.8Hz),7.60(d,1H,J＝2.4 Hz),7.44(d,1H,J＝8.4Hz),7.28(dd,1H,J＝4.4,7.2Hz),7.05(t,1H,J＝73.2Hz), 7.10-7.06(m,1H),6.74-6.72(m,2H),6.60(d,1H,J＝7.2Hz),4.25(t,2H,J＝6.0Hz),3.66(s,2H),2.94(t,2H,J＝6.0Hz).

Example 328

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(4-hydroxyphenyl)methyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

A solution of 4-hydroxybenzaldehyde (55 mg, 0.45 mmol) and N-[1-(2-aminoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.22 mmol) in THF (10 mL) was stirred at room temperature for 2 h, then NaBH ₃ CN (28 mg, 0.45 mmol) was added. The resulting solution was stirred at room temperature overnight. The reaction was then quenched by the addition of 0.1 mL of water. The resulting mixture was concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with 3% MeOH in DCM. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase: Water containing 0.05% FA and MeCN (34.0% MeCN to 38.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 34.0% in 2 minutes); Detector: UV 254/220 nm. This yielded 31.4 mg (23%) of the formate salt of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(4-hydroxyphenyl)methyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 20) [M+H] ⁺ = 554.2, R _T =1.58min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.74 (s, 1H), 9.34 (dd, 1H,J＝1.6,7.2Hz),8.69-8.67(m,2H),8.37(s,1H),7.62(dd,1H,J＝2.8,8.8Hz), 7.60(d,1H,J＝2.4Hz),7.44(d,1H,J＝8.4Hz),7.28(dd,1H,J＝4.4,7.2Hz),7.10 (d,2H,J＝8.4Hz),7.06(t,1H,J＝73.2Hz),6.68(d,2H,J＝8.4Hz),4.25(t,2H,J＝6.4Hz),3.63(s,2H),2.95(t,2H,J＝6.0Hz).

Example 329

2-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl]amino]acetic acid methyl ester formate

To a DMF (10 mL) solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (500 mg) was added tert-butyl N-(piperidin-4-yl)carbamate (208 mg, 1.04 mmol), DIEA (224 mg, 1.73 mmol), HATU (395.9 mg, 1.04 mmol). The resulting solution was stirred at room temperature overnight. Water (50 mL) was added. The precipitate was collected by filtration and dried. This gave 600 mg of tert-butyl N-[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl]carbamate as a yellow crude solid. LCMS (Method 25) [M+H] ⁺ = 645.1, _RT = 1.02 min.

A solution of the crude product from the previous step (600 mg) and saturated HCl in dioxane (15 mL) was stirred at room temperature overnight. The resulting solution was diluted with 100 mL of methanol, neutralized with saturated aqueous Na ₂ CO ₃ solution and concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with 30% MeOH in DCM. The appropriate fractions were collected and the solvent evaporated to give N-[1-[2-(4-aminopiperidin-1-yl)-2-oxoethyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (520 mg) as a yellow solid. LCMS (Method 25) [M+H] ⁺ = 545.1, _RT = 0.62 min.

Methyl 2-bromoacetate (30.74 mg, 0.20 mmol) was added to a mixture of N-[1-[2-(4-aminopiperidin-1-yl)-2-oxoethyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.18 mmol) in DMF (5 mL) and potassium carbonate (38.1 mg, 0.28 mmol). The resulting mixture was stirred at room temperature overnight. An additional amount of methyl 2-bromoacetate (30.74 mg, 0.20 mmol) was added, and the resulting solution was stirred at 60° C. for 2 h. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase: Water containing 0.05% FA and MeCN (25.0% MeCN to 32.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 25.0% in 2 minutes); Detector: UV 254/220 nm. This yielded 68.9 mg (57%) of methyl 2-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl]amino]-acetic acid ester as an off-white solid. LCMS (Method 24) [M+H] ⁺ =617.2, R _T =1.53min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s,1H),9.34dd,1H,J＝1.6,7.2Hz),8.69-8.67(m,2H),8.30(s,1H),7.62(dd,1H,J ＝2.4,8.8Hz),7.55(d,1H,J＝2.8Hz),7.45(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.0, 6.8Hz),7.08(t,1H,J＝73.2Hz),5.23-5.20(m,2H),4.11-4.07(m,1H),3.83-3.80 (m,1H),3.63(s,3H),3.39-3.31(m,2H),3.15-3.12(m,1H),2.87-2.83(m,1H), 2.71-2.67(m,1H),1.83-1.77(m,2H),1.29-1.14(m,2H).

Example 330

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(5-hydroxy-2-oxopiperidin-1-yl)piperidin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide formate

DIEA (71 mg, 0.55 mmol) was added to a solution of N-[1-[2-(4-aminopiperidin-1-yl)-2-oxoethyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.18 mmol) and (5-oxoxolan-2-yl)methyl trifluoromethanesulfonate (69 mg, 0.28 mmol) in CH ₃ CN (30 mL). The resulting solution was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep _C18 OBD Column, 5 μm, 19×150 mm; Mobile phase: Water containing 0.05% FA and MeCN (38.0% to 46.0% in 10 minutes, to 95.0% in 1 minute, maintained at 95.0% for 1 minute, and decreased to 38.0% in 2 minutes); Detector: UV 254/220 nm. This yielded 41.6 mg (33%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(5-hydroxy-2-oxopiperidin-1-yl)piperidin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 25) [M+H] ⁺ =643.2, R _T =1.49min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.35 (dd, 1H, J = 1.6, 6.8Hz), 8.69-8.68 (m, 2H), 8.32 (s, 1H), 7.63(dd,1H,J＝2.8,8.8Hz),7.56(d,1H,J＝2.8Hz),7.46(d,1H,J＝8.8Hz),7.29 (dd,1H,J＝4.4,7.2Hz),7.09(t,1H,J＝73.2Hz),5.26-5.24(m,2H),4.89(br,1H), 4.59-4.45(m,2H),4.08-3.90(m,2H),3.25-2.97(m,3H),2.68-2.58(m,1H), 2.46-2.21(m,2H),1.86-1.65(m,3H),1.57-1.50(m,3H).

Example 331

Methyl 3-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]propanoate

To a solution of tert-butyl piperazine-1-carboxylate (1 g, 5.37 mmol) in DMF (20 mL) were added Cs ₂ CO ₃ (589 mg, 1.81 mmol) and methyl 3-bromopropionate (744 mg, 4.46 mmol). The resulting mixture was stirred at room temperature overnight. Water (50 mL) and EtOAc (50 mL) were added. The phases were separated. The aqueous phase was extracted with EtOAc (50 mL). The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by flash chromatography on silica gel, eluting with 3.5% MeOH in DCM. Appropriate fractions were collected and the solvent was evaporated to give tert-butyl 4-(3-methoxy-3-oxopropyl)piperazine-1-carboxylate (895 mg, 74%) as a yellow oil. TLC: R _f = 0.3; ethyl acetate/petroleum ether = 1/2.

A solution of tert-butyl 4-(3-methoxy-3-oxopropyl)piperazine-1-carboxylate (895 mg, 3.29 mmol) and saturated HCl in dioxane (15 mL) was stirred at room temperature overnight. The solid was collected by filtration. This gave 710 mg of methyl 3-(piperazin-1-yl)propanoate hydrochloride as a white crude solid.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) in DMF (5 mL) was added methyl 3-(piperazin-1-yl)propanoate hydrochloride (44 mg, 0.21 mmol), DIEA (90 mg, 0.70 mmol), and HATU (79.2 mg, 0.21 mmol). The resulting solution was stirred at room temperature for 4 h and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep _C18 OBD Column, 5 μm, 19 x 150 mm; Mobile phase: Water containing 0.05% FA and MeCN (40.0% to 51.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 40.0% in 2 minutes); Detector: UV 254/220 nm. This yielded 56.1 mg (49%) of methyl 3-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]propanoate formate as a light yellow solid. LCMS (Method 25) [M+H] ⁺ =617.1, R _T =1.51min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J = 1.6, 7.2Hz), 8.68-8.67 (m, 2H), 8.31(s,1H),7.62(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J＝2.8Hz),7.45(d,1H,J＝ 8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.08(t,1H,J＝73.2Hz),5.23(s,2H),3.60(s, 3H), 3.49-3.41 (m, 4H), 2.59 (t, 2H, J = 6.4Hz), 2.51 (t, 2H, J = 6.4Hz), 2.39-2.33 (m, 4H).

Example 332

3-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]propanoate

The title compound was prepared from ethyl 3-bromopropionate using a synthetic method similar to that of methyl 3-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]propanoate. LCMS (Method 28) [M+H] ⁺ =631.2, R _T =0.89min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (d, 1H, J = 6.9Hz), 8.69-8.67(m,2H),8.30(s,1H),7.62(dd,1H,J＝2.7,8.7Hz),7.55(d,1H,J＝2.4 Hz),7.45(d,1H,J＝9.0Hz),7.28(dd,1H,J＝4.2,6.9Hz),7.07(t,1H,J＝72.9Hz), 5.23(s,2H),4.03(q,2H,J＝7.2Hz),3.59-3.47(m,4H),2.61-2.59(m,2H), 2.50-2.33(m,6H),1.16(t,3H,J＝7.2Hz).

Example 333

1-[2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]ethyl]piperidine-4-carboxylic acid methyl ester

To a stirred solution of tert-butyl 4-(2-hydroxyethyl)piperazine-1-carboxylate (1.5 g, 6.51 mmol) and DIEA (2.5 g, 19.34 mmol) in DCM (50 mL) was added dropwise MsCl (817 mg, 7.13 mmol) at 0 ° C. The resulting solution was stirred at room temperature for 1 h. Water (100 mL) was added. The phases were separated. The aqueous phase was extracted with dichloromethane and the organic layers were combined. The organic phase was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. This gave 1.8 g (crude) of tert-butyl 4-[2-(methylsulfonyloxy)ethyl]piperazine-1-carboxylate as a light yellow oil.

To a solution of tert-butyl 4-[2-(methylsulfonyloxy)ethyl]piperazine-1-carboxylate (431 mg, 1.39 mmol) in DMF (5 mL) was added methyl piperidine-4-carboxylate (300 mg, 2.095 mmol) and DIEA (361 mg, 2.79 mmol). The resulting solution was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The residue was purified by flash chromatography on silica gel, eluting with 3% MeOH in DCM. This gave 285 mg (57%) of tert-butyl 4-[2-[4-(methoxycarbonyl)piperidin-1-yl]ethyl]piperazine-1-carboxylate as a yellow oil. LCMS (Method 20) [M+H] ⁺ = 356.2, _RT = 0.99 min.

A solution of tert-butyl 4-[2-[4-(methoxycarbonyl)piperidin-1-yl]ethyl]piperazine-1-carboxylate (285 mg, 0.80 mmol) and saturated HCl in dioxane (15 mL) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. This gave 205 mg (88%) of methyl 1-[2-(piperazin-1-yl)ethyl]piperidine-4-carboxylate hydrochloride as a yellow solid. LCMS (Method 20) [M+H] ⁺ = 256.2, _RT = 0.44 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (150 mg, 0.32 mmol) in DMF (10 mL) was added methyl 1-[2-(piperazin-1-yl)ethyl]piperidine-4-carboxylate hydrochloride (190 mg, 0.65 mmol), DIEA (126 mg, 0.97 mmol), and HATU (148 mg, 0.39 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with a 4% MeOH solution in DCM. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep _C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing 0.05% FA and MeCN (22.0% to 31.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 22.0% in 2 minutes); Detector, UV 254/220 nm. This yielded 96.5 mg (40%) of the formate salt of methyl 1-[2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]ethyl]piperidine-4-carboxylate as a light yellow solid. LCMS (Method 24) [M+H] ⁺ =700.2, R _T = 2.21min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.6, 6.8Hz),8.69-8.68(m,2H),8.30(s,1H),7.62(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J ＝2.4Hz),7.46(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.08(t,1H,J＝ 73.2Hz),5.23(s,2H),3.59(s,3H),3.49-3.46(m,4H),2.84-2.81(m,2H),2.54-2.52 (m,6H),2.50-2.40(m,2H),2.33-2.27(m,1H),2.04-1.99(m,2H),1.88-1.70(m,2H), 1.55-1.50(m,2H).

Example 334

1-[2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]ethyl]piperidine-4-carboxylic acid ethyl ester

To a solution of tert-butyl 4-[2-(methylsulfonyloxy)ethyl]piperazine-1-carboxylate (392 mg, 1.27 mmol) in DMF (5 mL) was added DIEA (329 mg, 2.55 mmol) and ethyl piperidine-4-carboxylate (300 mg, 1.91 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was purified by flash chromatography on silica gel, eluting with 5% MeOH in DCM. This gave 310 mg (66%) of tert-butyl 4-[2-[4-(ethoxycarbonyl)piperidin-1-yl]ethyl]piperazine-1-carboxylate as a yellow solid. LCMS (Method 28) [M+H] ⁺ = 370.3, _RT = 0.50 min.

A mixture of tert-butyl 4-[2-[4-(ethoxycarbonyl)piperidin-1-yl]ethyl]piperazine-1-carboxylate (310 mg, 0.84 mmol) and saturated HCl in dioxane (15 mL) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. This gave 220 mg (86%) of ethyl 1-[2-(piperazin-1-yl)ethyl]piperidine-4-carboxylate hydrochloride as a light yellow solid. LCMS (Method 28) [M+H] ⁺ = 270.3, _RT = 0.89 min.

To a solution of 2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]acetic acid (150 mg, 0.32 mmol) in DMF (10 mL) was added ethyl 1-[2-(piperazin-1-yl)ethyl]piperidine-4-carboxylate hydrochloride (199 mg, 0.65 mmol), DIEA (126 mg, 0.97 mmol), and HATU (148 mg, 0.39 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was passed through a short pad of silica gel and eluted with a 3% MeOH solution in DCM. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase: Water containing 0.05% FA and MeCN (18.0% MeCN to 27.0% over 10 minutes, to 95.0% over 1 minute, hold at 95.0% for 1 minute, decrease to 18.0% over 2 minutes); Detector: UV 254/220 nm. This yielded 66.1 mg (27%) of ethyl 1-[2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]ethyl]piperidine-4-carboxylate as a formate salt. LCMS (Method 24) [M+H] ⁺ =714.2, R _T =2.42 min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J = 1.6, 6.8 Hz),8.69-8.68(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.8,8.8Hz),7.56(d,1H,J ＝2.4Hz),7.46(d,1H,J＝8.8Hz),7.29(dd,1H,J＝4.4,6.8Hz),7.08(t,1H,J＝73.2 Hz),5.23(m,2H),4.03(q,2H,J＝6.8Hz),3.50-3.47(m,4H),2.88-2.85(m,2H), 2.52-2.51(m,6H),2.47-2.26(m,3H),2.12-2.07(m,2H),1.80-1.78(m,2H), 1.60-1.55(m,2H),1.16(t,3H,J＝7.2Hz).

Example 335

1-[2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]ethyl]-4-methylpiperidine-4-carboxylic acid methyl ester formate

The title compound was prepared from methyl 4-methylpiperidine-4-carboxylate hydrochloride using a method similar to the synthesis of ethyl 1-[2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]ethyl]piperidine-4-carboxylate.

LCMS (Method 24) [M+H] ⁺ =714.2, R _T =1.94min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.6, 6.8Hz), 8.69-8.68 (m, 2H), 8.30(s,1H),7.65-7.62(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J＝2.8Hz),7.45(d,1H, J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.08(t,1H,J＝73.6Hz),5.22(s,2H), 3.62(s,3H),3.48-3.45(m,4H),2.57-2.51(m,2H),2.49-2.39(m,8H),2.01-1.93(m,4H),1.41-1.35(m,2H),1.10(s,3H).

Example 336

2-(Morpholin-4-yl)ethyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylate

2-(morpholin-4-yl) ethane-1-ol (858mg, 6.54mmol), EDC.HCl (1.00g, 5.22mmol), 4-dimethylaminopyridine (53.3mg, 0.44mmol) were added to a DCM (15mL) solution of 1-[(tert-butoxy)carbonyl]piperidine-4-carboxylic acid (1.00g, 4.36mmol). The resulting solution was stirred at room temperature overnight. Water (30mL) and DCM (50mL) were added. The phases were separated. The aqueous phase was extracted with DCM, and the organic layers were combined. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluted with dichloromethane/ethyl acetate (2:3). This gave 1.4g (crude) piperidine-1,4-dicarboxylic acid 1-tert-butyl ester 4-[2-(morpholin-4-yl)ethyl] ester as a yellow oil. TLC: R _f = 0.3; ethyl acetate.

A mixture of 1-tert-butyl 4-[2-(morpholin-4-yl)ethyl]piperidine-1,4-dicarboxylate (1.4 g, 4.09 mmol) and saturated HCl in dioxane (20 mL) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. This gave 1.2 g (crude) of 2-(morpholin-4-yl)ethyl piperidine-4-carboxylate hydrochloride as a yellow oil.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (150 mg, 0.26 mmol) in DMF (2 mL) was added 2-(morpholin-4-yl)ethyl piperidine-4-carboxylate hydrochloride (140 mg, 0.50 mmol), DIEA (101 mg, 0.78 mmol), and HATU (119 mg, 0.31 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was passed through a short pad of silica gel and eluted with a 10% MeOH solution in DCM. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep C18 OBD column, 5 μm, 19 x 150 mm; Mobile phase: Water containing 0.05% FA and MeCN (31.0% MeCN to 38.0% over 10 minutes, to 95.0% over 1 minute, hold at 95.0% for 1 minute, decrease to 31.0% over 2 minutes); Detector: UV 254/220 nm. This yielded 35.6 mg (20%) of the formate salt of 2-(morpholin-4-yl)ethyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylate as an off-white solid. LCMS (Method 28) [M+H] ⁺ =687.2, R _T =0.86min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J = 1.6, 7.2Hz), 8.69-8.68 (m,2H),8.32(s,1H),7.63(dd,1H,J＝2.4,8.8Hz),7.55(d,1H,J＝2.8Hz),7.46(d, 1H,J＝8.8Hz),7.29(dd,1H,J＝4.4,6.8Hz),7.08(t,1H,J＝73.2Hz),5.27-5.22(m, 2H),4.20-4.15(m,3H),3.89-3.82(m,1H),3.58-3.55(m,4H),3.26-3.17(m,1H), 2.89-2.78(m,1H),2.69-2.59(m,1H),2.53-2.51(m,2H),2.40-2.32(m,4H), 1.91-1.80(m,2H),1.69-1.41(m,2H).

Example 337

1-[2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]ethyl]-4-methylpiperidine-4-carboxylic acid ethyl ester formate

The title compound was prepared from ethyl 4-methylpiperidine-4-carboxylate hydrochloride using a method similar to the synthesis of ethyl 1-[2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]ethyl]piperidine-4-carboxylate. LCMS (Method 24) [M+H] ⁺ =728.2, R _T =1.87min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34(dd,1H,J＝1.2,6.8Hz),8.69-8.68(m,2H),8.30(s,1H),7.62(dd,1H,J＝2.8, 8.8Hz),7.55(d,1H,J＝2.8Hz),7.46(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8 Hz),7.08(t,1H,J＝73.2Hz),5.22(s,2H),4.06(q,2H,J＝7.2Hz),2.67-2.60(m, 2H),2.44-2.38(m,8H),2.09-2.04(m,2H),1.97-1.93(m,2H),1.42-1.41(m,2H), 1.16(t,3H,J=7.2Hz),1.10(s,3H).

Example 338

2-(Dimethylamino)ethyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylate

The title compound was prepared from 2-(dimethylamino)ethan-1-ol using a method analogous to the synthesis of 2-(morpholin-4-yl)ethyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylate. LCMS (Method 24) [M+H] ⁺ =645.2, R _T =2.97min. ¹ HNMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J ＝1.6,6.8Hz),8.68-8.67(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.4,8.8Hz),7.55(d, 1H, J＝2.4Hz), 7.45 (d, 1H, J＝8.8Hz), 7.28 (dd, 1H, J＝4.4, 7.2Hz), 7.21 (t, J＝ 73.1Hz,1H)5.30-5.14(m,2H),4.21-4.10(m,3H),3.89-3.78(m,1H),3.24-3.14(m, 1H),2.89-2.60(m,2H),2.49-2.45(m,2H),2.16(s,6H),1.89-1.81(m,2H),1.68-1.37(m,2H).

