Treatment of b-cell malignancies with combined JAK and PI3k inhibitors

Abstract

Methods of treating B-cell malignancies using combinations of inhibitors of JAK1 and / or JAK2 and inhibitors of PI3K δ are provided.SOLUTION: (3R) - 3-cyclopentyl-3 - [4 - (7H - pynolo [2, 3-d] pyrimidin-4-yl) - 1H - pyrazol-l-yl] propanenitrile or pharmaceutically acceptable salts thereof, to a subject in need thereof; (JAK1) - 3-cyclopentyl-3 - [4 - (JAK2 - pynolo [2, 3-d] pyrimidin-4-yl) - LA - pyrazol-l-yl] propanenitrile or pharmaceutically acceptable salts thereof. 4 - {3 - [1 - (4-amino-3-methyl-pyrazolo [3, 4-d] pyrimidin-1-yl) ethyl] - 5-chloro-2-ethoxy-6-fluorophenyl} pyrrolidin-2-one or pharmaceutically acceptable salts thereof, for use in the treatment or prevention of 1H delta in a subject in need of such treatment or prevention of PI3K delta in a subject.SELECTED DRAWING: None

Description

[Technical Field]

[0001] This application claims the benefit of priority to 61 / 976,815, filed April 8, 2014, which is incorporated herein by reference in its entirety.

[0002] The present invention relates to methods of treating B-cell malignancies using a combination of an inhibitor of JAK1 and / or JAK2 and an inhibitor of PI3Kδ. [Background technology]

[0003] The B cell receptor (BCR) is present on both normal and most malignant B cells. BCR engagement provides a critical survival signal, and disruption of BCR signaling can lead to B cell death. Studies using siRNA to inhibit BCR expression have shown that constitutive signaling by the BCR is crucial for the survival and proliferation of human B cell lymphomas. The primary role of BCR signaling in these cells appears to be activation of spleen tyrosine kinase (Syk), which in turn leads to several downstream events that promote cell survival, including activation of Bruton's tyrosine kinase (BTK), phosphatidylinositol 3-kinase (PI3K), and AKT. Many B cell malignancies, including diffuse large B cell lymphoma (DLBCL), have been shown to be particularly dependent on BCR survival signals, as evidenced by their sensitivity to genetic and pharmacological inhibition of BCR signaling components in vitro. DLBCL cells engage PI3K, which enhances anti-apoptotic NF-kB signaling and survival signals, and inhibition of the PI3K / AKT pathway has been shown to cooperate with NF-kB inhibition in killing DLBCL cell lines in vitro.

[0004] Abnormal activation of JAKs through the production of cytokines and growth factors has also been associated with increased malignant cell proliferation and survival in many tumor types. JAKs activate many downstream pathways involved in malignant cell proliferation and survival, including STATs, a family of important latent transcription factors. Clinically relevant, serum IL-10 and IL-6 levels, which signal through JAKs, have been found to be elevated in patients with DLBCL compared to normal controls (Gupta et al., 2012). Furthermore, patients with elevated serum IL-10 levels have been shown to have shorter event-free survival (Gupta et al., 2012). Among the JAK family kinases, JAK1, in cooperation with JAK2, JAK3, and TYK2, has been shown to play an exclusive role in mediating the signal transduction of many inflammatory cytokines, including IL-6, IL-10, and interferons.

[0005] In DLBCL, JAK pathway activation occurs through both autocrine and paracrine mechanisms. In tumor cells, BCR signaling leads to increased IL-6 and IL-10 production through activation of the NF-kB pathway (Lam et al., 2008). A subset of DLBCL is characterized by high expression of STAT3, IL-6, and / or IL-10, and JAK inhibition has been shown to be cytotoxic in these DLBCL cell lines and to synergize with NF-kB inhibitors. In addition to JAK / STAT pathway activation through the autocrine pathway, the stromal compartment may also provide a source of these cytokines in a paracrine manner (Hodge et al., 2005).

[0006] For these reasons, there is a need to develop new therapies that can be used to treat B-cell malignancies, such as DLBCL. This invention addresses this need and others. [Brief explanation of the drawings]

[0007] [Figure 1]Figure 1A shows Western blot analysis probing for IL6 and IL10 in various DLBCL cell lines, and Figure 1B shows Western blot analysis for actin and p-Stat3 in Pfeiffer cells treated with IL6 or IL10. [Figure 2] Figure 2B shows the percent inhibition in a cell proliferation assay in Pfeiffer cells as a function of the concentration of vehicle (DMSO), DMSO + IL10, and DMSO + IL10 + ruxolitinib and Compound 28. Figure 2B shows the percent inhibition in a cell proliferation assay in Pfeiffer cells as a function of the concentration of vehicle (DMSO), DMSO + IL10, and DMSO + IL10 + Compound 7 and Compound 28. [Figure 3] Figure 1 shows the % inhibition in a cell proliferation assay in HBL-1 cells as a function of concentration for vehicle (DMSO), DMSO + IL10, and DMSO + IL10 + ruxolitinib and compound 28. [Figure 4] 1 shows a Western blot analysis of Pfeiffer cells after treatment with vehicle (DMSO), ruxolitinib, Compound 28, or Compound 28 and ruxolitinib, with or without IL10. [Figure 5] 1 shows a Western blot analysis of Pfeiffer cells after treatment with vehicle (DMSO), Compound 7, Compound 28, or Compound 28 and Compound 7, with or without IL10. [Figure 6] Figure 1 shows the % inhibition in a cell proliferation assay in Pfeiffer cells as a function of concentration for vehicle (DMSO), DMSO + IL10, and DMSO + IL10 + Compound 16 and Compound 28. [Figure 7] 1 shows Annexin-V staining of Pfeiffer cells treated with Compound 28 + / - Compound 16, indicating synergistic apoptosis induction from the combination therapy. [Figure 8] 1 shows a Western blot analysis of Pfeiffer cells after treatment with Compound 28 + / - Compound 16, showing the effects on STAT3 and pAKT. Summary of the Invention

[0008] The present application provides a method of treating a B-cell malignancy in a patient in need thereof, comprising administering to the patient: (a) an inhibitor of JAK1 and / or JAK2; and (b) an inhibitor of PI3Kδ.

[0009] The present application provides for the treatment of diffuse large B-cell lymphoma, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Hodgkin's lymphoma, Burkitt's lymphoma, Waldenstrom's macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosa-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, primary effusion lymphoma, and thyroid cancer in patients in need thereof. Further provided is a method of treating a disease selected from effusion lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammopathy of undetermined significance (MGUS), activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL), and germinal center B-cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL), comprising administering to the patient: (a) an inhibitor of JAK1 and / or JAK2; and (b) an inhibitor of PI3Kδ.

[0010] In some embodiments of the method, the inhibitor of JAK1 and / or JAK2 is: 3-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; 4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-[4-fluoro-2-(trifluoromethyl)phenyl]piperidine-1-carboxamide; [3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(1-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)azetidin-3-yl]acetonitrile; [trans-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-3-(4-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperazin-1-yl)cyclobutyl]acetonitrile; {trans-3-(4-{[4-[(3-hydroxyazetidin-1-yl)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-{[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-{[(2R)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; 4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile; 5-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide; 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide; 5-{3-(cyanomethyl)-3-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide; {1-(cis-4-{[6-(2-hydroxyethyl)-2-(trifluoromethyl)pyrimidin-4-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-[(ethylamino)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-(1-hydroxy-1-methylethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-{[(3R)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-{[(3S)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {trans-3-(4-{[4-({[(1S)-2-hydroxy-1-methylethyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-({[(2R)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-({[(2S)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-(2-hydroxyethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile; 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide; and pharmaceutically acceptable salts of any of the foregoing.

[0011] In some embodiments of the method, the inhibitor of PI3Kδ is: 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one; (S)-7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one; 4-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-{1-[(2S)-2-hydroxypropyl]azetidin-3-yl}-3-methoxybenzonitrile; 4-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-[1-(2-hydroxyethyl)azetidin-3-yl]-3-methoxybenzonitrile; 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-cyano-2-ethoxy-5-methylphenyl}-N,N-dimethylpyridine-2-carboxamide; 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one; and N-{1-[5-chloro-8-(3-fluorophenyl)cinnolin-7-yl]ethyl}-9H-purin-6-amine; 4-chloro-3'-fluoro-3-methyl-6-[1-(9H-purin-6-ylamino)ethyl]biphenyl-2-carbonitrile; and pharmaceutically acceptable salts of any of the foregoing.

[0012] The present application also provides inhibitors of JAK1 and / or JAK2 for use in combination with a PI3Kδ inhibitor to treat a B-cell malignancy or any of the diseases embodied herein.

[0013] The present application further provides the use of an inhibitor of JAK1 and / or JAK2 and a PI3Kδ inhibitor for the preparation of a medicament for the treatment of a B-cell malignancy or any of the diseases embodied herein. DETAILED DESCRIPTION OF THE INVENTION

[0014] The present application relates to the treatment of, among other things, diffuse large B-cell lymphoma, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Hodgkin's lymphoma, Burkitt's lymphoma, Waldenstrom's macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosa-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, genotype 1 lymphoma, genotype 2 lymphoma, genotype 3 lymphoma, genotype 4 lymphoma, genotype 5 lymphoma, genotype 6 lymphoma, genotype 7 lymphoma, genotype 8 lymphoma, genotype 9 lymphoma, genotype 10 lymphoma, genotype 11 lymphoma, genotype 12 lymphoma, genotype 13 lymphoma, genotype 14 lymphoma, genotype 15 lymphoma, genotype 16 lymphoma, genotype 17 lymphoma, genotype 18 lymphoma, genotype 19 lymphoma, genotype 20 lymphoma, genotype 21 lymphoma, genotype 22 lymphoma, genotype 23 lymphoma, genotype 24 lymphoma, genotype 25 lymphoma, genotype 26 lymphoma, genotype 27 lymphoma, genotype 28 lymphoma, genotype 29 lymphoma, genotype 30 lymphoma, genotype 31 lymphoma, genotype 32 lymphoma, gen and (b) an inhibitor of PI3Kδ.

[0015] In some embodiments, the non-Hodgkin's lymphoma is a non-Hodgkin's lymphoma (NHL) that is relapsed or refractory NHL or recucurrent follicular NHL.

[0016] In some embodiments, the disease is diffuse large B-cell lymphoma (DLBCL).

[0017] In some embodiments, the disease is activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL) or germinal center B-cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL).

[0018] In some embodiments, the inhibitor of JAK1 and / or JAK2 and the inhibitor of PI3Kδ are administered simultaneously.

