Methods of treating and preventing alloantibody driven chronic graft versus host disease

Abstract

To provide a pharmaceutical for the treatment of bronchiolitis obliterans syndrome.SOLUTION: Provided is a pharmaceutical containing (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (ibrutinib).SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 61 / 910,944, filed December 2, 2013; and U.S. Provisional Patent Application No. 61 / 973,178, filed March 31, 2014; each of which is incorporated by reference herein in its entirety. [Background technology]

[0002] Chronic graft-versus-host disease (cGVHD) is the most common long-term complication after allogeneic stem cell transplantation (SCT), affecting 30–70% of patients who survive beyond the first 100 days. cGVHD and its associated immune defects have been identified as the leading cause of nonrelapse mortality (NRM) in allogeneic SCT survivors. SCT survivors with cGVHD are 4.7 times more likely to develop severe or life-threatening health conditions compared with healthy siblings, and patients with active cGVHD are more likely to report adverse health conditions, mental health, functional impairment, activity limitations, and pain than allo-SCT survivors without a history of cGVHD. Any organ system can be affected, and further morbidity is frequently caused by long-term exposure to corticosteroids and calcineurin inhibitors, which are required to treat the condition. In addition to specific CD4 T cell subsets, alloreactive B cells are important mediators of cGVHD. B cells and the deposition of pathogenic alloantibodies are unusually hyperactive in human cGVHD. Summary of the Invention

[0003] Disclosed herein, in some embodiments, are methods of treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the method comprising administering a therapeutically effective amount of an ACK inhibitor (e.g., an ITK or BTK inhibitor). In some embodiments, provided are methods of treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the method comprising administering to the patient in need thereof a therapeutically effective amount of a compound of Formula (A), having the following structure:

[0004] [ka] During the ceremony: A is N; R1 is phenyl-O-phenyl or phenyl-S-phenyl; R2 and R3 are independently H; R4 is L3-X-L4-G, where: L3 is optional and, when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, when present, is a single bond, -O-, -C(=O)-, -S-, -S(=O)-, -S(=O)2-, -NH-, -NR9-, -NHC(O)-, -C(O)NH-, -NR9C(O)-, -C(O)NR9-, -S(=O)2NH-, -NHS(=O)2-, -S(=O)2NR9-, -NR9S(=O)2-, -OC(O)NH-, -NHC(O)O-, -OC(O)NR9-, -NR9C(O)O-, -CH=NO-, -ON=CH-, -NR 10 C(O)NR 10 -, heteroaryl-, aryl-, -NR 10 C(=NR 11 )NR 10 -, -NR 10 C(=NR 11 )-, -C(=NR 11 )NR 10-, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0005] [ka] where: R6, R7, and R8 are independently selected from H, halogen, CN, OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; each R9 is independently selected from H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R 10 are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or The Two R's 10 the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R 10 and R 11 can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R 11 is independently selected from H or substituted or unsubstituted alkyl. In some embodiments, L3, X, and L4 taken together form a nitrogen-containing heterocycle. In some embodiments, the nitrogen-containing heterocycle is a piperidine group. In some embodiments, G is

[0006] [ka] In some embodiments, the compound of Formula (A) is (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (ibrutinib), or a pharmaceutically acceptable salt thereof.

[0007] [ka] In some embodiments, the patient exhibits one or more symptoms of cGVHD. In some embodiments, the cGVHD is untreated cGVHD. In some embodiments, the cGVHD is non-scleroderma cGVHD. In some embodiments, the cGVHD is multi-organ cGVHD. In some embodiments, the cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the cGVHD is pulmonary cGVHD. In some embodiments, the cGVHD is hepatic cGVHD. In some embodiments, the cGVHD is renal cGVHD. In some embodiments, the cGVHD is esophageal cGVHD. In some embodiments, the cGVHD is gastric cGVHD. In some embodiments, fibrosis is reduced. In some embodiments, pulmonary fibrosis is reduced. In some embodiments, liver fibrosis is reduced. In some embodiments, immunoglobulin (Ig) deposition in tissues is reduced. In some embodiments, the patient is suffering from cancer. In some embodiments, the patient is suffering from a hematological malignancy. In some embodiments, the patient is suffering from a relapsed or refractory hematological malignancy. In some embodiments, the patient is suffering from a B-cell malignancy. In some embodiments, the patient is suffering from a T-cell malignancy. In some embodiments, the patient is suffering from leukemia, lymphoma, or myeloma. In some embodiments, the B-cell malignancy is non-Hodgkin's lymphoma. In some embodiments, the B-cell malignancy is chronic lymphocytic leukemia (CLL). In some embodiments, the B-cell malignancy is a relapsed or refractory B-cell malignancy. In some embodiments, the B-cell malignancy is a relapsed or refractory non-Hodgkin's lymphoma. In some embodiments, the B-cell malignancy is relapsed or refractory CLL. In some embodiments, the patient is suffering from high-risk CLL. In some embodiments, the patient is suffering from a chromosome 17p deletion. In some embodiments, the patient has 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more CLL as determined by bone marrow biopsy, hi some embodiments, the patient has previously received one or more anti-cancer agents.In some embodiments, the patient has undergone a cell transplant. In some embodiments, the cell transplant is a hematopoietic cell transplant. In some embodiments, the cell transplant is an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, the compound of Formula (A) is administered simultaneously with an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, the compound of Formula (A) is administered after an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, the amount of an ACK inhibitor compound (e.g., a compound of Formula (A)) prevents or reduces cGVHD while maintaining a graft-versus-leukemia (GVL) response effective to reduce or eliminate the number of cancer cells in the patient's blood. In some embodiments, the compound of Formula (A) is administered at a dosage of between about 0.1 mg / kg per day and about 100 mg / kg per day. In some embodiments, the amount of the compound of Formula (A) administered is about 40 mg / day, about 140 mg / day, about 420 mg / day, about 560 mg / day, or about 840 mg / day. In some embodiments, the compound of Formula (A) is administered from day 1 to about day 1000 after allogeneic bone marrow or hematopoietic stem cell transplantation. In some embodiments, the compound of Formula (A) is administered from the onset of symptoms of alloantibody-driven cGVHD to about day 1000 after allogeneic bone marrow or hematopoietic stem cell transplantation. In some embodiments, the compound of Formula (A) is administered orally. In some embodiments, the compound of Formula (A) is administered in combination with one or more additional therapeutic agents.

[0008] In some embodiments, disclosed herein are methods for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of an alloantibody-driven cGVHD episode in a patient in need of a cell transplant, the methods comprising administering a therapeutically effective amount of an ACK inhibitor (e.g., an ITK or BTK inhibitor). In some embodiments, disclosed herein are methods for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of an alloantibody-driven cGVHD episode in a patient in need of a cell transplant, the methods comprising administering a therapeutically effective amount of a compound of Formula (A), having the structure:

[0009] [ka] During the ceremony: A is N; R1 is phenyl-O-phenyl or phenyl-S-phenyl; R2 and R3 are independently H; R4 is L3-X-L4-G, where: L3 is optional and, when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, when present, is a single bond, -O-, -C(=O)-, -S-, -S(=O)-, -S(=O)2-, -NH-, -NR9-, -NHC(O)-, -C(O)NH-, -NR9C(O)-, -C(O)NR9-, -S(=O)2NH-, -NHS(=O)2-, -S(=O)2NR9-, -NR9S(=O)2-, -OC(O)NH-, -NHC(O)O-, -OC(O)NR9-, -NR9C(O)O-, -CH=NO-, -ON=CH-, -NR 10 C(O)NR 10 -, heteroaryl-, aryl-, -NR 10 C(=NR 11 )NR 10 -, -NR 10 C(=NR11 )-, -C(=NR 11 )NR 10 -, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0010] [ka] where: R6, R7, and R8 are independently selected from H, halogen, CN, OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; each R9 is independently selected from H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R 10 are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or The Two R's 10 the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R 10 and R 11 can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R 11 is independently selected from H or substituted or unsubstituted alkyl. In some embodiments, L3, X, and L4 taken together form a nitrogen-containing heterocycle. In some embodiments, the nitrogen-containing heterocycle is a piperidine group. In some embodiments, G is

[0011] [ka] is.

[0012] In some embodiments, disclosed herein are methods for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of an alloantibody-driven cGVHD episode in a patient in need of a cell transplant, the methods comprising administering a therapeutically effective amount of an ACK inhibitor (e.g., an ITK or BTK inhibitor). In some embodiments, disclosed herein are methods for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of an alloantibody-driven cGVHD episode in a patient in need of a cell transplant, the methods comprising administering a therapeutically effective amount of a compound of formula (A) or a pharmaceutically acceptable salt thereof:

[0013] [ka] During the ceremony: A is N; R1 is phenyl-O-phenyl or phenyl-S-phenyl; R2 and R3 are independently H; R4 is L3-X-L4-G, where: L3 is optional and, when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, when present, is a single bond, -O-, -C(=O)-, -S-, -S(=O)-, -S(=O)2-, -NH-, -NR9-, -NHC(O)-, -C(O)NH-, -NR9C(O)-, -C(O)NR9-, -S(=O)2NH-, -NHS(=O)2-, -S(=O)2NR9-, -NR9S(=O)2-, -OC(O)NH-, -NHC(O)O-, -OC(O)NR9-, -NR9C(O)O-, -CH=NO-, -ON=CH-, -NR 10 C(O)NR10 -, heteroaryl-, aryl-, -NR 10 C(=NR 11 )NR 10 -, -NR 10 C(=NR 11 )-, -C(=NR 11 )NR 10 -, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0014] [ka] where: R6, R7, and R8 are independently selected from H, halogen, CN, OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; each R9 is independently selected from H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R 10 are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or The Two R's 10 the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R 10 and R 11 can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R 11is independently selected from H or substituted or unsubstituted alkyl; the compound is administered prior to or simultaneously with the allogeneic hematopoietic stem cells and / or allogeneic T cells. In some embodiments, L3, X, and L4 taken together form a nitrogen-containing heterocycle. In some embodiments, the nitrogen-containing heterocycle is a piperidine group. In some embodiments, G is

[0015] [ka] In some embodiments, disclosed herein are methods of preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of the onset of alloantibody-driven cGVHD in a patient in need of cell transplantation, the methods comprising administering a therapeutically effective amount of (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (ibrutinib).

[0016] [ka] In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the cGVHD is hepatic cGVHD. In some embodiments, the cGVHD is renal cGVHD. In some embodiments, the cGVHD is esophageal cGVHD. In some embodiments, the cGVHD is gastric cGVHD. In some embodiments, the patient is suffering from cancer. In some embodiments, the patient is suffering from a hematological malignancy. In some embodiments, the patient is suffering from a B-cell malignancy. In some embodiments, the cell transplant is a hematopoietic cell transplant. In some embodiments, the patient has received or will receive an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered simultaneously with an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered prior to an allogeneic bone marrow or hematopoietic stem cell transplant.

[0017] Disclosed herein, in some embodiments, is a method of treating a patient for the alleviation of an alloantibody response and consequently the alleviation of developed chronic graft-versus-host disease (cGVHD), comprising administering to the patient allogeneic hematopoietic stem cells and / or allogeneic T cells together with a therapeutically effective amount of an ACK inhibitor (e.g., an ITK or BTK inhibitor). Disclosed herein, in some embodiments, is a method of treating a patient for the alleviation of an alloantibody response and consequently the alleviation of developed chronic graft-versus-host disease (cGVHD), comprising administering to the patient allogeneic hematopoietic stem cells and / or allogeneic T cells and a therapeutically effective amount of a compound of Formula (A) or a pharmaceutically acceptable salt thereof:

[0018] [ka] During the ceremony: A is N; R1 is phenyl-O-phenyl or phenyl-S-phenyl; R2 and R3 are independently H; R4 is L3-X-L4-G, where: L3 is optional and, when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, when present, is a single bond, -O-, -C(=O)-, -S-, -S(=O)-, -S(=O)2-, -NH-, -NR9-, -NHC(O)-, -C(O)NH-, -NR9C(O)-, -C(O)NR9-, -S(=O)2NH-, -NHS(=O)2-, -S(=O)2NR9-, -NR9S(=O)2-, -OC(O)NH-, -NHC(O)O-, -OC(O)NR9-, -NR9C(O)O-, -CH=NO-, -ON=CH-, -NR 10 C(O)NR 10 -, heteroaryl-, aryl-, -NR 10 C(=NR 11 )NR 10 -, -NR 10 C(=NR 11 )-, -C(=NR 11 )NR 10 -, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0019] [ka] where: R6, R7, and R8 are independently selected from H, halogen, CN, OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; each R9 is independently selected from H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R 10 are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or The Two R's 10 the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R 10 and R 11 can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R 11 is independently selected from H, or substituted or unsubstituted alkyl. Disclosed herein, in some embodiments, are methods of treating a patient for attenuation of an alloantibody response, with consequent attenuation of developed chronic graft-versus-host disease (cGVHD), the methods comprising administering to the patient allogeneic hematopoietic stem cells and / or allogeneic T cells and a therapeutically effective amount of (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (ibrutinib).

[0020] [ka] In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the cGVHD is hepatic cGVHD. In some embodiments, the cGVHD is renal cGVHD. In some embodiments, the cGVHD is esophageal cGVHD. In some embodiments, the cGVHD is gastric cGVHD. In some embodiments, the patient is suffering from cancer. In some embodiments, the patient is suffering from a hematological malignancy. In some embodiments, the patient is suffering from a B-cell malignancy. In some embodiments, the cell transplant is a hematopoietic cell transplant. In some embodiments, the patient has received or will receive an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered simultaneously with an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered prior to an allogeneic bone marrow or hematopoietic stem cell transplant.