Example 339

2-(Morpholin-4-yl)ethyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)-4-methylpiperidine-4-carboxylate

To a solution of 1-[(tert-butoxy)carbonyl]-4-methylpiperidine-4-carboxylic acid (1.00 g, 4.11 mmol) in DCM (10 mL) was added 2-(morpholin-4-yl)ethan-1-ol (1.08 g, 8.23 mmol) and HOBt (667 mg, 4.94 mmol). The mixture was stirred at room temperature for 2 hours, then EDC.HCl (943 mg, 4.92 mmol) was added. The resulting solution was stirred at room temperature overnight and diluted with 30 mL of DCM. The resulting mixture was washed with water and brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with a 4% MeOH solution in DCM. This gave 800 mg (55%) of 1-tert-butyl 4-[2-(morpholin-4-yl)ethyl]-4-methylpiperidine-1,4-dicarboxylate as a yellow oil. LCMS (Method 28) [M+H] ⁺ = 357.2, _RT = 0.64 min.

A solution of 1-tert-butyl 4-[2-(morpholin-4-yl)ethyl]-4-methylpiperidine-1,4-dicarboxylate (800 mg, 2.24 mmol) and saturated HCl in dioxane (30 mL) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. This gave 600 mg (crude) of 2-(morpholin-4-yl)ethyl 4-methylpiperidine-4-carboxylate hydrochloride as a white, crude solid. LCMS (Method 28) [M+H] ⁺ = 257.0, _RT = 0.86 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) in DMF (3 mL) was added 2-(morpholin-4-yl)ethyl 4-methylpiperidine-4-carboxylate hydrochloride (76.2 mg, 0.26 mmol), DIEA (67.2 mg, 0.52 mmol), and HATU (79.2 mg, 0.21 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep _C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing 0.05% FA and MeCN (34.0% to 38.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 34.0% in 2 minutes); Detector, UV 254/220 nm. This yielded 43.0 mg (33%) of 2-(morpholin-4-yl)ethyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]-acetyl)-4-methylpiperidine-4-carboxylate as formate. LCMS (Method 20) [M+H] ⁺ =701.2, R _T =0.92min.1H NMR (400MHz, DMSO-d6) δ: (ppm) 9.75 (s, 1H), 9.35 (dd, 1H, J = 1.6, 7.2Hz), 8.69-8.68(m,2H),8.31(s,1H),7.63(dd,1H,J＝2.4,8.8Hz),7.55(d,1H,J＝2.8 Hz),7.46(d,1H,J＝9.6Hz),7.29(dd,1H,J＝4.4,6.8Hz),7.09(t,1H,J＝73.2Hz), 5.23(s,2H),4.21(t,2H,J=5.6Hz),4.02-3.98(m,1H),3.77-3.74(m,1H),3.53-3.51 (m,4H),3.27-3.22(m,1H),3.00-2.94(m,1H),2.55-2.52(m,2H),2.49-2.39(m,4H), 2.04-1.97(m,2H),1.18(s,3H).

Example 340

2-(Dimethylamino)ethyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)-4-methylpiperidine-4-carboxylate

To a solution of 1-[(tert-butoxy)carbonyl]-4-methylpiperidine-4-carboxylic acid (1g, 4.11mmol) in DCM (10 mL) was added 2-(dimethylamino)ethane-1-ol (733mg, 8.22mmol) and HOBt (667mg, 4.94mmol). The mixture was stirred at room temperature for 2h, and then EDC.HCl (943mg, 4.92mmol) was added. The resulting solution was stirred at room temperature overnight and water (30mL) was added. The resulting solution was extracted with DCM (x2), and the organic layers were combined. The organic phase was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with a 4% MeOH solution in DCM. This gave 800mg (62%) of 4-methylpiperidine-1,4-dicarboxylic acid 1-tert-butyl 4-[2-(dimethylamino)ethyl] ester as a yellow oil. TLC: R _f = 0.3; dichloromethane/methanol = 10/1.

A mixture of 1-tert-butyl 4-[2-(dimethylamino)ethyl]-4-methylpiperidine-1,4-dicarboxylate (800 mg, 2.54 mmol) and saturated HCl in dioxane (20 mL) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. This gave 340 mg (53%) of 2-(dimethylamino)ethyl 4-methylpiperidine-4-carboxylate hydrochloride as a white solid.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) in DMF (3 mL) was added 2-(dimethylamino)ethyl 4-methylpiperidine-4-carboxylate hydrochloride (87 mg, 0.35 mmol), DIEA (67.2 mg, 0.52 mmol), and HATU (79.2 mg, 0.21 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep _C18 OBD column, 5 μm, 19×150 mm; Mobile phase: Water containing 0.05% FA and MeCN (38.0% MeCN to 42.0% over 10 minutes, to 95.0% over 1 minute, hold at 95.0% for 1 minute, decrease to 38.0% over 2 minutes); Detector: UV 254/220 nm. This yielded 49.7 mg (41%) of 2-(dimethylamino)ethyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)-4-methylpiperidine-4-carboxylate as a formate salt. LCMS (Method 28) [M+H] ⁺ =659.2, R _T =1.23 min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.35 (dd, 1H, J = 1.2, 7.2 Hz),8.68-8.64(m,2H),8.31(s,1H),7.63(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J＝ 2.8Hz),7.46(d,1H,J＝9.2Hz),7.29(dd,1H,J＝4.4,6.8Hz),7.09(t,1H,J＝73.2 Hz),5.23(s,2H),4.16(t,2H,J=5.6Hz),3.97-3.93(m,1H),3.75-3.71(m,1H), 3.25-3.20(m,1H),3.00-2.94(m,1H),2.48-2.47(m,2H),2.33(s,6H),2.16-1.95(m,2H),1.49-1.44(m,2H),1.18(s,3H).

Example 341

1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-carboxylic acid piperidin-4-ylmethyl ester

To a solution of 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylic acid (150 mg, 0.26 mmol) in DMF (5 mL) was added tert-butyl 4-(hydroxymethyl)piperidine-1-carboxylate (281 mg, 1.31 mmol), 4-dimethylaminopyridine (10 mg, 0.08 mmol), and HATU (120 mg, 0.32 mmol). The resulting solution was stirred at room temperature overnight. DIEA (68 mg, 0.53 mmol) was then added. The resulting solution was stirred at room temperature for an additional 3 h and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 3% MeOH in DCM. This gave 150 mg (74%) of [1-[(tert-butoxy)carbonyl]piperidin-4-yl]methyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylate as a yellow oil. LCMS (Method 28) [M+H] ⁺ = 771.2, _RT = 1.10 min.

A solution of [1-[(tert-butoxy)carbonyl]piperidin-4-yl]methyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylate (150 mg, 0.19 mmol) and saturated HCl in dioxane (10 mL) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The resulting solution was diluted with 10 mL of methanol. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep C18 OBD column, 5 μm, 19 x 150 mm; Mobile phase: Water containing 0.05% FA and MeCN (30.0% to 33.0% over 10 minutes, to 95.0% over 1 minute, hold at 95.0% for 1 minute, decrease to 30.0% over 2 minutes); Detector: UV 254/220 nm. This yielded 82.3 mg (59%) of the formate salt of 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylic acid piperidin-4-ylmethyl ester as a light yellow solid. LCMS (Method 20) [M+H] ⁺ =671.3, R _T =1.58min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.33 (dd,1H,J＝1.2,6.9Hz),8.68-8.67(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.7,8.7 Hz),7.55(d,1H,J＝2.7Hz),7.45(d,1H,J＝8.7Hz),7.28(dd,1H,J＝4.5,7.2Hz), 7.01(t,1H,J=73.5),5.25-5.23(m,2H),4.24-4.19(m,1H),3.93-3.85(m,3H), 2.83-2.80(m,1H),2.72-2.64(m,3H),1.92-1.87(m,3H),1.71-1.67(m,3H), 1.59-1.48(m,1H),1.44-1.23(m,2H).

Example 342

1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylic acid

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (500 mg, 1.08 mmol) in DMF (5 mL) was added methyl piperidine-4-carboxylate (232.1 mg, 1.62 mmol), DIEA (279.2 mg, 2.16 mmol), and HATU (493.5 mg, 1.30 mmol). The resulting solution was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 3% MeOH in DCM. This gave 500 mg (79%) of methyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylate as a yellow oil. LCMS (Method 25) [M+H] ⁺ = 588.1, _RT = 0.87 min.

A solution of K ₂ CO ₃ (500 mg, 3.59 mmol) in H ₂ O (5 mL) was added to a solution of methyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylate (500 mg, 0.85 mmol) in methanol (10 mL). The resulting solution was stirred at 50° C. overnight and acidified to pH 2 with 1N HCl. The solid was collected by filtration. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase: Water containing 0.05% FA and MeCN (30.0% MeCN to 34.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 30.0% in 2 minutes); Detector: UV 254/220 nm. This yielded 390 mg (80%) of 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]-acetyl)piperidine-4-carboxylic acid as a light yellow solid. LCMS (Method 28) [M+H] ⁺ =574.1, R _T =1.11min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 12.34(s,1H),9.76(s,1H),9.34(dd,1H,J＝1.6,7.2Hz),8.69-8.67(m,2H),8.31(s, 1H),7.62(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J＝2.4Hz),7.46(d,1H,J＝8.8Hz), 7.28(dd,1H,J＝4.4,6.8Hz),7.08(t,1H,J＝73.2Hz),5.29-5.19(m,2H),4.28-4.18 (m,1H),3.87-3.84(m,1H),3.19-3.13(m,1H),2.83-2.77(m,1H),1.86-1.85(m,2H), 1.60-1.57(m,1H),1.43-1.41(m,2H).

Example 343

1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)-4-methylpiperidine-4-carboxylic acid

To a solution of 2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]acetic acid (500 mg, 1.08 mmol) in DMF (4 mL) was added methyl 4-methylpiperidine-4-carboxylate (275 mg, 1.75 mmol), DIEA (419 mg, 3.24 mmol), and HATU (494 mg, 1.30 mmol). The resulting solution was stirred at room temperature for 4 h. The resulting mixture was concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 3% MeOH in DCM. This gave 500 mg (77%) of methyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)-4-methylpiperidine-4-carboxylate as a yellow oil. LCMS (Method 25) [M+H] ⁺ =602.1, _RT =0.92 min.

To a solution of methyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)-4-methylpiperidine-4-carboxylate (500 mg, 0.83 mmol) in methanol (10 mL) was added a solution of potassium carbonate (700 mg, 5.06 mmol) in water (5 mL). The resulting solution was stirred at 60° C. overnight and acidified to pH 2 with 1N HCl. The solid was collected by filtration. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase: Water containing 0.05% FA and MeCN (30.0% MeCN to 35.0% over 10 minutes, to 95.0% over 1 minute, hold at 95.0% for 1 minute, decrease to 30.0% over 2 minutes); Detector: UV 254/220 nm. This yielded 26.6 mg of 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)-4-methylpiperidine-4-carboxylic acid as a white solid. LCMS (Method 20) [M+H] ⁺ =588.2, R _T =2.79min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 12.5 (br, 1H), 9.76 (s, 1H), 9.34 (d, 1H, J = 6.8Hz), 8.69-8.68(m,2H),8.32(s,1H),7.63(dd,1H,J＝2.8,8.8Hz),7.56(d,1H,J＝2.8 Hz),7.46(d,1H,J＝8.8Hz),7.29(dd,1H,J＝4.4,7.2Hz),7.08(t,1H,J＝73.2Hz), 5.28-5.18(m,2H),3.96-3.93(m,1H),3.74-3.70(m,1H),3.26-3.23(m,1H), 3.01-2.95(m,1H),1.98-1.93(m,2H),1.46-1.30(m,2H),1.18(s,3H).

Example 344

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(4-[methyl[(5-oxooxolan-2-yl)methyl]amino]piperidin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a cold solution of 5-(hydroxymethyl)oxolane-2-one (100 mg, 0.86 mmol) and DIEA (340 mg, 2.63 mmol) in DCM (5 mL) was added Tf ₂ O (247 mg, 0.88 mmol) at 0° C. The resulting solution was stirred at room temperature for 3 h and quenched with a saturated Na ₂ CO ₃ solution. The resulting solution was extracted with dichloromethane (x3) and the organic layers were combined. The combined organic layers were washed with brine. Dried over anhydrous sodium sulfate and concentrated under vacuum. This gave 180 mg (84%) of (5-oxooxolane-2-yl)methyl trifluoromethanesulfonate as a yellow oil.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(methylamino)piperidin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.18 mmol) in CH ₃ CN (5 mL) was added DIEA (70 mg, 0.54 mmol) and (5-oxoxolan-2-yl)methyl trifluoromethanesulfonate (66.5 mg, 0.27 mmol). The resulting solution was stirred at room temperature overnight. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing 0.05% FA and MeCN (30.0% to 36.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 30.0% in 2 minutes); Detector, UV 254/220 nm. This yielded 37 mg (31%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(4-[methyl[(5-oxooxolan-2-yl)methyl]amino]piperidin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. LCMS (Method 24) [M+H] ⁺ = 657.2, R _T =1.68min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H,J＝1.2,7.2Hz),8.69-8.68(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.4,8.8Hz), 7.55(d,1H,J＝2.8Hz),7.46(d,1H,J＝8.8Hz),7.29(dd,1H,J＝4.0,6.8Hz),7.09 (t,1H,J＝73.2Hz),5.24-5.17(m,2H),4.58-4.54(m,1H),4.41-4.38(m,1H), 3.98-3.95(m,1H),3.16-3.03(m,1H),2.69-2.55(m,4H),2.46-2.44(m,2H),2.26(s, 3H),2.24-2.16(m,1H),1.88-1.83(m,1H),1.79-1.71(m,2H),1.48-1.44(m,1H), 1.29-1.21(m,1H).

Example 345

Methyl 3-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl](methyl)amino]propanoate

To a solution of tert-butyl 4-aminopiperidine-1-carboxylate (1.00 g, 4.99 mmol) in CH ₃ CN (20 mL) were added potassium carbonate (1.38 g, 9.99 mmol) and methyl 3-bromopropionate (3.34 g, 20.00 mmol). The resulting solution was heated to reflux for 20 h and cooled to room temperature. Water (100 mL) and DCM (100 mL) were added. The phases were separated. The aqueous phase was extracted with DCM. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 4% MeOH in DCM. This gave 300 mg (21%) of tert-butyl 4-[(3-methoxy-3-oxopropyl)amino]piperidine-1-carboxylate as a yellow oil. LCMS (Method 20) [M+H] ⁺ = 287.0, _RT = 1.06 min.

To a solution of tert-butyl 4-[(3-methoxy-3-oxopropyl)amino]piperidine-1-carboxylate (300 mg, 1.05 mmol) in methanol (10 mL) was added paraformaldehyde (HCHO)n (90 mg, 3.00 mmol). The reaction mixture was stirred at room temperature for 6 hours, then _NaBH3CN (130 mg, 2.07 mmol) was added. The resulting solution was stirred at room temperature overnight and quenched with water (0.5 mL). The resulting mixture was concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 4% MeOH in DCM. This gave 300 mg (crude) of tert-butyl 4-[(3-methoxy-3-oxopropyl)(methyl)amino]piperidine-1-carboxylate as a yellow oil. LCMS (Method 20) [M+H] ⁺ = 301.0, _RT = 1.08 min.

A solution of tert-butyl 4-[(3-methoxy-3-oxopropyl)(methyl)amino]piperidine-1-carboxylate (300 mg, 1.00 mmol) and saturated HCl in dioxane (10 mL) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. This gave 300 mg (crude) of methyl 3-[methyl(piperidin-4-yl)amino]propanoate hydrochloride as a white solid. LCMS (Method 20) [M+H] ⁺ = 201.0, _RT = 0.31 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid trifluoroacetate (100 mg, 0.17 mmol) in DMF (3 mL) was added methyl 3-[methyl(piperidin-4-yl)amino]propanoate hydrochloride (62 mg, 0.26 mmol), DIEA (67.2 mg, 0.52 mmol), and HATU (79.2 mg, 0.21 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was passed through a short pad of silica gel and eluted with ethyl acetate/petroleum ether (10:1). The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep _C18 OBD Column, 5 μm, 19×150 mm; Mobile phase: Water containing 0.05% FA and MeCN (37.0% to 43.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 37.0% in 2 minutes); Detector: UV 254/220 nm. This yielded 45.2 mg (38%) of methyl 3-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl](methyl)amino]propanoate as an off-white solid. LCMS (Method 24) [M+H] ⁺ = 645.2, R _T = 1.64min. ¹ HNMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.35 (dd, 1H, J = 1.5, 6.9Hz),8.69-8.68(m,2H),8.31(s,1H),7.63(dd,1H,J＝2.7,8.7Hz),7.55(d,1H,J ＝2.7Hz),7.46(d,1H,J＝8.7Hz),7.29(dd,1H,J＝4.5,7.2Hz),7.04(t,1H,J＝ 73.2Hz),5.33-5.27(m,2H),4.52-4.48(m,1H),4.10-4.06(m,1H),3.66(s,3H), 3.51-3.34(m,2H),3.16-3.08(m,2H),2.86-2.51(m,6H),2.28-1.99(m,2H), 1.73-1.70(m,1H),1.55-1.45(m,1H).

Example 346

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-([[4-(methylsulfanyl)phenyl]methyl]amino)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide formate

To a solution of N-[1-(2-aminoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (130 mg, 0.29 mmol) in methanol (5 mL) was added 4-(methylsulfanyl)benzaldehyde (66.3 mg, 0.44 mmol). The mixture was stirred for 2 hours, then NaBH ₃ CN (27.5 mg, 0.44 mmol) was added. The resulting solution was stirred at room temperature overnight and quenched with water (1 mL). The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing 0.05% FA and MeCN (30.0% to 37.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 30.0% in 2 minutes); Detector, UV 254/220 nm. This gave 39.6 mg (23%) of the formate salt of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-([[4-(methylsulfanyl)phenyl]methyl]amino)-ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 20) [M+H] ⁺ =584.2, R _T =2.23min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.74 (s, 1H), 9.34(dd,1H,J＝1.6,7.2Hz),8.69-8.67(m,2H),8.37(s,1H),7.61(dd,1H,J＝2.8, 8.8Hz),7.59(d,1H,J＝2.8Hz),7.44(d,1H,J＝8.8Hz),7.31-7.27(m,3H),7.18(d, 2H,J＝8.0Hz),7.07(t,1H,J＝73.6Hz),4.25(t,2H,J＝5.2Hz),3.70(s,2H), 2.95-2.92(m,2H),2.50(s,3H).

Example 347

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-[4-(2-oxoxan-4-yl)piperazin-1-yl]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of tert-butyl piperazine-1-carboxylate (500 mg, 2.68 mmol) in MeOH (20 mL) was added 5,6-dihydro-2H-pyran-2-one (530 mg, 5.40 mmol). The resulting solution was stirred at room temperature for 20 h. The resulting mixture was concentrated under vacuum. The residue was purified by flash chromatography on silica gel, eluting with 3% MeOH in DCM. This gave 550 mg (72%) of tert-butyl 4-(2-oxooxan-4-yl)piperazine-1-carboxylate as a yellow oil. LCMS (Method 28) [M+H] ⁺ = 285.0, _RT = 0.53 min.

A mixture of tert-butyl 4-(2-oxooxan-4-yl)piperazine-1-carboxylate (550 mg, 1.93 mmol) and saturated HCl in dioxane (10 mL) was stirred at room temperature overnight. The solid was collected by filtration. This gave 335 mg (78%) of 4-(piperazin-1-yl)oxan-2-one hydrochloride as a white solid.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid trifluoroacetate (100 mg, 0.17 mmol) in DMF (3 mL) was added 4-(piperazin-1-yl)oxan-2-one hydrochloride (76 mg, 0.34 mmol), DIEA (67.2 mg, 0.52 mmol), and HATU (79.2 mg, 0.21 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep _C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing 0.05% FA and MeCN (35.0% to 39.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 35.0% in 2 minutes); Detector, UV 254/220 nm. This yielded 14.5 mg (13%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-[4-(2-oxooxan-4-yl)piperazin-1-yl]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 28) [M+H] ⁺ = 629.1, R _T = 1.44min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J = 2.0, 7.2Hz),8.69-8.68(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J ＝2.8Hz),7.45(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,7.2Hz),7.08(t,1H,J＝ 73.2Hz),5.24(s,2H),4.35-4.29(m,1H),4.19-4.16(m,1H),3.50-3.32(m,6H), 3.02-2.99(m,1H),2.69-2.51(m,4H),2.08-1.98(m,1H),1.77-1.75(m,1H).

Example 348

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(morpholin-4-ylmethyl)piperidin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of tert-butyl 4-formylpiperidine-1-carboxylate (1.00 g, 4.69 mmol) in MeOH (10 mL) was added morpholine (613 mg, 7.04 mmol). The resulting mixture was stirred for 6 hr, then NaBH ₃ CN (444 mg, 7.07 mmol) was added. The resulting solution was stirred at room temperature overnight and quenched with H ₂ O (1 mL). The resulting mixture was concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 3% MeOH in DCM. This gave 1.0 g (75%) of tert-butyl 4-(morpholin-4-ylmethyl)piperidine-1-carboxylate as a yellow oil. LCMS (Method 25) [M+H] ⁺ = 285.0, _RT = 0.59 min.