[0019] In some embodiments, the inhibitor of JAK1 and / or JAK2 and the inhibitor of PI3Kδ are administered sequentially.

[0020] In some embodiments, inhibitors of JAK1 and / or JAK2 are selective for JAK1 and JAK1 over JAK3 and TYK2. In some embodiments, inhibitors of JAK1 and / or JAK2 are selective for JAK1 over JAK2, JAK3, and TYK2. For example, some compounds described herein, or pharmaceutically acceptable salts thereof, preferentially inhibit JAK1 over one or more of JAK2, JAK3, and TYK2. In some embodiments, the compounds preferentially inhibit JAK1 over JAK2 (e.g., IC of JAK1 / JAK2). 50 In some embodiments, the compound or salt is about 10-fold selective for JAK1 over JAK2. In some embodiments, the compound or salt is about 3-fold, about 5-fold, about 10-fold, about 15-fold, or about 20-fold selective for JAK1 over JAK2, which is an IC50 ratio greater than 1 at 1 mM ATP. 50 (See, for example, Example A).

[0021] In some embodiments, the inhibitor of JAK1 and / or JAK2 is 3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile. In some embodiments, the inhibitor of JAK1 and / or JAK2 is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile (also known as ruxolitinib; INCB018424). Ruxolitinib has an IC of less than 10 nM at 1 mM ATP (Assay A) for JAK1 and JAK2. 503-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile and ruxolitinib can be made by the methods described in U.S. Pat. No. 7,598,257, filed December 12, 2006 (Example 67), which is incorporated herein by reference in its entirety. In some embodiments, the inhibitor of JAK1 and / or JAK2 is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile phosphate.

[0022] In some embodiments, the inhibitor of JAK1 and / or JAK2 is a compound in Table 1, or a pharmaceutically acceptable salt thereof. The compounds in Table 1 are selective JAK1 inhibitors (more selective than JAK2, JAK3, and TYK2). The IC obtained by the method of Assay A at 1 mM ATP was 50 is shown in Table 1. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] [Table 1-13] + means less than 10 nM (see Example A for assay conditions) ++ means 100 nM or less (see Example A for assay conditions) +++ means 300 nM or less (see Example A for assay conditions) a Data for Enantiomer 1 b Data for Enantiomer 2

[0023] In some embodiments, the inhibitor of JAK1 and / or JAK2 is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharmaceutically acceptable salt thereof.

[0024] In some embodiments, the inhibitor of JAK1 and / or JAK2 is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate.

[0025] In some embodiments, the inhibitor of JAK1 and / or JAK2 is 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide, or a pharmaceutically acceptable salt thereof.

[0026] In some embodiments, the inhibitor of JAK1 and / or JAK2 is (R)-3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, (R)-3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, (R)-4-[(4-{3-cyano-2-[4 -(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile, (R)-4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile, or (R)-4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3 -d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile, (S)-3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, (S)-3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, (S)-4-[(4-{3-cyano- 2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile, (S)-4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile, (S)-4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile; and pharmaceutically acceptable salts of any of the foregoing.

[0027] In some embodiments, the compounds in Table 1 are selected from the group consisting of U.S. Patent Publication Nos. 2010 / 0298334, filed May 21, 2010, 2011 / 0059951, filed August 31, 2010, 2011 / 0224190, filed March 9, 2011, and 2011 / 018 / 18, each of which is incorporated by reference in its entirety. and U.S. Patent Publication Nos. 2012 / 0149681, filed November 18, 2011, 2012 / 0149682, filed June 19, 2012, 2013 / 0018034, filed August 17, 2012, 2013 / 0045963, filed August 17, 2012, and 2014 / 0005166, filed May 17, 2013.

[0028] In some embodiments, the inhibitors of JAK1 and / or JAK2 are disclosed in U.S. Patent Publication Nos. 2010 / 0298334, filed May 21, 2010, 2011 / 0059951, filed August 31, 2010, 2011 / 0224190, filed March 9, 2011, and 2011 / 0224190, filed November 18, 2011, each of which is incorporated herein by reference in its entirety. No. 2012 / 0149681 filed Nov. 18, 2011, U.S. Patent Publication No. 2012 / 0149682 filed Jun. 19, 2012, U.S. Patent Publication No. 2013 / 0018034 filed Jun. 19, 2012, U.S. Patent Publication No. 2013 / 0045963 filed Aug. 17, 2012, and U.S. Patent Publication No. 2014 / 0005166 filed May 17, 2013.

[0029] Inhibitors of PI3Kδ described herein can be selective. "Selective" means that the compound binds to or inhibits a kinase with greater affinity or potency, respectively, compared to at least one other kinase. In some embodiments, the compounds described herein are selective inhibitors of PI3Kδ (e.g., over PI3Kα, PI3Kβ, and PI3Kγ). In some embodiments, the selectivity can be at least about 2-fold, 5-fold, 10-fold, at least about 20-fold, at least about 50-fold, at least about 100-fold, at least about 200-fold, at least about 500-fold, or at least about 1000-fold. Selectivity can be measured by methods routine in the art. In some embodiments, selectivity is measured by measuring the K m ATP concentration can be tested. In some embodiments, the selectivity of the compounds described herein can be determined by cellular assays associated with specific PI3K kinase activity.

[0030] In some embodiments, the inhibitor of PI3Kδ is a compound shown in Table 2. The compounds in Table 2 have been tested in Assay B and have an IC of Table 2 50 It has been shown to be an inhibitor of PI3Kδ with [Table 2-1] [Table 2-2] [Table 2-3] + means less than 50 nM ++ means 50nM to 200nM +++ means 50nM to 100nM

[0031] In some embodiments, the inhibitor of PI3Kδ is: (S)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (S)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (R)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; N-{(1S)-1-[5-chloro-8-(3-fluorophenyl)cinnolin-7-yl]ethyl}-9H-purin-6-amine; and pharmaceutically acceptable salts of any of the foregoing.

[0032] In some embodiments, the inhibitor of PI3Kδ is (S)-7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

[0033] In some embodiments, the inhibitor of PI3Kδ is 4-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-{1-[(2S)-2-hydroxypropyl]azetidin-3-yl}-3-methoxybenzonitrile, or a pharmaceutically acceptable salt thereof.

[0034] In some embodiments, the inhibitor of PI3Kδ is 4-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-[1-(2-hydroxyethyl)azetidin-3-yl]-3-methoxybenzonitrile, or a pharmaceutically acceptable salt thereof.

[0035] In some embodiments, the inhibitor of PI3Kδ is 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-cyano-2-ethoxy-5-methylphenyl}-N,N-dimethylpyridine-2-carboxamide, or a pharmaceutically acceptable salt thereof.

[0036] In some embodiments, the inhibitor of PI3Kδ is: 4-[(R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-{1-[(2S)-2-hydroxypropyl]azetidin-3-yl}-3-methoxybenzonitrile; 4-[1(R)-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-[1-(2-hydroxyethyl)azetidin-3-yl]-3-methoxybenzonitrile; 5-{3-[1(R)-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-cyano-2-ethoxy-5-methylphenyl}-N,N-dimethylpyridine-2-carboxamide; 4-[(S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-{1-[(2S)-2-hydroxypropyl]azetidin-3-yl}-3-methoxybenzonitrile; 4-[1(S)-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-[1-(2-hydroxyethyl)azetidin-3-yl]-3-methoxybenzonitrile; 5-{3-[1(S)-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-cyano-2-ethoxy-5-methylphenyl}-N,N-dimethylpyridine-2-carboxamide; and pharmaceutically acceptable salts of any of the foregoing.

[0037] In some embodiments, the inhibitor of PI3Kδ is a compound of U.S. Patent Publication No. US2011 / 0015212, filed June 28, 2010, U.S. Patent Publication No. 2013 / 0059835, filed August 31, 2013, U.S. Patent Publication No. 2011 / 0183985, filed December 17, 2010, or U.S. Patent Publication No. 2012 / 0157430, filed December 19, 2011, each of which is incorporated by reference herein in its entirety.

[0038] In some embodiments, the compounds in Table 2 are prepared by the method of U.S. Patent Publication No. US2011 / 0015212, filed June 28, 2010, U.S. Patent Publication No. 2013 / 0059835, filed August 31, 2013, U.S. Patent Publication No. 2011 / 0183985, filed December 17, 2010, or U.S. Patent Publication No. 2012 / 0157430, filed December 19, 2011, each of which is incorporated by reference herein in its entirety.

[0039] In some embodiments, the inhibitor of JAK1 and / or JAK2 is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, or a pharmaceutically acceptable salt thereof; and (7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

[0040] In some embodiments, the inhibitor of JAK1 and / or JAK2 is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharmaceutically acceptable salt thereof; and the inhibitor of PI3Kδ is 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

[0041] In some embodiments, the inhibitor of JAK1 and / or JAK2 is 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide, or a pharmaceutically acceptable salt thereof; and the inhibitor of PI3Kδ is 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

[0042] In some embodiments, the present application provides a method of treating diffuse large B-cell lymphoma in a patient in need thereof, comprising administering to the patient (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, or a pharmaceutically acceptable salt thereof; and the inhibitor of PI3Kδ is (7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

[0043] In some embodiments, the present application provides a method of treating diffuse large B-cell lymphoma in a patient in need thereof, comprising administering to the patient {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharmaceutically acceptable salt thereof; and 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

[0044] In some embodiments, the present application provides a method of treating diffuse large B-cell lymphoma in a patient in need thereof, comprising administering to the patient 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide, or a pharmaceutically acceptable salt thereof; and 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

[0045] The compounds described herein can be asymmetric (e.g., having one or more stereocenters). Unless otherwise specified, all stereoisomers, such as enantiomers and diastereomers, are intended. Compounds containing asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically inactive starting materials are known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like, can also be present in the compounds described herein, and all such stable isomers are contemplated within the present invention. Cis and trans geometric isomers of the compounds of the present invention are described and may be isolated as a mixture of isomers or as separate isomers.

[0046] In some embodiments, the compound has the (R) configuration. In some embodiments, the compound has the (S) configuration.

[0047] Resolution of racemic mixtures of compounds can be carried out by any of a number of methods known in the art. Exemplary methods include fractional recrystallization using chiral resolving acids, which are optically active, salt-forming organic acids. Suitable resolving agents for fractional recrystallization are optically active acids, such as the D- and L-forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, or various optically active camphorsulfonic acids, such as β-camphorsulfonic acid. Other resolving agents suitable for fractional crystallization include stereoisomerically pure forms (e.g., S- and R-forms, or diastereomerically pure forms) of α-methylbenzylamine, 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, and the like.