[0021] <Incorporated by reference> All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. [Brief explanation of the drawings]

[0022] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings. [Figure 1]These figures demonstrate that collagen deposition and lung function were therapeutically improved in a murine model of allo-HSCT-induced cGVHD with bronchiolitis obliterans. A-C) PFTs were performed on anesthetized animals 60 days after transplantation. Animals were artificially ventilated, and A) tolerance, B) elasticity, and C) compliance were measured as parameters of lung function distress in animals receiving bone marrow (BM) plus a low dose of splenocytes (S). Error bars = sem. D and E) Collagen deposition in lung tissue was determined using a Masson trichrome staining kit; blue indicates collagen deposition. D) Representative images of collagen deposition observed in each treatment cohort. Blue staining represents Mason trichrome-stained collagen. E) Quantification of collagen deposition as a ratio of blue area to total tissue area was performed using the analysis tool in Photoshop CS3. [Figure 2] Illustrating survival of cGVHD mice in the C57BL / 6→B10.BR model. Kaplan Meier plot of crude survival for bone marrow (BM)-free cGVHD mice, mice transplanted with BM+spleen cells (S) and treated with a non-cGVHD vehicle, or BM+S-transplanted mice treated with ibrutinib. [Figure 3] Illustrates the body weight of cGVHD mice in the C57BL / 6→B10.BR model. Body weight measurements for bone marrow (BM)-free cGVHD mice, BM+spleen cell (S)-transplanted mice treated with a non-cGVHD vehicle, or BM+S-transplanted mice treated with ibrutinib. [Figure 4] This figure illustrates that germinal center reactions and pulmonary immunoglobulin deposition are therapeutically attenuated by administration of ibrutinib. A) Germinal centers were imaged by staining 6 μm spleen sections with rhodamine-conjugated PNA. B) Splenocytes were purified from day 60 transplanted mice, and the frequency of germinal center B cells was quantified. C) 6 μm lung sections from day 60 transplanted mice were stained with FITC-conjugated anti-mouse Ig. D) Quantification was performed using Adobe Photoshop CS3. [Figure 5]Figure 1 illustrates that BTK expression in donor-derived B cells is required for the development of BO. A) Pulmonary function tests at day 60 from mice transplanted with low levels of WT T cells and WT or XID (kinase-inactive BTK) bone marrow. B and C) Histopathology scores of lung, liver, and spleen from transplanted mice at day 60. n = 5 mice per group from two independent experiments. [Figure 6] Figure 1 illustrates that the development of BO depends on the expression of ITK in donor mature T cells. A) Pulmonary function tests at day 60 of mice transplanted with WT bone marrow and either low numbers of WT T cells or ITK-deficient T cells. B and C) Histopathological scores of lungs, liver, and spleen of transplanted mice at day 60. n = 5 mice per group from two independent experiments. DETAILED DESCRIPTION OF THE INVENTION

[0023] Disclosed herein, in some embodiments, is a method of treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the method comprising administering a therapeutically effective amount of an ACK inhibitor (e.g., an ITK or BTK inhibitor). In some embodiments, a method of treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient is provided, the method comprising administering to a patient in need thereof a therapeutically effective amount of a compound of Formula (A) having the following structure:

[0024] [ka] During the ceremony: A is N; R1 is phenyl-O-phenyl or phenyl-S-phenyl; R2 and R3 are independently H; R4 is L3-X-L4-G, where: L3 is optional and, when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, when present, is a single bond, -O-, -C(=O)-, -S-, -S(=O)-, -S(=O)2-, -NH-, -NR9-, -NHC(O)-, -C(O)NH-, -NR9C(O)-, -C(O)NR9-, -S(=O)2NH-, -NHS(=O)2-, -S(=O)2NR9-, -NR9S(=O)2-, -OC(O)NH-, -NHC(O)O-, -OC(O)NR9-, -NR9C(O)O-, -CH=NO-, -ON=CH-, -NR 10 C(O)NR 10 -, heteroaryl-, aryl-, -NR 10 C(=NR 11 )NR 10 -, -NR 10 C(=NR 11 )-, -C(=NR 11 )NR 10 -, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0025] [ka] where: R6, R7, and R8 are independently selected from H, halogen, CN, OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; each R9 is independently selected from H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R 10 are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or The Two R's 10 the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R 10 and R 11 can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R 11 is independently selected from H or substituted or unsubstituted alkyl. In some embodiments, L3, X, and L4 taken together form a nitrogen-containing heterocycle. In some embodiments, the nitrogen-containing heterocycle is a piperidine group. In some embodiments, G is

[0026] [ka] In some embodiments, the compound of Formula (A) is (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (ibrutinib), or a pharmaceutically acceptable salt thereof.

[0027] [ka] In some embodiments, the patient exhibits one or more symptoms of alloantibody-driven cGVHD. In some embodiments, the alloantibody-driven cGVHD is untreated cGVHD. In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multi-organ cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the cGVHD is hepatic cGVHD. In some embodiments, the cGVHD is renal cGVHD. In some embodiments, the cGVHD is esophageal cGVHD. In some embodiments, the cGVHD is gastric cGVHD. In some embodiments, fibrosis is reduced. In some embodiments, pulmonary fibrosis is reduced. In some embodiments, liver fibrosis is reduced. In some embodiments, immunoglobulin (Ig) deposition in tissues is reduced. In some embodiments, the patient is suffering from cancer. In some embodiments, the patient is suffering from a hematological malignancy. In some embodiments, the patient is suffering from a relapsed or refractory hematological malignancy. In some embodiments, the patient is suffering from a B-cell malignancy. In some embodiments, the patient is suffering from a T-cell malignancy. In some embodiments, the patient is suffering from leukemia, lymphoma, or myeloma. In some embodiments, the B-cell malignancy is non-Hodgkin's lymphoma. In some embodiments, the B-cell malignancy is chronic lymphocytic leukemia (CLL). In some embodiments, the B-cell malignancy is a relapsed or refractory B-cell malignancy. In some embodiments, the B-cell malignancy is a relapsed or refractory non-Hodgkin's lymphoma. In some embodiments, the B-cell malignancy is relapsed or refractory CLL. In some embodiments, the patient is suffering from high-risk CLL. In some embodiments, the patient has a chromosome 17p deletion.In some embodiments, the patient has 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more CLL as determined by bone marrow biopsy. In some embodiments, the patient has previously received one or more anti-cancer agents. In some embodiments, the patient has previously undergone a cell transplant. In some embodiments, the cell transplant is a hematopoietic cell transplant. In some embodiments, the cell transplant is an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, the compound of Formula (A) is administered simultaneously with an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, the compound of Formula (A) is administered after an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, the amount of the ACK inhibitor compound (e.g., a compound of Formula (A)) prevents or reduces cGVHD while maintaining a graft-versus-leukemia (GVL) response effective to reduce or eliminate the number of cancer cells in the patient's blood. In some embodiments, the compound of Formula (A) is administered at a dosage of between about 0.1 mg / kg per day and about 100 mg / kg per day. In some embodiments, the amount of the compound of Formula (A) administered is about 40 mg / day, about 140 mg / day, about 420 mg / day, about 560 mg / day, or about 840 mg / day. In some embodiments, the compound of Formula (A) is administered from day 1 to about day 1000 after allogeneic bone marrow or hematopoietic stem cell transplantation. In some embodiments, the compound of Formula (A) is administered from the onset of symptoms of alloantibody-driven cGVHD to about day 1000 after allogeneic bone marrow or hematopoietic stem cell transplantation. In some embodiments, the compound of Formula (A) is administered orally. In some embodiments, the compound of Formula (A) is administered in combination with one or more additional therapeutic agents.

[0028] In some embodiments, disclosed herein are methods for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of an alloantibody-driven cGVHD episode in a patient in need of a cell transplant, the methods comprising administering a therapeutically effective amount of an ACK inhibitor (e.g., an ITK or BTK inhibitor). In some embodiments, disclosed herein are methods for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of an alloantibody-driven cGVHD episode in a patient in need of a cell transplant, the methods comprising administering a therapeutically effective amount of a compound of Formula (A), having the structure:

[0029] [ka] During the ceremony: A is N; R1 is phenyl-O-phenyl or phenyl-S-phenyl; R2 and R3 are independently H; R4 is L3-X-L4-G, where: L3 is optional and, when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, when present, is a single bond, -O-, -C(=O)-, -S-, -S(=O)-, -S(=O)2-, -NH-, -NR9-, -NHC(O)-, -C(O)NH-, -NR9C(O)-, -C(O)NR9-, -S(=O)2NH-, -NHS(=O)2-, -S(=O)2NR9-, -NR9S(=O)2-, -OC(O)NH-, -NHC(O)O-, -OC(O)NR9-, -NR9C(O)O-, -CH=NO-, -ON=CH-, -NR 10 C(O)NR 10 -, heteroaryl-, aryl-, -NR 10 C(=NR 11 )NR 10 -, -NR 10 C(=NR11 )-, -C(=NR 11 )NR 10 -, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0030] [ka] where: R6, R7, and R8 are independently selected from H, halogen, CN, OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; each R9 is independently selected from H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R 10 are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or The Two R's 10 the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R 10 and R 11 can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R 11 is independently selected from H or substituted or unsubstituted alkyl. In some embodiments, L3, X, and L4 taken together form a nitrogen-containing heterocycle. In some embodiments, the nitrogen-containing heterocycle is a piperidine group. In some embodiments, G is

[0031] [ka] In some embodiments, disclosed herein are methods of preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of the onset of alloantibody-driven cGVHD in a patient in need of cell transplantation, the method comprising administering to the patient a therapeutically effective amount of (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (ibrutinib).

[0032] [ka] In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the patient is suffering from cancer. In some embodiments, the patient is suffering from a hematological malignancy. In some embodiments, the patient is suffering from a B-cell malignancy. In some embodiments, the patient is suffering from a T-cell malignancy. In some embodiments, the patient is suffering from leukemia, lymphoma, or myeloma. In some embodiments, the amount of ibrutinib prevents or reduces alloantibody-driven cGVHD while maintaining a graft-versus-leukemia (GVL) response effective to reduce or eliminate the number of cancer cells in the patient's blood. In some embodiments, the cell transplant is a hematopoietic cell transplant. In some embodiments, the patient has undergone or will undergo an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered simultaneously with the allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered prior to the allogeneic bone marrow or hematopoietic stem cell transplant.

[0033] Disclosed herein, in some embodiments, is a method of treating a patient for the alleviation of an alloantibody response and consequently the alleviation of developed chronic graft-versus-host disease (cGVHD), comprising administering to the patient a therapeutically effective amount of allogeneic hematopoietic stem cells and / or allogeneic T cells, and an ACK inhibitor (e.g., an ITK or BTK inhibitor). Disclosed herein, in some embodiments, is a method of treating a patient for the alleviation of an alloantibody response and consequently the alleviation of developed chronic graft-versus-host disease (cGVHD), comprising administering to the patient a therapeutically effective amount of allogeneic hematopoietic stem cells and / or allogeneic T cells, and a compound of Formula (A) having the following structure, or a pharmaceutically acceptable salt thereof:

[0034] [ka] During the ceremony: A is N; R1 is phenyl-O-phenyl or phenyl-S-phenyl; R2 and R3 are independently H; R4 is L3-X-L4-G, where: L3 is optional and, when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, when present, is a single bond, -O-, -C(=O)-, -S-, -S(=O)-, -S(=O)2-, -NH-, -NR9-, -NHC(O)-, -C(O)NH-, -NR9C(O)-, -C(O)NR9-, -S(=O)2NH-, -NHS(=O)2-, -S(=O)2NR9-, -NR9S(=O)2-, -OC(O)NH-, -NHC(O)O-, -OC(O)NR9-, -NR9C(O)O-, -CH=NO-, -ON=CH-, -NR 10 C(O)NR 10 -, heteroaryl-, aryl-, -NR 10 C(=NR 11 )NR 10-, -NR 10 C(=NR 11 )-, -C(=NR 11 )NR 10 -, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0035] [ka] where: R6, R7, and R8 are independently selected from H, halogen, CN, OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; each R9 is independently selected from H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R 10 are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or The Two R's 10 the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R 10 and R 11 can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R 11 is independently selected from H or substituted or unsubstituted alkyl. In some embodiments, L3, X, and L4 taken together form a nitrogen-containing heterocycle. In some embodiments, the nitrogen-containing heterocycle is a piperidine group. In some embodiments, G is

[0036] [ka] Disclosed herein, in some embodiments, are methods of treating a patient for alleviation of an alloantibody response, with consequent alleviation of advanced chronic graft-versus-host disease (cGVHD), comprising administering allogeneic hematopoietic stem cells and / or allogeneic T cells and a therapeutically effective amount of (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (ibrutinib).

[0037] [ka] In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the patient is suffering from cancer. In some embodiments, the patient is suffering from a hematological malignancy. In some embodiments, the patient is suffering from a B-cell malignancy. In some embodiments, the patient is suffering from a T-cell malignancy. In some embodiments, the patient is suffering from leukemia, lymphoma, or myeloma. In some embodiments, ibrutinib prevents or reduces alloantibody-driven cGVHD while maintaining an effective graft-versus-leukemia (GVL) response to reduce or eliminate the number of cancer cells in the patient's blood. In some embodiments, the cell transplant is a hematopoietic cell transplant. In some embodiments, the patient has undergone or will undergo an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered simultaneously with the allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered prior to the allogeneic bone marrow or hematopoietic stem cell transplant.