A solution of tert-butyl 4-(morpholin-4-ylmethyl)piperidine-1-carboxylate (1.0 g, 3.52 mmol) and saturated HCl in dioxane (20 mL) was stirred at room temperature overnight. This gave 660 mg (85%) of 4-(piperidin-4-ylmethyl)morpholine hydrochloride as a white solid. LCMS (Method 20) [M+H] ⁺ = 185.0, _RT = 0.30 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (150 mg, 0.26 mmol) in DMF (2 mL) was added 4-(piperidin-4-ylmethyl)morpholine hydrochloride (69 mg, 0.31 mmol), DIEA (101.2 mg, 0.78 mmol), and HATU (119.2 mg, 0.31 mmol). The resulting solution was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing ₁₀ mmol _NH₄HCO₃ and MeCN (25.0% MeCN to 36.0% in 10 min, to 95.0% in 1 min, hold at 95.0% for 1 min, decrease to 25.0% in 2 min); Detector, UV 254/220 nm. This yielded 61 mg (37%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(morpholin-4-ylmethyl)piperidin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a solid. LCMS (Method 20) [M+H] ⁺ =629.2, R _T =1.55min. ¹ HNMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J＝ 1.6,7.2Hz),8.69-8.68(m,2H),8.30(s,1H),7.62(dd,1H,J=2.8,8.8Hz),7.55(d, 1H,J＝2.4),7.45(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.0,6.8Hz),7.08(t,1H,J＝ 73.2Hz),5.27-5.16(m,2H),4.34-4.30(m,1H),3.92-3.89(m,1H),3.56(t,4H,J＝ 4.2Hz),3.17-3.06(m,1H),2.67-2.59(m,1H),2.39-2.25(m,4H),2.13-2.12(m,2H), 1.90-1.71(m,3H),1.20-1.08(m,1H),0.97-0.95(m,1H).

Example 349

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of tert-butyl 4-oxopiperidine-1-carboxylate (1.00 g, 5.02 mmol) in MeOH (50 mL) was added 1-methylpiperazine (1.0 g, 9.98 mmol) and AcOH (0.1 mL, 1.75 mmol). The reaction was stirred for 3 h, and NaBH ₃ CN (378 mg, 6.02 mmol) was added. The resulting solution was stirred at room temperature for 12 h. The reaction was then quenched with saturated NH ₄ Cl (2 mL). The resulting mixture was concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 8% MeOH in DCM. This gave 800 mg (39%) of tert-butyl 4-(4-methylpiperazin-1-yl)piperidine-1-carboxylate as a light yellow oil. LCMS (Method 20) [M+H] ⁺ = 284.0, _RT = 0.97 min.

A solution of tert-butyl 4-(4-methylpiperazin-1-yl)piperazine-1-carboxylate (800 mg, 1.97 mmol) and saturated HCl in dioxane (15 mL) was stirred at room temperature for 3 h. The solid was collected by filtration and dried. This gave 600 mg (83%) of 1-methyl-4-(piperazin-1-yl)piperazine hydrochloride as a white solid. LCMS (Method 20) [M+H] ⁺ = 184.0, _RT = 0.32 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (200 mg, 0.35 mmol) in DMF (4 mL) was added 1-methyl-4-(piperidin-4-yl)piperazine hydrochloride (170 mg, 0.77 mmol), DIEA (134.4 mg, 1.04 mmol), and HATU (160 mg, 0.42 mmol). The resulting solution was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase: Water containing 0.05% FA and MeCN (25.0% to 34.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 25.0% in 2 minutes); Detector: UV 254/220 nm. This yielded 74.3 mg (32%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide formate as an off-white solid. LCMS (Method 20) [M+H] ⁺ =628.2, R _T =2.20min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.36-9.34 (dd, 1H,J＝1.6,7.2Hz),8.69-8.68(m,2H),8.31(s,1H),7.63(dd,1H,J＝2.4,8.8Hz), 7.55(d,1H,J＝2.4Hz),7.46(d,1H,J＝9.2Hz),7.29(dd,1H,J＝4.0,6.8Hz),7.09 (t,1H,J＝73.2Hz),5.25-5.21(m,2H),4.36-4.29(m,1H),3.98-3.79(m,1H), 3.10-3.00(m,1H),2.69-2.51(m,1H),2.49-2.32(m,8H),2.18(s,3H),1.91-1.86(m,2H),1.47-1.41(m,1H),1.39-1.25(m,1H).

Example 350

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(4-methylpiperazin-1-yl)methyl]piperidin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

A solution of tert-butyl 4-formylpiperidine-1-carboxylate (1 g, 4.69 mmol) and 1-methylpiperazine (703 mg, 7.02 mmol) in methanol (10 mL) was stirred for 6 h, then NaBH ₃ CN (444 mg, 7.07 mmol) was added. The resulting solution was stirred at room temperature overnight and quenched with water (1 mL). The resulting mixture was concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 5% MeOH in DCM. This gave 1.1 g (79%) of tert-butyl 4-[(4-methylpiperazin-1-yl)methyl]piperidine-1-carboxylate as a yellow oil. LCMS (Method 28) [M+H] ⁺ = 298.0, _RT = 0.46 min.

A mixture of tert-butyl 4-[(4-methylpiperazin-1-yl)methyl]piperidine-1-carboxylate (1.1 g, 3.70 mmol) and saturated HCl in dioxane (20 mL) was stirred at room temperature overnight. The solid was collected by filtration. This gave 560 mg (65%) of 1-methyl-4-(piperidin-4-ylmethyl)piperazine hydrochloride as a white solid. LCMS (Method 20) [M+H] ⁺ = 198.0, _RT = 0.32 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) in DMF (2 mL) was added 1-methyl-4-(piperidin-4-ylmethyl)piperazine hydrochloride (49 mg, 0.21 mmol), DIEA (67.2 mg, 0.52 mmol), and HATU (79.2 mg, 0.21 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing ₁₀ mmol of _NH₄HCO₃ and MeCN (20.0% MeCN to 30.0% in 10 min, to 95.0% in 1 min, hold at 95.0% for 1 min, decrease to 20.0% in 2 min); Detector, UV 254/220 nm. This yielded 52.7 mg (47%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(4-methylpiperazin-1-yl)methyl]piperidin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 24) [M+H] ⁺ = 642.2, R _T =1.55min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H,J＝1.6,7.2Hz),8.69-8.68(m,2H),8.30(s,1H),7.62(dd,1H,J＝2.4,8.8Hz), 7.54(d,1H,J＝2.8Hz),7.45(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.08 (t,1H,J＝73.2Hz),5.26-5.16(m,2H),4.37-4.30(m,1H),3.93-3.87(m,1H), 3.11-3.05(m,1H),2.67-2.59(m,1H),2.49-2.23(m,8H),2.16-2.12(m,5H), 1.83-1.71(m,3H),1.10-1.07(m,1H),0.97-0.88(m,1H).

Example 351

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-oxo-2-[4-[(5-oxo-2,5-dihydrofuran-3-yl)methyl]piperazin-1-yl]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(piperazin-1-yl)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.19 mmol) in DMF (3 mL) was added DIEA (48.8 mg, 0.38 mmol) and 4-(bromomethyl)-2,5-dihydrofuran-2-one (66.4 mg, 0.38 mmol). The resulting solution was stirred at room temperature for 4 h and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing 0.05% FA and MeCN (34.0% to 39.0% in 10 minutes, to 95.0% in 1 minute, maintained at 95.0% for 1 minute, and decreased to 34.0% in 2 minutes); Detector, UV 254/220 nm. This yielded 53.4 mg (45%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-oxo-2-[4-[(5-oxo-2,5-dihydrofuran-3-yl)methyl]piperazin-1-yl]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 24) [M+H] ⁺ =627.1, R _T =1.55min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.35 (d, 1H, J = 6.8 Hz),8.69-8.68(m,2H),8.31(s,1H),7.63(dd,1H,J＝2.4,8.8Hz),7.55(d,1H,J＝ 2.4Hz),7.46(d,1H,J＝9.2Hz),7.29(dd,1H,J＝4.2,7.0Hz),7.09(t,1H,J＝73.2 Hz), 6.12 (s, 1H), 5.25 (s, 2H), 4.91 (s, 2H), 3.51 (t, 4H, J = 5.4Hz), 3.41 (s, 2H), 2.50-2.40 (m, 4H).

Example 352

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(4-[methyl[(5-oxo-2,5-dihydrofuran-3-yl)methyl]amino]piperidin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(methylamino)piperidin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.18 mmol) in DMF (4 mL) was added DIEA (46 mg, 0.36 mmol) and 4-(bromomethyl)-2,5-dihydrofuran-2-one (63 mg, 0.36 mmol). The resulting solution was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column, XBridge Prep C18 OBD column, 5 μm, 19×150 mm; Mobile phase, water containing 0.05% FA and MeCN (40.0% to 48.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 40.0% in 2 minutes); Detector, UV 254/220 nm. This yielded 15.5 mg (13%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(4-[methyl[(5-oxo-2,5-dihydrofuran-3-yl)methyl]amino]piperidin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 20) [M+H] ⁺ =655.2, R _T =1.91min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34(dd,1H,J＝1.6,7.2Hz),8.69-8.68(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.8, 8.8Hz),7.55(d,1H,J＝2.8Hz),7.46(d,1H,J＝9.2Hz),7.28(dd,1H,J＝4.4,7.2 Hz),7.09(t,1H,J＝73.2Hz),6.04(s,1H),5.24-5.22(m,2H),4.86(d,2H,J＝1.2 Hz),4.39-3.37(m,1H),4.01-3.93(m,1H),3.47(s,2H),3.10-3.01(m,1H),2.67-2.62 (m,2H),2.20(s,3H),1.76-1.73(m,2H),1.56-1.42(m,1H),1.40-1.29(m,1H).

Example 353

3-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]propanoic acid

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (200 mg, 0.35 mmol) in DMF (4 mL) was added ethyl 3-(piperazin-1-yl)propanoate hydrochloride (90 mg, 0.40 mmol), DIEA (179.25 mg, 1.39 mmol), and HATU (158.20 mg, 0.42 mmol). The resulting solution was stirred at room temperature for 6 h. The resulting mixture was concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 10% MeOH in DCM. This gave 270 mg (crude) of ethyl 3-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]propanoate as a yellow solid. LCMS (Method 28) [M+H] ⁺ =631.2, _RT =0.66 min.

To a solution of ethyl 3-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]propanoate (270 mg, 0.43 mmol) in ethanol (10 mL) was added KOH (200 mg, 3.56 mmol) and water (4 mL). The resulting solution was stirred at room temperature for 1 h and neutralized with 1N HCl solution. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (Prep-HPLC-005): Column: XBridge Prep C18 OBD Column, 5 μm, 19×150 mm; Mobile phase: Water containing 0.05% FA and MeCN (27.0% to 35.0% in 10 minutes, to 95.0% in 1 minute, hold at 95.0% for 1 minute, decrease to 27.0% in 2 minutes); Detector: UV 254/220 nm. This yielded 31.5 mg (11%) of the formate salt of 3-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]-acetyl)piperazin-1-yl]propanoic acid as an off-white solid. LCMS (Method 28) [M+H] ⁺ =603.1, R _T =0.81min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J = 1.4, 7.0Hz), 8.69-8.67 (m,2H),8.31(s,1H),7.62(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J＝2.4Hz),7.46(d, 1H,J＝8.4Hz),7.28(dd,1H,J＝4.2,7.0Hz),7.08(t,1H,J＝73.6Hz),5.24(s,1H), 3.50-3.47(m,4H),2.57(t,2H,J＝7.2Hz),2.49-2.39(m,6H).

Example 354

tert-Butyl (3R,4R)-4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]-3-hydroxypiperidine-1-carboxylate

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]-pyrazolo[1,5-a]pyrimidine-3-carboxamide (600 mg, 1.482 mmol) in 1,2-dichloroethane (15 mL) was added Yb(OTf) ₃ (300 mg, 0.484 mmol) and tert-butyl 7-oxa-3-azabicyclo[4.1.0]heptane-3-carboxylate (1.20 g, 6.02 mmol). The resulting mixture was stirred at 65° C. overnight and concentrated under vacuum. The residue was purified by flash silica gel chromatography eluting with ethyl acetate/petroleum ether (4/1). The regioisomer/stereoisomer mixture (500 mg) was separated by Chiral-Prep-HPLC using the following conditions: column, Phenomenex Lux 5u Packed 250*21.2 mm, 5 um; mobile phase, mobile phase A: hexane, mobile phase B: ethanol; flow rate: 20 mL/min; gradient: 35B to 35B in 28 min; RT 1:10; RT 2:13; detector, 254/220 nm. 30 mg of product was obtained, Alpha (25 degree C, Hex:EtOH=60:40), giving 31.3 mg (3.5%) of (3R,4R)-4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]-3-hydroxypiperidine-1-carboxylic acid tert-butyl ester as a solid. (This single isomer is the first peak in Chiral-HPLC using the above conditions.) LCMS (Method 20) [M+H] ⁺ = 604.3, _RT = 2.08 min. ¹ H NMR (300 MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.10 (dd, 1H, J = 1.4, 6.9 Hz), 8.69-8.66 (m, 2H), 8.40 (s, 1H), 7.70 (d, 1H, J = 2.4 Hz), 7.55 (dd, 1H, J = 2.7, 8.7 Hz), 7.40 (d, 1H, J = 8.7 Hz), 7.21 (dd, 1H, J = 4.2, 6.9 Hz), 6.59 (t, 1H, J = 73.7 Hz),4.37-4.03(m,4H),3.03(m,1H),2.15(m,1H),1.59(m,2H),1.52(m,9H).

Example 355

(3S,4S)-tert-Butyl 4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]-3-hydroxypiperidine-1-carboxylate

(This single isomer was the second peak in Chiral-HPLC using the above conditions) to give 30.5 mg (3.5%) of (3S,4S)-tert-butyl 4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]-3-hydroxypiperidine-1-carboxylate as a solid. LCMS (Method 20) [M+H] ⁺ =604.3, R _T =2.08min. ¹ H NMR (300MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.10 (dd, 1H, J＝ 1.2,6.9Hz),8.69-8.66(m,2H),8.40(s,1H),7.71(s,1H),7.55(d,1H,J＝8.4Hz), 7.40(d,1H,J＝8.4Hz),7.26-7.22(m,1H),6.59(t,1H,J＝73.5Hz),4.37-4.03(m,4H),3.03(m,1H),2.15(m,1H),1.59(m,2H),1.52(m,9H).

Example 356

tert-Butyl (3R,4R)-3-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]-4-hydroxypiperidine-1-carboxylate

(This single isomer was the third peak in Chiral-HPLC using the above conditions) to give 24.8 mg (2.8%) of (3R,4R)-tert-butyl 3-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]-4-hydroxypiperidine-1-carboxylate as a solid. LCMS (Method 20) [M+H] ⁺ =604.3, R _T =2.08min. ¹ H NMR (300MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.10 (dd, 1H, J＝ 1.2,6.9Hz),8.69-8.65(m,2H),8.38(s,1H),7.69(s,1H),7.55(d,1H,J＝7.8Hz), 7.39(d,1H,J＝8.7Hz),7.25-7.22(m,1H),6.83(t,1H,J＝73.5Hz),4.38-4.35(m, 1H),4.25-4.15(m,2H),4.01-3.93(m,1H),2.97(m,1H),2.77(m,1H),2.17-2.13(m,2H),1.53(m,9H).

Example 357

(3S,4S)-tert-Butyl 3-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]-4-hydroxypiperidine-1-carboxylate

(This single isomer was the fourth peak in Chiral-HPLC using the above conditions) to give 24.9 mg (2.8%) of (3S,4S)-3-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]-4-hydroxypiperidine-1-carboxylic acid tert-butyl ester as a solid. LCMS (Method 20) [M+H] ⁺ = 604.3, R _T =2.08min. ¹ H NMR (300MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.10 (dd, 1H, J = 1.2, 6.9Hz),8.69-8.65(m,2H),8.38(s,1H),7.69(s,1H),7.55(d,1H,J＝8.4Hz), 7.42-7.39(d,1H,J＝8.1Hz),7.25-7.22(m,1H),6.58(t,1H,J＝73.5Hz),4.38-4.35 (m,1H),4.25-4.15(m,2H),4.01-3.93(m,1H),2.97(m,1H),2.77(m,1H),2.15-2.14 (m,2H),1.53(m,9H).

Example 358

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(3R,4R)-3-hydroxypiperidin-4-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (presumed)

A mixture of the first peak from the chiral separation of Example 357 (20 mg, 0.033 mmol) and saturated HCl in dioxane (4 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum. This afforded 18 mg (100%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(3R,4R)-3-hydroxypiperidin-4-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide hydrochloride (presumed) as a solid. LCMS (Method 20) [M+H] ⁺ =504.2, R _T =1.38min. ¹ HNMR (300MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.10(dd,1H,J＝1.5,6.9Hz),8.67-8.65(m,2H),8.44(s,1H),7.71(d,1H,J＝2.4 Hz),7.56(dd,1H,J＝2.5,8.8Hz),7.40(d,1H,J＝9Hz),7.21(dd,1H,J＝4.4,7.1 Hz),6.85(t,1H,J=73.4Hz),4.45-4.43(m,1H),4.28-4.23(m,1H),3.77-3.50(m, 4H),2.32-2.27(m,1H),1.91-1.86(m,1H).

Example 359

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(3S,4S)-3-hydroxypiperidin-4-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (presumed)

A mixture of the second peak (Example 358) from the chiral separation of Example 357 and saturated HCl in dioxane (4 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum. This gave 20 mg (86%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(3S,4S)-3-hydroxypiperidin-4-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide hydrochloride (presumed) as a yellow solid. LCMS (Method 20) [M+H] ⁺ =504.2, R _T =1.71min. ¹ HNMR (300MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.10 (d, 1H,J＝6.9Hz),8.66-8.64(m,2H),8.43(s,1H),7.71(d,1H,J＝2.7Hz),7.55(dd, 1H,J＝2.7,8.7Hz),7.39(d,1H,J＝9.0Hz),7.21(dd,1H,J＝4.1,6.9Hz),6.60(t, 1H,J=73.4Hz),4.47-4.44(m,1H),4.28-4.21(m,1H),3.77-3.50(m,4H),2.31-2.26(m,1H),1.93-1.82(m,1H).

Example 360

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(3R,4R)-4-hydroxypiperidin-3-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (assumed absolute)

A mixture of the third peak from the chiral separation of Example 357 (Example 359) (20 mg, 0.033 mmol) and saturated HCl in dioxane was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum. This yielded 11.3 mg (63%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(3R,4R)-4-hydroxypiperidin-3-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide hydrochloride as a solid. LCMS (Method 20) [M+H] ⁺ =504.2, R _T =1.38min. ¹ HNMR (300MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.12 (d,1H,J＝6.0Hz),8.73-8.66(m,2H),8.41(s,1H),7.69(d,1H,J＝1.5Hz),7.57(dd, 1H,J＝2.4,8.7Hz),7.42(d,1H,J＝5.7Hz),7.23(dd,1H,J＝4.4,7.1Hz),6.62(t, 1H, J＝73.4Hz), 4.46-4.4.28(m,2H), 3.69-3.61(m,2H), 3.26(m,1H), 3.11-3.04(m, 1H), 2.72-2.24(m,2H).

Example 361

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(3S,4S)-4-hydroxypiperidin-3-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

A mixture of the fourth peak from Example 357 (Example 360) (26 mg) and saturated HCl in dioxane (4 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum. This gave 22.4 mg (96%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(3S,4S)-4-hydroxypiperidin-3-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide hydrochloride as a yellow solid. LCMS (Method 20) [M+H] ⁺ = 504.2, R _T =1.73min. ¹ H NMR (300MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.08 (d, 1H, J = 7.2 Hz),8.64-8.63(m,2H),8.37(s,1H),7.65(d,1H,J＝2.1Hz),7.53(dd,1H,J＝2.1, 9.0Hz),7.37(d,1H,J＝8.7Hz),7.19(dd,1H,J＝4.8,6.3Hz),6.58(t,1H,J＝73.5 Hz),4.39-4.4.27(m,2H),3.74-3.40(m,2H),3.31(m,1H),3.07-3.00(m,1H), 2.44-2.43(m,2H).

Example 362

Ethyl 2-[(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-ylamino]-1H-pyrazol-1-yl]ethyl)(pyridin-3-ylmethyl)amino]acetate

To a solution of N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (510 mg, 1.00 mmol) in EtOH (30 mL) was added triethylamine (1.01 g, 9.98 mmol) and ethyl 2-aminoacetate hydrochloride (681 mg, 4.88 mmol). The resulting solution was stirred at 80° C. for 24 h. The resulting mixture was concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 3% MeOH in DCM. This gave 420 mg (79%) of ethyl 2-[(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]ethyl)amino]acetate as a light yellow oil. LCMS (Method 20) [M+H] ⁺ = 534.0, _RT = 1.18 min.