[0048] Resolution of racemic mixtures can also be carried out by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). Suitable elution solvent compositions can be determined by those skilled in the art.

[0049] The compounds described herein also include tautomeric forms. Tautomeric forms result from the interchange of adjacent double and single bonds with the concomitant migration of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs, and rings in which protons can occupy more than one position in the heterocyclic ring system, such as 1H- and 3H-imidazole, 1H-, 2H-, and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomeric forms can be sterically locked into one form by equilibrium or appropriate substitution.

[0050] The compounds described herein may also include all isotopes of atoms occurring in the intermediates or final compounds. Isotopes include atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium.

[0051] As used herein, the term "compound" is intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the depicted structure. A compound identified herein by name or structure as one particular tautomeric form is intended to include other tautomeric forms unless otherwise specified.

[0052] All compounds, and pharmaceutically acceptable salts thereof, may be found together with other substances such as water and solvents (eg, hydrates and solvates) or may be isolated.

[0053] In some embodiments, the compounds described herein, or salts thereof, are substantially isolated. "Substantially isolated" means that the compound is at least partially or substantially separated from the environment in which it was formed or detected. Partial isolation can include, for example, a composition enriched in the compounds described herein. Substantial isolation can include a composition containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of the compounds described herein, or salts thereof. Methods for isolating compounds and their salts are routine in the art.

[0054] The phrase "pharmaceutically acceptable" is employed herein to refer to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit-risk ratio.

[0055] As used herein, the expressions "ambient temperature" and "room temperature" or "rt" are art-recognized and generally refer to a temperature, e.g., a reaction temperature, relative to the temperature of the room in which the reaction is carried out, e.g., a temperature of about 20°C to about 30°C.

[0056] The present invention also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the compounds of the present disclosure, in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts of the present invention include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, alcohols (e.g., methanol, ethanol, isopropanol, or butanol), or acetonitrile (ACN) are preferred. Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

[0057] As used herein, the terms "individual" or "patient" are used interchangeably and refer to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, or primate, and most preferably a human.

[0058] In some embodiments, the inhibitor is administered in a therapeutically effective amount. As used herein, the phrase "therapeutically effective amount" refers to an amount of an active compound or pharmaceutical agent that elicits the biological or medicinal response desired in a tissue, system, animal, individual, or human by a researcher, veterinarian, physician, or other clinician. In some embodiments, the dosage of the compound, or a pharmaceutically acceptable salt thereof, administered to a patient or individual is about 1 mg to about 2 g, about 1 mg to about 1000 mg, about 1 mg to about 500 mg, about 1 mg to about 200 mg, about 1 mg to about 100 mg, about 1 mg to 50 mg, or about 50 mg to about 500 mg.

[0059] As used herein, the term "treat" or "treatment" refers to one or more of: (1) inhibiting the disease; e.g., inhibiting the disease, condition, or disorder (i.e., halting further development of the pathology and / or symptomology) in an individual experiencing or exhibiting the pathology or symptomology of the disease, condition, or disorder; and (2) ameliorating the disease; e.g., ameliorating the disease, condition, or disorder (i.e., ameliorating the pathology and / or symptomology), e.g., reducing the severity of the disease, in an individual experiencing or exhibiting the pathology or symptomology of the disease, condition, or disorder.

[0060] Combination therapy For example, one or more additional pharmaceutical agents, such as chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressants, and other agents, such as Bcr-Abl, Flt-3, EGFR, HER2, c-MET, VEGFR, PDGFR, cKit, IGF-1R, RAF, FAK, AktmTOR, PIM, and AKT (e.g., AKT1, AKT2, or AKT3) kinase inhibitors, or therapeutic antibodies, such as those described in WO2006 / 056399, can be used in combination with the compounds of the present invention to treat PI3K-related diseases, disorders, or conditions. The one or more additional pharmaceutical agents can be administered to a patient simultaneously or sequentially.

[0061] Examples of antibodies for use in combination therapy include, but are not limited to, trastuzumab (e.g., anti-HER2), ranibizumab (e.g., anti-VEGF-A), bevacizumab (trade name Avastin, e.g., anti-VEGF), panitumumab (e.g., anti-EGFR), cetuximab (e.g., anti-EGFR), Rituxan (anti-CD20), and antibodies directed against c-MET.

[0062] One or more of the following agents may be used in combination with the compounds of the invention, and are presented as a non-limiting list: cytostatic agents, cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan, camptostar, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen, 5-fluorouracil, methoxtrexate, temozolomide, cyclophosphamide, SCH 66336, R115777, L778, 123, BMS 214662, Iressa, Tarceva, antibodies against EGFR, Gleevec™, Intron, ara-C, adriamycin, cytoxan, gemcitabine, uracil mustard, chlormethine, ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-methyl Lecaptopurine, 6-thioguanine, fludarabine phosphate, oxaliplatin, leucovirin, ELOXATIN™, pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin C, L-asparaginase, teniposide 17 alpha.Ethinyl estradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone, megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide, flutamide, toremifene, goserelin, cisplatin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, navelbene, anastrazole, letrazole, capecitabine, reloxafine, droloxafine, hexafluorophosphate, Samethylmelamine, Avastin, Herceptin, Vexar, Velcade, Zevalin, Trisenox, Xeloda, Vinorelbine, Porfimer, Erbitux, Liposomal, Thiotepa, Altretamine, Melphalan, Trastuzumab, Lerozole, Fulvestrant, Exemestane, Fulvestrant, Ifosfomide, Rituximab, C225 , campath, clofarabine, cladribine, aphidicolon, rituxan, sunitinib, dasatinib, tezacitabine, Sml1, fludarabine, pentostatin, triaprine, didox, trimidox, amidox, 3-AP, MDL-101, 731, bendamustine (Treanda), ofatumumab, or GS-1101 (also known as CAL-101).

[0063] Examples of chemotherapeutic agents include proteosome inhibitors (eg, bortezomib), thalidomide, revlimid, and DNA damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, carmustine, and the like.

[0064] Examples of steroids include corticosteroids such as dexamethasone or prednisone.

[0065] Examples of Bcr-Abl inhibitors include compounds of the genera and species described in US Pat. No. 5,521,184, WO 04 / 005281, and US Provisional Patent Application No. 60 / 578,491, and pharmaceutically acceptable salts thereof.

[0066] Examples of suitable Flt-3 inhibitors include compounds disclosed in WO03 / 037347, WO03 / 099771, and WO04 / 046120, and pharmaceutically acceptable salts thereof.

[0067] Examples of suitable RAF inhibitors include the compounds disclosed in WO00 / 09495 and WO05 / 028444, and pharmaceutically acceptable salts thereof.

[0068] Examples of suitable FAK inhibitors include compounds disclosed in WO04 / 080980, WO04 / 056786, WO03 / 024967, WO01 / 064655, WO00 / 053595, and WO01 / 014402, and pharmaceutically acceptable salts thereof.

[0069] Examples of suitable mTOR inhibitors include the compounds disclosed in WO2011 / 025889, and pharmaceutically acceptable salts thereof.

[0070] In some embodiments, the compounds of the present invention can be used in combination with one or more other kinase inhibitors, including imatinib, particularly for treating patients who are resistant to imatinib or other kinase inhibitors.

[0071] In some embodiments, the compounds of the present invention can be used in combination with chemotherapeutic agents in the treatment of cancers such as multiple myeloma, potentially improving treatment response compared to the response to the chemotherapeutic agent alone, without exacerbating its toxic effects. Examples of additional pharmaceutical agents used in the treatment of multiple myeloma include, but are not limited to, melphalan, melphalan plus prednisone [MP], doxorubicin, dexamethasone, and Velcade (bortezomib). Further additional agents used in the treatment of multiple myeloma include Bcr-Ab1, Flt-3, RAF, and FAK kinase inhibitors. Additive or synergistic effects are desirable results of combining the PI3K inhibitors of the present invention with additional agents. Furthermore, resistance of multiple myeloma cells to agents such as dexamethasone can be reversible upon treatment with the PI3K inhibitors of the present invention. The agent can be used in combination with the compound in a single or sequential dosage form, or the agent can be administered simultaneously or sequentially in separate dosage forms.

[0072] In some embodiments, a corticosteroid, such as dexamethasone, is administered to a patient in combination with a compound of the invention when the dexamethasone is administered intermittently as opposed to continuously.

[0073] In some further embodiments, the combination of the compounds of the invention and other therapeutic agents may be administered to the patient before, during, and / or after bone marrow or stem cell transplantation.

[0074] Pharmaceutical Preparations and Dosage Forms When employed as pharmaceuticals, the compounds described herein can be administered in the form of pharmaceutical compositions. These compositions can be prepared in a manner well known in the pharmaceutical arts and can be administered by a variety of routes, depending on whether local or systemic treatment is desired and the area to be treated. Administration can be topical (including transdermal, epithelial, ocular, and mucosal, including intranasal, intravaginal, and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or sprays, including by nebulizer; intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular, or injection or infusion; or intracranial, e.g., intrathecal or intraventricular, administration. Parenteral administration can be in the form of a single bolus dose or can be, for example, by a continuous infusion pump. Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners, and the like may be necessary or desirable. The present invention also includes pharmaceutical compositions containing the compounds described herein or pharmaceutically acceptable salts thereof as active ingredients in combination with one or more pharmaceutically acceptable carriers (excipients). In some embodiments, the compositions are suitable for topical administration. When preparing compositions of the present invention, the active ingredient is typically mixed with an excipient, diluted by the excipient, or enclosed within such a carrier, for example, in the form of a capsule, sachet, paper, or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid substance that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions can take the form of tablets, pills, powders, troches, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, sprays (in a solid or liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile water for injection, and powders in sterile containers.

[0075] When preparing formulations, active compound can be milled to obtain suitable particle size before being mixed with other components.If active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh.If active compound is substantially water-soluble, particle size can be adjusted by milling to obtain substantially uniform distribution in formulation, for example, about 40 mesh.

[0076] The compounds described herein can be milled using known milling methods, such as wet milling, to obtain suitable particle sizes for tablet formulations and other formulation types.Finely divided (nanoparticulate) preparations of the compounds described herein can be prepared by methods known in the art, see, for example, International Patent Application No. WO2002 / 000196.

[0077] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulation may additionally contain: lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl benzoate and propylhydroxybenzoate; sweeteners; and flavoring agents. The compositions of the present invention can be formulated to provide quick, sustained, or delayed release of the active ingredient after administration to a patient using methods known in the art.