[0038] In some embodiments, there is provided a use of a compound of Formula (A), or a pharmaceutically acceptable salt thereof, for treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient, wherein Formula (A) has the following structure:

[0039] [ka] During the ceremony: A is N; R1 is phenyl-O-phenyl or phenyl-S-phenyl; R2 and R3 are independently H; R4 is L3-X-L4-G, where: L3 is optional and, when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, when present, is a single bond, -O-, -C(=O)-, -S-, -S(=O)-, -S(=O)2-, -NH-, -NR9-, -NHC(O)-, -C(O)NH-, -NR9C(O)-, -C(O)NR9-, -S(=O)2NH-, -NHS(=O)2-, -S(=O)2NR9-, -NR9S(=O)2-, -OC(O)NH-, -NHC(O)O-, -OC(O)NR9-, -NR9C(O)O-, -CH=NO-, -ON=CH-, -NR 10 C(O)NR 10 -, heteroaryl-, aryl-, -NR 10 C(=NR 11 )NR 10 -, -NR 10 C(=NR 11 )-, -C(=NR 11 )NR 10 -, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0040] [ka] where: R6, R7, and R8 are independently selected from H, halogen, CN, OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; each R9 is independently selected from H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R 10 are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or The Two R's 10 the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R 10 and R 11 can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R 11 is independently selected from H or substituted or unsubstituted alkyl. In some embodiments, L3, X, and L4 taken together form a nitrogen-containing heterocycle. In some embodiments, the nitrogen-containing heterocycle is a piperidine group. In some embodiments, G is

[0041] [ka] In some embodiments, the compound of Formula (A) is (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (ibrutinib), or a pharmaceutically acceptable salt thereof.

[0042] [ka] In some embodiments, the patient exhibits one or more symptoms of cGVHD. In some embodiments, the cGVHD is untreated cGVHD. In some embodiments, the cGVHD is non-scleroderma cGVHD. In some embodiments, the cGVHD is multi-organ cGVHD. In some embodiments, the cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the cGVHD is pulmonary cGVHD. In some embodiments, fibrosis is reduced. In some embodiments, pulmonary fibrosis is reduced. In some embodiments, liver fibrosis is reduced. In some embodiments, immunoglobulin (Ig) deposition in tissues is reduced. In some embodiments, the patient is suffering from cancer. In some embodiments, the patient is suffering from a hematological malignancy. In some embodiments, the patient is suffering from a relapsed or refractory hematological malignancy. In some embodiments, the patient is suffering from a B-cell malignancy. In some embodiments, the patient is suffering from a T-cell malignancy. In some embodiments, the patient has leukemia, lymphoma, or myeloma. In some embodiments, the B cell malignancy is non-Hodgkin's lymphoma. In some embodiments, the B cell malignancy is chronic lymphocytic leukemia (CLL). In some embodiments, the B cell malignancy is a relapsed or refractory B cell malignancy. In some embodiments, the B cell malignancy is a relapsed or refractory non-Hodgkin's lymphoma. In some embodiments, the B cell malignancy is relapsed or refractory CLL. In some embodiments, the patient has high-risk CLL. In some embodiments, the patient has a chromosome 17p deletion. In some embodiments, the patient has 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more CLL as determined by bone marrow biopsy. In some embodiments, the patient has previously received one or more anti-cancer agents. In some embodiments, the patient has undergone a cell transplant. In some embodiments, the cell transplant is a hematopoietic cell transplant. In some embodiments, the cell transplant is an allogeneic bone marrow or hematopoietic stem cell transplant.In some embodiments, the compound of Formula (A) is administered simultaneously with allogeneic bone marrow or hematopoietic stem cell transplantation. In some embodiments, the compound of Formula (A) is administered after allogeneic bone marrow or hematopoietic stem cell transplantation. In some embodiments, the amount of ACK inhibitor compound (e.g., a compound of Formula (A)) prevents or reduces cGVHD while maintaining a graft-versus-leukemia (GVL) response effective to reduce or eliminate the number of cancer cells in the patient's blood. In some embodiments, the compound of Formula (A) is in an amount corresponding to a dosage of between about 0.1 mg / kg per day and about 100 mg / kg per day. In some embodiments, the compound of Formula (A) is in an amount of about 40 mg / day, about 140 mg / day, about 420 mg / day, about 560 mg / day, or about 840 mg / day. In some embodiments, the compound of Formula (A) is administered from day 1 to about 1000 after allogeneic bone marrow or hematopoietic stem cell transplantation. In some embodiments, the compound of Formula (A) is administered after allogeneic bone marrow or hematopoietic stem cell transplantation until about day 1000 from the onset of symptoms of alloantibody-driven cGVHD. In some embodiments, the compound of Formula (A) is suitable for oral administration. In some embodiments, the compound of Formula (A) is administered in combination with one or more additional therapeutic agents.

[0043] <Specific terminology> It is to be understood that the foregoing general description and the following detailed description are exemplary and illustrative only and are not restrictive of any claimed subject matter. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, as used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise. In this application, the use of "or" means "and / or" unless specifically stated otherwise. Furthermore, the use of the term "including," as well as other forms such as "include," "includes," and "included," is open-ended.

[0044] As used herein, "amelioration" refers to any reduction in severity, delay in onset, delay in progression, or reduction in duration of alloantibody-driven cGVHD, whether permanent, temporary, persistent, or instantaneous, that may result from or be associated with the administration of a compound or composition.

[0045] As used herein, "ACK" and "accessible cysteine kinase" are synonyms. They refer to kinases with accessible cysteine residues. ACKs include, but are not limited to, BTK, ITK, Bmx / ETK, TEC, EFGR, HER4, HER4, LCK, BLK, C-src, FGR, Fyn, HCK, Lyn, YES, ABL, Brk, CSK, FER, JAK3, and SYK. In some embodiments, the ACK is a TEC family kinase. In some embodiments, the ACK is HER4. In some embodiments, the ACK is BTK. In some embodiments, the ACK is ITK.

[0046] As used herein, the term "Bruton's tyrosine kinase" refers to Bruton's tyrosine kinase from Homo sapiens, as disclosed, for example, in U.S. Pat. No. 6,326,469 (GenBank Accession No. NP_000052).

[0047] The term "Bruton's tyrosine kinase homolog," as used herein, refers to an ortholog of Bruton's tyrosine kinase, e.g., a mouse (GenBank Accession No. AAB47246), dog (GenBank Accession No. XP_549139), rat (GenBank Accession No. NP_001007799), chicken (GenBank Accession No. NP_989564), or zebrafish (GenBank Accession No. XP_698117) ortholog, or a fusion protein of any of the foregoing that exhibits kinase activity for one or more substrates of Bruton's tyrosine kinase (e.g., a peptide substrate having the amino acid sequence "AVLESEEELYSSARQ" SEQ ID NO:1).

[0048] As used herein, the term "cognate cysteine" refers to a cysteine residue found in a sequence position homologous to cysteine 481 of Bruton's tyrosine kinase, as defined herein. For example, cysteine 482 is the cognate cysteine in the rat ortholog of Bruton's tyrosine kinase; cysteine 479 is the cognate cysteine in the chicken ortholog; and cysteine 481 is the cognate cysteine in the zebrafish ortholog. In another example, the cognate cysteine in TXK, a member of the Tec kinase family related to the Bruton's tyrosine kinase, is Cys350.

[0049] As used herein, the term "irreversible BTK inhibitor" refers to an inhibitor of BTK that can form a covalent bond with an amino acid residue of BTK. In one embodiment, an irreversible inhibitor of BTK can form a covalent bond with a cysteine residue of BTK; in a specific embodiment, an irreversible inhibitor can form a covalent bond with the Cys481 residue of BTK (or a homolog thereof) or a cysteine residue at the corresponding position in a homolog of another tyrosine kinase.

[0050] The terms "individual," "patient," and "subject" are used interchangeably. They refer to a mammal (e.g., a human) who is the object of treatment or observation. This term should not be construed as requiring the supervision of a medical professional (e.g., a physician, physician assistant, nurse, janitor, or hospice care worker).

[0051] The terms "treat," "treating," or "treatment," as used herein, include reducing the severity of alloantibody-driven cGVHD, slowing the onset of cGVHD, causing regression of cGVHD, alleviating the disease caused by cGVHD, or halting symptoms resulting from GVHD. The terms "treat," "treating," or "treatment" include, but are not limited to, prophylactic and / or therapeutic treatments.

[0052] As used herein, "alloantibody-driven chronic graft-versus-host disease" refers to chronic GVHD driven in part by alloantibody production after allogeneic transplantation, such as hematopoietic stem cell transplantation. In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD.

[0053] <Graft-versus-host disease> Disclosed herein, in some embodiments, are methods for treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the method comprising administering to the patient a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib), thereby treating the alloantibody-driven cGVHD. In some embodiments, the alloantibody-driven cGVHD is untreated cGVHD. In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the cGVHD is hepatic cGVHD. In some embodiments, the cGVHD is renal cGVHD. In some embodiments, the cGVHD is esophageal cGVHD. In some embodiments, the cGVHD is gastric cGVHD. In some embodiments, the patient has undergone a hematopoietic cell transplant. In some embodiments, the patient has undergone a peripheral blood stem cell transplant. In some embodiments, the patient has undergone a bone marrow transplant. In some embodiments, the ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered prior to administration of the cell transplant. In some embodiments, the ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered after administration of the cell transplant. In some embodiments, the ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered simultaneously with administration of the cell transplant. In some embodiments, the ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered after the onset of symptoms of alloantibody-driven cGVHD. In some embodiments, the patient exhibits one or more symptoms of alloantibody-driven cGVHD.

[0054] Further described herein, in some embodiments, are methods for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of alloantibody-driven cGVHD in a patient in need of cell transplantation, the methods comprising administering to the patient a composition comprising a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, ibrutinib, etc.). In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the patient requires a hematopoietic cell transplant. In some embodiments, the patient requires a peripheral blood stem cell transplant. In some embodiments, the patient requires a bone marrow transplant. In some embodiments, the ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered prior to administration of the cell transplant. In some embodiments, the ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered after administration of the cell transplant. In some embodiments, the ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered simultaneously with administration of the cell transplant. In some embodiments, the patient exhibits one or more symptoms of alloantibody-driven cGVHD.

[0055] Disclosed herein, in some embodiments, are methods for treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the method comprising administering a therapeutically effective amount of ibrutinib to the patient, thereby treating the alloantibody-driven cGVHD. In some embodiments, the alloantibody-driven cGVHD is untreated cGVHD. In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the patient has undergone hematopoietic cell transplantation. In some embodiments, the patient has undergone peripheral blood stem cell transplantation. In some embodiments, the patient has undergone bone marrow transplantation. In some embodiments, ibrutinib is administered prior to administration of the cell transplant. In some embodiments, ibrutinib is administered after administration of the cell transplant. In some embodiments, ibrutinib is administered simultaneously with administration of the cell transplant. In some embodiments, ibrutinib is administered after the onset of symptoms of alloantibody-driven cGVHD. In some embodiments, the patient exhibits one or more symptoms of alloantibody-driven cGVHD.

[0056] Described herein are methods for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of alloantibody-driven cGVHD in a patient requiring stem cell transplantation, the methods comprising administering to the patient a composition comprising a therapeutically effective amount of ibrutinib. In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the patient requires hematopoietic stem cell transplantation. In some embodiments, the patient requires peripheral blood stem cell transplantation. In some embodiments, the patient requires bone marrow transplantation. In some embodiments, ibrutinib is administered prior to administration of the stem cell transplant. In some embodiments, ibrutinib is administered after administration of the stem cell transplant. In some embodiments, ibrutinib is administered simultaneously with the administration of stem cell transplantation. In some embodiments, ibrutinib is administered before, after, or simultaneously with the administration of allogeneic hematopoietic stem cells and / or allogeneic T cells.

[0057] Further described herein are methods of treating a patient for the alleviation of an alloantibody response, with consequent alleviation of advanced chronic graft-versus-host disease (cGVHD), comprising administering allogeneic hematopoietic stem cells and / or allogeneic T cells to the patient, wherein a therapeutically effective amount of an ACK inhibitory compound (e.g., a BTK inhibitor such as ibrutinib) is administered before, after, or simultaneously with the administration of the allogeneic hematopoietic stem cells and / or allogeneic T cells.

[0058] Treatment of proliferative blood disorders, such as leukemia, lymphoma, and myeloma, typically involves one or more forms of chemotherapy and / or radiation therapy. These treatments not only destroy malignant cells but also healthy blood cells. Allogeneic hematopoietic cell transplantation is an effective therapy for treating many hematologic malignancies, including B-cell and T-cell malignancies. In allogeneic hematopoietic cell transplantation, bone marrow (or, in some cases, peripheral blood) from an unrelated or related (non-identical twin) donor is used to replace healthy blood cells destroyed in cancer patients. Bone marrow (or peripheral blood) contains stem cells, which are precursors for all heterogeneous cell types found in blood (e.g., red blood cells, phagocytes, platelets, and lymphocytes). Allogeneic hematopoietic cell transplantation is known to have both restorative and therapeutic effects. The restorative effect results from the ability of stem cells to repopulate the cellular components of blood. The therapeutic properties of allogeneic hematopoietic cell transplantation are primarily due to the graft-versus-leukemia (GVL) effect. Hematopoietic cells (specifically T lymphocytes) transplanted from the donor attack cancer cells, enhancing the palliative effects of other forms of treatment. Essentially, the GVL effect involves the attack of cancer cells by blood cells obtained from the transplant, reducing the likelihood of malignant tumors returning after transplantation. Controlling the GVL effect prevents its progression to GVHD. Similar effects on tumors (graft versus tumor) are also known.

[0059] Allogeneic hematopoietic cell transplants are often toxic to patients, resulting from the difficulty in separating the effects of GVL or GVT from graft-versus-host disease (GVHD), the most often fatal complication of allogeneic BMT.