To a solution of ethyl 2-[(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)amino]acetate (420 mg, 0.79 mmol) in EtOH (30 mL) was added AcOH (0.1 mL, 1.75 mmol) and pyridine-3-carboxaldehyde (126 mg, 1.18 mmol). The mixture was stirred at room temperature for 3 h, and NaBH ₃ CN (99 mg, 1.58 mmol) was added. The resulting solution was stirred at 60° C. for 12 h. The solid was filtered off. The resulting mixture was concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 3% MeOH in DCM. The crude product (100 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-2): column, _C18 silica gel; mobile phase, _CH3CN : _H2O = 5:95 increasing to _CH3CN : _H2O = 24:40 over 12 min; detector, UV 254 nm. 26 mg of product was obtained. This gave 25.6 mg (5%) of ethyl 2-[(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]-ethyl)(pyridin-3-ylmethyl)amino]acetate as an off-white solid. LCMS (Method 20) [M+H] ⁺ =625.3, R _T =1.68min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.73(s,1H),9.35(d,1H,J＝6.8Hz),8.69-8.68(m,2H),8.39-8.37(m,3H), 7.63-7.61(m,2H),7.51(s,1H),7.43(d,1H,J=8.8Hz),7.31-7.25(m,2H),7.04(t, 1H,J＝73.6Hz),4.30-4.27(m,2H),4.07(q,2H,J＝6.8Hz),3.82(s,2H),3.44(s, 2H),3.15-3.11(m,2H),1.17(t,3H,J＝7.2Hz).

Example 363

2-[(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)(pyridin-3-ylmethyl)amino]acetic acid

A mixture of ethyl 2-[(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)(pyridin-3-ylmethyl)amino]acetate (180 mg, 0.29 mmol), ethanol (10 mL), and 1N sodium hydroxide (2 mL) was stirred at room temperature for 30 min. The mixture was acidified to pH 5 with 1N HCl. The resulting mixture was concentrated under vacuum. The crude product (200 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, C18 silica gel; mobile phase, CH3CN:H2O = 5:95 increasing to _CH3CN : _H2O = 24:40 over 12 min; detector, UV 254 nm. 8.6 mg of product was obtained. This gave 8.6 mg (5%) of 2-[(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)(pyridin-3-ylmethyl)amino]acetic acid formate as a white solid. LCMS (Method 20) [M+H] ⁺ =597.2, R _T =2.72min. ¹ H NMR (300MHz, DMSO-d ₆ )δ: (ppm) 9.72 (s, 1H), 9.34 (d, 1H, J = 7.2Hz), 8.69-8.68 (m, 2H), 8.39-8.37 (m, 3H), 7.63-7.60(m,2H),7.51(d,1H,J＝2.1Hz),7.43(d,1H,J＝9.0Hz),7.32-7.22(m, 2H),6.99(t,1H,J=73.2Hz),4.33-4.22(m,2H),3.83(s,2H),3.334(s,2H), 3.15-3.07 (m, 2H).

Example 364

Ethyl 2-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl]amino]acetate

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (665 mg, 1.44 mmol) in DMF (7 mL) was added tert-butyl N-(piperidin-4-yl)carbamate (300 mg, 1.50 mmol), triethylamine (700 mg, 6.92 mmol), and HATU (670 g, 2.78 mol). The resulting solution was stirred at room temperature for 1.5 h. EtOAc (100 mL) and water (50 mL) were added. The phases were separated. The organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with ethyl acetate/hexane (1/1). This gave 600 mg (65%) of tert-butyl N-[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl]carbamate as a yellow solid. TLC: R _f = 0.3; ethyl acetate/petroleum ether = 1/1.

TFA (2.0 mL, 26.93 mmol) was added to a solution of tert-butyl N-[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl]carbamate (600 mg, 0.93 mmol) in dichloromethane (8 mL). The resulting solution was stirred at room temperature for 2 h and concentrated under vacuum. The residue was dissolved in ethyl acetate (100 mL) and saturated _NaHCO₃ (50 mL) was added. The phases were separated. The organic layer was washed with saturated _NaHCO₃ , brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. This gave 450 mg (89%) of N-[1-[2-(4-aminopiperidin-1-yl)-2-oxoethyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. TLC: R _f = 0.3; MeOH/DCM = 1/5.

To a solution of N-[1-[2-(4-aminopiperidin-1-yl)-2-oxoethyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (126 mg, 0.24 mmol) in DMF (3 mL) was added potassium carbonate (76 mg, 0.55 mmol) and ethyl 2-bromoacetate (0.03 mL, 0.27 mmol). The resulting mixture was stirred at 25°C for 1 h and diluted with 100 mL of ethyl acetate. Water (50 mL) was added. The phases were separated. The organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. The crude product was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): Column: Xbridge C18, 19*150 mm, 5 μm; Mobile phase A: Water/0.05% NH ₄ HCO ₃ , Mobile phase B: ACN; Flow rate: 30 mL/min; Gradient: 20% B to 85% B in 10 min; 254 nm. This gave 29.3 mg (98%) of ethyl 2-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl]amino]acetate as a white solid. LCMS (Method 25) [M+H] ⁺ = 631.2, R _T = 1.89min. ¹ H NMR (400MHz, CD ₃ OD-d ₄ ) δ: (ppm) 8.98 (d, 1H, J = 6.8Hz), 8.53-8.52(m,2H),8.25(s,1H),7.58(d,1H,J＝2.4Hz),7.44(dd,1H,J＝2.4,8.8 Hz), 7.29 (d, 1H, J = 8.8Hz), 7.09 (dd, 1H, J = 4.4, 6.8Hz), 6.51 (t, 1H, J = 73.6Hz), 5.20-5.13(m,2H),4.32(d,1H,J＝13.2Hz),4.07(q,2H,J＝7.2Hz),3.91(d,1H,J ＝13.2Hz),3.36(s,2H),3.21-3.09(m,1H),2.78-2.68(m,2H),1.94-1.83(m,2H), 1.34-1.22(m,2H),1.16(t,3H,J＝7.2Hz).

Example 365

2-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl]amino]acetic acid

A mixture of ethyl 2-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl]amino]-acetate (200 mg, 0.32 mmol) and KOH (100 mg, 1.78 mmol) in methanol (5 mL) was stirred at 25° C. for 12 h. The crude product was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): Column: Xbridge C ₁₈ , 19*150 mm, 5 μm; Mobile phase A: water/0.05% NH ₄ HCO ₃ , Mobile phase B: ACN; Flow rate: 30 mL/min; Gradient: 20% B to 85% B in 10 min; HPLC: 254 nm. This gave 28.5 mg (15%) of 2-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]-acetyl)piperidin-4-yl]amino]acetic acid as a white solid. LCMS (Method 25) [M+H] ⁺ =603.1, R _T =1.39min. ¹ H NMR (400MHz, CD3OD-d ₄ ) δ: (ppm) 8.98 (dd, 1H, J = 1.2, 6.8Hz), 8.54-8.53 (m, 2H),8.24(s,1H),7.57(d,1H,J＝2.4Hz),7.45(dd,1H,J＝2.8,8.8Hz),7.30(d,1H, J＝8.8Hz),7.09(dd,1H,J＝4.4,6.8Hz),6.70(t,1H,J＝73.4Hz),5.23-5.10(m, 2H),4.54(d,1H,J＝13.2Hz),4.05(d,1H,J＝13.6Hz),3.45(s,2H),3.30-3.11(m, 4H),2.71-2.65(m,1H),2.11-2.05(m,1H),1.58-1.43(m,2H).

Example 366

Ethyl 2-(4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]piperidin-1-yl)acetate

To a solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (1 g, 4.97 mmol) in DCM (50 mL) was added DIEA (2.00 g, 15.47 mmol), 4-dimethylaminopyridine (10 mg, 0.08 mmol) and MsCl (862 mg, 7.53 mmol). The resulting solution was stirred at room temperature for 12 h, washed with water and brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with ethyl acetate/petroleum ether (1/1). This gave 860 mg (62%) of tert-butyl 4-(methylsulfonyloxy)piperidine-1-carboxylate as a light yellow solid. LCMS (Method 20) [M+H] ⁺ = 224.0, _RT = 1.42 min.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (500 mg, 1.24 mmol) in MeCN (50 mL) was added _{CsCO (} 1.21 g, 3.71 mmol) and tert-butyl 4-(methylsulfonyloxy)piperidine-1-carboxylate (828 mg, 2.96 mmol). The resulting solution was stirred at 80°C for 18 h. The solid was filtered off. The filtrate was concentrated under vacuum. The residue was purified by flash chromatography on silica gel, eluting with 2% MeOH in DCM. This gave 1.1 g (crude) of tert-butyl 4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]piperidine-1-carboxylate as a yellow solid. LCMS (Method 25) [M+H] ⁺ =588.0, _RT =1.17 min.

A mixture of the crude product 4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]-piperidine-1-carboxylic acid tert-butyl ester (1.1 g) from the previous step and saturated HCl dioxane solution (20 mL) was stirred at room temperature for 8 h. The resulting mixture was concentrated under vacuum. The residue was dissolved in ethanol (20 mL) and neutralized to pH ~8 with 2N NaOH. The resulting mixture was concentrated under vacuum. The residue was purified by flash silica gel chromatography eluting with 15% MeOH in DCM. This gave 320 mg (35%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(piperidin-4-yl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(piperidin-4-yl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (320 mg, 0.66 mmol) in ethanol (20 mL) was added 50% ethyl 2-oxoacetate hydrate (268 mg, 1.31 mmol), AcOH (0.1 mL, 1.75 mmol) and NaBH ₃ CN (83 mg, 1.32 mmol). The resulting solution was stirred at 60° C. for 12 h and concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with 2% MeOH in DCM. The crude product (100 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, silica gel; mobile phase, CH ₃ CN:H ₂ O = 5:95 increasing to CH ₃ CN:H ₂ O = 55:75 over 12 min; detector, UV 254 nm. 25.3 mg of product was obtained. This gave 25.3 mg (7%) of ethyl 2-(4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]piperidin-1-yl)acetate as an off-white solid. LCMS (Method 20) [M+H] ⁺ =574.2, R _T =2.70min. ¹ H NMR (300MHz, DMSO-d ₆ )δ: (ppm) 9.73(s,1H),9.36(s,1H),8.69-8.67(m,2H),8.36(s,1H),7.63-7.25(m,5H), 4.25-4.23(m,1H),4.13-4.10(m,2H),3.28-3.27(m,2H),3.01-2.98(m,2H), 2.42-2.40(m,2H),2.12-1.98(m,4H),1.25-1.20(m,3H).

Example 367

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[(1,3-oxazol-2-ylmethyl)amino]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of N-[1-(2-aminoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.22 mmol) in MeOH (15 mL) was added 1,3-oxazole-2-carbaldehyde (21.7 mg, 0.22 mmol) followed by NaBH ₃ CN (16 mg, 0.25 mmol). The resulting solution was stirred at room temperature for 12 h. The resulting mixture was concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with 5% MeOH in DCM. The crude product was purified by Prep-HPLC using the following conditions (2#-AnalyseHPLC-SHIMADZU (HPLC-10)): Column, XBridge BEH130 Prep C18 OBD column, 19×, 100 mm, 5 μm, 13 nm; Mobile phase, water containing 10 mM NH₄CO₃ and ACN (20% ACN to 50% over 6 minutes); Detector, UV 254/220 nm. 52.7 mg of product was obtained. This yielded 52.7 mg (45%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[(1,3-oxazol-2-ylmethyl)amino]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 20) [M+H] ⁺ =529.2, R _T =2.42min. ¹ H NMR (300MHz, CD ₃ OD-d ₄ ) δ: (ppm) 9.08 (dd, 1H, J = 1.7, 7.1Hz), 8.66-8.63 (m, 2H), 8.34 (s, 1H), 7.87(d,1H,J＝0.6Hz),7.67(d,1H,J＝2.7Hz),7.53(dd,1H,J＝2.4,8.7Hz),7.38 (d,1H,J＝8.7Hz),7.19(dd,1H,J＝4.2,7.2Hz),7.13(d,1H,J＝0.6Hz),6.55(t, 1H, J＝73.5Hz), 4.33 (t, 2H, J＝6.0Hz), 3.95 (s, 2H), 3.14 (t, 2H, J＝6.0Hz).

Example 368

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[(pyridin-3-ylmethyl)amino]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

A mixture of N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (150 mg, 0.29 mmol) and pyridin-3-ylmethanamine (158 mg, 1.46 mmol) in MeCN (3 mL) was stirred at 70°C for 12 h. The resulting mixture was concentrated under vacuum. The crude product was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-2): column, _C18 silica gel; mobile phase, _CH3CN : _H2O = 5:95 increasing to _CH3CN : _H2O = 30:42 over 12 min; detector, UV 254 nm. This gave 57.7 mg (37%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[(pyridin-3-ylmethyl)amino]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 25) [M+H] ⁺ =539.1, R _T =0.87min. ¹ H NMR (300MHz, DMSO-d ₆ )δ: (ppm) 9.76(s,1H),9.33(dd,1H,J＝1.5,6.9Hz),8.69-8.67(m,2H),8.50(d,1H,J＝1.8H z),8.41(dd,1H,J＝1.5,4.8Hz),8.38(s,1H),7.73-7.70(m,1H),7.61(dd,1H,J＝ 2.7,8.7Hz),7.59(d,1H,J＝2.4Hz),7.43(d,1H,J＝8.7Hz),7.33-7.28(m,2H), 6.99(t,1H,J＝73.2Hz), 4.24(t,2H,J＝6.0Hz), 3.75-3.72(m,2H), 3.01-2.94(m, 2H), 2.27(br,1H).

Example 369

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2S)-2-(methylamino)propyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of (2S)-2-aminopropan-1-ol (1.0 g, 13.31 mmol) in MeOH (30 mL) was added 4-methoxybenzaldehyde (1.8 g, 13.22 mmol). NaBH ₃ CN (600 mg) was then added. The resulting solution was stirred at room temperature for 12 h. The solid was filtered off. The resulting mixture was concentrated under vacuum. This gave 2.1 g (crude) of (2S)-2-[[(4-methoxyphenyl)methyl]amino]propan-1-ol as a light yellow oil. LCMS (Method 27) [M+H] ⁺ = 196.0, _RT = 0.95 min.

To a solution of (2S)-2-[[(4-methoxyphenyl)methyl]amino]propan-1-ol (2.1 g, 10.75 mmol) and 37% formaldehyde (900 mg) in methanol (50 mL) was added AcOH (0.1 mL, 1.75 mmol) and NaBH ₃ CN (1.8 g, 28.64 mmol). The resulting solution was stirred at room temperature for 5 h, quenched with water (10 mL) and concentrated under vacuum. The residue was dissolved in dichloromethane (100 mL), washed with water and brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 10% MeOH in DCM. This gave 1.0 g (44%) of (2S)-2-[[(4-methoxyphenyl)methyl](methyl)amino]propan-1-ol as a colorless oil. LCMS (Method 21) [M+H] ⁺ = 210.0, _RT = 0.92 min.

MsCl (250 mg, 2.18 mmol) was added dropwise to a stirred solution of (2S)-2-[[(4-methoxyphenyl)methyl](methyl)amino]propan-1-ol (400 mg, 1.91 mmol) and DIEA (516 mg, 3.99 mmol) in DCM (15 mL) at 0°C. The resulting solution was stirred at room temperature for 5 h and quenched with water (50 mL). The phases were separated. The aqueous phase was extracted with DCM (x2) and the organic layers were combined. The organic phase was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. This gave 510 mg (crude) [(2S)-1-chloroprop-2-yl][(4-methoxyphenyl)methyl]methylamine as a light red oil. LCMS (Method 25) [M+H] ⁺ = 228.0, _RT = 0.55 min.

A 20-mL microwave tube was charged with N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]-pyrimidine-3-carboxamide (400 mg, 0.99 mmol), Cs ₂ CO ₃ (652 mg, 2.00 mmol), DMF (10 mL) and [(2S)-1-chloroprop-2-yl][(4-methoxyphenyl)methyl]methylamine (454 mg, 1.99 mmol). The tube was evacuated and refilled with nitrogen three times. The final reaction mixture was irradiated with microwaves at 120° C. for 30 min. The reaction was then quenched by the addition of 50 mL of water. The resulting solution was extracted with dichloromethane (×2), and the organic layers were combined. The organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with dichloromethane/ethyl acetate (1:1). This gave 420 m of a mixture of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2S)-2-[[(4-methoxyphenyl)methyl](methyl)amino]propyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide and N-[5-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2S)-2-[[(4-methoxyphenyl)methyl](methyl)amino]propyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 25) [M+H] ⁺ =596.0, _RT =0.81 min.

To a solution of the regioisomer mixture from the previous step (200 mg) in CH ₃ CN (20 mL) was added chloro(1-chloroethoxy)methanone (240 mg, 1.68 mmol). The resulting solution was stirred at 80° C. for 5 h. The resulting mixture was concentrated under vacuum and dissolved in 20 mL of methanol. The resulting solution was stirred at 80° C. for 12 h and concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with 5% MeOH in DCM. The crude product (120 mg) was purified by Prep-HPLC using the following conditions (3#-Pre-HPLC-006 (Waters)): column, XSelect CSH Prep C ₁₈ OBD column, 5 μm, 19*150 mm; mobile phase, water containing 10 mmol NH ₄ HCO ₃ and MeCN (26.0% MeCN to 40.0% in 8 min); detector, 254/220. Two fractions containing the main isomer were obtained (36.3 mg), which was the title compound. LCMS (Method 20) [M+H] ⁺ =476.2, R _T =2.53min. ¹ H NMR (300MHz, DMSO-d ₆ )δ: (ppm) 9.73 (s, 1H), 9.33 (dd, 1H, J = 1.2, 6.9Hz), 8.68-8.66 (m, 2H), 8.33(s,1H),7.65-7.61(m,2H),7.43(d,1H,J＝7.2Hz),7.27(dd,1H,J＝4.5,7.2 Hz), 6.97 (t, 1H, J = 73.5Hz), 4.11 (dd, 1H, J = 6.3, 13.5Hz), 4.01 (dd, 1H, J = 6.0, 13.5Hz),2.97-2.91(m,1H),2.29(s,3H),1.76(br,1H),0.95(d,3H,J＝6.3Hz).

Example 370

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2S)-2-(dimethylamino)propyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of N-[1-[(2S)-2-aminopropyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (170 mg, 0.37 mmol) in MeOH (15 mL) was added 37% formaldehyde solution (16.6 mg, 0.55 mmol) and NaBH ₃ CN (23.3 mg, 0.37 mmol). The resulting solution was stirred at room temperature for 12 h and concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with 10% MeOH in DCM. The crude product (80 mg) was purified by Prep-HPLC using the following conditions (#1-Pre-HPLC-006 (Waters)): Column, XSelect CSH Prep C18 OBD column, 5 μm, 19*150 mm; Mobile phase, water containing 10 mM NH4CO3 and MeCN (20.0% MeCN to 35.0% over 8 minutes); Detector, 254/220. This yielded 29 mg (16%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2S)-2-(dimethylamino)propyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 10) [M+H] ⁺ =490.1, R _T =0.84min. ¹ H NMR (300MHz, DMSO-d ₆ )δ: (ppm) 9.73 (s, 1H), 9.34 (dd, 1H, J = 1.5, 6.9Hz), 8.69-8.66 (m, 2H), 8.34(s,1H),7.61(dd,1H,J＝2.7,8.7Hz),7.60(d,1H,J＝2.1Hz),7.44(d,1H,J＝ 8.4Hz),7.28(dd,1H,J＝4.5,7.2Hz),6.99(t,1H,J＝73.5Hz),4.22(dd,1H,J＝7.2, 13.8Hz), 4.02 (dd, 1H, J＝6.9, 13.8Hz), 3.20-3.18 (m, 1H), 2.22 (s, 6H), 0.88 (d, 3H, J＝6.6Hz).