[0078] The compositions can be formulated in a unit dosage form, each dosage containing from about 5 to about 1000 mg (1 g), more usually about 100 to about 500 mg, of the active ingredient. The term "unit dosage form" refers to physically discrete units suitable for use in human subjects and other mammals as unit dosages, each unit containing a predetermined quantity of active material calculated to produce a desired therapeutic effect, in association with a suitable pharmaceutical excipient.

[0079] In some embodiments, the compositions of the present invention contain about 5 to about 50 mg of the active ingredient. One of skill in the art will recognize that this specifically refers to compositions containing about 5 to about 10, about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50 mg of the active ingredient.

[0080] In some embodiments, the compositions of the present invention contain about 50 to about 500 mg of the active ingredient. One of skill in the art will recognize that this specifically refers to compositions containing about 50 to about 100, about 100 to about 150, about 150 to about 200, about 200 to about 250, about 250 to about 300, about 350 to about 400, or about 450 to about 500 mg of the active ingredient.

[0081] In some embodiments, the compositions of the present invention comprise about 500 to about 1000 mg of the active ingredient. One of skill in the art will recognize that this specifically refers to compositions containing about 500 to about 550, about 550 to about 600, about 600 to about 650, about 650 to about 700, about 700 to about 750, about 750 to about 800, about 800 to about 850, about 850 to about 900, about 900 to about 950, or about 950 to about 1000 mg of the active ingredient.

[0082] Similar dosages may be used for the compounds described herein in the methods and uses of the present invention.

[0083] The active compounds can be effective over a wide dosage range and are generally administered in a pharmaceutically effective amount. However, it will be understood that the amount of compound actually administered will usually be determined by the physician according to the relevant circumstances, including the condition to be treated, the selected route of administration, the actual compound administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, etc.

[0084] To prepare solid compositions such as tablets, the primary active ingredient is mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogeneous mixture of the compound of the present invention. When these preformulation compositions are referred to as homogeneous, the active ingredient is typically dispersed evenly throughout the composition so that the composition can be easily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules. This solid preformulation is then subdivided into the type of unit dosage form described above, containing, for example, about 0.1 to about 1000 mg of the active ingredient of the present invention.

[0085] The tablets or pills of the present invention can be coated or otherwise compounded to provide a dosage form with the advantage of prolonged action. For example, the tablets or pills can comprise an inner medicinal and an outer medicinal component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, including a number of polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0086] Liquid forms into which the compounds and compositions of the present invention may be incorporated for oral or injectable administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0087] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the compositions are administered by oral or nasal respiratory routes for local or systemic effect. The compositions can be nebulized using an inert gas. Nebulized solutions can be inhaled directly from the nebulizing device, or the nebulizing device can be attached to a face mask, tent, or intermittent positive pressure respirator. Solution, suspension, or powder compositions can be administered orally or intranasally from a device that delivers the formulation in an appropriate manner.

[0088] Topical formulations may contain one or more conventional carriers. In some embodiments, ointments may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, etc. Cream carrier compositions may be based on water in combination with glycerol and one or more other ingredients, such as glycerol monostearate, PEG-glycerol monostearate, and cetylstearyl alcohol. Gels may be formulated using isopropyl alcohol and water, suitably combined with other ingredients such as glycerol and hydroxyethylcellulose. In some embodiments, topical formulations contain at least about 0.1, at least about 0.25, at least about 0.5, at least about 1, at least about 2, or at least about 5 wt.% of a compound described herein. Topical formulations may be suitably packaged, for example, in 100 g tubes, optionally accompanied by instructions for use for the treatment of a selected indication, such as psoriasis or other skin conditions.

[0089] The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of the administration, such as prophylaxis or therapy, the condition of the patient, the mode of administration, etc. In therapeutic applications, compositions may be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose will depend on the disease state being treated and on the judgment of the attending clinician, which will depend on factors such as the severity of the disease, the age, weight, and general condition of the patient.

[0090] The compositions administered to patients can be in the form of pharmaceutical compositions described above. These compositions can be sterilized by conventional sterilization techniques or sterile filtered. Aqueous solutions can be packaged for immediate use or lyophilized, with the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the compound preparations will typically be between 3 and 11, more preferably 5-9, and most preferably 7-8. It will be understood that the use of certain of the aforementioned excipients, carriers, or stabilizers may result in the formation of pharmaceutical salts.

[0091] Therapeutic dosages of the compounds of the invention can vary according to, for example, the particular use for which the treatment is made, the mode of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of a compound described herein in a pharmaceutical composition can vary depending on many factors, including dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, for parenteral administration, the compounds described herein can be provided in an aqueous physiological buffer containing about 0.1 to about 10% (w / v) compound. Some typical dosage ranges are about 1 μg / kg to about 1 g / kg body weight per day. In some embodiments, the dosage range is about 0.01 mg / kg to about 100 mg / kg body weight per day. Dosages tend to depend on variables such as the type and extent of progression of the disease or disorder, the overall health of the particular patient, the relative biological efficacy of the selected compound, the excipient formulation, and its route of administration. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0092] The compositions of the present invention may further comprise one or more additional pharmaceutical agents, such as chemotherapeutic agents, steroids, anti-inflammatory compounds, or immunosuppressants, examples of which are listed herein.

[0093] kit The present invention also includes pharmaceutical kits useful for treating or preventing PI3K-related diseases or disorders, such as cancer, which include one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound described herein. As will be readily apparent to one of skill in the art, such kits can optionally further include one or more of a variety of conventional pharmaceutical kit components, such as, for example, a container with one or more pharmaceutically acceptable carriers, additional containers, etc. Instructions for use, either as an insert or label, indicating the amounts of the components to be administered, administration guidelines, and / or guidelines for mixing the components, can also be included in the kit. [Example]

[0094] Example 1. ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile [ka] Step 1. tert-Butyl (4S)-2,2-dimethyl-4-vinyl-1,3-oxazolidine-3-carboxylate To a suspension of methyltriphenylphosphonium bromide (5.63 g, 15.8 mmol) in tetrahydrofuran (140 mL) was added 2.5 M n-butyllithium in hexane (7.35 mL, 18.4 mmol). The deep red solution was stirred at 0 °C for 1 h. Then, a solution of tert-butyl (4R)-4-formyl-2,2-dimethyl-1,3-oxazolidine-3-carboxylate (obtained from Aldrich, 3.01 g, 13.1 mmol) in tetrahydrofuran (7.3 mL) was added dropwise at 0 °C. The red solution was allowed to warm to room temperature and stirred for 12 h. Hexane was added to the reaction mixture in a 4:1 (v / v) ratio. The suspension was filtered through Celite, and the filtrate was concentrated. The resulting residue was purified by flash chromatography (eluting with 10% ethyl acetate in hexane) to give the desired compound as a colorless oil (1.92 g, 64%).

[0095] Step 2. tert-Butyl [(1S)-1-(hydroxymethyl)prop-2-en-1-yl]carbamate To a solution of tert-butyl (4S)-2,2-dimethyl-4-vinyl-1,3-oxazolidine-3-carboxylate (1.90 g, 8.36 mmol) in methanol (83 mL) was added p-toluenesulfonic acid monohydrate (0.80 g, 4.2 mmol) at 0 °C. The mixture was allowed to warm slowly to room temperature overnight. The reaction mixture was diluted with saturated aqueous NaHCO, concentrated, and then diluted with ethyl acetate. The organic layer was washed with saturated aqueous NaHCO (2x) and brine, dried over NaSO, filtered, and concentrated to give the desired product as a colorless oil (1.187 g, 76%). 1 H NMR (400 MHz, CDCl3) δ 5.81 (1H, m), 5.25 (2H, m), 4.90 (1H, m), 4.25 (1H, br s), 3.67 (2H, m), 1.45 (9H, s)ppm.

[0096] Step 3. tert-Butyl [(1S)-1-({[1-(hydroxymethyl)prop-2-en-1-yl]oxy}methyl)prop-2-en-1-yl]carbamate A flask was charged with tert-butyl [(1S)-1-(hydroxymethyl)prop-2-en-1-yl]carbamate (0.401 g, 2.14 mmol), tris(dibenzylideneacetone)dipalladium(0) (59 mg, 0.064 mmol), N,N'-(1S,2S)-cyclohexane-1,2-diylbis[2-(diphenylphosphino)-1-naphthamide] (150 mg, 0.19 mmol), and 4-dimethylaminopyridine (78 mg, 0.64 mmol). The reaction mixture was purged with N three times, followed by the addition of methylene chloride (21.3 mL) and 1.0 M triethylborane in THF (130 μL, 0.13 mmol). After stirring for 10 minutes, 2-vinyloxirane (0.150 g, 2.14 mmol) was added, and the resulting mixture was stirred overnight. The reaction was diluted with dichloromethane and saturated aqueous NaHCO3. The organic layer was separated, dried over Na2SO4, filtered, and concentrated. The crude residue was purified by flash chromatography (eluting with 0-50% ethyl acetate / hexanes) to give the desired product (0.271 g, 49%). 1 H NMR (300 MHz, CDCl3) δ 5.85 (1H, m), 5.67 (1H, m), 5.84–5.17 (4H, m), 4.83 (1H, m), 4.30 (1H, br s), 3.83 (1H, m), 3.69 (1H, dd, J = 4.5 and 6.9 Hz), 3.54 (2H, m), 3.36 (1H, dd, J = 4.5 and 6.9 Hz), 1.45 (9H, s) ppm.

[0097] Step 4. 2-({(2S)-2-[(tert-butoxycarbonyl)amino]but-3-en-1-yl}oxy)but-3-en-1-yl acetate To a mixture of tert-butyl [(1S)-1-({[1-(hydroxymethyl)prop-2-en-1-yl]oxy}methyl)prop-2-en-1-yl]carbamate (268 mg, 1.04 mmol) in methylene chloride (10 mL) was added triethylamine (435 μL, 3.12 mmol). The mixture was cooled to 0° C. and acetyl chloride (150 μL, 2.1 mmol) was added dropwise. The reaction was stirred at room temperature for 2 hours and then quenched with water. The organic layer was concentrated and the resulting residue was purified on silica gel (eluted with 20% ethyl acetate / hexanes) to give the desired product (0.26 g, 85%). LCMS C 10 H 18 NO3 (M-100+H) + Measured value for: m / z = 200.1; Found: 200.1.