[0060] Graft-versus-host disease (GVHD) is a major complication of allogeneic hematopoietic cell transplantation (hCT). GVHD is an inflammatory disease initiated by T cells in the donor graft that recognize host histocompatible and other tissue antigens. GVHD is mediated by various effector cells and inflammatory cytokines. GVHD manifests in both acute and chronic forms. The most commonly affected organs are the skin, liver, and gastrointestinal tract. GVHD may also involve other organs, such as the lungs. Treatment of GVHD is usually successful in only 50–75% of cases; the remaining patients usually do not survive. The risk and severity of this immune-mediated disease are directly related to the degree of mismatch between the host and the hematopoietic cell donor. For example, GVHD develops in up to 30% of recipients of human leukocyte antigen (HLA)-matched sibling bone marrow, up to 60% of recipients of HLA-matched unrelated donor bone marrow, and a higher percentage of recipients of HLA-mismatched bone marrow. Patients with mild intestinal GVHD experience loss of appetite, nausea, vomiting, abdominal pain, and diarrhea, while those with severe GVHD are disabling. If untreated, symptoms of intestinal GVHD persist and frequently progress; spontaneous remission is rare. In its most severe form, GVHD leads to necrosis and desquamation of most epithelial cells of the intestinal mucosa and frequently fatal disease. Symptoms of acute GVHD are usually present within 100 days of transplantation. Symptoms of chronic GVHD are usually present later, up to 3 years after allogeneic HCT, and are frequently preceded by a history of acute GVHD.

[0061] Described herein are methods for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of alloantibody-driven cGVHD in a patient requiring a cell transplant, the methods comprising administering to the patient a composition comprising a therapeutically effective amount of ibrutinib. In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the patient requires a hematopoietic cell transplant. Further described herein are methods of treating a patient for the alleviation of bone marrow-mediated disease, with consequent alleviation of advanced graft-versus-host disease (GVHD), comprising administering to the patient allogeneic hematopoietic stem cells and / or allogeneic T cells, wherein a therapeutically effective amount of an inhibitor (e.g., ibrutinib) precedes, follows, or is administered simultaneously with the administration of the allogeneic hematopoietic stem cells and / or allogeneic T cells. In some embodiments, the patient is suffering from cancer. In some embodiments, the patient is suffering from a hematological malignancy. In some embodiments, the patient is suffering from a B-cell malignancy. In some embodiments, the patient is suffering from a T-cell malignancy. In some embodiments, the patient is suffering from leukemia, lymphoma, or myeloma. In some embodiments, the compounds disclosed herein prevent or reduce cGVHD while maintaining a graft-versus-leukemia (GVL) response effective to reduce or eliminate the number of cancer cells in the patient's blood. In some embodiments, the patient has undergone or will undergo an allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered simultaneously with the allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered prior to the allogeneic bone marrow or hematopoietic stem cell transplant. In some embodiments, ibrutinib is administered after the allogeneic bone marrow or hematopoietic stem cell transplant.

[0062] In some embodiments, the patient has non-Hodgkin's lymphoma. In some embodiments, the patient has Hodgkin's lymphoma. In some embodiments, the patient has a B-cell malignancy.

[0063] Disclosed herein, in some embodiments, is a method of treating a patient for alleviation of an alloantibody response, with consequent alleviation of advanced chronic graft-versus-host disease (cGVHD), comprising administering to the patient allogeneic hematopoietic stem cells and / or allogeneic T cells together with a therapeutically effective amount of BK inhibitor A.

[0064] Disclosed herein, in some embodiments, are methods for treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the method comprising administering to the patient a therapeutically effective amount of a BTK inhibitor, thereby treating the alloantibody-driven cGVHD. In some embodiments, the alloantibody-driven cGVHD is untreated cGVHD. In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, fibrosis is reduced. In some embodiments, pulmonary fibrosis is reduced. In some embodiments, liver fibrosis is reduced. In some embodiments, immunoglobulin (Ig) deposition in tissues is reduced. In some embodiments, the patient has undergone a hematopoietic cell transplant. In some embodiments, the patient has undergone a peripheral blood stem cell transplant. In some embodiments, the patient has undergone a bone marrow transplant. In some embodiments, the BTK inhibitor is administered prior to administration of the cell transplant. In some embodiments, the BTK inhibitor is administered after administration of the cell transplant. In some embodiments, the BTK inhibitor is administered simultaneously with administration of the cell transplant. In some embodiments, the BTK inhibitor is administered after the onset of symptoms of alloantibody-driven cGVHD. In some embodiments, the patient exhibits one or more symptoms of alloantibody-driven cGVHD.

[0065] In some embodiments, described herein are methods for preventing the onset of or reducing the severity of alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient requiring a cell transplant, the methods comprising administering to the patient a composition comprising a therapeutically effective amount of a BTK inhibitor. In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the patient requires a hematopoietic cell transplant. In some embodiments, the patient requires a peripheral blood stem cell transplant. In some embodiments, the patient requires a bone marrow transplant. In some embodiments, the BTK inhibitor is administered prior to administration of the cell transplant. In some embodiments, the BTK inhibitor is administered after administration of the cell transplant. In some embodiments, the BTK inhibitor is administered simultaneously with the administration of the cell transplant. In some embodiments, the BTK inhibitor is administered before, after, or simultaneously with the administration of allogeneic hematopoietic stem cells and / or allogeneic T cells. In some embodiments, the patient exhibits one or more symptoms of alloantibody-driven cGVHD.

[0066] Disclosed herein, in some embodiments, are methods of treating a patient for alleviation of an alloantibody response, with consequent alleviation of advanced chronic graft-versus-host disease (cGVHD), comprising administering to the patient allogeneic hematopoietic stem cells and / or allogeneic T cells together with a therapeutically effective amount of an ITK inhibitor.

[0067] Disclosed herein, in some embodiments, are methods for treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the method comprising administering a therapeutically effective amount of an ITK inhibitor to the patient, thereby treating the alloantibody-driven cGVHD. In some embodiments, the alloantibody-driven cGVHD is untreated cGVHD. In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the patient has undergone hematopoietic cell transplantation. In some embodiments, the patient has undergone peripheral blood stem cell transplantation. In some embodiments, the patient has undergone bone marrow transplantation. In some embodiments, the ITK inhibitor is administered prior to administration of the cell transplant. In some embodiments, the ITK inhibitor is administered after administration of the cell transplant. In some embodiments, the ITK inhibitor is administered simultaneously with administration of the cell transplant. In some embodiments, the ITK inhibitor is administered after the onset of symptoms of alloantibody-driven cGVHD. In some embodiments, the patient exhibits one or more symptoms of alloantibody-driven cGVHD.

[0068] In some embodiments, described herein are methods for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of alloantibody-driven cGVHD in a patient requiring cell transplantation, the methods comprising administering to the patient a composition comprising a therapeutically effective amount of an ITK inhibitor. In some embodiments, the alloantibody-driven cGVHD is non-scleroderma cGVHD. In some embodiments, the alloantibody-driven cGVHD is multiorgan cGVHD. In some embodiments, the alloantibody-driven cGVHD is bronchiolitis obliterans syndrome. In some embodiments, the alloantibody-driven cGVHD is pulmonary cGVHD. In some embodiments, the patient requires hematopoietic cell transplantation. In some embodiments, the patient requires peripheral blood stem cell transplantation. In some embodiments, the patient requires bone marrow transplantation. In some embodiments, the ITK inhibitor is administered before administration of the cell transplant. In some embodiments, the ITK inhibitor is administered after administration of the cell transplant. In some embodiments, the ITK inhibitor is administered simultaneously with the administration of a cell transplant. In some embodiments, the ITK inhibitor is administered before, after, or simultaneously with the administration of allogeneic hematopoietic stem cells and / or allogeneic T cells. In some embodiments, the patient exhibits one or more symptoms of alloantibody-driven cGVHD.

[0069] <Combination therapy> Disclosed herein, in some embodiments, is a method of treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the method comprising administering a therapeutically effective amount of an ACK inhibitor (e.g., an ITK or BTK inhibitor) and an additional therapeutic agent.

[0070] Further described herein is a method for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of an alloantibody-driven cGVHD episode in a patient in need of cell transplantation, the method comprising administering to the patient a composition comprising a therapeutically effective amount of an ACK inhibitor compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) and an additional therapeutic agent.

[0071] Further described herein, in some embodiments, are methods of treating a patient for alleviation of an alloantibody response, with consequent alleviation of advanced chronic graft-versus-host disease (cGVHD), comprising administering to the patient allogeneic hematopoietic stem cells and / or allogeneic T cells, wherein a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) and an additional therapeutic agent precedes, follows, or is administered simultaneously with the administration of the allogeneic hematopoietic stem cells and / or allogeneic T cells. In some embodiments, the individual is administered an additional treatment, such as, but not limited to, extracorporeal photopheresis, or infusion of mesenchymal stem cells or donor lymphocytes.

[0072] In some embodiments, the additional therapeutic agent is an anti-GVHD therapeutic agent. In some embodiments, the anti-GVHD therapeutic agent is an immunosuppressant. In some embodiments, the immunosuppressant comprises cyclosporine, tacrolimus, methotrexate, mycophenolate mofetil, corticosteroids, azathioprine, or antithymocyte globulin (ATG). In some embodiments, the immunosuppressant is a monoclonal antibody (e.g., anti-CD3, anti-CD5, and anti-IL-2 antibodies). In some embodiments, the immunosuppressant is mycophenolate mofetil, alemtuzumab, antithymocyte globulin (ATG), sirolimus, tacrolimus, thalidomide, daclizumab, infliximab, or clofazimine, which are used to treat chronic GVHD. In some embodiments, the additional therapeutic agent is denileukin diftitox, defibrotide, budesonide, beclomethasone dipropionate, or pentostatin.

[0073] In some embodiments, the additional therapeutic agent is an IL-6 receptor inhibitor. In some embodiments, the additional therapeutic agent is an IL-6 receptor antibody.

[0074] In some embodiments, the additional therapeutic agent is a TLR5 agonist.

[0075] In some embodiments, the patient undergoes additional treatments such as extracorporeal photopheresis or infusion of mesenchymal stem cells or donor lymphocytes.

[0076] In some embodiments, the additional therapeutic agent is a locally acting corticosteroid (TAC), ie, beclomethasone dipropionate, alclometasone dipropionate, budesonide, 22S-budesonide, 22R-budesonide, beclomethasone-17-monopropionate, betamethasone, clobetasol propionate, dexamethasone, diflorasone acetate, flunisolide, fluocinonide, fludroxycortide, fluticasone propionate, halobetasol propionate, halcinonide, mometasone furoate, triamcinalone acetonide, or a combination thereof.

[0077] In some embodiments, the additional therapeutic agent is an antifungal agent, hi some embodiments, the additional therapeutic agent is nystatin, clotrimazole, amphotericin, fluconazole, itraconazole, or a combination thereof.

[0078] In some embodiments, the additional therapeutic agent is a sialagogue, hi some embodiments, the additional therapeutic agent is cevimeline, pilocarpine, bethanecol, or a combination thereof.

[0079] In some embodiments, the additional therapeutic agent is a local anesthetic, hi some embodiments, the additional therapeutic agent is lidocaine, dyclonine, diphenhydramine, doxepin, or a combination thereof.

[0080] The methods described herein may employ appropriate techniques for chemotherapy, biotherapy, immunosuppression, and radiation therapy known in the art. For example, the chemotherapeutic agent may be any agent that exhibits oncolytic effects on cancer or tumor cells in a subject. For example, the chemotherapeutic agent may be, but is not limited to, an anthracycline, an alkylating agent, an alkyl sulfonate, an aziridine, an ethyleneimine, a methyleneamine, a nitrogen mustard, a nitrosourea, an antibiotic, an antimetabolite, a folic acid analog, a purine analog, a pyrimidine analog, an enzyme, a podophyllotoxin, a platinum-containing agent, or a cytokine. Preferably, the chemotherapeutic agent is one known to be effective against the specific cancerous or neoplastic cell type. In some embodiments, the chemotherapeutic agent is effective in treating hematological malignancies, such as thiotepa, cisplatin-based compounds, and cyclophosphamide. Cytokines include interferons, G-CSF, erythropoietin, GM-CSF, interleukins, parathyroid hormone, and the like. Biotherapeutics include alemtuzumab, rituximab, bevacizumab, vascular disrupting agents, lenalidomide, etc. Radiosensitizers include nicotinamide, etc.