Example 371

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2S)-2-(ethylamino)propyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of N-[1-[(2S)-2-aminopropyl]-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (200 mg, 0.43 mmol) in EtOH (20 mL) was added acetaldehyde (48 mg, 40% aqueous solution) and NaBH ₃ CN (32.7 mg, 0.52 mmol). The resulting solution was stirred at room temperature for 12 h and concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with 5% MeOH in DCM. The crude product (60 mg) was purified by Prep-HPLC using the following conditions (3#-Pre-HPLC-006 (Waters)): Column, XSelect CSH Prep C ₁₈ OBD column, 5 μm, 19 x 150 mm; Mobile phase, water containing 10 mM NH 4 CO 3 and MeCN (26.0% MeCN to 40.0% over 8 minutes); Detector, 254/220. 24.2 mg of product was obtained. This yielded 24.2 mg (11%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2S)-2-(ethylamino)propyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 28) [M+H] ⁺ =490.1, R _T =0.86min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.74 (s, 1H), 9.34 (dd, 1H, J = 1.6, 7.2Hz), 8.68-8.67(m,2H),8.34(s,1H),7.64(d,1H,J＝2.1Hz),7.60(dd,1H,J＝2.8,8.4 Hz),7.44(d,1H,J＝8.4Hz),7.28(dd,1H,J＝4.0,7.2Hz),7.04(t,1H,J＝73.2Hz), 4.11(dd,1H,J＝6.0,13.6Hz),4.01(dd,1H,J＝6.2,13.6Hz),3.08-3.00(m,1H), 2.68-2.50(m,2H),1.52(br,1H),0.97(t,3H,J＝7.0Hz),0.95(d,3H,J＝6.4Hz).

Example 372

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[2-(methylsulfanyl)ethyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of tert-butyl piperazine-1-carboxylate (1.00 g, 5.37 mmol) in DMF (30 mL) was added 1-chloro-2-(methylsulfanyl)ethane (710 mg, 6.42 mmol) and Cs ₂ CO ₃ (2.6 g, 7.98 mmol). The resulting mixture was stirred at 50° C. for 12 h. Water (50 mL) and EtOAc (30 mL) were added. The phases were separated. The aqueous phase was extracted with ethyl acetate (×2), and the organic layers were combined. The organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with ethyl acetate/petroleum ether (1:2). This gave 180 mg (13%) of tert-butyl 4-[2-(methylsulfanyl)ethyl]piperazine-1-carboxylate as a light yellow oil. LCMS (Method 25) [M+H] ⁺ = 261, _RT = 0.63 min.

A mixture of tert-butyl 4-[2-(methylsulfanyl)ethyl]piperazine-1-carboxylate (180 mg, 0.69 mmol) in DCM (5 mL) and trifluoroacetic acid (10 mL) was stirred at room temperature for 12 h. The resulting mixture was concentrated under vacuum. The residue was dissolved in 5 mL of dioxane-HCl. The resulting mixture was concentrated under vacuum. This gave 120 mg (88%) of 1-[2-(methylsulfanyl)ethyl]piperazine hydrochloride as a light yellow solid. LCMS (Method 25) [M+H] ⁺ = 161.1, _RT = 0.21 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (120 mg, 0.26 mmol) in DMF (10 mL) was added 1-[2-(methylsulfanyl)ethyl]piperazine hydrochloride (102 mg, 0.52 mmol), HATU (119 mg, 0.31 mmol), and DIEA (168 mg, 1.30 mmol). The resulting solution was stirred at room temperature for 12 h. Water (50 mL) was added. The resulting solution was extracted with DCM (x3) and the organic layers were combined. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated under vacuum. The crude product (150 mg) was purified by Prep-HPLC using the following conditions (1#-Pre-HPLC-006 (Waters)): Column, XSelect CSH Prep C18 OBD column, 5 μm, 19*150 mm; Mobile phase, water containing ₁₀ mmol _NH4HCO3 and MeCN (20.0% MeCN to 35.0% in 8 minutes); Detector, 254/220. This gave 44.2 mg (28%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[2-(methylsulfanyl)ethyl]-piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]-pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 20) [M+H] ⁺ =605.2, R _T =1.61 min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.6, 7.2 Hz),8.69-8.68(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.4,8.8Hz),7.55(d,1H,J＝ 2.4Hz),7.45(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.08(t,1H,J＝73.2 Hz),5.23(s,2H),3.50-3.47(m,4H),2.67-2.56(m,4H),2.2.47-2.33(m,4H),2.08(s, 3H).

Example 373

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-[4-(propan-2-yl)piperazin-1-yl]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (150 mg, 0.32 mmol) in DMF (10 mL) was added DIEA (126 mg, 0.97 mmol), 1-(propan-2-yl)piperazine (126 mg, 0.98 mmol), and HATU (185 mg, 0.49 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The crude product (200 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, C18 silica gel; mobile phase, CH3CN:H2O = 5:95 increasing to _CH3CN : _H2O = 40:60 over 14 minutes; detector, UV 254 nm. This gave 51.6 mg (28%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-[4-(propan-2-yl)piperazin-1-yl]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 20) [M+H] ⁺ =573.2, R _T =1.80min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.76(s,1H),9.34(dd,1H,J＝1.2,6.9Hz),8.69-8.67(m,2H),8.31(s,1H),7.62(dd, 1H,J＝2.7,9.0Hz),7.55(d,1H,J＝2.7Hz),7.45(d,1H,J＝8.7Hz),7.28(dd,1H,J ＝4.2,6.9Hz),7.02(t,1H,J＝73.2Hz),5.23(s,2H),3.52-3.48(m,4H),2.78-2.65 (m,1H),2.46-2.39(m,4H),1.09-0.97(d,6H,J＝6.6Hz).

Example 374

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(cyclopropylmethyl)piperazin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) in DMF (10 mL) was added DIEA (83 mg, 0.64 mmol), 1-(cyclopropylmethyl)piperazine (60.6 mg, 0.43 mmol), and HATU. The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The crude product (120 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, _C18 silica gel; mobile phase, _CH3CN : _H2O = 5:95 increasing to _CH3CN : _H2O = 24:40 over 14 minutes; detector, UV 254 nm. This gave 47.6 mg (38%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(cyclopropylmethyl)piperazin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 20) [M+H] ⁺ =585.2, R _T =2.40 min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.5, 6.9 Hz),8.69-8.67(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.7,8.7Hz),7.55(d,1H,J ＝2.7Hz),7.45(d,1H,J＝8.7Hz),7.28(dd,1H,J＝4.2,6.9Hz),7.02(t,1H,J＝73.2 Hz),5.23(s,2H),3.51-3.49(m,4H),2.49-2.41(m,4H),2.23-2.21(m,2H),0.87-0.82 (m,1H),0.50-0.44(m,2H),0.13-0.08(m,2H).

Example 375

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(2,4-dimethylpiperazin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of tert-butyl 2-methylpiperazine-1-carboxylate (500 mg, 2.50 mmol) in EtOH (20 mL) was added 37% aqueous formaldehyde (405 mg) and AcOH (0.02 mL, 0.35 mmol). The mixture was stirred for 2 h, and NaBH ₃ CN (315 mg, 5.01 mmol) was added. The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 4% MeOH in DCM. This gave 430 mg (80%) of tert-butyl 2,4-dimethylpiperazine-1-carboxylate as a colorless oil. LCMS (Method 25) [M+H] ⁺ = 215.0, _RT = 1.45 min.

A mixture of tert-butyl 2-methylpiperazine-1-carboxylate (430 mg, 2.15 mmol) and saturated HCl in dioxane (15 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum. This gave 520 mg (crude) of 1,3-dimethylpiperazine hydrochloride as a light yellow solid. LCMS (Method 24) [M+H] ⁺ = 115.0, _RT = 0.25 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (150 mg, 0.32 mmol) in DMF (10 mL) was added DIEA (209 mg, 1.62 mmol), 1,3-dimethylpiperazine hydrochloride (146 mg, 0.97 mmol) and HATU (16 mg, 0.04 mmol). The resulting solution was stirred at room temperature overnight. Water (50 mL) and DCM (30 mL) were added. The phases were separated. The aqueous phase was extracted with DCM, and the organic layers were combined. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated under vacuum. The residue was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-2): column, C18 silica gel; mobile phase, CH ₃ CN:H ₂ O=5:95 increasing to CH ₃ CN:H ₂ O=24:40 over 12 min; detector, UV 254 nm. This gave 59.1 mg (33%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(2,4-dimethylpiperazin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. LCMS (Method 25) [M+H] ⁺ =559.2, R _T =1.63 min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.5, 7.2 Hz),8.69-8.67(m,2H),8.34-8.31(m,1H),7.62(dd,1H,J＝2.7,8.7Hz),7.55(d,1H, J＝2.7Hz),7.45(d,1H,J＝8.7Hz),7.28(dd,1H,J＝4.2,7.2Hz),7.02(t,1H,J＝ 73.2Hz),5.31-5.11(m,2H),4.52-4.47(m,0.5H),4.20-4.11(m,1H),3.78-3.69(m, 0.5H),3.32-3.21(m,0.5H),2.97-2.83(m,0.5H),2.75(d,1H,J＝11.1Hz),2.63(d, 1H,J＝11.4Hz),2.20-1.71(m,5H),1.38-1.12(m,3H).

Example 376

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(3,4-dimethylpiperazin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

The title compound was prepared from tert-butyl 3-methylpiperazine-1-carboxylate using a method analogous to the synthesis of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(2,4-dimethylpiperazin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 20) [M+H] ⁺ =559.2, R _T = 2.23min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.5 Hz,7.2Hz),8.69-8.67(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.7,8.7Hz),7.55(d, 1H,J＝2.7Hz),7.45(d,1H,J＝8.7Hz),7.28(dd,1H,J＝4.2,6.9Hz),7.02(t,1H,J ＝73.2Hz),5.35-5.13(m,2H),4.18-4.02(m,1H),3.86-3.78(m,1H),3.26-3.10(m, 1H),2.84-2.73(m,2H),2.26-1.91(m,5H),1.01(d,3H,J＝6.0Hz).

Example 377

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(2,2,4-trimethylpiperazin-1-yl)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

The title compound was prepared from tert-butyl 2,2-dimethylpiperazine-1-carboxylate using a method analogous to the synthesis of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(2,4-dimethylpiperazin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 25) [M+H] ⁺ =573.2, R _T =0.97min. ¹ HNMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J＝ 1.2,6.9Hz),8.69-8.68(m,2H),8.27(s,1H),7.62(dd,1H,J=2.7,8.7Hz),7.56(d, 1H,J＝2.4Hz),7.45(d,1H,J＝8.7Hz),7.28(dd,1H,J＝4.5,6.9Hz),7.03(t,1H, 73.1Hz),5.17(s,2H),3.48-3.42(m,2H),2.28-2.22(m,2H),2.17(s,3H),2.08(s,2H),1.39(s,6H).

Example 378

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(3,3,4-trimethylpiperazin-1-yl)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

The title compound was prepared from tert-butyl 3,3-dimethylpiperazine-1-carboxylate using a method analogous to the synthesis of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(2,4-dimethylpiperazin-1-yl)-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 20) [M+H] ⁺ =573.2, R _T =2.51min. ¹ HNMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (dd, 1H, J ＝1.5,7.2Hz),8.69-8.67(m,2H),8.33(s,1H),7.62(dd,1H,J＝2.7,8.7Hz),7.55(d, 1H,J＝2.1Hz),7.45(d,1H,J＝9.0Hz),7.28(dd,1H,J＝4.2,6.9Hz),7.02(t,1H,J ＝73.2Hz),5.26(s,1H),5.17(s,1H),3.51-3.48(m,2H),3.22(d,2H,J＝11.4Hz), 2.45-2.36(m,2H),2.13(s,3H),0.99(s,3H),0.92(s,3H).

Example 379

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[2-(dimethylamino)ethyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of 2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]-pyrimidin-3-amido]-1H-pyrazol-1-yl]acetic acid (100 mg, 0.22 mmol) in DMF (10 mL) was added dimethyl[2-(piperazin-1-yl)ethyl]amine (68 mg, 0.43 mmol), DIEA (125 mg, 0.97 mmol) and HATU (123 mg, 0.32 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (2#-AnalyseHPLC-SHIMADZU (HPLC-10)): Column, XBridge BEH130 Prep C18 OBD column, 19×, 100 mm, 5 μm, 13 nm; Mobile phase, water containing 10 mM NH₄CO₃ and ACN (15% ACN to 40% over 6 minutes); Detector, UV 254/220 nm. This yielded 79.1 mg (61%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[2-(dimethylamino)ethyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 25) [M+H] ⁺ =602.2, R _T =0.89min. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.7, 7.1Hz), 8.69-8.67 (m, 2H),8.30(s,1H),7.62(dd,1H,J＝2.7,8.7Hz),7.55(d,1H,J＝2.7Hz),7.45(d,1H, J＝9.0Hz),7.28(dd,1H,J＝4.2,6.9Hz),7.02(t,1H,J＝73.2Hz),5.23(s,2H), 3.48-3.42(m,4H),2.45-2.32(m,8H),2.13(s,6H).

Example 380

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(1,3-oxazol-2-ylmethyl)piperazin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of tert-butyl piperazine-1-carboxylate (300 mg, 1.61 mmol) in MeOH (15 mL) was added 1,3-oxazole-2-carbaldehyde (172 mg, 1.77 mmol), AcOH (0.01 mL, 0.17 mmol), and NaBH ₃ CN (152 mg, 2.42 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 5% MeOH in DCM. This afforded 480 mg (89%) of tert-butyl 4-(1,3-oxazol-2-ylmethyl)piperazine-1-carboxylate as a colorless oil. TLC: R _f = 0.4; MeOH/DCM = 1/10.

A mixture of tert-butyl 4-(1,3-oxazol-2-ylmethyl)piperazine-1-carboxylate (480 mg, 1.44 mmol) and a saturated HCl solution in dioxane (15 mL) was stirred at room temperature for 3 h. This gave 300 mg (82%) of 1-(1,3-oxazol-2-ylmethyl)piperazine hydrochloride as a white solid. LCMS (Method 20) [M+H] ⁺ = 168.0, _RT = 0.32 min.

To a solution of 2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]-pyrimidine-3-amido]-1H-pyrazol-1-yl]acetic acid (100 mg, 0.22 mmol) in DMF (10 mL) was added 1-(1,3-oxazol-2-ylmethyl)piperazine hydrochloride (132 mg, 0.65 mmol), HATU (123 mg, 0.32 mmol), and DIEA (139 mg, 1.08 mmol). The resulting solution was stirred at room temperature overnight. Water (50 mL) and DCM (30 mL) were added. The phases were separated. The aqueous phase was extracted with DCM (x2), and the organic layers were combined. The organic layer was washed with brine, dried over Na ₂ SO ₄ , and concentrated under vacuum. The crude product (150 mg) was purified by Prep-HPLC using the following conditions (2#-AnalyseHPLC-SHIMADZU (HPLC-10)): Column, XBridge BEH130 Prep C18 OBD column, 19×, 100 mm, 5 μm, 13 nm; Mobile phase, water containing 10 mM NH₄CO₃ and ACN (28% ACN to 38% over 6 min); Detector, UV 254/220 nm. This yielded 68.2 mg (52%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(1,3-oxazol-2-ylmethyl)piperazin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 25) [M+H] ⁺ =612.1, R _T =1.70min. ¹ H NMR (300MHz, DMSO-d ₆ )δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.7, 7.1Hz), 8.69-8.67 (m, 2H), 8.30 (s, 1H), 8.09(d,1H,J＝0.6Hz),7.62(dd,1H,J＝2.7,8.7Hz),7.55(d,1H,J＝2.7Hz),7.45 (d,1H,J＝9.0Hz),7.28(dd,1H,J＝4.2,7.2Hz),7.18(d,1H,J＝0.6Hz),7.01(t, 1H, J＝73.2Hz), 5.23 (s, 2H), 3.76 (s, 2H), 3.54-3.48 (m, 4H), 2.45-2.42 (m, 4H).

Example 381

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(5-oxooxolan-2-yl)methyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

Methanesulfonyl chloride (2.5 g, 21.8 mmol) was added dropwise to a stirred solution of tert-butyl N-(2-hydroxyethyl)carbamate (3.0 g, 18.61 mmol) and DIEA (7.2 g, 55.71 mmol) in DCM (50 mL) at 0 ° C. The resulting solution was stirred at room temperature for 8 h. The reaction was then quenched by adding 100 mL of sodium bicarbonate. The phases were separated. The aqueous phase was extracted with DCM (x2) and the organic layers were combined. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with ethyl acetate/petroleum ether (1:2). This gave 3.4 g (76%) of tert-butyl N-[2-(methylsulfonyloxy)-ethyl]carbamate as a red solid. LCMS (Method 25) [M+H] ⁺ = 225.0, _RT = 0.77 min.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (1.00 g, 2.47 mmol) in CH ₃ CN (30 mL) was added Cs ₂ CO ₃ (2.00 g, 6.14 mmol) and tert-butyl N-[2-(methylsulfonyloxy)-ethyl]carbamate (1.20 g, 5.01 mmol). The resulting solution was stirred at 70° C. overnight. The solid was filtered off. The filtrate was concentrated under vacuum. The residue was purified by flash chromatography on silica gel, eluting with DCM/EtOAc (3:1). This gave 680 mg (50%) of tert-butyl N-(2-[3-[5-chloro-2-(difluoromethoxy)pyridin-3-yl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)carbamate as a yellow oil. LCMS (Method 28) [M+H] ⁺ =548.0, _RT =1.33 min

A mixture of tert-butyl N-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]ethyl)carbamate (420 mg, 0.77 mmol) and saturated HCl dioxane solution (10 mL) was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum. The residue was dissolved in 15 mL of ethanol. The pH value of the solution was adjusted to 8 with sodium hydroxide (2 mol/L). The resulting mixture was concentrated under vacuum. The residue was applied to a silica gel column with dichloromethane/methanol (85/15). This gave 260 mg (76%) of N-[1-(2-aminoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 20) [M+H] ⁺ = 448.0, _RT = 0.76 min.

To a solution of N-[1-(2-aminoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (200 mg, 0.45 mmol) in CH 3 CN (20 mL) was added methyl trifluoromethanesulfonate (5-oxooxolan-2-yl) (166 mg, 0.67 mmol) and DIEA (173 mg, 1.34 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was passed through a short pad of silica gel and eluted with a 5% MeOH solution in DCM. The crude product (100 mg) was purified by Prep-HPLC using the following conditions (#1-Pre-HPLC-006 (Waters)): Column, XSelect CSH Prep C ₁₈ OBD column, 5 μm, 19 x 150 mm; Mobile phase, water containing 0.05% FA and MeCN (24.0% MeCN to 40.0% over 8 minutes); Detector, 254/220. 31.6 mg of product was obtained. This yielded 32 mg (12%) of the formate salt of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(5-oxooxolan-2-yl)methyl]-amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 28) [M+H] ⁺ =546.1, R _T =0.82min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.76(s,1H),9.34(dd,1H,J＝1.6,7.2Hz),8.68-8.66(m,2H),8.31(s,1H),7.64-7.62 (m,2H),7.44(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.06(t,1H,J＝73.2 Hz),4.56-4.53(m,1H),4.23(t,2H,J＝6.4Hz),3.01(t,2H,J＝6.0Hz),2.77-2.75 (m,2H),2.48-2.44(m,2H),2.19-2.16(m,1H),1.92-1.88(m,1H).

Example 382

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-oxo-2-[4-[(5-oxooxolan-2-yl)methyl]piperazin-1-yl]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

Neat trifluoromethanesulfonic anhydride _Tf2O (486 mg, 1.72 mmol) was added dropwise to a stirred solution of 5-(hydroxymethyl)oxolan-2-one (200 mg, 1.72 mmol) and DIEA (665 mg, 5.15 mmol) in DCM (20 mL) at -5°C. The resulting solution was stirred at room temperature for 3 h and quenched with saturated _NaHCO3 solution (50 mL). The resulting solution was extracted with DCM (x2), and the organic layers were combined. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. This yielded 280 mg (crude) of (5-oxooxolan-2-yl)methyl trifluoromethanesulfonate as a yellow liquid. TLC: _Rf = 0.5; ethyl acetate/petroleum ether = 1/2.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(piperazin-1-yl)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.19 mmol) in CH ₃ CN (15 mL) was added DIEA (73.03 mg, 0.57 mmol) and (5-oxoxolan-2-yl)methyl trifluoromethanesulfonate (93.49 mg, 0.38 mmol). The resulting solution was stirred at room temperature for 12 h, stirred for 3 h, and concentrated under vacuum. The crude product (180 mg) was purified by Prep-HPLC using the following conditions (2#-AnalyseHPLC-SHIMADZU (HPLC-10)): Column, XBridge BEH130 Prep C18 OBD column, 19×, 100 mm, 5 μm, 13 nm; Mobile phase, water containing 10 mM NH₄CO₃ and ACN (30% ACN to 40% over 6 min); Detector, UV 254/220 nm. 65 mg of product was obtained. The residue was applied to a silica gel column using 5% MeOH in DCM. This gave 29.5 mg of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-oxo-2-[4-[(5-oxoxolan-2-yl)methyl]piperazin-1-yl]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]-pyrimidine-3-carboxamide as a white solid. LCMS (Method 28) [M+H] ⁺ =629.1, R _T =0.81min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H),9.34(dd,1H,J＝1.5,6.9Hz),8.69-8.67(m,2H),8.31(s,1H),7.62(dd,1H,J＝ 2.7,8.7Hz),7.55(d,1H,J＝2.7Hz),7.45(d,1H,J＝9.0Hz),7.28(dd,1H,J＝4.2, 6.9Hz),7.02(t,1H,J＝73.2Hz),5.24(s,2H),4.71-4.66(m,1H),3.50-3.33(m,4H), 2.58(d,2H,J＝5.4Hz),2.60-2.46(m,6H),2.48-2.42(m,4H),2.30-2.21(m,1H), 1.90-1.83(m,1H).