[0098] Step 5. {(5S)-5-[(tert-butoxycarbonyl)amino]-5,6-dihydro-2H-pyran-2-yl}methyl acetate To a 500 mL two-necked round-bottom flask was added benzylidene(dichloro)(1,3-dimesitylimidazolidin-2-id-2-yl)(tricyclohexylphosphoranyl)ruthenium (38 mg, 0.044 mmol). After purging with nitrogen three times, dichloromethane (anhydrous, 8 mL) was added, followed by 2-({(2S)-2-[(tert-butoxycarbonyl)amino]but-3-en-1-yl}oxy)but-3-en-1-yl acetate (265 mg, 0.885 mmol). The reaction mixture was stirred at room temperature for 15 hours. The mixture was concentrated in vacuo. The residue was purified via flash chromatography (eluting with hexane to 25% EtOAc in hexane) to give the desired product as a brown oil (0.205 g, 85%). LCMS C9H 14 NO5 (M+H-Bu+H) + Measured value for: m / z = 216.1; Found: 216.1. 1H NMR (300 MHz, CDCl) δ 5.94 (0.17H, m), 5.84 (0.83H, m), 5.69 (1H, m), 4.89 (0.13H, m), 4.70 (0.83H, m), 4.25 (1H, m), 4.05 (4H, m), 3.56 (0.13H, m), 3.38 (0.87H, m), 2.04 (2.49H, s), 2.03 (0.51H, m), 1.38 (9H, s) ppm (the product was a ca. 5:1 mixture of trans and cis isomers).

[0099] Step 6. [(5S)-5-Amino-5,6-dihydro-2H-pyran-2-yl]methyl acetate To a solution of {(5S)-5-[(tert-butoxycarbonyl)amino]-5,6-dihydro-2H-pyran-2-yl}methyl acetate (205 mg, 0.756 mmol) in methylene chloride (5.2 mL) was added 4.0 M hydrogen chloride in dioxane (1.5 mL, 6.0 mmol). The reaction solution was stirred at room temperature for 6 hours. The solvent was removed under reduced pressure to give the desired product as a white solid. LCMS C8H 14 NO3(M+H) + Measured value for: m / z = 172.1; Found: 172.1.

[0100] Step 7. {(5S)-5-[(6-nitrothieno[3,2-b]pyridin-7-yl)amino]-5,6-dihydro-2H-pyran-2-yl}methyl acetate A mixture of 7-chloro-6-nitrothieno[3,2-b]pyridine (156 mg, 0.727 mmol), [(5S)-5-amino-5,6-dihydro-2H-pyran-2-yl]methyl acetate (129 mg, 0.754 mmol) and N,N-diisopropylethylamine (0.26 mL, 1.5 mmol) in isopropyl alcohol (1.7 mL) was heated at 90° C. for 2 hours. The reaction mixture was concentrated and purified by flash chromatography to give the desired product (0.21 g, 83%). LCMS C 15 H 16 N3O5S (M+ H)+ Measured value for: m / z = 350.1; Found value: 350.0.

[0101] Step 8. {(5S)-5-[(6-aminothieno[3,2-b]pyridin-7-yl)amino]tetrahydro-2H-pyran-2-yl}methyl acetate A mixture of {(5S)-5-[(6-nitrothieno[3,2-b]pyridin-7-yl)amino]-5,6-dihydro-2H-pyran-2-yl}methyl acetate (210 mg, 0.600 mmol) and 10% palladium on carbon catalyst (0.21 g) in methanol (4.0 mL) was subjected to balloon pressure of H at room temperature for 2 hours. The mixture was filtered, and the filtrate was concentrated and purified by flash chromatography (eluting with 15% methanol in dichloromethane) to give the desired product (145 mg, 75%). LCMS C 15 H 20 N3O3S (M+ H) + Measured value for: m / z = 322.1; Found: 322.0.

[0102] Step 9. (1R)-1-{1-[(3S)-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-2-yl}ethanol A mixture of (2R)-2-hydroxypropanamide (131 mg, 1.47 mmol) and triethyloxonium tetrafluoroborate (263 mg, 1.38 mmol) in THF (2 mL) was stirred at room temperature for 2 hours. The solvent was removed, and the residue was dissolved in ethanol (0.85 mL) and added to a suspension of {(5S)-5-[(6-aminothieno[3,2-b]pyridin-7-yl)amino]tetrahydro-2H-pyran-2-yl}methyl acetate (145 mg, 0.451 mmol) in ethanol (3.1 mL). The mixture was stirred at 80°C for 1 hour. The reaction was cooled to room temperature and diluted with water (1.0 mL). Lithium hydroxide (32.4 mg, 1.35 mmol) was added, and the mixture was stirred for 2 hours. The reaction mixture was diluted with methanol and purified by preparative LCMS (XBridge C18 column, eluting with a gradient of acetonitrile / water containing 0.1% ammonium hydroxide, flow rate 60 mL / min) to give the desired product as a white solid (95 mg, 63%). 16 H 20 N3O3S (M+ H) + Measured value for: m / z = 334.1; Found: 334.0.

[0103] Step 10: ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)methyl 4-methylbenzenesulfonate and ((2S,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)methyl 4-methylbenzenesulfonate To a solution of (1R)-1-{1-[(3S)-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-2-yl}ethanol (100 mg, 0.300 mmol) (previous step) in methylene chloride (3.4 mL) and pyridine (0.146 mL, 1.80 mmol) was added p-toluenesulfonyl chloride (57.2 mg, 0.300 mmol) and 4-dimethylaminopyridine (1.8 mg, 0.015 mmol) at 0 °C. The reaction mixture was allowed to warm to room temperature overnight. The reaction mixture was concentrated, diluted with methanol, and purified by preparative LCMS (XBridge C18 column, eluting with a gradient of acetonitrile / water containing 0.1% ammonium hydroxide, flow rate 60 mL / min) to give two peaks. Analytical HPLC (Waters SunFire C18, 2.1 × 50 mm, 5 μM; flow rate 3 mL / min; injection volume 2 μL; gradient 2 to 80% B in 3 min (A = water with 0.025% TFA, B = acetonitrile)): first peak (45.3 mg, 31%) retention time 1.81 min, LCMS C 23 H 26 N3O5S2(M+ H) + Measured value for m / z = 488.1; Found: 488.1. Second peak (8.5 mg, 5.8%), retention time 1.88 min, LCMS C 23 H 26 N3O5S2(M+ H) + Measured value for: m / z = 488.1; Found: 488.1.

[0104] Step 11. ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile A mixture of ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)methyl 4-methylbenzenesulfonate (from the first peak of the previous step, 27 mg, 0.055 mmol) and sodium cyanide (4.5 mg, 0.092 mmol) in dimethyl sulfoxide (0.4 mL) was stirred at 50 °C for 4 h. After cooling, the mixture was diluted with methanol and purified by preparative LCMS (XBridge C18 column, eluted with a gradient of acetonitrile / water containing 0.1% ammonium hydroxide, flow rate 30 mL / min) to give the desired product (14.5 mg, 76%). LCMS C 17 H 19 N4O2S (M+ H) + Measured value for: m / z = 343.1; Found: 343.0. 1 H NMR (DMSO-d6, 500 MHz) δ 9.51 (1H, s), 8.45 (1H, d, J = 5.5 Hz), 7.97 (1H, d, J = 5..5 Hz), 5.31 (1H, m), 5.20 (1H, m), 4.31 (1H, m), 4.23 (1H, m), 4.02 (1H, m), 2.96 (1H, dd, J = 17.0 and 4.5 Hz), 2.85 (1H, dd, J = 17.0 and 4.5 Hz), 2.66 (1H, m), 2.26 (1H, m), 2.09 (1H, m), 1.73 (1H, m), 1.69 (3H, d, J = 6.5 Hz) ppm.

[0105] Example 1a. ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile hydrate [ka] ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile (52 mg, 0.15 mmol) from Example 25 was crystallized from a mixture of acetonitrile (8 mL) and water (4 mL). The resulting recovered colorless prisms were suitable for X-ray crystallography.

[0106] Crystal data are: approximately 0.520×0.180×0.100mm, orthorhombic, P212121, a=6.962(3)Å, b=11.531(4)Å, c=20.799(7)Å, volume=1669.6(10)Å 3 , Z=4, T=-100.℃, formula weight=359.42, density=1.430g / cm 3 , μ(Mo)=0.22mm -1 Shows.

[0107] Data collection was performed using a Bruker SMART APEX-II CCD system, MoKalpha radiation, standard focal tube, anode power = 50 kV x 42 mA, crystal-to-plate distance = 5.0 cm, 512 x 512 pixels / frame, beam center = (256.13, 253.14), total frames = 1151, oscillation / frame = 0.50°, exposure / frame = 10.1 s / frame, SAINT integration, hkl min / max = (-9, 9, -15, 15, -27, 27), data input to shelx = 17025, unique data = 3975, 2theta range = 3.92–55.72°, completeness for 2theta 55.72 = 99.80%, R(int-xl) = 0.0681, SADABS correction applied.

[0108] The structure was solved using XS (Shelxtl) and refined using the shelxtl software package. 2 Refinement by full matrix least squares based on the scattering factors from Int.Tab.Vol C Table 4.2.6.8 and 6.1.1.4, number of data = 3975, number of constraints = 0, number of parameters = 235, data / parameter ratio = 16.91, F 2Goodness of fit = 1.04, R-index [I>4 sigma (I)] R1 = 0.0505, wR2 = 0.1242, R-index (all data) R1 = 0.0769, wR2 = 0.1401, maximum residual peak and hole = 0.724 and -0.277 e / Å 3 The refinement Flack parameter was −0.12 (13). All CH hydrogen atoms were refined using the riding model. OH hydrogens were found from the difference map and fully refined.

[0109] The results showed that the asymmetric unit contains one α- and one water molecule, as indicated by the thermal ellipsoids at a 50% probability level. The stereochemistry at each of the three stereocenters (as indicated in the compound names and structures above) was confirmed. The Flack parameter was refined to 0.28 (24), suggesting accurate enantiomeric assignment.