[0081] In some embodiments, the ACK inhibitor is administered in combination with a chemotherapeutic or biologic agent selected from an antibody, a B cell receptor pathway inhibitor, a T cell receptor inhibitor, a PI3K inhibitor, an IAP inhibitor, an mTOR inhibitor, a radioimmunotherapeutic agent, a DNA damaging agent, a histone deacetylase inhibitor, a protein kinase inhibitor, a hedgehog inhibitor, an Hsp90 inhibitor, a telomerase inhibitor, a Jak1 / 2 inhibitor, a protease inhibitor, an IRAK inhibitor, a PKC inhibitor, a PARP inhibitor, a CYP3A4 inhibitor, an AKT inhibitor, an Erk inhibitor, a proteosome inhibitor, an alkylating agent, an antimetabolite, a plant alkaloid, a terpenoid, a cytotoxin, a topoisomerase inhibitor, or a combination thereof. In some embodiments, the B cell receptor pathway inhibitor is a CD79A inhibitor, a CD79B inhibitor, a CD19 inhibitor, a Lyn inhibitor, a Syk inhibitor, a PI3K inhibitor, a Blk inhibitor, a PLCγ inhibitor, a PKCβ inhibitor, a CD22 inhibitor, a Bcl-2 inhibitor, an IRAK 1 / 4 inhibitor, a JAK inhibitor (e.g., ruxolitinib, baricitinib, CYT387, lestaurtinib, pacritinib, TG101348, SAR302503, tofacitinib (Xeljanz), etanercept (Enbrel), GLPG0634, R256), a microtubule inhibitor, a Topoisomerase inhibitor, a ribonucleotide analogue (RIA), ... II inhibitors, anti-TWEAK antibodies, anti-IL17 bispecific antibodies, CK2 inhibitors, anaplastic lymphoma kinase (ALK) and c-Met inhibitors, demethylase enzyme inhibitors such as demethylases, HDM, LSDI, and KDM, fatty acid synthase inhibitors such as spirocyclic piperidine derivatives, glucocorticosteroid receptor agonists, fused anti-CD19-cytotoxic agent conjugates, antimetabolites, p70S6K inhibitors, immunomodulatory components, AKT / PKB inhibitors, procaspase-3 activator PAC-1, BRAF inhibitors, lactate dehydrogenase A (LDH-A) inhibitors, CCR2 inhibitors, CXCR4 inhibitors, chemokine receptor antagonists, DNA double stranded break repair inhibitors, NOR202, GA-101, TLR2 inhibitors, or combinations thereof. In some embodiments, the T cell receptor inhibitor is muromonab-CD3. In some embodiments, the chemotherapeutic agent isRituximab (Rituxan), carfilzomib, fludarabine, cyclophosphamide, vincristine, prednisolone, chlorambucil, ifosfamide, doxorubicin, mesalazine, thalidomide, Revlimid, lenalidomide, temsirolimus, everolimus, fostamatinib, paclitaxel, docetaxel, ofatumumab, dexamethasone, bendamustine, prednisone, CAL-101, ibritumomab, tositumomab, bortezomib, pentostatin, endostatin, ritonavir, ketoconazole, anti-VEGF antibodies, Herceptin, cetuximab, cisplatin, carboplatin, docetaxel, erlotinib, etoposide e), 5-fluorouracil, gemcitabine, ifosfamide, imatinib mesylate (Gleevec), gefitinib, erlotinib, procarbazine, prednisone, irinotecan, leucovorin, mechlorethamine, methotrexate, oxaliplatin, paclitaxel, sorafenib, sunitinib, topotecan, vinblastine, GA-1 101, dasatinib, sipuleucel-T, disulfiram, epigallocatechin-3-gallate, salinosporamide A, ONX0912, CEP-18770, MLN9708, R-406, lenarinomide, spirocyclic piperidine derivatives, quinazoline-carboxamide-azetidine compounds, thiotepa, DWA2114R, NK121, IS 3 295, 254-S, alkyl sulfonates such as busulfan, improperfan, and piposulfan; aziridines such as benzodepa, carboquone, meturedepa, and uredepa; ethyleneimines, methylmelamines such as altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylmelamine; chlornaphazine; estramustine; ifosfamide; mechlorethamine; oxide hydrochloride; novobiocin; phenesterine; prednimustine; tofosfamide; uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; aclacinomycin, actinomycin, anthramycin, azaserine, bleomycin, cactinomycin, calicheamicin,Antibiotics such as carubicin, carminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, queramycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin, etc.; methotrexate Anti-metabolites such as tetracycline and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, and trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and floxuridine; androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testolactone; aminoglutethimide, mitotane,Anti-adrenals such as trilostane; folic acid supplements such as folinic acid; aceglatone; aldophosphamide glycosides; aminolevulinic acid; amsacrine; bestravcil; bisantrene; edatrexate; defosfamide; demecolcine; diaziconazole; eflornithine; elliptinium acetate; etoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllic acid; 2-ethylhydrazide; procarbazine; polysaccharide-K; razoxane; sizofiran; spirogermanium; tenuazonic acid ;Triaziquone;2,2',2"-Trichlorotriethylamine;Urethane;Vindesine;Dacarbazine;Mannomustine;Mitobronitol;Mitolactol;Pipobroman;Gacytosine;Cytosine arabinoside;Taxoids (e.g., paclitaxel and docetaxel);6-Thioguanine;Mercaptopurine;Methotrexate;Platinum analogs;Platinum;Etoposide (VP-16);Ifosfamide;Mitomycin C;Mitoxantrone;Vincristine;Vinorelbine;Navelbine;Novantrone;Teniposide;Daunomycin;Aminopterin;Xeloda;Ibandronate;CPT1 1; the topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO) retinoic acid; esperamycins; capecitabine; and pharmaceutically acceptable salts, acids, or derivatives thereof; antihormonal agents such as antiestrogens, including, for example, tamoxifen, raloxifene, the aromatase inhibitor 4(5)-imidazole, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene (Fareston); antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; AVL-263 (Avila Therapeutics / Celgene Corporation), AVL-292 (Avila Therapeutics / Celgene Corporation),AVL-291 (Avila Therapeutics / Celgene Corporation), BMS-488516 (Bristol-Myers Squibb), BMS-509744 (Bristol-Myers Squibb), CGI-1746 (CGI Pharma / Gilead Sciences), CTA-056, GDC-0834 (Genentech), HY-11066 (also CTK4I7891, HMS3265G21, HMS3265G22, HMS3265H21, HMS3265H22, 439574-61-5, AG-F-54930), ONO-4059 (Ono Pharmaceutical Co., Ltd.), ONO-WG37 (Ono Pharmaceutical Co., Ltd.), PLS-123 (Peking University), RN486 (Hoffmann-La ACK inhibitors such as HM71224 (Hanmi Pharmaceutical Company Limited), HM71224 (Roche), or a combination thereof.

[0082] When an additional agent is co-administered with an ACK inhibitor, the additional agent and the ACK inhibitor need not be administered in the same pharmaceutical composition, and optionally, due to different physical and chemical properties, are administered by different routes. Initial administration can be, for example, according to an established protocol, with subsequent changes in dosage, dosage form, and frequency of administration based on the observed effects.

[0083] By way of example only, if nausea is a side effect experienced by an individual after receiving an ACK inhibitor, it may then be appropriate to administer an anti-emetic drug in combination with the ACK inhibitor.

[0084] Alternatively, by way of example only, the therapeutic effect of an ACK inhibitor described herein may be enhanced by administration of an adjuvant (i.e., the adjuvant itself may have minimal therapeutic effect, but when combined with another therapeutic agent, the overall therapeutic effect on the patient may be enhanced). Alternatively, by way of example only, the benefit experienced by an individual may be increased by administering an ACK inhibitor described herein along with another therapeutic agent (including treatment regimen) that also has a therapeutic effect. In all cases, regardless of the disease or disorder being treated, the overall benefit experienced by the patient may, in some embodiments, be the mere addition of the two therapeutic agents, or in other embodiments, the patient may experience a synergistic effect.

[0085] The particular choice of compounds used will depend on the diagnosis of the attending physician and their judgment regarding the patient's condition and the appropriate treatment protocol. The compounds are optionally administered simultaneously (e.g., simultaneously, near simultaneously, or within the same treatment protocol) or sequentially, depending on the nature of the patient's disorder or disease and the actual choice of compounds used. The determination of the order of administration of each therapeutic agent during a treatment protocol, and the number of repeated administrations, is based on an evaluation of the disease being treated and the condition of the patient.

[0086] In some embodiments, when drugs are used in a combination treatment, the therapeutically effective amounts are different. Methods for experimentally determining the therapeutically effective amounts of drugs and other agents used in combination therapy regimens are described in the literature. For example, the use of metered dosing, i.e., providing more frequent, lower doses to minimize toxic side effects, has been extensively described in the literature. Combination treatments also include cyclical treatments that begin and end at different times to aid in the clinical management of patients.

[0087] With respect to the combination therapies described herein, the dosage of the co-administered compounds will, of course, vary depending on the type of drug being combined, the particular drug being utilized, the disorder being treated, etc. Additionally, when co-administered with an additional therapeutic agent, the ACK inhibitors described herein may be administered simultaneously with the additional therapeutic agent or sequentially. If administered sequentially, the attending physician will determine the appropriate sequence of proteins to administer in combination with the biologically active agent(s).

[0088] When the additional therapeutic agent and the ACK inhibitor are administered simultaneously, the multiple therapeutic agents are optionally provided in a single, combined form or in multiple forms (by way of example only, as a single pill or two separate pills). In some embodiments, one of the therapeutic agents is given in multiple doses, or both are given in multiple doses. If not simultaneous, the timing between the multiple doses is from about 0 weeks or more to about 4 weeks or less. In addition, combination methods, compositions, and formulations are not limited to the use of only two agents; the use of multiple therapeutic combinations is also contemplated.

[0089] It will be understood that dosage regimens to treat, prevent, or ameliorate the condition for which relief is sought may be modified in accordance with a variety of factors. These factors include the age, weight, sex, diet, and medical condition of the subject, as well as the disorder from which the subject suffers. Thus, the dosage regimen actually utilized may vary widely and, therefore, may deviate from the administration regimens set forth herein.

[0090] In some embodiments, the pharmaceutical agents comprising the combination therapy disclosed herein are administered in a combined dosage form or in separate dosage forms intended for substantially simultaneous administration. In some embodiments, the pharmaceutical agents comprising the combination therapy are administered sequentially, with any therapeutic compounds being administered in a regimen requiring two-step administration. In some embodiments, the two-step administration regimen requires sequential administration of the active agents or spaced-apart administration of the separate active agents. The time between multiple administration steps ranges from minutes to hours, depending on the properties of each pharmaceutical agent, such as potency, solubility, bioavailability, plasma half-life, and kinetics. In some embodiments, circadian variations in target molecule concentrations determine the optimal administration interval.

[0091] In some embodiments, the ACK inhibitor compound and the additional therapeutic agent are administered in a single dosage form. In some embodiments, the ACK inhibitor compound and the additional therapeutic agent are administered in separate dosage forms. In some embodiments, the ACK inhibitor compound and the additional therapeutic agent are administered simultaneously or sequentially.

[0092] <Administration> Described herein, in some embodiments, are methods for treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the methods comprising administering a therapeutically effective amount of an ACK inhibitor (e.g., an ITK or BTK inhibitor).

[0093] Further described herein is a method for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of an alloantibody-driven cGVHD onset in a patient in need of cell transplantation, the method comprising administering to the patient a composition comprising a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib).

[0094] Further described herein, in some embodiments, are methods of treating a patient for mitigation of an alloantibody response, with consequent mitigation of developed chronic graft-versus-host disease (cGVHD), comprising administering to the patient allogeneic hematopoietic stem cells and / or allogeneic T cells, wherein a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered before, after, or simultaneously with the administration of the allogeneic hematopoietic stem cells and / or allogeneic T cells.

[0095] In some embodiments, an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered before, during, or after the development of cGVHD. In some embodiments, an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is used as a prophylactic and is administered continuously to a subject (e.g., an allogeneic transplant recipient) who is predisposed to developing cGVHD. In some embodiments, an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered to an individual during or as soon as possible after the development of alloantibody-driven cGVHD. In some embodiments, administration of the ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is initiated within the first 48 hours of the onset of symptoms, within the first 6 hours of the onset of symptoms, or within 3 hours of the onset of symptoms. In some embodiments, the initial administration of an ACK inhibitor compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is via any route, such as intravenous injection, bolus injection, infusion over 5 minutes to about 5 hours, pill, capsule, tablet, transdermal patch, buccal delivery, or a combination thereof. The ACK inhibitor compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) should be administered as soon as practicable after the onset of a disorder is detected or suspected, and for as long as necessary to treat the disease, such as about one month to about three months. The length of treatment may vary for each subject, and can be determined using known classification criteria. In some embodiments, the ACK inhibitor compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered for at least two weeks, for about one month to about five years, or for about one month to about three years.

[0096] A therapeutically effective amount will depend on the severity and course of the disorder, previous treatment, the patient's health status, weight, and response to the drugs, and the judgment of the treating physician. A prophylactically effective amount will depend on the patient's condition, weight, severity and course of the disorder, previous treatment, response to the drugs, and the judgment of the treating physician.

[0097] In some embodiments, an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered to a patient periodically, for example, three times a day, twice a day, once a day, every other day, or every third day. In other embodiments, an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered to a patient intermittently, for example, twice a day, then once a day, then three times a day, or on the first two days of each week, or on the first, second, and third days of each week. In some embodiments, intermittent dosing is as effective as standard dosing. In further or alternative embodiments, an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered only when a patient exhibits a particular symptom, such as the onset of pain, fever, inflammation, or skin disorder. The dosing schedule for each compound may or may not be dependent on the others.

[0098] If the patient's disease does not improve, at the physician's discretion, the compound may be administered chronically, i.e., for an extended period of time, including for the entire lifespan of the patient, to ameliorate or otherwise control or limit the symptoms of the patient's disorder.

[0099] If the patient's condition improves, after the physician's judgment, the compound may be given continuously; alternatively, the administered dose of drug may be temporarily reduced or temporarily discontinued for a specified period of time (i.e., a "drug holiday"). The length of the drug holiday may vary between 2 days and 1 year (including, by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days). Dose reductions during drug holidays can be 10%-100% (including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%).

[0100] Once improvement in the patient's disease occurs, a maintenance regimen is administered if necessary. Thereafter, the dosage or frequency of administration of the ACK inhibitor compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) may be reduced as a function of symptoms to a level at which the individual's disease improvement is maintained. However, the individual may require intermittent treatment long-term after any recurrence of symptoms.

[0101] The amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) will vary depending on factors such as the particular compound, the disorder and its severity, the identity (e.g., weight) of the subject or host requiring treatment, and will be determined according to the particular circumstances surrounding the case, including, for example, the particular agent being administered, the route of administration, and the subject or host being treated. In general, however, doses utilized for adult human treatment will typically be in the range of 0.02-5000 mg per day, or 1-1500 mg per day. The desired dose may conveniently be provided in a single dose or in divided doses administered simultaneously (or over a short period of time) or at appropriate intervals (e.g., two, three, four, or more subdoses per day).