Example 383

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(methylcarbamoyl)methyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of 2-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]acetic acid (500 mg, 2.05 mmol) in DMF (20 mL) was added methylamine hydrochloride (406 mg, 6.01 mmol), DIEA (793 mg, 6.14 mmol), and HATU (1.17 g, 3.08 mmol). The resulting solution was stirred at room temperature for 12 h. Water (50 mL) was added. The resulting solution was extracted with DCM (x3), and the organic layers were combined. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 5% MeOH in DCM. This gave 500 mg of tert-butyl 4-[(methylcarbamoyl)methyl]piperazine-1-carboxylate as a yellow oil. LCMS (Method 25) [M+H] ⁺ = 258.0, _RT = 0.52 min.

A mixture of tert-butyl 4-[(methylcarbamoyl)methyl]piperazine-1-carboxylate (500 mg, 1.94 mmol) and saturated HCl in dioxane was stirred at room temperature for 3 h. The resulting solution was concentrated under vacuum. This gave 300 mg (crude) of N-methyl-2-(piperazin-1-yl)acetamide hydrochloride. LCMS (Method 20) [M+H] ⁺ = 158.0, _RT = 0.37 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (150 mg, 0.32 mmol) in DMF (10 mL) was added N-methyl-2-(piperazin-1-yl)acetamide hydrochloride (125 mg, 0.65 mmol), HATU (185 mg, 0.49 mmol), and DIEA (126 mg, 0.97 mmol). The resulting solution was stirred at room temperature for 12 h and concentrated under vacuum. The residue was passed through a short pad of silica gel and eluted with 8% MeOH in DCM. The crude product (180 mg) was purified by Prep-HPLC using the following conditions (3#-Pre-HPLC-006 (Waters)): Column, XSelect CSH Prep C18 OBD column, 5 μm, 19*150 mm; Mobile phase, water containing 10 mM NH4CO3 and MeCN (26.0% MeCN to 40.0% over 8 minutes); Detector, 254/220. This yielded 73.9 mg (38%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(methylcarbamoyl)methyl]piperazin-1-yl]-2-oxoethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 20) [M+H] ⁺ =602.2, R _T =3.66min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H),9.34(dd,1H,J＝1.6,6.8Hz),8.69-8.68(m,2H),8.32(s,1H),7.75(q,1H,J＝ 4.8Hz),7.63(dd,1H,J＝2.6,8.6Hz),7.56(d,1H,J＝2.8Hz),7.46(d,1H,J＝8.8 Hz),7.29(dd,1H,J＝4.4,7.2Hz),7.08(t,1H,J＝73.2Hz),5.24(s,2H),3.54(t,4H, J＝5.0Hz),2.95(s,2H),2.63(d,3H,J＝4.8Hz),2.48-2.45(m,2H),2.44-2.41(m,2H).

Example 384

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(1R)-1-phenylethyl]amino]ethyl)-1H-pyrazole-4-

1,5-a]pyrazolo[1,5-a]pyrimidine-3-carboxamide

A mixture of N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.20 mmol) and (1R)-1-phenylethan-1-amine (242 mg, 2.00 mmol) in MeCN (3 mL, 57.07 mmol) was stirred at 80° C. for 12 h. The crude product (120 mg) was purified by Prep-HPLC using the following conditions (2#-Analyse HPLC-SHIMADZU (HPLC-10)): Column, XBridge BEH130 Prep C18 OBD column, 19×, 100 mm, 5 μm, 13 nm; Mobile phase, water containing 10 mM NH₄CO₃ and ACN (40% ACN to 58% over 6 min); Detector, UV 254/220 nm. This yielded 38 mg of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(1R)-1-phenylethyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 25) [M+H] ⁺ =552.1, R _T =1.16min. ¹ H NMR (300MHz, DMSO-d ₆ )δ: (ppm) 9.72(s,1H),9.34(dd,1H,J＝1.5,6.9Hz),8.69-8.67(m,2H),8.34(s,1H),7.60(dd, 1H,J＝2.7,9.0Hz),7.55(d,1H,J＝2.7Hz),7.43(d,1H,J＝8.7Hz),7.31-7.17(m, 6H), 6.98 (t, 1H, J = 73.2Hz), 4.18 (t, 2H, J = 6.0Hz), 3.70 (q, 1H, J = 6.6Hz), 2.83-2.73(m,2H),1.22(d,3H,J＝6.6Hz).

Example 385

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(1S)-1-phenylethyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

The title compound was prepared from (1S)-1-phenylethan-1-amine using a method analogous to the synthesis of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(1R)-1-phenylethyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 25) [M+H] ⁺ =552.1, R _T =1.15 min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.72 (s, 1H), 9.33 (dd, 1H, J = 1.5, 6.9 Hz),8.69-8.67(m,2H),8.34(s,1H),7.60(dd,1H,J＝2.9,8.9Hz),7.55(d,1H,J＝ 2.7Hz),7.43(d,1H,J＝8.7Hz),7.31-7.16(m,6H),6.98(t,1H,J＝73.2Hz),4.18(t, 2H, J＝6.0Hz), 3.70 (q, 1H, J＝6.6Hz), 2.83-2.73 (m, 2H), 1.22 (d, 3H, J＝6.6Hz).

Example 386

Methyl 2-[(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)amino]acetate

The title compound was prepared from methyl 2-aminoacetate hydrochloride and DIEA using a method analogous to the synthesis of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(1R)-1-phenylethyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 28) [M+H] ⁺ =520.1, R _T =0.85min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.73 (s, 1H), 9.33 (dd, 1H, J＝ 1.8,6.9Hz),8.69-8.67(m,2H),8.36(s,1H),7.65-7.61(m,2H),7.44(d,1H,J＝9.3 Hz),7.28(dd,1H,J＝4.2,7.2Hz),7.00(t,1H,J＝73.5Hz),4.22(t,2H,J＝6.0Hz), 3.62(s,3H),3.37(s,2H),2.98(t,2H,J=6.0Hz),2.27(br,1H).

Example 387

(2S)-2-amino-4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]butanoic acid methyl ester

To a solution of (2S)-2-[[(tert-butoxy)carbonyl]amino]-4-hydroxybutanoic acid (2.20 g, 10.03 mmol) in DCM (80 mL) was added DIEA (2.58 g, 19.96 mmol), followed by the addition of TBDMS-Cl (1.65 g, 10.95 mmol) in portions at 0 ° C. The resulting solution was stirred at room temperature for 12 h. Water (100 mL) was added. The phases were separated. The aqueous phase was extracted with DCM (x2) and the organic layers were combined. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with ethyl acetate/petroleum ether (4:1). This gave 800 mg of (2S)-2-[[(tert-butoxy)carbonyl]amino]-4-[(tert-butyldimethylsilyl)oxy]butanoic acid as a light yellow oil. LCMS (Method 28) [M+H] ⁺ = 275.0, _RT = 1.16 min.

To a solution of (2S)-2-[[(tert-butoxy)carbonyl]amino]-4-[(tert-butyldimethylsilyl)oxy]butanoic acid (800 mg, 2.40 mmol) in DCM (30 mL) was added 4-dimethylaminopyridine (29 mg, 0.24 mmol), EDC.HCl (550 mg, 2.87 mmol), and methanol (5 mL). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was purified by flash chromatography on silica gel, eluting with ethyl acetate/petroleum ether (1:4). This gave 600 mg (72%) of methyl (2S)-2-[[(tert-butoxy)carbonyl]amino]-4-[(tert-butyldimethylsilyl)oxy]butanoate as a light yellow oil. LCMS (Method 25) [M+H] ⁺ = 348.0, _RT = 1.21 min.

A solution of methyl 2-[[(tert-butoxy)carbonyl]amino]-4-[(tert-butyldimethylsilyl)oxy]butanoate (600 mg, 1.73 mmol) in THF (2 mL), water (2 mL), and AcOH (8 mL, 139.61 mmol, 80.90 equiv) was stirred for 2 h. The reaction mixture was neutralized with saturated sodium bicarbonate solution, extracted with DCM (x3), and the organic layers were combined. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. This gave 400 mg of methyl 2-[[(tert-butoxy)carbonyl]amino]-4-hydroxybutanoate as a light yellow oil. LCMS (Method 28) [M+H] ⁺ = 175, _RT = 0.65 min.

To a stirred solution of crude methyl 2-[[(tert-butoxy)carbonyl]amino]-4-hydroxybutanoate (400 mg) in DCM (20 mL) was added DIEA (387 mg, 2.99 mmol), followed by the dropwise addition of MsCl (114 mg, 1.00 mmol) with stirring at 0°C. The resulting solution was stirred at room temperature for 3 h. Saturated _NaHCO₃ (50 mL) was added. The resulting solution was extracted with DCM (x2) and the organic layers were combined. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. This gave 510 mg (80%) of methyl 2-[[(tert-butoxy)carbonyl]amino]-4-(methylsulfonyloxy)butanoate as a yellow oil. LCMS (Method 25) [M+H] ^⁺ = 212.0, _RT = 0.76 min.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (200 mg, 0.49 mmol) in CH ₃ CN (30 mL) was added Cs ₂ CO ₃ (326 mg, 1.00 mmol) and methyl (2S)-2-[[(tert-butoxy)carbonyl]amino]-4-(methylsulfonyloxy)butanoate (400 mg, crude). The resulting solution was stirred at room temperature overnight. The solid was filtered off. The filtrate was concentrated under vacuum. The residue was purified by flash silica gel chromatography eluting with dichloromethane/ethyl acetate (70/30). This gave 280 mg (73%) of methyl 2-[[(tert-butoxy)carbonyl]amino]-4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]butanoate as an off-white solid. LCMS (Method 25) [M+H] ⁺ =620.0, _RT =0.98 min.

A mixture of (2S)-2-[[(tert-butoxy)carbonyl]amino]-4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]-butanoic acid methyl ester (280 mg) in DCM/TFA (15 mL, 2:1) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The crude product (200 mg) was purified by Prep-HPLC using the following conditions (#1-Pre-HPLC-006 (Waters)): column, XSelect CSH Prep C18 OBD column, 5 μm, 19 x 150 mm; mobile phase, water containing 0.05% FA and MeCN (24.0% MeCN to 40.0% over 8 minutes); detector, 254/220. 150 mg of product was obtained. This gave 148.4 mg (73%) of (2S)-methyl 2-amino-4-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]butanoate formate as an off-white solid. LCMS (Method 28) [M+H] ⁺ =520.1, R _T =0.82min. ¹ H NMR (300MHz, DMSO-d ₆ )δ: (ppm) 9.75(s,1H),9.34(dd,1H,J＝1.5,6.9Hz),8.69-8.68(m,2H),8.36(s,1H),7.65-7.61 (m,2H),7.45(d,1H,J＝8.4Hz),7.28(dd,1H,J＝4.2,7.2Hz),7.01(t,1H,J＝73.2 Hz),4.40-4.26(m,2H),3.64(s,3H),3.50-3.43(m,1H),2.32-2.18(m,1H),2.11-1.99(m,1H).

Example 388

1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylic acid (1-methylpiperidin-4-yl)methyl ester

To a DCM (50 mL) solution of 1-[(tert-butoxy)carbonyl]piperidine-4-carboxylic acid (1.00 g, 4.36 mmol) was added (1-methylpiperidin-4-yl)methanol (1.13 g, 8.75 mmol), EDC.HCl (1.0 g, 5.22 mmol), 4-dimethylaminopyridine (54 mg, 0.44 mmol) and the resulting solution was stirred at room temperature overnight. Water (50 mL) was added. The phases were separated. The aqueous phase was extracted with DCM (x2) and the organic layers were combined. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluted with a DCM solution of 6% MeOH. This gave 1.2 g of piperidine-1,4-dicarboxylic acid 1-tert-butyl 4-(1-methylpiperidin-4-yl)methyl ester as a light yellow oil. LCMS (Method 28) [M+H] ⁺ = 341.0, _RT = 0.58 min.

A mixture of 1-tert-butyl 4-(1-methylpiperidin-4-yl)methyl piperidine-1,4-dicarboxylate (1.2 g, 3.52 mmol) and saturated HCl in dioxane (15 mL) was stirred at room temperature overnight. The resulting solution was concentrated under vacuum. This gave 1.5 g (crude) of (1-methylpiperidin-4-yl)methyl piperidine-4-carboxylate hydrochloride as a white solid. LCMS (Method 25) [M+H] ⁺ = 241.0, _RT = 0.19 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (150 mg, 0.32 mmol) in DMF (10 mL) was added piperidine-4-carboxylic acid (1-methylpiperidin-4-yl)methyl ester hydrochloride (178 mg, 0.64 mmol), DIEA (126 mg, 0.97 mmol), HATU (160 mg, 0.42 mmol). The resulting solution was stirred at room temperature overnight. Water (50 mL) was added. The resulting solution was extracted with DCM (x3) and the organic layers were combined. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was passed through a short silica gel pad and eluted with 8% MeOH in DCM. The crude product (80 mg) was purified by Prep-HPLC using the following conditions (3#-Pre-HPLC-006 (Waters)): Column, XSelect CSH Prep C ₁₈ OBD column, 5 μm, 19*150 mm; Mobile phase, water containing 0.05% FA and MeCN (32.0% MeCN to 50.0% over 8 minutes); Detector, 254/220. This yielded 37 mg (16%) of the formate salt of (1-methylpiperidin-4-yl)methyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylate as an off-white solid. LCMS (Method 25) [M+H] ⁺ =685.2, R _T =1.05min. ¹ H NMR (400MHz, DMSO-d6) δ: (ppm) 9.75 (s, 1H), 9.35 (dd, 1H, J = 1.2, 6.8Hz), 8.69-8.68(m,2H),8.32(s,1H),7.63(dd,1H,J＝2.8,8.8Hz),7.55(d,1H,J＝2.4 Hz),7.46(d,1H,J＝8.8Hz),7.29(dd,1H,J＝4.4,7.2Hz),7.08(t,1H,J＝73.2Hz), 5.36-5.18(m,2H),4.28-4.21(m,1H),3.96-3.88(m,3H),3.25-3.12(m,1H),2.8 6-2.74(m,3H),2.73-2.66(m,1H),2.15(s,3H),1.92-1.83(m,4H),1.65-1.52(m, 4H),1.48-1.38(m,1H),1.29-1.16(m,2H).

Example 389

1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)-4-methylpiperidine-4-carboxylic acid (1-methylpiperidin-4-yl)methyl ester

The title compound was prepared from 1-[(tert-butoxy)carbonyl]-4-methylpiperidine-4-carboxylic acid using a method analogous to the synthesis of (1-methylpiperidin-4-yl)methyl 1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidine-4-carboxylate. LCMS (Method 28) [M+H] ⁺ =699.2, R _T =0.95min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s,1H),9.34(dd,1H,J＝1.6,7.2Hz),8.69-8.68(m,2H),8.31(s,1H),7.63(dd,1H, J＝2.4,8.8Hz),7.55(d,1H,J＝2.8Hz),7.46(d,1H,J＝8.8Hz),7.29(dd,1H,J＝ 4.0,6.8Hz),7.08(t,1H,J＝73.2Hz),5.24(s,2H),3.99-3.96(m,2H),3.94-3.88(m, 1H),3.78-3.71(m,1H),3.26-3.20(m,1H),3.08-3.00(m,1H),2.85-2.80(m,2H), 2.20(s,3H),2.02-1.94(m,4H),1.68-1.59(m,4H),1.54-1.48(m,1H),1.41-1.34(m,1H),1.28-1.23(m,2H),1.18(s,3H).

Example 390

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[methyl[(5-oxooxolan-2-yl)methyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[[(5-oxooxolan-2-yl)methyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (140 mg, 0.26 mmol) in MeOH (20 mL) was added 37% formaldehyde in water (23 mg) followed by NaBH ₃ CN (24.6 mg, 0.39 mmol). The resulting solution was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with 8% MeOH in DCM. The crude product (60 mg) was purified by Prep-HPLC using the following conditions (#1-Pre-HPLC-006 (Waters)): Column, XSelect CSH Prep C18 OBD column, 5 μm, 19*150 mm; Mobile phase, water containing 0.05% FA and MeCN (18.0% MeCN to 36.0% over 8 minutes); Detector, 254/220. This yielded 22.4 mg of the formate salt of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[methyl[(5-oxooxolan-2-yl)methyl]amino]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 28) [M+H] ⁺ =560.2, R _T =0.94min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H),9.34(dd,1H,J＝1.6,7.2Hz),8.69-8.67(m,2H),8.38(s,1H),7.62(dd,1H,J＝ 2.8,8.8Hz),7.60(d,1H,J＝2.4Hz),7.45(d,1H,J＝8.8Hz),7.29(dd,1H,J＝4.0, 6.8Hz),7.08(t,1H,J=73.2Hz),4.59-4.55(m,1H),4.30-4.27(m,2H),2.94-2.88 (m,2H),2.62(d,2H,J＝5.6Hz),2.44-2.41(m,2H),2.32(s,3H),2.14-2.11(m,1H), 1.83-1.77(m,1H).

Example 391

Ethyl 3-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl](methyl)amino]propanoate

To a solution of tert-butyl 4-(methylamino)piperidine-1-carboxylate (1.00 g, 4.67 mmol) in ethanol (50 mL) was added AcOH (0.1 mL, 1.75 mmol) and ethyl 3-oxopropanoate (1.63 g, 14.04 mmol). The mixture was stirred at room temperature for 5 h, and NaBH ₃ CN (882 mg, 14.04 mmol) was added. The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with 4% MeOH in DCM. This gave 580 mg of tert-butyl 4-[(3-ethoxy-3-oxopropyl)(methyl)amino]piperidine-1-carboxylate as a colorless oil. LCMS (Method 20) [M+H] ⁺ = 315.0, _RT = 1.14 min.

A mixture of tert-butyl 4-[(3-ethoxy-3-oxopropyl)(methyl)amino]piperidine-1-carboxylate (580 mg, 1.84 mmol) and saturated HCl in dioxane was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum. The residue was triturated with EtOAc. The solid was collected by filtration. This gave 200 mg (crude) of ethyl 3-[methyl(piperidin-4-yl)amino]propanoate hydrochloride as an off-white solid. LCMS (Method 20) [M+H] ⁺ = 215.0, _RT = 0.45 min.

To a DMF (10 mL) solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg, 0.22 mmol) was added HATU (123 mg, 0.32 mmol), DIEA (83 mg, 0.64 mmol) and ethyl 3-[methyl(piperidin-4-yl)amino]propanoate hydrochloride (200 mg). The resulting solution was stirred at room temperature overnight. Water (50 mL) was added. The resulting mixture was extracted with DCM (x3) and the organic layers were combined. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The crude product (82 mg) was purified by Prep-HPLC using the following conditions (#1-Pre-HPLC-006 (Waters)): Column, XSelect CSH Prep C18 OBD column, 5 μm, 19 x 150 mm; Mobile phase, water containing 0.05% FA and MeCN (18.0% MeCN to 36.0% over 8 minutes); Detector, 254/220. 20.3 mg of product was obtained. This yielded 20.3 mg (13%) of ethyl 3-[[1-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperidin-4-yl](methyl)amino]propanoate as an off-white solid, as a formate salt. LCMS (Method 28) [M+H] ⁺ = 659.2, R _T =0.90min. ¹ HNMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (dd,1H,J＝1.5,7.2Hz),8.68-8.66(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.7,8.7 Hz),7.55(d,1H,J＝2.4Hz),7.45(d,1H,J＝8.7Hz),7.28(dd,1H,J＝4.2,7.2Hz), 7.03(t,1H,J＝73.2Hz),5.36-5.15(m,2H),4.48-4.33(m,1H),4.02(q,2H,J＝7.2 Hz),4.00-3.96(m,1H),3.18-3.00(m,1H),2.89-2.80(m,3H),2.68-2.54(m,3H), 2.31(s,3H),1.85-1.76(m,2H),1.61-1.25(m,2H),1.19(t,3H,J＝6.9Hz).