[0110] Example 2. 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide [ka] Step 1: 2,4,5-trifluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide To a solution of 2,4,5-trifluorobenzoic acid (5.00 g, 28.4 mmol) in acetonitrile (50 mL) was added N,N-dimethylformamide (40 μL), followed by oxalyl chloride (3.60 mL, 42.6 mmol). After 90 minutes, the volatiles were removed under reduced pressure. The residue was coevaporated with acetonitrile (50 mL). The residue was then dissolved in methylene chloride (50 mL). This solution was added dropwise to a cooled (ice bath) mixture of (2S)-1,1,1-trifluoropropan-2-amine hydrochloride (5.52 g, 36.9 mmol) (Synquest, 98% ee) and 0.5 M aqueous sodium hydroxide (142 mL, 71.0 mmol) in toluene (100 mL). After the addition, the ice bath was removed, and the reaction was allowed to warm to room temperature. The reaction was stirred overnight. The organic layer was separated. The aqueous layer was extracted with methylene chloride (50 mL). The combined organic layers were washed with 20% saturated brine (75 mL) and water (2 x 75 mL), dried over MgSO, filtered, and concentrated under reduced pressure to give the desired product (6.49 g, 84%), which was used directly in the next step without further purification. 1 H NMR (300 MHz, DMSO-d6) δ 9.01 (d, J = 7.6 Hz, 1H), 7.92 - 7.50 (m, 2H), 4.76 (m, 1H), 1.31 (d, J = 7.0 Hz, 3H) ppm. LCMS C 10 H8F6NO (M+1) + Measured value for: m / z = 272.0; Found: 272.0.

[0111] Step 2: 2,5-Difluoro-4-(3-hydroxyazetidin-1-yl)-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide A mixture of 2,4,5-trifluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide (6.39 g, 23.6 mmol), azetidin-3-ol hydrochloride (3.19 g, 28.3 mmol), and 1,8-diazabicyclo[5.4.0]undec-7-ene (8.81 mL, 58.9 mmol) in acetonitrile (25 mL) was stirred at 80 °C for 2 h. The reaction mixture was diluted with EtOAc (75 mL) and washed with 1 N HCl (50 mL), 1 N NaHCO (60 mL), 20% saturated brine (50 mL), and water (75 mL). The aqueous layer was extracted with EtOAc (100 mL). The organic layers were combined, dried over MgSO, filtered, and concentrated under reduced pressure to give the desired product (7.59 g, 91.8%). 1 H NMR (300 MHz, DMSO-d6) δ 8.38 (dd, J = 8.9, 1.9 Hz, 1H), 7.27 (dd, J = 12.8, 6.5 Hz, 1H), 6.38 (dd, J = 12.3, 7.5 Hz, 1H), 5.71 (d, J = 6.4 ppm. LCMS C 13 H 14 F5N2O2(M+1) + Measured value for: m / z = 325.1; Found: 325.1.

[0112] Step 3: 2,5-Difluoro-4-(3-oxoazetidin-1-yl)-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide To a solution of 2,5-difluoro-4-(3-hydroxyazetidin-1-yl)-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide (7.57 g, 23.3 mmol) in methylene chloride (93 mL) was added iodobenzene diacetate (9.40 g, 29.2 mmol) and 2,2,6,6-tetramethyl-1-piperidinyloxy free radical (1.82 g, 11.7 mmol) (TEMPO) at room temperature. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc (100 mL) and washed with 0.5 N NaHCO (2 × 80 mL), 20% saturated brine (100 mL), and water (100 mL). The aqueous layer was extracted with ethyl acetate (75 mL). The organic extracts were combined, dried over MgSO, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography on a silica gel column eluting with 0% to 5% ethyl acetate in methylene chloride to give the crude product, which was recrystallized from MTBE (50 mL) and heptane (100 mL) to give the desired product (5.44 g, 72%) as a colorless solid. 1 H NMR (300 MHz, DMSO-d6) δ 8.52 (d, J = 8.0 Hz, 1H), 7.36 (dd, J = 12.5, 6.5 Hz, 1H), 6.63 (dd, J = 12.1, 7.6 Hz, 1H), 4.90 (d, J = 2.1 Hz, 4H), 4.86 - 4.68 (m, 1H), 1.31 (d, J = 7.1 Hz, 3H) ppm. LCMS C 13 H 12 F5N2O2(M+1) + Measured value for: m / z = 323.1; Found: 323.0.

[0113] Step 4: 4-[3-(cyanomethylene)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide Diethyl cyanomethylphosphonate (1.95 mL, 11.8 mmol) was added dropwise to a cooled (ice bath) solution of 1.0 M potassium tert-butoxide in THF (11.8 mL, 11.8 mmol) diluted with tetrahydrofuran (12 mL). The ice bath was removed, and the reaction was allowed to warm to room temperature and stirred for 90 minutes. The reaction solution was cooled again in an ice bath. The above prepared solution was then added over 12 minutes to a cooled (ice bath) solution of 2,5-difluoro-4-(3-oxoazetidin-1-yl)-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide (4.00 g, 12.4 mmol) in tetrahydrofuran (50 mL). The reaction mixture was stirred for 30 minutes. The ice bath was removed, and the reaction was stirred at room temperature overnight, then quenched by the addition of 20% saturated brine (75 mL) and ethyl acetate (75 mL). The organic layer was separated. The aqueous layer was extracted with ethyl acetate (50 mL). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography on a silica gel column using ethyl acetate in hexanes (0% to 30%) to give the desired product (2.6 g). 1 H NMR (400 MHz, DMSO-d6) δ 8.59 - 8.37 (m, 1H), 7.33 (dd, J = 12.5, 6.4 Hz, 1H), 6.59 (dd, J = 12.0, 7.4 Hz, 1H), 5.88 (m, 1H), 4.94 - 4.75 (m, 4H), 4.76 (m, 1H), 1.31 (d, J = 7.1 Hz, 3H) ppm. LCMS C 15 H 13 F5N3O (M+1) + Measured value for: m / z = 346.1; Found: 346.1.

[0114] Step 5: 4-{3-(cyanomethyl)-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide A mixture of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1.00 g, 5.15 mmol), 4-[3-(cyanomethylene)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide (1.78 g, 5.15 mmol), and 1,8-diazabicyclo[5.4.0]undec-7-ene (0.31 mL, 2.1 mmol) in acetonitrile (20.2 mL) was heated at 50° C. overnight. After cooling, the solvent was removed under reduced pressure. The residue was used in the next step without further purification. LCMS C 24 H 28 BF5N5O3(M+1) + Measured value for: m / z = 540.2; Found: 540.1.

[0115] Step 6: 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide A mixture of 4-{3-(cyanomethyl)-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide (329 mg, 0.610 mmol), 4-bromo-3,5-dimethyl-1H-pyrazole (206 mg, 1.18 mmol), tetrakis(triphenylphosphine)palladium(0) (110 mg, 0.098 mmol), and sodium carbonate (320 mg, 3.0 mmol) in 1,4-dioxane (10 mL) / water (5 mL) was purged with nitrogen and stirred at 110° C. for 1 h. The reaction mixture was diluted with EtOAc, washed with water and saturated brine, and concentrated. The residue was purified first on silica gel (eluting with 0–100% EtOAc / hexanes, then 10% methanol / dichloromethane) and then by preparative LCMS (XBridge C18 column, eluting with a gradient of acetonitrile / water containing 0.1% ammonium hydroxide, flow rate 60 mL / min) to give the desired product (30 mg, 9.7%). 1 H NMR (500 MHz, DMSO-d6) δ 12.17 (1H, s), 8.45 (1H, d, J = 8.0 Hz), 8.10 (1H, s), 7.70 (1H, s), 7.34 (1H, m), 6.61 (1H, s), 4.77 (1H, m), 4.62 (2H, d, J = 9.0 Hz), 4.39 (1H, d, J = 9.0 Hz), 3.64 (2H, s), 2.22 (6H, s), 1.31 (6H, d, J = 7.0 Hz) ppm. LCMS C 23 H 23 F5N7O (M+H) + Measured value for: m / z = 508.2; Found: 508.0.

[0116] Example A: In vitro JAK kinase assay The compounds herein were tested for inhibitory activity against JAK targets according to the following in vitro assay described by Park et al., Analytical Biochemistry 1999, 269, 94-104. The catalytic domains of human JAK1 (aa 837-1142), JAK2 (aa 828-1132), and JAK3 (aa 781-1124) were expressed in insect cells using baculovirus and purified. The catalytic activity of JAK1, JAK2, or JAK3 was assessed by measuring the phosphorylation of a biotinylated peptide. The phosphorylated peptide was detected by homogeneous time-resolved fluorescence (HTRF). The IC of the compounds was 50 was measured for each kinase in 40 μL reactions containing enzyme, ATP, and 500 nM peptide in 50 mM Tris (pH 7.8) buffer with 100 mM NaCl, 5 mM DTT, and 0.1 mg / mL (0.01%) BSA. An IC of 1 mM was obtained. 50 For measurements, the ATP concentration in the reaction was 1 mM. Reactions were carried out at room temperature for 1 hour and then stopped with 20 μL of 45 mM EDTA, 300 nM SA-APC, 6 nM Eu-Py20 in assay buffer (Perkin Elmer, Boston, MA). Binding to the europium-labeled antibody occurred for 40 minutes, and the HTRF signal was measured on a PHERA star plate reader (BMG, Cary, NC). Data for JAK1 and / or JAK2 inhibitors were obtained by testing the compounds in the Example A assay with 1 mM ATP.

[0117] Example B: PI3Kδ Scintillation Proximity Assay material [γ- 33[P]ATP (10 mCi / mL) was purchased from Perkin-Elmer (Waltham, MA). The lipid kinase substrate, D-myo-phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) D(+)-sn-1,2-di-O-octanoylglyceryl, 3-O-phosphorus linked (PIP2), CAS 204858-53-7, was purchased from Echelon Biosciences (Salt Lake City, UT). PI3Kδ (p110δ / p85α) was purchased from Millipore (Bedford, MA). ATP, MgCl2, DTT, EDTA, MOPS, and CHAPS were purchased from Sigma-Aldrich (St. Louis, MO). Wheat Germ Agglutinin (WGA) YSi SPA scintillation beads were purchased from GE Healthcare Life Sciences (Piscataway, NJ).

[0118] Kinase reactions were performed in a polystyrene 384-well matrix white plate (Thermo Fisher Scientific) in a final volume of 25 μL. Inhibitors were first serially diluted in DMSO and added to the plate wells before the addition of other reaction components. The final concentration of DMSO in the assay was 0.5%. PI3K assays were performed at room temperature in 20 mM MOPS, pH 6.7, 10 mM MgCl2, 5 mM DTT, and 0.03% CHAPS. Reactions were initiated by the addition of ATP, and the final reaction mixture contained 20 μM PIP2, 20 μM ATP, 0.2 μCi [γ- 33The reaction consisted of [P]ATP, 4 nM PI3Kδ. Reactions were incubated for 210 min and terminated by the addition of 40 μL SPA beads suspended in stop buffer: 150 mM potassium phosphate pH 8.0, 20% glycerol, 25 mM EDTA, 400 μM ATP. The final concentration of SPA beads was 1.0 mg / mL. After sealing, the plates were shaken overnight at room temperature and centrifuged at 1800 rpm for 10 min, and product radioactivity was determined by scintillation counting in a TopCount (Perkin-Elmer). IC was calculated by curve fitting of % control activity against the log of inhibitor concentration using GraphPad Prism 3.0 software. 50 The determination was made: Data for PI3Kδ inhibitors was obtained by testing the compounds in the Example B assay.