[0102] In some embodiments, the therapeutic amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is 100 mg / day to 2000 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is 140 mg / day to 840 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is 420 mg / day to 840 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 40 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 140 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 280 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 420 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 560 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 700 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 840 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 980 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 1120 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 1260 mg / day. In some embodiments, the amount of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is about 1400 mg / day. In some embodiments, the compound of Formula (A) is administered at a dosage of between about 0.1 mg / kg per day and about 100 mg / kg per day.

[0103] In some embodiments, the dosage of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is escalated over time. In some embodiments, the dosage of the ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) is escalated, for example, to about 1.25 mg / kg / day to 12.5 mg / kg / day over a predetermined period of time. In some embodiments, the predetermined period of time is 1 month or more, 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, 7 months or more, 8 months or more, 9 months or more, 10 months or more, 11 months or more, 12 months or more, 18 months or more, or 24 months or more.

[0104] ACK inhibitor compounds (e.g., ITK or BTK inhibitors, such as ibrutinib) may be formulated in unit dosage forms suitable for single administration of precise dosage amounts. In unit dosage forms, the formulation is divided into unit doses containing appropriate amounts of one or both compounds. The unit dosages may be in the form of packages containing discrete amounts of the formulation. Non-limiting examples are packaged tablets or capsules, and powders in vials or ampoules. Aqueous suspension compositions may be packaged in single-dose non-reclosable containers. Alternatively, multi-dose reclosable containers may be used, in which case it is common to include a preservative in the composition. By way of example only, formulations for parenteral injection may be provided in unit dosage forms, including, but not limited to, ampoules, or in multi-dose containers with added preservatives.

[0105] It is understood that a medical professional will determine the dosing regimen according to a variety of factors, including the severity of the subject's GVHD, as well as the subject's age, weight, sex, diet, and medical condition.

[0106] <Compound> Described herein, in some embodiments, are methods for treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the methods comprising administering a therapeutically effective amount of an ACK inhibitor (e.g., an ITK or BTK inhibitor).

[0107] Further described herein is a method for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of an alloantibody-driven cGVHD onset in a patient in need of cell transplantation, the method comprising administering to the patient a composition comprising a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib).

[0108] Further described herein, in some embodiments, are methods of treating a patient for mitigation of an alloantibody response, with consequent mitigation of developed chronic graft-versus-host disease (cGVHD), comprising administering to the patient allogeneic hematopoietic stem cells and / or allogeneic T cells, wherein a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered before, after, or simultaneously with the administration of the allogeneic hematopoietic stem cells and / or allogeneic T cells.

[0109] In the following description of irreversible BTK compounds suitable for use in the methods described herein, definitions referring to standard chemical terms (unless defined herein) can be found in reference materials, including Carey and Sundberg "Advanced Organic Chemistry 4th Ed." Vols. A (2000) and B (2001), Plenum Press, New York. Unless otherwise indicated, conventional methods of mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA technology, and pharmacology, within the skill of those in the art, will be used. In addition, nucleic acid and amino acid sequences for Btk (e.g., human Btk) are known in the art, as disclosed, for example, in U.S. Pat. No. 6,326,469. Unless specific definitions are given, the nomenclature utilized in conjunction with analytical chemistry, synthetic organic chemistry, medicinal chemistry, and pharmaceutical chemistry described herein, and the laboratory methods and techniques thereof, are known to those of skill in the art. Standard techniques may be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, delivery, and treatment of patients.

[0110] The BTK inhibitor compounds described herein are selective for BTK and kinases that have a cysteine residue at the amino acid sequence position of a tyrosine kinase homologous to the amino acid sequence position of cysteine 481 in BTK. Generally, irreversible inhibitors of BTK used in the methods described herein are identified or characterized in an in vitro assay, such as a cellular biochemical assay or a cellular functional assay. Such assays are useful for determining the in vitro IC50 for irreversible BTK inhibitor compounds.

[0111] For example, a cellular kinase assay can be used to determine BTK activity after incubation of the kinase with or without various concentrations of a candidate irreversible BTK inhibitor compound. If the candidate compound is indeed an irreversible BTK inhibitor, BTK kinase activity will not be restored by repeated washing with inhibitor-free medium. See, e.g., J.B. Small et al. (1999), J. Med. Chem. 42(10):1803-1815. Furthermore, the formation of a covalent complex between BTK and a candidate irreversible BTK inhibitor is a useful indicator of irreversible inhibition of BTK, which can be readily determined by numerous methods known in the art (e.g., mass spectrometry). For example, some irreversible BTK inhibitor compounds can form a covalent bond with Cys481 of BTK (e.g., via the Michael reaction).

[0112] Cellular functional assays for BTK inhibition involve measuring one or more cellular endpoints in response to stimulation of a BTK-mediated pathway in a cell line (e.g., BCR activation in Ramos cells) in the presence or absence of varying concentrations of a candidate irreversible BTK inhibitor compound. Useful endpoints for determining response to BCR activation include, for example, BTK autophosphorylation, phosphorylation of a BTK target protein (e.g., PLC-γ), and cytosolic calcium flux.

[0113] High-throughput assays for many cellular biochemical assays (e.g., kinase assays) and cellular functional assays (e.g., calcium flux) are well known to those of skill in the art. In addition, high-throughput screening systems are commercially available (see, e.g., Zymark Corp., Hopkinton, MA; Air Technical Industries, Mentor, OH; Beckman Instruments, Inc., Fullerton, CA; Precision Systems, Inc., Natick, MA, etc.). These systems typically automate the entire procedure, including all sample and reagent pipetting, liquid dispensing, timed incubation, and final reading of the microplate in a detector appropriate for the assay. Automated systems thereby enable the identification and characterization of a large number of irreversible BTK compounds without undue labor.

[0114] In some embodiments, the BTK inhibitor is selected from the group consisting of a small organic molecule, a macromolecule, a peptide, or a non-peptide.

[0115] In some embodiments, the BTK inhibitors provided herein are reversible or irreversible inhibitors. In certain embodiments, the BTK inhibitors are irreversible inhibitors.

[0116] In some embodiments, the irreversible BTK inhibitor forms a covalent bond with a cysteine side chain of Bruton's tyrosine kinase, a Bruton's tyrosine kinase homolog, or a BTK tyrosine kinase cysteine homolog.

[0117] Irreversible BTK inhibitor compounds can be used in the manufacture of a medicament for treating any of the aforementioned diseases (e.g., an autoimmune disease, an inflammatory disease, an allergic disorder, a B-cell proliferative disorder, or a thromboembolic disorder).

[0118] In some embodiments, the irreversible BTK inhibitor compound used for the methods described herein has a concentration of less than 10 μM (e.g., less than 1 μM, less than 0.5 μM, less than 0.4 μM, less than 0.3 μM, less than 0.1 μM, less than 0.08 μM, less than 0.06 μM, less than 0.05 μM, less than 0.04 μM, less than 0.03 μM, less than 0.02 μM, less than 0.01 μM, less than 0.008 μM, less than 0.006 μM, less than 0.005 μM). Inhibits the kinase activity of BTK or a BTK homolog with an in vitro IC50 of less than 0.004 μM, less than 0.003 μM, less than 0.002 μM, less than 0.001 μM, less than 0.00099 μM, less than 0.00098 μM, less than 0.00097 μM, less than 0.00096 μM, less than 0.00095 μM, less than 0.00094 μM, less than 0.00093 μM, less than 0.00092 μM, or less than 0.00090 μM).

[0119] In some embodiments, the irreversible BTK inhibitor compound is selected from ibrutinib (PCI-32765), PCI-45292, PCI-45466, AVL-101, AVL-291, AVL-292, or ONO-WG-37. In some embodiments, the irreversible BTK inhibitor compound is ibrutinib.

[0120] In one embodiment, an irreversible BTK inhibitor compound selectively and irreversibly inhibits the activated form of its target tyrosine kinase (e.g., the phosphorylated form of the tyrosine kinase). For example, activated BTK is transphosphorylated at tyrosine 551. Thus, in these embodiments, the irreversible BTK inhibitor inhibits the target kinase in a cell once the target kinase has been activated by a signaling event.

[0121] In other embodiments, the BTK inhibitor used in the methods described herein has the structure of any of Formula (A). Also described herein are pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically active metabolites, and pharmaceutically acceptable prodrugs of said compounds. Pharmaceutical compositions comprising at least one such compound, or a pharmaceutically acceptable salt, pharmaceutically acceptable solvate, pharmaceutically active metabolite, or pharmaceutically acceptable prodrug of such a compound, are provided.

[0122] Definitions of standard chemical terms can be found in reference sources, including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4TH ED." Vols. A (2000) and B (2001), Plenum Press, New York. Unless otherwise indicated, conventional methods of mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA technology, and pharmacology are used, within the scope of those skilled in the art. Unless specific definitions are given, the nomenclature utilized in conjunction with analytical chemistry, synthetic organic chemistry, medicinal chemistry, and pharmaceutical chemistry described herein, and the laboratory procedures and techniques therefor, are known to those of skill in the art. Standard techniques are optionally used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, and delivery, and patient treatment. Standard techniques are optionally used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipofection). Reaction and purification techniques are performed using documented methods or those described herein.

[0123] It is understood that the methods and compositions described herein are not limited to the particular methodology, protocols, cell lines, constructs, and reagents described herein, as such may vary. It is also understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the methods and compositions described herein, which will be limited only by the appended claims.

[0124] Unless otherwise specified, terms used in composite moieties (i.e., multiple linkages of moieties) are to be read equally from left to right or right to left. For example, an alkylenecycloalkylene group refers to both an alkylene group followed by a cycloalkylene group, or a cycloalkylene group followed by an alkylene group.

[0125] The suffix "ene" added to a group indicates that the group is a diradical. As just one example, methylene is the diradical of a methyl group, i.e., a "-CH-" group. And ethylene is the diradical of an ethyl group, i.e., a "-CHCH-" group.

[0126] An "alkyl" group refers to an aliphatic hydrocarbon group. The alkyl moiety includes "saturated alkyl" groups, which means that they do not contain any alkene or alkyne moieties. The alkyl moiety also includes "unsaturated alkyl" moieties, which means that they contain at least one alkene or alkyne moiety. An "alkene" moiety refers to a group having at least one carbon-carbon double bond, and an "alkyne" moiety refers to a group having at least one carbon-carbon triple bond. The alkyl moiety includes branched, straight-chain, or cyclic moieties, whether saturated or unsaturated. Depending on the structure, alkyl groups include monoradicals or diradicals (i.e., alkylene groups), and in the case of "lower alkyls," they have from 1 to 6 carbon atoms.

[0127] As used herein, C1-C x is C1-C2, C1-C3...C1-C x Includes.

[0128] An "alkyl" moiety optionally has 1 to 10 carbon atoms (wherever herein, a numerical range such as "1 to 10" refers to each integer in the specified range; for example, "1 to 10 carbon atoms" means that the alkyl group is selected from 1 carbon atom, 2 carbon atoms, 3 carbon atoms, and moieties containing up to 10 carbon atoms, but this definition also covers occurrences of the term "alkyl" where no numerical range is specified). The alkyl group of the compounds described herein may be designated as "C1-C4 alkyl" or similar designation. By way of example only, "C1-C4 alkyl" indicates that 1 to 4 carbon atoms are in the alkyl chain, i.e., the alkyl chain is selected from among methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and t-butyl. Thus, C1-C4 alkyl includes C1-C2 alkyl and C1-C3 alkyl. Alkyl groups are optionally substituted or unsubstituted. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl, ethenyl, propenyl, butenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0129] The term "alkenyl" refers to a type of alkyl group in which the first two atoms of the alkyl group form a double bond that is not part of an aromatic group. That is, an alkenyl group begins with the atom -C(R)=C(R)-R, where R refers to the remainder of the alkenyl group, which may be the same or different. The alkenyl portion is optionally branched, straight-chain, or cyclic (in which case it is also known as a "cycloalkenyl" group). Depending on the structure, an alkenyl group comprises a monoradical or a diradical (i.e., an alkenylene group). An alkenyl group is optionally substituted. Non-limiting examples of alkenyl groups include -CH=CH, -C(CH)=CH, -CH=CHCH, and -C(CH)=CHCH. Alkenylene groups include, but are not limited to, -CH=CH-, -C(CH)=CH-, -CH=CHCH-, -CH=CHCHCH-, -CH=CHCHCH-, and -C(CH)=CHCH-. Alkenyl groups optionally have from 2 to 10 carbons, and if "lower alkenyl," have from 2 to 6 carbon atoms.

[0130] The term "alkynyl" refers to a type of alkyl group in which the first two atoms of the alkyl group form a triple bond. That is, an alkynyl group begins with the atom -C≡CR, and R refers to the remainder of the alkynyl group, which may be the same or different. The "R" portion of the alkynyl moiety can be branched, straight-chain, or cyclic. Depending on the structure, alkynyl groups include monoradicals or diradicals (i.e., alkynylene groups). Alkynyl groups are optionally substituted. Non-limiting examples of alkynyl groups include, but are not limited to, -C≡CH, -C≡CCH, -C≡CCHCH, -C≡C-, and -C≡CCH-. Alkynyl groups have 2 to 10 carbons, and "lower alkynyl" groups have 2 to 6 carbon atoms.

[0131] An "alkoxy" group refers to a (alkyl)O- group, where alkyl is as defined herein.