Example 392

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-[4-(5-oxooxolan-3-yl)piperazin-1-yl]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

A mixture of tert-butyl piperazine-1-carboxylate (1.0 g, 5.37 mmol), oxolane-2,4-dione (1.07 g, 10.69 mmol), and AcOH (0.2 mL) in THF (50 mL) was stirred at room temperature for 3 h. NaBH ₃ CN (508 mg, 8.08 mmol) was added. The resulting solution was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The residue was passed through a short pad of silica gel, eluting with dichloromethane/ethyl acetate (1/4). This gave 400 mg (28%) of tert-butyl 4-(5-oxooxolane-3-yl)piperazine-1-carboxylate as a light yellow oil. LCMS (Method 10) [M+H] ⁺ = 271.0, _RT = 0.56 min.

A mixture of tert-butyl 4-(5-oxooxolan-3-yl)piperazine-1-carboxylate (400 mg, 1.48 mmol) and saturated HCl in dioxane (15 mL) was stirred at room temperature for 3 h. The solid was collected by filtration. This gave 380 mg (crude) of 4-(piperazin-1-yl)oxolan-2-one hydrochloride as a white solid. LCMS (Method 22) [M+H] ⁺ = 171.0, _RT = 0.38 min.

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (200 mg, 0.43 mmol) in DMF (10 mL) was added 4-(piperazine-1-yl)oxolane-2-one hydrochloride (250 mg, 1.21 mmol), HATU (247 mg, 0.65 mmol), and DIEA (167 mg, 1.29 mmol). The resulting solution was stirred at room temperature overnight. Water (50 mL) was added. The resulting solution was extracted with DCM (x3) and the organic layers were combined. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was passed through a short silica gel pad and eluted with an 8% MeOH solution in DCM. The crude product (80 mg) was purified by Prep-HPLC using the following conditions (#1-Pre-HPLC-006 (Waters)): Column, XSelect CSH Prep C18 OBD column, 5 μm, 19 x 150 mm; Mobile phase, water containing 0.05% FA and MeCN (32.0% MeCN to 60.0% over 10 minutes); Detector, 254/220. 37 mg of product was obtained. This yielded 36.7 mg (14%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-[4-(5-oxooxolan-3-yl)piperazin-1-yl]ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a white solid. LCMS (Method 28) [M+H] ⁺ = 615.0, R _T = 0.99min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.76 (s, 1H), 9.34 (d, 1H, J = 6.9 Hz),8.69-8.67(m,2H),8.30(s,1H),7.62(dd,1H,J=2.4,9.0Hz),7.56(s,1H),7.45 (d,1H,J＝8.7Hz),7.28(dd,1H,J＝4.2,6.9Hz),7.02(t,1H,J＝73.2Hz),5.24(s, 2H),4.44-4.38(m,1H),4.26-4.18(m,1H),3.49(t,4H,J＝5.1Hz),3.40-3.37(m, 1H),2.78-2.68(m,1H),2.60-2.55(m,1H),2.47-2.35(m,4H).

Example 393

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(morpholin-4-yl)piperidin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (100 mg) in DMF (2 mL) was added HATU (100 mg, 0.26 mmol), DIEA (85 mg, 0.66 mmol), and 4-(piperidin-4-yl)morpholine hydrochloride (67 mg, 0.32 mmol). The resulting solution was stirred at room temperature overnight. The crude product (60 mg) was purified by Prep-HPLC using the following conditions (#1-Pre-HPLC-006 (Waters)): Column, XSelect CSH Prep C18 OBD column, 5 μm, 19 x 150 mm; Mobile phase, Water containing 10 mM NH₄CO₃ and MeCN (20.0% MeCN to 35.0% over 8 min); Detector, 254/220. This gave 28.3 mg (21%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-[4-(morpholin-4-yl)piperidin-1-yl]-2-oxoethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow solid. LCMS (Method 20) [M+H] ⁺ =615.2, R _T =1.84min. ¹ HNMR (300MHz, DMSO-d ₆ )δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.5, 6.9Hz), 8.69-8.67(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.7,8.7Hz),7.55(d,1H,J＝2.7 Hz),7.45(d,1H,J＝8.7Hz),7.28(dd,1H,J＝4.2,6.9Hz),7.02(t,1H,J＝73.2Hz), 5.24-5.21(m,2H),4.42-4.31(m,1H),4.02-3.95(m,1H),3.55(t,4H,J＝4.5Hz), 3.16-3.02(m,1H),2.78-2.64(m,1H),2.49-2.45(m,5H),1.90-1.76(m,2H),1.54- 1.21(m,2H).

Example 394

N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2E)-4-(morpholin-4-yl)but-2-en-1-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

A mixture of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (400 mg, 0.99 mmol), (2E)-1,4-dichlorobut-2-ene (370 mg, 2.96 mmol) and cesium carbonate (1 g, 3.07 mmol) was stirred at 65°C for 20 h. The reaction mixture was cooled. The resulting solution was diluted with ethyl acetate (100 mL) and washed with _H2O (x3). The organic layer was dried over sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with ethyl acetate/hexane (1:1). The appropriate fractions were combined and concentrated under vacuum. This gave 320 mg (66%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2E)-4-chlorobut-2-en-1-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. TLC: R _f = 0.4; ethyl acetate/petroleum ether = 1:1.

A solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2E)-4-chlorobut-2-en-1-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (160 mg, 0.324 mmol) and morpholine (0.5 mL) in DMF (4 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum. The crude product (200 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, silica gel; mobile phase, CH ₃ CN/H ₂ O = 20% increasing to CH ₃ CN/H ₂ O = 50% over 20 min; detector, UV 254 nm. 35.4 mg of product was obtained. This resulted in 35.4 mg (20%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2E)-4-(morpholin-4-yl)but-2-en-1-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 23) [M+H] ⁺ =544.0, R _T =2.37min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.74 (s, 1H), 9.34 (dd, 1H, J = 1.6, 6.8Hz), 8.68-8.66 (m, 2H),8.31(s,1H),7.62(dd,1H,J＝2.8,8.8Hz),7.58(d,1H,J＝2.8Hz),7.44(d, 1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.06(t,1H,J＝73.2Hz),5.94-5.84 (m,1H),5.79-5.69(m,1H),4.82(d,2H,J＝6.0Hz),3.56(t,4H,J＝4.6Hz),2.96 (d,2H,J＝6.4Hz),2.40-2.30(m,4H).

Example 395

N-[3-[5-Chloro-2-(difluoromethoxy)phenyl]-1-[(2E)-4-(4-methylpiperazin-1-yl)but-2-en-1-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

The title compound was prepared from 1-methylpiperazine using a method analogous to the synthesis of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[(2E)-4-(morpholin-4-yl)but-2-en-1-yl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide. LCMS (Method 23) [M+H] ⁺ =557.1, R _T =1.76min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.74 (s, 1H), 9.34 (d, 1H, J = 5.2Hz), 8.68-8.66 (m, 2H),8.31(s,1H),7.62(d,1H,J＝8.8Hz),7.58(d,1H,J＝2.4Hz),7.47(d,1H,J＝ 8.8Hz),7.28(dd,1H,J＝4.0,6.8Hz),7.06(t,1H,J＝73.2Hz),5.92-5.80(m,1H), 5.78-5.66(m,1H),4.81(d,2H,J＝6.0Hz),2.94(d,2H,J＝6.4Hz),2.45-2.20(m,8H),2.13(s,3H).

Example 396

2-(2-(3-(5-chloro-2-(difluoromethoxy)phenyl)-4-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)-1H-pyrazol-1-yl)ethylamino)acetate

To a solution of 2-(methylsulfanyl)ethan-1-ol (2.40 g, 26.04 mmol) in DMF (15 mL) was added 2-[(tert-butoxy)carbonyl]amino]acetic acid (3.19 g, 18.21 mmol), HOBt (2.82 g, 20.87 mmol), EDC-HCl (3.99 g, 20.81 mmol), and DIEA (2 _mL ). The resulting solution was stirred at room temperature overnight. The mixture was partitioned between ethyl acetate and water. The organic layer was washed with brine, dried over _Na₂SO₄ , and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with ethyl acetate/petroleum ether (3/1). This yielded 2.00 g (31%) of 2-(methylsulfanyl)ethyl 2-[[(tert-butoxy)carbonyl]amino]acetate as a light yellow oil. LCMS (Method 21): [M+H] ⁺ = 250, _RT = 1.37 min.

To a solution of 2-(methylsulfanyl)ethyl 2-[[(tert-butoxy)carbonyl]amino]acetate (2.00 g, 8.02 mmol) in dioxane (5 mL) was added saturated HCl in dioxane (10 mL) at room temperature. The resulting solution was stirred at room temperature for 2 h. The solid was collected by filtration. This gave 1.2 g of 2-(methylsulfanyl)ethyl 2-aminoacetate HCl salt as a white solid. LCMS (Method 25): [M+H] ⁺ = 150, _RT = 0.28 min.

To a solution of 2-(methylsulfanyl)ethyl 2-aminoacetate HCl salt (792 mg, 3.57 mmol) in CH ₃ CN was added N-[1-(2-bromoethyl)-3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (364 mg, 0.71 mmol) and DIEA (1.57 g, 12.15 mmol). The resulting solution was stirred at 75° C. overnight and concentrated under vacuum. The crude product was purified by Prep-HPLC using the following conditions (#2 - Waters 2767-2 (HPLC-08)): Column, Xbridge Prep Phenyl, 5 μm, 19 x 150 mm; Mobile phase, water containing 50 mmol of ammonium bicarbonate and acetonitrile (10.0% acetonitrile to 33.0% over 2 minutes, to 53.0% over 8 minutes, to 100.0% over 1 minute, decreasing to 10.0% over 1 minute); Detector, UV 220 nm. This yielded 53.1 mg (2%) of 2-(methylsulfanyl)ethyl 2-[(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethyl)amino]acetate, formate, as a light yellow oil. LCMS (Method 20): [M+H] ⁺ =580.15, R _T =2.76min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.73 (s, 1H), 9.33 (dd, 1H, J ＝1.5,6.9Hz),8.67(dd,1H,J＝1.5,4.2Hz),8.66(s,1H),8.36(s,1H),7.65-7.61(m, 2H),7.44(d,1H,J＝9.3Hz),7.23(dd,1H,J＝4.5,7.2Hz),6.99(t,1H,J＝73.5Hz), 4.27-4.18(m,4H),3.38(s,2H),2.99(t,2H,J＝6.0Hz),2.67(t,2H,J＝6.6Hz),2.07 (s,3H).

Example 397

Ethyl 2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]acetate

To a solution of ethyl 2-(piperazin-1-yl)acetate hydrochloride (400 mg, 1.92 mmol) in DMF (10 mL) was added {3-(5-chloro-2-difluoromethoxyphenyl)-4-[(pyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazol-1-yl}acetic acid (500 mg), DIEA (440 mg, 3.40 mmol), and HATU (460 mg, 1.21 mmol). The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was purified by flash chromatography on silica gel eluting with 4.5% MeOH in DCM to afford 59.4 mg (11%) of ethyl 2-[4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]acetyl)piperazin-1-yl]acetate as a white solid. LCMS (Method 20) [M+H] ⁺ =617.2, R _T =2.68 min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.75 (s, 1H), 9.34 (dd, 1H, J = 1.6, 6.8 Hz),8.69-8.67(m,2H),8.31(s,1H),7.62(dd,1H,J＝2.4,8.8Hz),7.56(d,1H,J ＝2.8Hz),7.45(d,1H,J＝8.8Hz),7.28(dd,1H,J＝4.4,6.8Hz),7.07(t,1H,J＝73.2 Hz), 5.24 (s, 2H), 4.07 (q, 2H, J = 7.2Hz), 3.52-3.49 (m, 4H), 3.28 (s, 2H), 3.17-3.16 (m, 1H), 2.59-2.50 (m, 4H), 1.18 (t, 3H, J = 7.2Hz).

Example 403

N-(3-(5-chloro-2-(difluoromethoxy)phenyl)-1-(2-hydroxy-3-morpholinopropyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (1.00 g, 2.47 mmol) in N,N-dimethylformamide (10 mL) was added Cs ₂ CO ₃ (970 mg, 2.977 mmol) and stirred at room temperature for 10 minutes. 2-(Chloromethyl)oxirane (2.28 g, 24.64 mmol) was then added dropwise at room temperature. The resulting mixture was stirred at room temperature for 14 hours, diluted with ethyl acetate (100 mL), and washed with water and brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash chromatography on silica gel using ethyl acetate/petroleum ether (60/40) as the eluent to afford 500 mg (44%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(oxiran-2-ylmethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 24), [M+H] ⁺ = 461.1, _RT = 1.30 min.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(oxiran-2-ylmethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (50 mg, 0.11 mmol) in N,N-dimethylformamide (1 mL) was added DIEA (67 mg, 0.518 mmol) and morpholine (14 mg, 0.161 mmol) at room temperature. The resulting solution was stirred at 60°C overnight, cooled, diluted with 30 mL of ethyl acetate, and washed with 2 x 10 mL of water and 2 x 10 mL of brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with dichloromethane/methanol (95/5). This gave 50 mg of crude product, which was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): Column: X Bridge C18, 19*150 mm, 5 um; Mobile phase A: water/0.05% NH ₄ HCO ₃ , Mobile phase B: ACN; Flow rate: 20 mL/min; Gradient: 30% B to 70% B in 10 min; At 254 nm, 40 mg (67%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-hydroxy-3-(morpholin-4-yl)propyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide was obtained as a white solid. LCMS (Method 35) [M+H] ⁺ = 548.2, _RT = 2.28 min. ¹ H-NMR (300 MHz, DMSO-d ₆ ): δ (ppm) 9.73 (s, 1H), 9.33 (dd, 1H, J = 1.5, 7.2 Hz), 8.68-8.67 (m, 2H), 8.34 (s, 1H), 7.65 (d, 1H, J = 2.7 Hz), 7.61 (s, 1H),7.44(d,1H,J＝8.7Hz),7.27(dd,1H,J＝4.2,6.9Hz),7.00(t,1H,J＝73.5H z),4.99(d,1H,J＝4.5Hz),4.31-4.28(m,1H),4.13-4.07(m,2H),3.56(t,4H,J =4.5Hz), 2.42 (t, 4H, J = 4.5Hz), 2.32 (d, 2H, J = 4.8Hz).

Example 414

N-(3-(5-chloro-2-(difluoromethoxy)phenyl)-1-(2-oxo-3-(piperidin-1-yl)propyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a stirred solution of oxalyl chloride (0.46 mL, 0.923 mmol, 2 M) in dichloromethane (6 mL) at –78°C was added a solution of DMSO (144 mg) in dichloromethane (1 mL) dropwise over 10 minutes. The mixture was stirred at this temperature for an additional 10 minutes, followed by the dropwise addition of a solution of N-(3-(5-chloro-2-(difluoromethoxy)phenyl)-1-(2-hydroxy-3-morpholinopropyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (253 mg, 0.461 mmol) in 2 mL of dichloromethane. The mixture was stirred at this temperature for an additional 30 minutes. DIEA (476 mg, 3.683 mmol) was then added dropwise at this temperature. The resulting solution was stirred at –78°C for 0.5 hours, then warmed to room temperature and stirred for an additional 30 minutes. The resulting solution was diluted with 50 mL of ethyl acetate and washed with 2 x 20 mL of water and ₂ x 20 mL of brine. The organic layer was dried over sodium sulfate and concentrated under vacuum. The crude product (200 mg) was purified by Flash-Prep-HPLC using the following conditions: Column: X Bridge RP, 19 x 150 mm, 5 μm; Mobile phase A: water/0.05% _NH₄HCO₃ , Mobile phase B: ACN; Flow rate: 30 mL/min; Gradient: 25% B to 44% B over 10 min; HPLC ... LCMS (Method 24) [M+H] ⁺ =546.2, R _T =1.68min. ¹ H-NMR (300MHz, CDCl ₃ -d): δ (ppm) 9.89 (s, 1H),8.77(dd,1H,J＝1.5,6.9Hz),8.72(s,1H),8.55(dd,1H,J＝1.5,4.2Hz), 8.40(s,1H),7.68(d,1H,J＝2.4Hz),7.42(dd,1H,J＝2.4,8.7Hz),7.28(d,1H,J =8.7Hz),6.99(dd,1H,J＝4.2,7.2Hz),6.47(t,1H,J＝73.8Hz),5.07(s,1H), 3.76(t,4H,J＝4.5Hz), 3.26(s,2H), 2.56(t,4H,J＝4.8Hz).

Example 415

N-(3-(5-chloro-2-(difluoromethoxy)phenyl)-1-(2-(1-methylpiperidin-4-ylidene)ethyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a mixture of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (215.20 mg, 0.532 mmol) and Cs ₂ CO ₃ (694.8 mg, 2.132 mmol) in N,N-dimethylformamide (15 mL) was added tert-butyl 4-[2-(methylsulfonyloxy)ethylidene]piperidine-1-carboxylate (650 mg, 2.128 mmol). The resulting mixture was stirred at 60° C. overnight and concentrated under vacuum. The residue was purified by flash silica gel chromatography eluting with ethyl acetate/petroleum ether (9/1). This gave 310 mg (95%) of tert-butyl 4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethylidene)piperidine-1-carboxylate as a yellow oil. LCMS (Method 25) [M+H] ⁺ = 614.2, _RT = 1.11 min.

To a solution of tert-butyl 4-(2-[3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]ethylidene)piperidine-1-carboxylate (310 mg, 0.505 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (6 mL) at room temperature. The resulting solution was stirred at room temperature for 5 h and concentrated under vacuum. The crude product was used without further purification. LCMS (Method 25) [M+H] ⁺ = 514.1, _RT = 0.91 min.

40% aqueous formaldehyde solution (210 mg) was added to a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(piperidin-4-ylidene)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (360.00 mg, 0.700 mmol) in methanol (30 mL). The resulting solution was stirred at ambient temperature for 2.5 h, and NaBH ₃ CN (44.02 mg, 0.700 mmol) was added. The resulting solution was stirred at this temperature for 3 h and concentrated under vacuum. The residue was applied to a short pad of silica gel and eluted with dichloromethane/MeOH (10/1). The crude product (60 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, silica gel; mobile phase, ACN/10 mmol NH ₄ HCO ₃ = 40% increasing to ACN/10 mmol NH ₄ HCO ₃ = 75% within 6 min; detector, UV 254 nm, to obtain 4.3 mg (1%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(1-methylpiperidin-4-ylidene)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 32): [M+H] ⁺ =528.1, R _T =2.21min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.73 (s, 1H), 9.34 (dd, 1H, J = 1.6, 6.8 Hz),8.68-8.66(m,2H),8.32(s,1H),7.67-7.61(m,2H),7.45(d,1H,J＝8.4Hz), 7.28(dd,1H,J＝4.0,6.8Hz),7.06(t,1H,J＝73.2Hz),5.44(t,1H,J＝6.8Hz),4.27 (d,2H,J＝6.8Hz),2.81-2.68(m,2H),2.52-2.50(m,2H),2.50-2.42(m,2H),2.19 (s,3H),2.12-2.04(m,2H).

Example 421

N-(3-(5-chloro-2-(difluoromethoxy)phenyl)-1-(2-(4-(2-cyanoethylamino)cyclohexylidene)ethyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

Methanesulfonyl chloride (1.87 g, 16.325 mmol) was added dropwise to a solution of 2-[1,4-dioxaspiro[4.5]dec-8-ylidene]ethan-1-ol (2.00 g, 10.86 mmol) and DIEA (4.21 g, 32.57 mmol) in dichloromethane (100 mL) at room temperature. The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with ethyl acetate/petroleum ether (1/9). This yielded 1.95 g (68%) of 2-[1,4-dioxaspiro[4.5]dec-8-ylidene]ethyl methanesulfonate as an off-white oil. TLC: R _f = 0.6; ethyl acetate/petroleum ether = 1/2.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (1.50 g, 3.71 mmol) in N,N-dimethylformamide (120 mL) was added Cs ₂ CO ₃ (3.62 g, 11.11 mmol) and 2-[1,4-dioxaspiro[4.5]decan-8-ylidene]ethyl methanesulfonate (1.94 g, 7.39 mmol) at room temperature. The resulting solution was stirred at room temperature for 4 h and concentrated under vacuum. The residue was purified by flash chromatography on silica gel using ethyl acetate/petroleum ether (3/2) as the eluent to afford 1.59 g (75%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[1,4-dioxaspiro[4.5]dec-8-ylidene]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 31) [M+H] ⁺ = 571.4, _RT = 1.21 min.