[0119] Example C: Pfeiffer Model of Lymphoma method: Female SCID mice (5–8 weeks old, Charles River Laboratories, Wilmington, MA) were inoculated with 1 × 10 tumor cells (Pfeiffer, ATCC No. CRL-2632, Manassas, VA) and Matrigel (BD Biosciences No. 354234) in 0.2 mL of sterile saline. Inoculations were performed subcutaneously in the flank. Tumor tissue fragments (approximately 3 mm × 3 mm) were harvested 3–6 weeks after inoculation with cultured cells and implanted subcutaneously in place of the cell inoculation. The tissue fragments were implanted as solid pieces using blunt-tip forceps. Treatment of tumor-bearing mice began 15–25 days after tumor inoculation, depending on tumor size. Animals were grouped to achieve approximately equal mean tumor volumes within each group. The minimum mean tumor volume across all groups was 150 mm on the first day of treatment, and each group consisted of 7 animals. The experimental treatment, Example 347, was administered orally (PO) to mice. Treatment frequency was twice daily for a minimum of 14 days for efficacy. Subcutaneous tumor size was measured two to three times weekly using digital calipers. Tumor volume was calculated by measuring the tumor in two dimensions and using the formula: volume = [length × (width 2)] / 2 (larger number = length, smaller number = width). When multiple tumors formed, the final volume was calculated as the sum of the individual tumor volumes, each applied to the same formula: e.g., for two tumors; volume = {[L1 × (W1)2] / 2} + {[L2 × (W2)2] / 2}. The effect on tumor growth was reported as percent tumor growth inhibition (TGI %). The TGI rate (%) was calculated using the formula: (1-(Tx volume / control volume)) x 100, where control volume was the vehicle or untreated tumor volume on a given day and Tx volume was the tumor volume of any treatment group on the same day. Statistical differences between treatments and vehicle controls were assessed using one-way analysis of variance.

[0120] result: Compound 32c (Table 2 above) was evaluated as a single agent in a Pfeiffer human tumor xenograft model of diffuse large B-cell lymphoma, a subtype of NHL. Pfeiffer cancer cells were shown to be sensitive to the antiproliferative effects of Example 347 in vitro. Therefore, a tumor model was established based on subcutaneous inoculation of tumor cells into immunocompromised SCID mice and tumor-bearing mice, which received oral administration of vehicle or 0.3, 1, 3, or 10 mg / kg of compound 32c twice daily for 14 days. Compound 32c treatment inhibited tumor growth by 22%, 24%, 36%, and 58% (tumor growth inhibition rate (%)) with increasing doses.

[0121] Example D. Western Blot Analysis The following materials and methods were used in the Western blot analysis described below. Cells (5 million) were lysed in 300 μl of lysis buffer. The soluble fraction was collected by centrifugation. 25 μl of cell lysate was loaded onto a Tris-glycine polyacrylamide gel and subjected to electrophoresis. Proteins were transferred to a nitrocellulose membrane and probed with antibodies from Cell Signaling Technology for the following proteins: phospho-Stat3 Y705, phospho-Akt S473, pim1, pim2, pim3, c-myc, phospho-p70S6K, phospho-S6, phospho-Bad S112, and actin.

[0122] Example E. IL6 and IL10 levels in cell lines High levels of IL6 and IL10 were found in various DLBCL cell lines (Figure 1A). IL6 and IL10 have also been shown to activate JAK / STAT signaling, with IL10 being a stronger activator of JAK / STAT signaling than IL6 across a panel of DLBCL cell lines (Figure 1B). High levels of IL6 and IL10 were present in serum from DLBCL patients and correlated with shorter event-free survival and higher International Prognostic Index scores.

[0123] Example F. IL10 renders Pfeiffer cells resistant to PI3Kδ inhibition and can be reversed by JAK1 / 2 or JAK1 blockade. Cell proliferation assay Diffuse large B-cell lymphoma cells were seeded at 2000 cells / well in 96-well culture plates in the absence or presence of 10 ng / ml IL10. Compounds were first diluted in DMSO before being added to the cells, and then diluted (4x) in culture medium. Cells were cultured in an incubator at 5% CO for 3 days. Cell proliferation was assessed using a cell titer-glow assay (Promega, Madison, WI). Cell proliferation assays were first performed in Pfeiffer cells (germinal center B-cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL) cells) and HBL-1 cells (activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL)).

[0124] Figure 2A shows the percent inhibition in a cell proliferation assay using Pfeiffer cells as a function of concentration of vehicle (DMSO), DMSO + IL10, and DMSO + IL10 + ruxolitinib (a JAK1 / JAK2 inhibitor) and Compound 28 (a PI3Kδ inhibitor). Figure 2B shows the percent inhibition in a cell proliferation assay using Pfeiffer cells as a function of concentration of vehicle (DMSO), DMSO + IL10, and DMSO + IL10 + Compound 7 (a selective JAK1 inhibitor) and Compound 28 (a PI3Kδ inhibitor). The results indicate that IL10 renders Pfeiffer cells resistant to PI3Kδ inhibition, but this resistance can be reversed by blocking JAK1 and / or JAK2 signaling. Thus, a synergistic effect on Pfeiffer cell proliferation was observed when a PI3Kδ inhibitor and a JAK1 and / or JAK2 inhibitor were used in combination. Synergy was also observed without IL10. Induction of apoptosis was also observed with this combination.

[0125] Similar results were observed in HBL-1 cells with ruxolitinib. Thus, Figure 3 shows the percent inhibition in a cell proliferation assay in HBL-1 cells as a function of concentration for vehicle (DMSO), DMSO + IL10, and DMSO + IL10 + ruxolitinib (JAK1 / JAK2 inhibitor) and compound 28 (PI3Kδ inhibitor).

[0126] Example G. IL10-induced expression of Pim2 is blocked by JAK1 / 2 inhibitors Pfeiffer cells were treated with vehicle (DMSO), ruxolitinib (18424), Compound 28, or Compound 28 and ruxolitinib (18424) for 24 hours, with or without IL10, and then subjected to Western blot analysis to probe for the following proteins: phospho-Stat3 Y705, phospho-Akt S473, Pim2, c-Myc, phospho-p70S6K, phospho-S6, phospho-Bad S112, and actin. Figure 4 shows that IL10-induced expression of Pim2 is blocked by ruxolitinib (a JAK1 / JAK2 inhibitor). IL6 and IL10 promote cell survival through the expression of Pim2, which is dependent on JAK1 activity. Figure 4 also shows a synergistic decrease in c-Myc and P-S6 in the presence of combined Compound 28 and ruxolitinib treatment. The reduction of c-Myc protein may be involved in the synergistic effect of the combined treatment.

[0127] Example H. IL10-induced expression of Pim2 is blocked by a selective JAK1 inhibitor Pfeiffer cells were treated with vehicle (DMSO), Compound 7, Compound 28, or Compound 28 and Compound 7, with or without IL10, for 24 hours and then subjected to Western blot analysis to probe for the following proteins: phospho-Stat3 Y705, phospho-Akt S473, Pim2, c-Myc, phospho-p70S6K, phospho-S6, phospho-Bad S112, and actin. Figure 5 shows that IL10-induced expression of Pim2 is blocked by a selective JAK1 inhibitor (Compound 7).

[0128] Example I. Enhancement of PI3Kδ inhibitor potency by selective JAK1 inhibitors To test the effect of IL-10 on cell growth and sensitivity to BCR pathway inhibition, the Pfeiffer cell line was used as a model system for DLBCL. Pfeiffer cells are of the germinal center B cell (GCB) subtype of DLBCL and have been shown to express PI3Kδ, be sensitive to PI3Kδ inhibition, and activate the JAK / STAT pathway in response to multiple cytokines, as described above. Pfeiffer cells were treated with various concentrations of compound 28 in the presence or absence of IL-10 and 1 μM compound 16 for 3 days, and cell growth was measured using ATP readings (see table below). As shown in Figure 6, the presence of IL-10 altered the potency of compound 28 by approximately 10-fold (IC 50 = 0.67 μM, -IL-10; IC 50 = 6.36 μM, + IL-10). Addition of the JAK1 inhibitor, Compound 16, reversed this effect, so the combination was approximately 50-fold more potent. The JAK1 inhibitor alone had no effect in this system (IC 50 Furthermore, as shown in Figure 7 (showing Annexin-V staining of Pfeiffer cells treated for 3 days in 10% FBS + IL10; compound 16 was tested at 1 µM), inhibition of PI3Kδ together with JAK1 signaling resulted in increased apoptosis, whereas neither agent alone had a significant effect. [Table 3]

[0129] Example J. Effect of JAK1 and PI3Kδ combined treatment on STAT3 phosphorylation and pAKT inhibition To evaluate the effects on downstream signaling pathways, Pfeiffer cells were treated with Compound 28 + / - Compound 16 for 4 hours and then stimulated with IL-10 for 15 minutes. Extracts were analyzed for pAKT and pSTAT3 by Western blotting. As shown in Figure 8, the AKT pathway was constitutively activated in Pfeiffer cells. Blockade of PI3Kδ signaling resulted in complete inhibition of pAKT, while treatment with a JAK1 inhibitor had no effect. In contrast, Compound 16 inhibited STAT3 phosphorylation, while the PI3Kδ inhibitor did not. A combination of both compounds was required to block both pathways.