[0132] "Hydroxyalkyl" refers to an alkyl radical, as defined herein, substituted with at least one hydroxyl group. Non-limiting examples of hydroxyalkyl include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl, 2,3-dihydroxypropyl, 1-(hydroxymethyl)-2-hydroxyethyl, 2,3-dihydroxybutyl, 3,4-dihydroxybutyl, and 2-(hydroxymethyl)-3-hydroxypropyl.

[0133] "Alkoxyalkyl" refers to an alkyl radical, as defined herein, substituted by an alkoxy group, as defined herein.

[0134] The term "alkylamine" refers to an -N(alkyl) x H y refers to a group where x and y are selected from among x=1, y=1, and x=2, y=0. When x=2, the alkyl group taken together with the N atom to which it is attached optionally forms a cyclic ring structure.

[0135] "Alkylaminoalkyl" refers to an alkyl radical, as defined herein, substituted with an alkylamine, as defined herein.

[0136] "Hydroxyalkylaminoalkyl" refers to an alkyl radical, as defined herein, substituted with an alkylamine and alkylhydroxy, as defined herein.

[0137] "Alkoxyalkylaminoalkyl" refers to an alkyl radical, as defined herein, substituted with an alkylamine and substituted by alkylalkoxy, as defined herein.

[0138] An "amide" is a chemical moiety with the formula -C(O)NHR or -NHC(O)R, where R is selected from alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heteroalicyclic (bonded through a ring carbon). In some embodiments, the amide moiety forms a linkage between an amino acid or peptide molecule and a compound described herein, thereby forming a prodrug. An amine or carboxyl side chain on a compound described herein can be aminated. Procedures and specific groups for making such amides can be found in sources such as Greene and Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley & Sons, New York, NY, 1999, the disclosure of which is incorporated herein by reference.

[0139] The term "ester" refers to a chemical moiety with the formula -COOR, where R is selected from among alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heteroalicyclic (bonded through a ring carbon). Any hydroxy or carboxyl side chain on the compounds described herein can be esterified. Procedures and specific groups for making such esters can be found in sources such as Greene and Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley & Sons, New York, NY, 1999, the disclosure of which is incorporated herein by reference.

[0140] As used herein, the term "ring" refers to any covalently closed structure. Rings include, for example, carbocycles (e.g., aryls and cycloalkyls), heterocycles (e.g., heteroaryls and non-aromatic heterocycles), aromatics (e.g., aryls and heteroaryls), and non-aromatics (e.g., cycloalkyls and non-aromatic heterocycles). Rings can be optionally substituted. Rings can be monocyclic or polycyclic.

[0141] As used herein, the term "ring system" refers to one or more rings.

[0142] The term "membered" can include any cyclic structure. The term "membered" refers to the number of skeletal atoms that make up the ring. Thus, for example, cyclohexyl, pyridine, pyran, and thiopyran are six-membered, and cyclopentyl, pyrrole, furan, and thiophene are five-membered.

[0143] The term "fused" refers to a structure in which two or more rings share one or more bonds.

[0144] The term "carbocyclic" or "carbocycle" refers to a ring in which each of the atoms forming the ring is a carbon atom. Carbocycle includes aryl and cycloalkyl. This term thus distinguishes heterocycle from heterocycle ("heterocyclic") in which the backbone of the ring contains at least one atom other than carbon (i.e., a heteroatom). Heterocycle includes heteroaryl and heterocycloalkyl. Carbocycles and heterocycles can be optionally substituted.

[0145] The term "aromatic" refers to a planar ring having a delocalized π-electron system containing 4n+2 π-electrons, where n is an integer. An aromatic ring can be formed from 5, 6, 7, 8, 9, or more than 9 atoms. An aromatic can be optionally substituted. The term "aromatic" includes both carbocyclic aryl (e.g., phenyl) and heterocyclic aryl (or "heteroaryl" or "heteroaromatic") groups (e.g., pyridine). The term includes monocyclic or fused polycyclic (i.e., rings that share adjacent paired carbon atoms) groups.

[0146] As used herein, the term "aryl" refers to an aromatic ring in which each of the atoms forming the ring is a carbon atom. The aryl ring can be formed by 5, 6, 7, 8, 9, or more than 9 carbon atoms. The aryl group can be optionally substituted. Examples of aryl groups include, but are not limited to, phenyl, naphthalenyl, phenanthrenyl, anthracenyl, fluorenyl, and indenyl. Depending on the structure, the aryl group can be a monoradical or a diradical (i.e., an arylene group).

[0147] An "aryloxy" group refers to an (aryl)O- group, where aryl is as defined herein.

[0148] As used herein, the term "carbonyl" refers to a group containing a moiety selected from the group consisting of -C(O)-, -S(O)-, -S(O)2-, and -C(S)-, including, but not limited to, at least one ketone group, at least one aldehyde group, at least one ester group, at least one carboxylic acid group, and / or at least one thioester group. The carbonyl group includes ketones, aldehydes, carboxylic acids, esters, and thioesters. In some embodiments, the group is part of a branched, straight, or cyclic molecule.

[0149] The term "cycloalkyl" refers to a monocyclic or polycyclic radical containing only carbon and hydrogen, and which is optionally saturated, partially unsaturated, or fully unsaturated. Cycloalkyl groups include groups having from 3 to 10 ring atoms. Illustrative examples of cycloalkyl groups include the following moieties:

[0150] [ka] Depending on the structure, a cycloalkyl group can be either a monoradical or a diradical (ie, a cycloalkylene group), and, if "lower cycloalkyl," has from 3 to 8 carbon atoms.

[0151] "Cycloalkylalkyl" means an alkyl radical, as defined herein, substituted with a cycloalkyl group. Non-limiting cycloalkylalkyl groups include cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, and the like.

[0152] The term "heterocycle" refers to heteroaromatic and heteroalicyclic groups containing 1 to 4 heteroatoms, each selected from O, S, and N, where each heterocycle group has 4 to 10 atoms in its ring system, provided that the ring of the group does not contain two adjacent O or S atoms. As used herein, whenever the number of carbon atoms in a heterocycle is indicated (e.g., a C1-C6 heterocycle), at least one other atom (heteroatom) is also present in the ring. A designation such as "C1-C6 heterocycle" refers only to the number of carbon atoms in the ring, not the total number of atoms in the ring. It is understood that a heterocycle may have additional heteroatoms in the ring. A designation such as "4- to 6-membered heterocycle" refers to the total number of atoms contained in the ring (i.e., 4-, 5-, or 6-membered, in which at least one atom is a carbon atom, at least one atom is a heteroatom, and the remaining 2 to 4 atoms are carbon atoms or heteroatoms). In heterocycles having two or more heteroatoms, the two or more heteroatoms can be the same or different. The heterocycle can be optionally substituted. Attachment to the heterocycle can be at a heteroatom or through a carbon atom. Non-aromatic heterocyclic groups include groups having only four atoms in their ring system, while aromatic heterocyclic groups must have at least five atoms in their ring system. Heterocyclic groups include benzo-fused ring systems. An example of a 4-membered heterocyclic group is azetidinyl (derived from azetidine). An example of a 5-membered heterocyclic group is thiazolyl. An example of a 6-membered heterocyclic group is pyridyl, and an example of a 10-membered heterocyclic group is quinolinyl.Examples of non-aromatic heterocyclic groups are pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyranyl, and 1,2,3,6-tetrahydropyranyl. dioxanyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, 3H-indolyl, and quinolizinyl. Examples of aromatic heterocyclic groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. The foregoing groups, as derived from the above groups, are optionally C-attached or N-attached where possible. For example, groups derived from pyrrole include pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached). Furthermore, groups derived from imidazole include imidazol-1-yl or imidazol-3-yl (both N-attached), or imidazol-2-yl, imidazol-4-yl, or imidazol-5-yl (all C-attached). Heterocyclic groups include benzo-fused ring systems and ring systems substituted with one or two oxo (=O) moieties, such as pyrrolidin-2-one. Depending on the structure, heterocyclic groups can be monoradicals or diradicals (i.e., heterocyclene groups).

[0153] The term "heteroaryl," or alternatively, "heteroaromatic," refers to an aromatic group containing one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. An N-containing "heteroaromatic" or "heteroaryl" moiety refers to an aromatic group in which at least one of the skeletal atoms of the ring is a nitrogen atom. Illustrative examples of heteroaryl groups include the following moieties:

[0154] [ka] Depending on the structure, a heteroaryl group can be a monoradical or a diradical (i.e., a heteroarylene group).

[0155] As used herein, the terms "non-aromatic heterocycle," "heterocycloalkyl," or "heteroalicyclic" refer to a non-aromatic ring in which one or more atoms forming the ring are heteroatoms. A "non-aromatic heterocycle" or "heterocycloalkyl" group refers to a cycloalkyl group containing at least one heteroatom selected from nitrogen, oxygen, and sulfur. In some embodiments, the radical is fused to an aryl or heteroaryl. A heterocycloalkyl ring can be formed by 3, 4, 5, 6, 7, 8, 9, or more atoms. A heterocycloalkyl ring can be optionally substituted. In certain embodiments, a non-aromatic heterocycle includes one or more carbonyl or thiocarbonyl groups, such as, for example, oxo- and thio-containing groups. Examples of heterocycloalkyls are lactams, lactones, cyclic imides, cyclic thioimides, cyclic carbamates, tetrahydrothiopyrans, 4H-pyrans, tetrahydropyrans, piperidines, 1,3-dioxins, 1,3-dioxanes, 1,4-dioxines, 1,4-dioxanes, piperazines, 1,3-oxathianes, 1,4-oxathiines, 1,4-oxathianes, tetrahydro-1,4-thiazines, 2H-1,2-oxazines, maleimides, succinimides, barbituric acids, thiobarbituric acids, dioxopiperazines, hydantoins, dihydro-1,4-thiazines, 2H-1,2-ox ... Examples of heterocycloalkyl groups include, but are not limited to, hydrouracil, morpholine, trioxane, hexahydro-1,3,5-triazine, tetrahydrothiophene, tetrahydrofuran, pyrroline, pyrrolidine, pyrrolidone, pyrrolidione, pyrazoline, pyrazolidine, imidazoline, imidazolidine, 1,3-dioxole, 1,3-dioxolane, 1,3-dithiol, 1,3-dithiolane, isoxazoline, isoxazolidine, oxazoline, oxazolidine, oxazolidinone, thiazoline, thiazolidine, and 1,3-oxathiolane. Illustrative examples of heterocycloalkyl groups, also referred to as non-aromatic heterocycles, include the following:

[0156] [ka] The term "heteroalicyclic" also includes all cyclic forms of carbohydrates, including but not limited to monosaccharides, disaccharides, and oligosaccharides. Depending on the structure, a heterocycloalkyl group can be a monoradical or a diradical (i.e., a heterocycloalkylene group).

[0157] The term "halo", or alternatively "halogen", or "halide", refers to fluoro, chloro, bromo, and iodo.

[0158] The term "haloalkyl" refers to an alkyl structure in which at least one hydrogen atom has been replaced with a halogen atom. In certain embodiments in which two or more hydrogen atoms have been replaced with halogen atoms, the halogen atoms are all the same as each other. In other embodiments in which two or more hydrogen atoms have been replaced with halogen atoms, the halogen atoms are not all the same as each other.

[0159] As used herein, the term "fluoroalkyl" refers to an alkyl group in which at least one hydrogen is replaced with a fluorine atom. Examples of fluoroalkyl groups include, but are not limited to, -CF, -CHCF, -CFCF, -CHCHCF, etc.

[0160] As used herein, the term "heteroalkyl" refers to an optionally substituted alkyl radical in which one or more skeletal atoms is a heteroatom (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, or a combination thereof). The heteroatom may be placed at any interior position of the heteroalkyl group or at the position at which the heteroalkyl group is attached to the remainder of the molecule. Examples include, but are not limited to, -CH2-O-CH3, -CH2-CH2-O-CH3, -CH2-NH-CH3, -CH2-CH2-NH-CH3, -CH2-N(CH3)-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2, -S(O)-CH3, -CH2-CH2-S(O)-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, and -CH=CH-N(CH3)-CH3. Additionally, in some embodiments, up to two heteroatoms are consecutive, such as, by way of example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3.

[0161] The term "heteroatom" refers to an atom other than carbon or hydrogen. Heteroatoms are typically independently selected from among oxygen, sulfur, nitrogen, silicon, and phosphorus, but are not limited to these atoms. In embodiments in which two or more heteroatoms are present, the two or more heteroatoms can all be the same as one another, or some or all of the two or more heteroatoms can be different from each other.

[0162] The term "bond" or "single bond" refers to a chemical bond between two atoms or two moieties when the atoms connected by the single bond are considered to be part of a larger substructure.

[0163] The term "moiety" refers to a specific segment or functional group of a molecule. Chemical moieties are often recognized as chemical entities embedded in or attached to a molecule.

[0164] The "thioalkoxy" group or "alkylthio" group refers to an -S-alkyl group.

[0165] The "SH" group is also referred to as a thiol group or a sulfhydryl group.

[0166] The terms "optionally substituted" or "substituted" mean that the reference group can be substituted by one or more additional groups individually and independently selected from alkyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, alkylthio, arylthio, alkylsulfoxide, arylsulfoxide, alkylsulfone, arylsulfone, cyano, halo, acyl, nitro, haloalkyl, fluoroalkyl, amino (including mono- and di-substituted amino groups), and their protected derivatives. As an example, an optional substituent can be L s R s where each L s is independently selected from a single bond, -O-, -C(=O)-, -S-, -S(=O)-, -S(=O)2-, -NH-, -NHC(O)-, -C(O)NH-, S(=O)2NH-, -NHS(=O)2, -OC(O)NH-, -NHC(O)O-, -(substituted or unsubstituted C1-C6 alkyl), or -(substituted or unsubstituted C2-C6 alkenyl), and each R s is independently selected from H, (substituted or unsubstituted C1-C4 alkyl), (substituted or unsubstituted C3-C6 cycloalkyl), heteroaryl, or heteroalkyl. The protecting groups that form the protected derivatives of the above substituents are found in sources such as the above "Greene and Wuts".