Concentrated aqueous HCl (16 mL) was added to a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[1,4-dioxaspiro[4.5]dec-8-ylidene]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (1.59 g, 2.785 mmol) in 1,4-dioxane (160 mL). The resulting solution was stirred at room temperature for 3 h and neutralized with saturated sodium bicarbonate solution. The resulting mixture was concentrated under vacuum. The residue was dissolved in dichloromethane, washed with water and brine. The organic layer was dried and concentrated under vacuum. The residue was purified by flash silica gel chromatography eluting with ethyl acetate/petroleum ether (3/2). This gave 930 mg (63%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(4-oxocyclohexylidene)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 25) [M+H] ⁺ =527.1, _RT =0.94 min.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(4-oxocyclohexylidene)ethyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (300.0 mg, 0.569 mmol) in methanol (30 mL) was added 3-aminopropionitrile (199.5 mg, 2.85 mmol). The resulting solution was stirred at room temperature for 3 minutes. NaBH ₃ CN (71.56 mg, 1.14 mmol) was then added. The resulting solution was stirred at room temperature overnight and concentrated under vacuum. The residue was applied to a short pad of silica gel and eluted with ethyl acetate. The crude product (60 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, silica gel; mobile phase, ACN/0.05% _NH4HCO3 = 30% increasing to ACN/0.05% _NH4HCO3 = ₅₅ % over ₈ min; detector, UV 254 nm. 6.7 mg of product was obtained. This gave 6.7 mg (2%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(2-cyanoethyl)amino]cyclohexylidene]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as an off-white solid. LCMS (Method 34) [M+H] ⁺ =581.4, R _T = 2.44min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.73 (s, 1H), 9.33 (dd, 1H, J ＝1.5,6.9Hz),8.68-8.66(m,2H),8.27(s,1H),7.64-7.60(m,2H),7.43(d,1H,J＝ 8.1Hz),7.27(dd,1H,J＝4.2,6.9Hz),7.00(t,1H,J＝73.5Hz),5.45(t,1H,J＝ 6.9Hz), 4.80 (d, 2H, J = 7.2Hz), 2.79-2.72 (m, 3H), 2.71-2.57 (m, 5H), 2.10-1.97 (m, 2H), 1.94-1.86 (m, 2H), 1.23-1.16 (m, 2H).

Example 420

N-(3-(5-chloro-2-(difluoromethoxy)phenyl)-1-(2-(4-((2-cyanoethyl)(methyl)amino)cyclohexylidene)ethyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(2-cyanoethyl)amino]cyclohexylidene]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (140.0 mg, 0.241 mmol) in methanol (20 mL) was added 40% aqueous HCHO (75 mg, 0.96 mmol). The resulting solution was stirred at room temperature overnight. NaBH ₃ CN (15.14 mg, 0.241 mmol) was then added. The resulting solution was stirred at room temperature for 4 h and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with dichloromethane/MeOH (10/1). This gave 34.7 mg (24%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(2-[4-[(2-cyanoethyl)(methyl)amino]cyclohexylidene]ethyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 33) [M+H] ⁺ =595.4, R _T =1.43min. ¹ H NMR (300MHz, DMSO-d ₆ ) δ: (ppm) 9.73 (s, 1H), 9.33 (dd, 1H, J＝ 1.5,6.9Hz),8.69-8.66(m,2H),8.28(s,1H),7.64-7.60(m,2H),7.43(d,1H,J＝ 8.4Hz),7.27(dd,1H,J＝4.5,7.5Hz),7.00(t,1H,J＝73.5Hz),5.43(t,1H,J＝ 7.2Hz),4.81(d,2H,J＝6.9Hz),2.88-2.82(m,1H),2.66-2.59(m,5H),2.33-2.29 (m,1H),2.22(s,3H),2.10-2.05(m,1H),1.93-1.80(m,3H),1.37-1.25(m,2H).

Example 452

N-(3-(5-chloro-2-(difluoromethoxy)phenyl)-1-((1-(piperidin-4-yl)-1H-1,2,3-triazol-4-yl)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

Sodium azide (246 mg, 2.79 mmol) was added to a solution of tert-butyl 4-bromopiperidine-1-carboxylate (1.00 g, 3.79 mmol) and sodium iodide (113 mg, 0.753 mmol) in N,N-dimethylformamide (20 mL) at room temperature. The resulting solution was stirred at 60°C overnight and cooled to room temperature. Note: The reaction should be performed behind a blast shield. The reaction was then quenched by adding 10 mL of saturated aqueous sodium bicarbonate. The resulting solution was extracted with 2 x 50 mL of ethyl acetate, and the organic layers were combined and concentrated under vacuum. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1/4). This yielded 800 mg (93%) of tert-butyl 4-azidopiperidine-1-carboxylate as a colorless oil. TLC: R _f = 0.6; ethyl acetate/petroleum ether = 1/4.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (2 g, 4.941 mmol) in N,N-dimethylformamide (50 mL) was added Cs ₂ CO ₃ (3.23 g, 9.913 mmol) and 3-chloroprop-1-yne (720 mg, 9.663 mmol). The resulting mixture was stirred at 50° C. for 5 h and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with ethyl acetate. This gave 1.74 g (80%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(prop-2-yn-1-yl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 23) [M+H] ⁺ = 443.0, _RT = 1.45 min.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-(prop-2-yn-1-yl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (1.68 g, 3.794 mmol) in N,N-dimethylformamide (40 mL) was added DIEA (980.6 mg, 7.587 mmol), CuI (143.7 mg, 0.755 mmol), and tert-butyl 4-azidopiperidine-1-carboxylate (858.8 mg, 3.795 mmol). The resulting solution was stirred at room temperature for 7 h and concentrated under vacuum. The residue was purified by flash silica gel chromatography, eluting with ethyl acetate. This gave 1.98 g (78%) of tert-butyl 4-[4-([3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidin-3-amido]-1H-pyrazol-1-yl]methyl)-1H-1,2,3-triazol-1-yl]piperidine-1-carboxylate as a light yellow solid. LCMS (Method 24) [M+H] ⁺ =669.2, _RT =1.34 min.

To a solution of tert-butyl 4-[4-([3-[5-chloro-2-(difluoromethoxy)phenyl]-4-[pyrazolo[1,5-a]pyrimidine-3-amido]-1H-pyrazol-1-yl]methyl)-1H-1,2,3-triazol-1-yl]piperidine-1-carboxylate (1.98 g, 2.96 mmol) in methanol (30 mL) was added concentrated aqueous HCl (15 mL). The resulting solution was stirred at room temperature for 5 h. The resulting mixture was concentrated under vacuum. This gave 1.85 g of crude product, which was sufficient for the next step without further purification. A small portion was purified for characterization and biological submission. The crude product (100 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, silica gel; mobile phase, ACN/ _H2O (10 _{mmol NH4HCO3} ) = 18% increasing to ACN/ _H2O (10 mmol _NH4HCO3 ) = 49% over ₉ min; detector, UV 254 nm. 42.1 mg of product was obtained. This gave 42.0 mg of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[[1-(piperidin-4-yl)-1H-1,2,3-triazol-4-yl]methyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 36) [M+H] ⁺ =569.2, R _T =2.59min. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: (ppm) 9.69 (s, 1H), 9.34 (d, 1H, J = 6.4Hz), 8.68-8.64 (m, 2H), 8.40 (s, 1H), 8.22(s,1H),7.67-7.61(m,2H),7.45(d,1H,J＝8.4Hz),7.28(dd,1H,J＝4.2,7.0 Hz),7.06(t,1H,J＝73.4Hz),5.49(s,2H),4.58-4.51(m,1H),3.05-3.02(m,2H), 2.62-2.57(m,1H),2.30-2.25(br,1H),2.01-1.98(m,2H),1.87-1.73(m,2H).

Example 457

N-(3-(5-chloro-2-(difluoromethoxy)phenyl)-1-((1-(1-(tetrahydro-2H-pyran-4-yl)piperidin-4-yl)-1H-1,2,3-triazol-4-yl)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[[1-(piperidin-4-yl)-1H-1,2,3-triazol-4-yl]methyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide hydrochloride (200 mg, 0.330 mmol) in dichloromethane (30 mL) was added oxan-4-one (264.9 mg, 2.646 mmol). The resulting solution was stirred at room temperature overnight. NaBH(OAc) ₃ (280.8 mg, 1.325 mmol) was then added. The resulting solution was stirred at room temperature for 6 h and concentrated under vacuum. The residue was applied to a silica gel column and eluted with DCM/MeOH (80:20). The crude product (150 mg) was purified by Flash-Prep-HPLC using the following conditions (IntelFlash-1): column, silica gel; mobile phase, ACN/ _H2O (10 mmol _NH4HCO3 ) = 20% increasing to ACN/ _H2O (10 _{mmol NH4HCO3} ) = 49% over ₈ min; detector, UV 254 nm. 56.9 mg of product was obtained. This gave 56.9 mg (26%) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-([1-[1-(oxan-4-yl)piperidin-4-yl]-1H-1,2,3-triazol-4-yl]methyl)-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a light yellow solid. LCMS (Method 20) [M+H] ⁺ =653.3, R _T =1.74min. ¹ H NMR (400MHz, DMSO-d ₆ )δ: (ppm) 9.73 (s, 1H), 9.34 (dd, 1H, J = 1.6, 7.2Hz), 8.67-8.66(m,2H),8.39(s,1H),8.26(s,1H),7.65-7.62(m,2H),7.44(d,1H,J＝8.4 Hz),7.28(dd,1H,J＝4.0,6.8Hz),7.06(t,1H,J＝73.4Hz),5.48(s,2H),4.58-4.45 (m,1H),3.90-3.87(m,2H),3.32-3.25(m,2H),3.04-2.98(m,2H),2.51-2.33(m,1H), 2.30-2.24(m,2H),2.09-2.06(m,2H),1.99-1.91(m,2H),1.70-1.67((m,2H), 1.49-1.40(m,2H).

Example 468

N-(3-(5-chloro-2-(difluoromethoxy)phenyl)-1-(2-(4-(((1-cyanocyclopropyl)methyl)amino)piperidin-1-yl)-2-oxoethyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

Under nitrogen, triethylamine (11.2 mL, 80.5 mmol) was added to a solution of 1,4-dioxa-8-azaspiro[4,5]decane (9.80 g, 67.1 mmol) in dichloromethane (150 mL). The mixture was then cooled to 0°C, and a solution of 2-chloroacetyl chloride (6.53 mL, 80.5 mmol) in dichloromethane (50 mL) was added dropwise with stirring over 1.5 hours. The mixture was then stirred overnight while warming to room temperature. The reaction was then quenched by the addition of 1N HCl (100 mL). The mixture was then extracted with ethyl acetate, and the ethyl acetate layer was dried over solid anhydrous magnesium sulfate powder, filtered, and concentrated to give 2-chloro-1-(1,4-dioxa-8-azaspiro[4,5]dec-8-yl)ethan-1-one as a brown oil (15.44 g). This was used in the next step without further purification.

To N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1H-pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (4.65 g, 11.5 mmol) and cesium carbonate (10.08 g, 30.94 mmol) was added N,N-dimethylformamide (40 mL) and stirred at room temperature for 5 minutes. A solution of 2-chloro-1-(1,4-dioxa-8-azaspiro[4,5]dec-8-yl)ethan-1-one (5.34 g, 20.2 mmol) in N,N-dimethylformamide (4.5 mL) was added and stirred at room temperature for 3.5 hours. LC/MS (Method 31) indicated the reaction was complete. The reaction was then filtered and the filtrate concentrated under reduced pressure to yield a red oil. The product was then dissolved in 30 mL of dichloromethane and purified by ISCO using a silica gel column eluted with 0-100% ethyl acetate in heptane over 5 minutes, followed by 0-15% methanol in dichloromethane, to afford 5.16 g (8.78 mmol) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(1,4-dioxa-8-azaspiro[4,5]dec-8-yl)-2-oxo-ethyl]pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a yellow foamy solid. LC/MS (Method 39) [M+H] ⁺ = 589.2, _RT = 1.91 min.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-(1,4-dioxa-8-azaspiro[4,5]dec-8-yl)-2-oxo-ethyl]pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (1.01 g, 1.72 mmol) in acetone (20 mL) was added p-toluenesulfonic acid (359 mg, 2.06 mmol) and refluxed for 6 hours. LC/MS (Method 31) showed the product with a small amount of starting material present. The reaction was cooled to room temperature and quenched with water. It was then extracted with ethyl acetate, dried over solid magnesium sulfate powder and filtered to give a red oil. This was then purified by ISCO using a silica gel column and eluting with 0-15% methanol in dichloromethane to afford 0.43 g (0.78 mmol) of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(4-oxo-1-piperidinyl)ethyl]pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide as a foamy, off-white solid. LC/MS (Method 39) [M+H] ⁺ = 544.1, _RT = 1.91 min.

To a solution of N-[3-[5-chloro-2-(difluoromethoxy)phenyl]-1-[2-oxo-2-(4-oxo-1-piperidinyl)ethyl]pyrazol-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (201.7 mg 0.38 mmol) in dichloromethane (1.0 mL) was added acetic acid (0.3 mL, 5.24 mmol). 1-(aminomethyl)cyclopropanecarbonitrile (47.2 mg, 0.466 mmol) was then added. The mixture was stirred at room temperature for 5 minutes. The reaction was then cooled to 0°C, and sodium triacetoxyborohydride (124.1 mg, 0.56 mmol) was added and stirred for 3 hours while the temperature rose to room temperature. LC/MS (Method 31) showed the product as the main peak. The reaction was quenched by adding 2 mL of water, followed by 10 mL of saturated sodium bicarbonate solution. This was extracted with dichloromethane, and the organic layer was dried over solid anhydrous magnesium sulfate powder, filtered, and concentrated to yield a white solid. 20.6 mg was then purified by reverse phase HPLC to yield 10.2 mg of N-(3-(5-chloro-2-(difluoromethoxy)phenyl)-1-(2-(4-(((1-cyanocyclopropyl)methyl)amino)piperidin-1-yl)-2-oxoethyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide. LC/MS (Method 38) [M+H] ⁺ = 624.2, R _T = 2.80min.1H NMR (400MHz, DMSO-d6) 9.75 (s, 1H), 9.34 (dd, J = 7.0, 1.7Hz, 1H), 8.68 (dd, J = 4.3, 1.6Hz, 1H), 8.67(s,1H),8.30(s,1H),7.63(dd,J＝8.8,2.7Hz,1H),7.56(d,J＝2.7Hz,1H),7.46 (d,J＝8.8Hz,1H),7.29(dd,J＝7.0,4.2Hz,1H),7.25(t,J＝73.0Hz,1H),5.23(d,J ＝4.7Hz,2H),4.12(d,J＝12.7Hz,1H),3.84(d,J＝13.7Hz,1H),3.14(t,J＝11.9 Hz,1H),2.85(t,J＝11.9Hz,1H),2.71-2.65(m,3H),2.02(s,1H),1.83(t,J＝16.2 Hz,2H),1.33 1.12(m,4H),0.94(m,2H).

Enzyme assay

The JAK enzyme assay was performed as follows :

The activity of isolated recombinant JAK1 and JAK2 kinase domains was determined using Caliper technology (Caliper Life Sciences, Hopkinton, MA) by monitoring phosphorylation of a JAK3-derived peptide (Val-Ala-Leu-Val-Asp-Gly-Tyr-Phe-Arg-Leu-Thr-Thr, fluorescently labeled at the N-terminus with 5-carboxyfluorescein). To determine the inhibition constant ( _Ki ), compounds were serially diluted in DMSO and added to a 50 μL kinase reaction containing purified enzyme (1.5 nM JAK1 or 0.2 nM JAK2), 100 mM HEPES buffer (pH 7.2), 0.015% Brij-35, 1.5 μM peptide substrate, ATP (25 μM), ₁₀ mM MgCl2, and 4 mM DTT, with a final DMSO concentration of 2%. The reaction was incubated at 22°C for 30 minutes in a 384-well polypropylene microtiter plate and then terminated by adding 25 μL of EDTA-containing solution (100 mM HEPES buffer (pH 7.2), 0.015% Brij-35, 50 mM EDTA) to a final EDTA concentration of 50 mM. After termination of the kinase reaction, the proportion of phosphorylated product was determined as a fraction of the total peptide substrate using Caliper 3000 according to the manufacturer's instructions. K values [Ki = Ki,app / (1 + [ _ATP] / Km _,app )] were then determined using a modified Morrison tight binding model for _ATP -competitive inhibition (Morrison, JF, Biochim. Biophys. Acta. 185: 269-296 (1969); William, JW and Morrison, JF, _Meth . Enzymol., 63: 437-467 (1979)).

JAK1 pathway assays in cell lines were performed as follows :

Inhibitor potency ( _EC50 ) was determined in a cell-based assay designed to measure JAK1-dependent STAT phosphorylation. As described above, inhibition of IL-4, IL-13, and IL-9 signaling by blocking the Jak/Stat signaling pathway can alleviate asthma symptoms in preclinical lung 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).

In one assay, TF-1 human erythroleukemia cells obtained from the American Type Culture Collection (ATCC; Manassas, VA) were used to measure JAK1-dependent STAT6 phosphorylation downstream of IL-13 stimulation. Prior to use in the assay, TF-1 cells were cultured overnight in OptiMEM medium (Life Technologies, Grand Island, NY) supplemented with 0.5% charcoal/dextran-stripped fetal bovine serum (FBS), 0.1 mM non-essential amino acids (NEAA), and 1 mM sodium pyruvate, lacking GM-CSF. The assay was performed in 384-well plates at 300,000 cells per well in serum-free OptiMEM medium. In a second assay, BEAS-2B human bronchial epithelial cells obtained from ATCC were plated at 100,000 cells per well in 96-well plates the day before the experiment. The BEAS-2B assay was performed in complete growth medium (Bronchoepithelial Basal Medium plus bulletkit; Lonza; Basel, Switzerland).

The test compounds were serially diluted 1:2 in DMSO and then diluted 1:50 in culture medium just before use. The diluted compounds were added to the cells to a final DMSO concentration of 0.2% and incubated at 37°C for 30 minutes (for TF-1 assay) or 1 hour (for BEAS-2B assay). The cells were then stimulated with human recombinant cytokines at their respective _EC90 concentrations, as previously determined for each individual batch. Cells were stimulated with IL-13 (R&D Systems, Minneapolis, MN) for 15 minutes at 37°C. The TF-1 cell reaction was stopped by directly adding 10x lysis buffer (Cell Signaling Technologies, Danvers, MA), while the BEAS-2B cell incubation was stopped by removing the culture medium and adding 1x lysis buffer. The resulting samples were frozen in plates at -80°C. Compound-mediated inhibition of STAT6 phosphorylation was measured in cell lysates using MesoScale Discovery (MSD) technology (Gaithersburg, MD). _EC50 values were determined as the concentration of compound required for 50% inhibition of STAT phosphorylation relative to that measured by DMSO controls.

Table 2 provides JAK1 _Kj , JAK2 _Kj , and IL-13-pSTAT6 _IC50 information and LCMS (ESI) information for the indicated examples of the indicated tables.

Table 2:

Blank: Not measured

Clearance determination

In some cases, high systemic clearance of a drug substance may be beneficial. For example, in the treatment of pulmonary disease, it may be beneficial to have high drug concentrations in the lungs and lower concentrations in the bloodstream or peripheral organs. In this case, it may be advantageous to administer a drug with high systemic clearance by an inhalation delivery route. This approach can maximize the pharmacological activity of the drug in the lungs while minimizing the possibility of target toxicity elsewhere in the body. The functional groups attached to the 5-chloro-2-difluoromethoxyphenylpyrazolopyrimidine skeleton affect systemic clearance. For example, compound i exhibited a mouse intravenous (IV) clearance value of 5.3 mL/min/kg, or a mouse liver blood flow of 6%. Representative mouse IV clearance values for embodiments of the present invention are shown in Table 3.

Table 3. Mouse IV clearance values of compounds of the invention.

Female BALB/c mice were obtained from Charles River Labs (Hollister, CA USA). A single IV dose of 1 mg/kg compound was administered to 12 animals via the tail vein. Representative dose solutions were prepared as follows: the compound was dissolved in a mixture of 5% dimethyl sulfoxide (DMSO), 35% polyethylene glycol (PEG 400), and 60% water (v/v/v) in a volume of 5 mL/kg. At the start of the study, mice weighed approximately 20-25 g. After intravenous administration of the compound, each animal was bled twice by retroorbital and cardiac puncture. PK time points were collected at 2 minutes, 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, and 8 hours after administration. Plasma samples were stored at approximately -80°C until thawed for LC-MS/MS analysis. PK data analysis was performed on mean plasma concentration-time data, and PK parameters were determined by non-compartmental methods using Pharsight® Enterprise, version 5.2.1 (Pharsight Corporation; Mountain View, CA).

Claims (8)

1. A compound or a salt thereof, said compound being selected from: 2. The compound of claim 1 or a salt thereof, wherein the compound is selected from: 3. The compound of claim 2 or a salt thereof, wherein the compound is selected from: 4. The compound of claim 3 or a salt thereof, wherein the compound is 5. The compound of claim 3 or a salt thereof, wherein said compound is 6. The compound of claim 3 or a salt thereof, wherein said compound is 7. The compound of claim 3 or a salt thereof, wherein said compound is 8. The compound of claim 3 or a salt thereof, wherein said compound is