[0130] All patents, patent publications, and journal articles referenced above are incorporated herein by reference in their entirety. The present invention also includes the following. [Invention 1] In patients who need it, it is used to treat diffuse large B-cell lymphoma, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Hodgkin's lymphoma, Burkitt's lymphoma, Waldenstrom's macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosa-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, 1. A method of treating a disease selected from primary effusion lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammopathy of undetermined significance (MGUS), activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL), and germinal center B-cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL), comprising administering to the patient: (a) an inhibitor of JAK1 and / or JAK2; and (b) an inhibitor of PI3Kδ; (a) the inhibitor of JAK1 and / or JAK2 is: 3-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; 4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-[4-fluoro-2-(trifluoromethyl)phenyl]piperidine-1-carboxamide; [3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(1-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)azetidin-3-yl]acetonitrile; [trans-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-3-(4-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperazin-1-yl)cyclobutyl]acetonitrile; {trans-3-(4-{[4-[(3-hydroxyazetidin-1-yl)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-{[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-{[(2R)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; 4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile; 5-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide; 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide; 5-{3-(cyanomethyl)-3-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide; {1-(cis-4-{[6-(2-hydroxyethyl)-2-(trifluoromethyl)pyrimidin-4-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-[(ethylamino)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-(1-hydroxy-1-methylethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-{[(3R)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-{[(3S)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {trans-3-(4-{[4-({[(1S)-2-hydroxy-1-methylethyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-({[(2R)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-({[(2S)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-(2-hydroxyethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile; 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide; and a pharmaceutically acceptable salt of any of the foregoing; (b) the inhibitor of PI3Kδ is: 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one; (S)-7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one; 4-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-{1-[(2S)-2-hydroxypropyl]azetidin-3-yl}-3-methoxybenzonitrile; 4-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-[1-(2-hydroxyethyl)azetidin-3-yl]-3-methoxybenzonitrile; 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-cyano-2-ethoxy-5-methylphenyl}-N,N-dimethylpyridine-2-carboxamide; 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one; and N-{1-[5-chloro-8-(3-fluorophenyl)cinnolin-7-yl]ethyl}-9H-purin-6-amine; 4-chloro-3'-fluoro-3-methyl-6-[1-(9H-purin-6-ylamino)ethyl]biphenyl-2carbonitrile; and a pharmaceutically acceptable salt of any of the foregoing. [Invention 2] The inhibitor of JAK1 and / or JAK2: (R)-3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; (R)-3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; (R)-4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; (R)-4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; (R)-4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile; (S)-3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; (S)-3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; (S)-4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; (S)-4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; (S)-4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile; and any pharmaceutically acceptable salt thereof. [Invention 3] The method according to claim 1, wherein the inhibitor of JAK1 and / or JAK2 is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, or a pharmaceutically acceptable salt thereof. [Invention 4] The method of invention 1, wherein the inhibitor of JAK1 and / or JAK2 is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile phosphate. [Invention 5] The method of invention 1, wherein the inhibitor of JAK1 and / or JAK2 is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharmaceutically acceptable salt thereof. [Invention 6] The method of invention 1, wherein the inhibitor of JAK1 and / or JAK2 is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate. [Invention 7] The method according to claim 1, wherein the inhibitor of JAK1 and / or JAK2 is 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide, or a pharmaceutically acceptable salt thereof. [Invention 8] The inhibitor of PI3Kδ: (S)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (S)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (R)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (N-{(1S)-1-[5-chloro-8-(3-fluorophenyl)cinnolin-7-yl]ethyl}-9H-purin-6-amine; and any pharmaceutically acceptable salt thereof. [Invention 9] The method of invention 1, wherein said inhibitor of PI3Kδ is (S)-7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof. [Invention 10] The method of invention 1, wherein said inhibitor of PI3Kδ is 4-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-[1-(2-hydroxyethyl)azetidin-3-yl]-3-methoxybenzonitrile, or a pharmaceutically acceptable salt thereof. [Invention 11] The method of invention 1, wherein the inhibitor of PI3Kδ is 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-cyano-2-ethoxy-5-methylphenyl}-N,N-dimethylpyridine-2-carboxamide, or a pharmaceutically acceptable salt thereof. [Invention 12] 2. The method of claim 1, wherein said inhibitor of PI3Kδ is (N-{(1S)-1-[5-chloro-8-(3-fluorophenyl)cinnolin-7-yl]ethyl}-9H-purin-6-amine, or a pharmaceutically acceptable salt thereof. [Invention 13] The method according to Invention 1, wherein the disease is diffuse large B-cell lymphoma. [Invention 14] The method according to invention 1, wherein the disease is activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL) or germinal center B-cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL). [Invention 15] The method according to claim 1, wherein said inhibitor of JAK1 and / or JAK2 and said inhibitor of PI3Kδ are administered simultaneously. [Invention 16] The method according to claim 1, wherein said inhibitor of JAK1 and / or JAK2 and said inhibitor of PI3Kδ are administered sequentially. [Invention 17] 1. A method of treating diffuse large B-cell lymphoma in a patient in need thereof, comprising administering to the patient (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, or a pharmaceutically acceptable salt thereof; and (7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof. [Invention 18] 1. A method of treating diffuse large B-cell lymphoma in a patient in need thereof, comprising administering to the patient {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharmaceutically acceptable salt thereof; and 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof. [Invention 19] 1. A method of treating diffuse large B-cell lymphoma in a patient in need thereof, comprising administering to the patient 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide, or a pharmaceutically acceptable salt thereof; and 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

Claims

1. In patients in need thereof, the treatment of diffuse large B-cell lymphoma, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Hodgkin's lymphoma, Burkitt's lymphoma, Waldenstrom's macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosa-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, 1. A method of treating a disease selected from primary effusion lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammopathy of undetermined significance (MGUS), activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL), and germinal center B-cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL), comprising administering to said patient: (a) an inhibitor of JAK1 and / or JAK2; and (b) an inhibitor of PI3Kδ; (a) the inhibitor of JAK1 and / or JAK2: 3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; 4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; 4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-[4-fluoro-2-(trifluoromethyl)phenyl]piperidine-1-carboxamide; [3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(1-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)azetidin-3-yl]acetonitrile; [trans-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-3-(4-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperazin-1-yl)cyclobutyl]acetonitrile; {trans-3-(4-{[4-[(3-hydroxyazetidin-1-yl)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-{[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-{[(2R)-2-(hydroxymethyl)pyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; 4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile; 5-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide; 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide; 5-{3-(cyanomethyl)-3-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-isopropylpyrazine-2-carboxamide; {1-(cis-4-{[6-(2-hydroxyethyl)-2-(trifluoromethyl)pyrimidin-4-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-[(ethylamino)methyl]-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-(1-hydroxy-1-methylethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-{[(3R)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {1-(cis-4-{[4-{[(3S)-3-hydroxypyrrolidin-1-yl]methyl}-6-(trifluoromethyl)pyridin-2-yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile; {trans-3-(4-{[4-({[(1S)-2-hydroxy-1-methylethyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-({[(2R)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-({[(2S)-2-hydroxypropyl]amino}methyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; {trans-3-(4-{[4-(2-hydroxyethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy}piperidin-1-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]cyclobutyl}acetonitrile; ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile; 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide; and a pharmaceutically acceptable salt of any of the foregoing; (b) the inhibitor of PI3Kδ is: 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one; (S)-7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one; 4-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-{1-[(2S)-2-hydroxypropyl]azetidin-3-yl}-3-methoxybenzonitrile; 4-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-[1-(2-hydroxyethyl)azetidin-3-yl]-3-methoxybenzonitrile; 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-cyano-2-ethoxy-5-methylphenyl}-N,N-dimethylpyridine-2-carboxamide; 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one; and N-{1-[5-chloro-8-(3-fluorophenyl)cinnolin-7-yl]ethyl}-9H-purin-6-amine; 4-chloro-3'-fluoro-3-methyl-6-[1-(9H-purin-6-ylamino)ethyl]biphenyl-2carbonitrile; and a pharmaceutically acceptable salt of any of the foregoing.

2. The inhibitor of JAK1 and / or JAK2 is: (R)-3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; (R)-3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; (R)-4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; (R)-4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; (R)-4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile; (S)-3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; (S)-3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile; (S)-4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; (S)-4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile; (S)-4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile; and a pharmaceutically acceptable salt of any of the foregoing.

3. 2. The method of claim 1, wherein the inhibitor of JAK1 and / or JAK2 is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, or a pharmaceutically acceptable salt thereof.

4. 2. The method of claim 1, wherein the inhibitor of JAK1 and / or JAK2 is (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile phosphate.

5. 2. The method of claim 1, wherein the inhibitor of JAK1 and / or JAK2 is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharmaceutically acceptable salt thereof.

6. 2. The method of claim 1, wherein the inhibitor of JAK1 and / or JAK2 is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate.

7. 2. The method of claim 1, wherein the inhibitor of JAK1 and / or JAK2 is 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide, or a pharmaceutically acceptable salt thereof.

8. 10. The inhibitor of PI3Kδ: (S)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (S)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (R)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (N-{(1S)-1-[5-chloro-8-(3-fluorophenyl)cinnolin-7-yl]ethyl}-9H-purin-6-amine; and a pharmaceutically acceptable salt of any of the foregoing.

9. 2. The method of claim 1, wherein the inhibitor of PI3Kδ is (S)-7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

10. 2. The method of claim 1, wherein the inhibitor of PI3Kδ is 4-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-chloro-2-[1-(2-hydroxyethyl)azetidin-3-yl]-3-methoxybenzonitrile, or a pharmaceutically acceptable salt thereof.

11. 2. The method of claim 1, wherein the inhibitor of PI3Kδ is 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-6-cyano-2-ethoxy-5-methylphenyl}-N,N-dimethylpyridine-2-carboxamide, or a pharmaceutically acceptable salt thereof.

12. 2. The method of claim 1, wherein the inhibitor of PI3Kδ is (N-{(1S)-1-[5-chloro-8-(3-fluorophenyl)cinnolin-7-yl]ethyl}-9H-purin-6-amine, or a pharmaceutically acceptable salt thereof.

13. 10. The method of claim 1, wherein the disease is diffuse large B-cell lymphoma.

14. 2. The method of claim 1, wherein the disease is activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL) or germinal center B-cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL).

15. 2. The method of claim 1, wherein the inhibitor of JAK1 and / or JAK2 and the inhibitor of PI3Kδ are administered simultaneously.

16. 2. The method of claim 1, wherein the inhibitor of JAK1 and / or JAK2 and the inhibitor of PI3Kδ are administered sequentially.

17. 1. A method of treating diffuse large B-cell lymphoma in a patient in need thereof, comprising administering to the patient (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, or a pharmaceutically acceptable salt thereof; and (7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

18. 1. A method of treating diffuse large B-cell lymphoma in a patient in need thereof, comprising administering to the patient {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile, or a pharmaceutically acceptable salt thereof; and 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

19. 1. A method of treating diffuse large B-cell lymphoma in a patient in need thereof, comprising administering to the patient 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide, or a pharmaceutically acceptable salt thereof; and 7-(1-(9H-purin-6-ylamino)ethyl)-6-(3-fluorophenyl)-3-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one, or a pharmaceutically acceptable salt thereof.

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