[0167] <ACK inhibitor compound> This specification describes, in some embodiments, a method for treating chronic graft-versus-host disease (cGVHD) driven by allogeneic antibodies in a patient in need thereof, the method comprising administering a therapeutically effective amount of an ACK inhibitor (e.g., an ITK or BTK inhibitor).

[0168] Further described herein is a method for preventing the onset of or reducing the severity of the onset of graft-versus-host disease (cGVHD) in a patient in need of cell transplantation, the method comprising administering to the patient a composition comprising a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib).

[0169] Further described herein are methods of treating a patient for the alleviation of bone marrow-mediated disease, with consequent alleviation of advanced graft-versus-host disease (cGVHD), comprising administering to the patient allogeneic hematopoietic stem cells and / or allogeneic T cells, wherein a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered before, after, or simultaneously with the administration of the allogeneic hematopoietic stem cells and / or allogeneic T cells.

[0170] The ACK inhibitor compounds described herein are selective for kinases that have an accessible cysteine on the inhibitor compound that can form a covalent bond with a Michael acceptor moiety. In some embodiments, the cysteine residue is accessible or becomes accessible when the binding site portion of the irreversible inhibitor binds to the kinase. That is, the binding site portion of the irreversible inhibitor binds to the active site of the ACK, and the Michael acceptor moiety of the irreversible inhibitor gains access (in one embodiment, the binding process leads to a conformational change in the ACK, thereby exposing the cysteine) or is otherwise exposed to the cysteine residue of the ACK; a covalent bond is then formed between the "S" of the cysteine residue and the Michael acceptor of the irreversible inhibitor. Consequently, the binding site portion of the irreversible inhibitor remains bound or otherwise blocks the active site of the ACK.

[0171] In some embodiments, the ACK is BTK, a BTK homolog, or a tyrosine kinase having a cysteine residue at an amino acid sequence position homologous to cysteine 481 in BTK. In some embodiments, the ACK is ITK. In some embodiments, the ACK is HER4. The inhibitor compounds described herein comprise a Michael acceptor moiety, a binding site moiety, and a linker, where the linker connects the binding site moiety and the Michael acceptor moiety (and, in some embodiments, the structure of the linker provides a conformation or otherwise orients the Michael acceptor moiety to improve the selectivity of the irreversible inhibitor for a particular ACK). In some embodiments, the ACK inhibitor inhibits ITK and BTK.

[0172] In some embodiments, the ACK inhibitor is a compound of Formula (A), and pharmaceutically acceptable metabolites, pharmaceutically acceptable solvates, pharmaceutically acceptable salts, or pharmaceutically acceptable prodrugs thereof:

[0173] [ka] During the ceremony A is independently selected from N or CR5; R1 is H, L2-(optionally substituted alkyl), L2-(optionally substituted cycloalkyl), L2-(optionally substituted alkenyl), L2-(optionally substituted cycloalkenyl), L2-(optionally substituted heterocycle), L2-(optionally substituted heteroaryl), L2-(optionally substituted aryl), where L2 is a single bond, O, S, -S(=O), -S(=O)2, C(=O), -(optionally substituted C1-C6 alkyl), or -(optionally substituted C2-C6 alkenyl); R2 and R3 are independently selected from H, lower alkyl, and substituted lower alkyl; R4 is L3-X-L4-G, where: L3 is optional and, when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, when present, may be a single bond, O, -C(=O), S, -S(=O), -S(=O), -NH, -NR, -NHC(O), -C(O)NH, -NRC(O), -C(O)NR, -S(=O)NH, -NHS(=O), -S(=O)NR, -NRS(=O), -OC(O)NH-, -NHC(O)O, -OC(O)NR, -NRC(O)O, -CH=NO-, -ON=CH-, -NR 10 C(O)NR 10 -, heteroaryl, aryl, -NR 10 C(=NR 11 )NR 10 -, -NR 10 C(=NR 11 )-, -C(=NR 11 )NR 10 -, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0174] [ka] where: R6, R7, and R8 are independently selected from H, lower alkyl or substituted lower alkyl, lower heteroalkyl or substituted lower heteroalkyl, substituted or unsubstituted lower cycloalkyl, substituted or unsubstituted lower heterocycloalkyl; R5 is H, halogen, -L6-(substituted or unsubstituted C1-C3 alkyl), -L6-(substituted or unsubstituted C2-C4 alkenyl), -L6-(substituted or unsubstituted heteroaryl), or -L6-(substituted or unsubstituted aryl), where L6 is a single bond, O, S, -S(=O), S(=O)2, NH, C(O), -NHC(O)O, -OC(O)NH, -NHC(O), or C(O)NH; each R9 is independently selected from H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R 10 are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or The Two R's 10 the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R 10 and R 11 can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R 11 is independently selected from H or alkyl.

[0175] In some embodiments, the compound of Formula (A) is a BTK inhibitor. In some embodiments, the compound of Formula (A) is an ITK inhibitor. In some embodiments, the compound of Formula (A) inhibits ITK and BTK. In some embodiments, the compound of Formula (A) has the following structure:

[0176] [ka] During the ceremony: A is N; R2 and R3 are each H; R1 is phenyl-O-phenyl or phenyl-S-phenyl; and R4 is L3-X-L4-G, where: L3 is optional and, when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, when present, may be a single bond, O, -C(=O), S, -S(=O), -S(=O), -NH, -NR, -NHC(O), -C(O)NH, -NRC(O), -C(O)NR, -S(=O)NH, -NHS(=O), -S(=O)NR, -NRS(=O), -OC(O)NH-, -NHC(O)O, -OC(O)NR, -NRC(O)O, -CH=NO-, -ON=CH-, -NR 10 C(O)NR 10 -, heteroaryl, aryl, -NR 10 C(=NR 11 )NR 10 -, -NR 10 C(=NR 11 )-, -C(=NR 11 )NR 10 -, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0177] [ka] where: R6, R7, and R8 are independently selected from H, lower alkyl or substituted lower alkyl, lower heteroalkyl or substituted lower heteroalkyl, substituted or unsubstituted lower cycloalkyl, and substituted or unsubstituted lower heterocycloalkyl.

[0178] In some embodiments, the ACK inhibitor is (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (i.e., PCI-32765 / ibrutinib).

[0179] [ka]

[0180] In some embodiments, the ACK inhibitor is ibrutinib, PCI-45292, PCI-45466, AVL-101 / CC-101 (Avila Therapeutics / Celgene Corporation), AVL-263 / CC-263 (Avila Therapeutics / Celgene Corporation), AVL-292 / CC-292 (Avila Therapeutics / Celgene Corporation), AVL-291 / CC-291 (Avila Therapeutics / Celgene Corporation), BMS-488516 (Bristol-Myers Squibb), BMS-509744 (Bristol-Myers Squibb), CGI-1746 (CGI Pharma / Gilead Sciences), CGI-560 (CGI Pharma / Gilead Sciences), Sciences), CTA-056, GDC-0834 (Genentech), HY-11066 (also CTK4I7891, HMS3265G21, HMS3265G22, HMS3265H21, HMS3265H22, 439574-61-5, AG-F-54930), ONO-4059 (Ono Pharmaceutical Co., Ltd.), ONO-WG37 (Ono Pharmaceutical Co., Ltd.), PLS-123 (Peking University), RN486 (Hoffmann-La Roche), HM71224 (Hanmi Pharmaceutical Company Limited), LFM-A13, BGB-3111 (Beigene), KBP-7536 (KBP BioSciences), ACP-196 (Acerta Pharma), or JTE-051(Japan Tobacco) Inc.

[0181] In some embodiments, the ACK inhibitor is 4-(tert-butyl)-N-(2-methyl-3-(4-methyl-6-((4-(morpholine-4-carbonyl)phenyl)amino)-5-oxo-4,5-dihydropyrazin-2-yl)phenyl)benzamide (CGI-1746); 7-benzyl-1-(3-(piperidin-1-yl)propyl)-2-(4-(pyridin-4-yl)phenyl)- 1H-Imidazo[4,5-g]quinoxalin-6(5H)-one (CTA-056); (R)-N-(3-(6-(4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenylamino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide (GDC-0834); 6-cyclo Propyl-8-fluoro-2-(2-hydroxymethyl-3-{1-methyl-5-[5-(4-methyl-piperazin-1-yl)-pyridin-2-ylamino]-6-oxo-1,6-dihydro-pyridin-3-yl}-phenyl)-2H-isoquinolin-1-one (RN-486); N-[5-[5-(4-acetylpiperazine-1-carbonyl)-4-methoxy-2-methylphenyl]sulfanyl- 1,3-thiazol-2-yl]-4-[(3,3-dimethylbutan-2-ylamino)methyl]benzamide (BMS-509744, HY-11092); or N-(5-((5-(4-acetylpiperazine-1-carbonyl)-4-methoxy-2-methylphenyl)thio)thiazol-2-yl)4-(((3-methylbutan-2-yl)amino)methyl)benzamide (HY11066).

[0182] In some embodiments, the ACK inhibitor is

[0183] [ka]

[0184] [ka]

[0185] [Chemical formula] is.

[0186] <BTK inhibitor> In some embodiments, the ACK inhibitor is a BTK inhibitor. The BTK inhibitory compounds described herein are selective for BTK and kinases having a cysteine residue at the amino acid sequence position homologous to the amino acid sequence position of cysteine 481 in BTK. The Btk inhibitory compound can form a covalent bond with Cys481 of Btk (e.g., via a Michael reaction).

[0187] In some embodiments, the BTK inhibitor is a compound of formula (A) having the following structure, or a pharmaceutically acceptable salt thereof:

[0188] [Chemical formula] Where: A is N; R1 is phenyl-O-phenyl or phenyl-S-phenyl; R2 and R3 are independently H; R4 is L3-X-L4-G, where L3 is optional, and when present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional, and when present, is a single bond, -O-, -C(=O)-, -S-, -S(=O)-, -S(=O)2-, -NH-, -NR9-, -NHC(O)-, -C(O)NH-, -NR9C(O)-, -C(O)NR9-, -S(=O)2NH-, -NHS(=O)2-, -S(=O)2NR9-, -NR9S(=O)2-, -OC(O)NH-, -NHC(O)O-, -OC(O)NR9-, -NR9C(O)O-, -CH=NO-, -ON=CH-, -NR 10 C(O)NR 10-, heteroaryl-, aryl-, -NR 10 C(=NR 11 )NR 10 -, -NR 10 C(=NR 11 )-, -C(=NR 11 )NR 10 -, -OC(=NR 11 )- or -C(=NR 11 )O-; L4 is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; or L3, X, and L4 taken together form a nitrogen-containing heterocycle; G is

[0189] [ka] where: R6, R7, and R8 are independently selected from H, halogen, CN, OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; each R9 is independently selected from H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R 10 are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or The Two R's 10 the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R 10 and R 11 can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R 11is independently selected from H or substituted or unsubstituted alkyl. In some embodiments, L3, X, and L4 taken together form a nitrogen-containing heterocycle. In some embodiments, the nitrogen-containing heterocycle is a piperidine group. In some embodiments, G is

[0190] [ka] In some embodiments, the compound of Formula (A) is 1-[(3R)-3-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl)prop-2-en-1-one.

[0191] In some embodiments, the BTK inhibitor compound of Formula (A) has Formula (B), and a pharmaceutically acceptable metabolite, pharmaceutically acceptable solvate, pharmaceutically acceptable salt, or pharmaceutically acceptable prodrug thereof, of the following structure:

[0192] [ka] During the ceremony: Y is alkyl or substituted alkyl, or 4-, 5-, or 6-membered cycloalkyl; Each Ra is independently H, halogen, —CF, —CN, —NO, OH, NH, —La-(substituted or unsubstituted alkyl), —La-(substituted or unsubstituted alkenyl), —La-(substituted or unsubstituted heteroaryl), or —La-(substituted or unsubstituted aryl), where L a is a single bond, O, S, -S(=O), -S(=O)2, NH, C(O), CH2, -NHC(O)O, -NHC(O), or -C(O)NH; G is

[0193] [ka] where: R6, R7, and R8 are independently selected from H, lower alkyl or substituted lower alkyl, lower heteroalkyl or substituted lower heteroalkyl, substituted or unsubstituted lower cycloalkyl, substituted or unsubstituted lower heterocycloalkyl; R 12 is H or lower alkyl; or Both Y and R obtained 12 form a 4-, 5-, or 6-membered heterocyclic ring.

[0194] In some embodiments, G is

[0195] [ka] In some embodiments,

[0196] [ka] teeth,

[0197] [ka] is selected from among:

[0198] In some embodiments, the BTK inhibitor compound of Formula (A) has the following structure: Formula (C), and a pharmaceutically acceptable metabolite, pharmaceutically acceptable solvate, pharmaceutically acceptable salt, or pharmaceutically acceptable prodrug thereof:

[0199] [ka] Y is alkyl or substituted alkyl, or 4-, 5-, or 6-membered cycloalkyl; R 12 is H or lower alkyl; or Y and R obtained together 12 forms a 4-, 5- or 6-membered heterocyclic ring; G is

[0200] [ka] where: R6, R7, and R8 are independently selected from H, lower alkyl or substituted lower alkyl, lower heteroalkyl or substituted lower heteroalkyl, substituted or unsubstituted lower cycloalkyl, and substituted or unsubstituted lower heterocycloalkyl.

[0201] In some embodiments, the "G" group in any of Formula (A), Formula (B), or Formula (C) is an optional group used to adjust the physical and biological properties of a molecule. Such adjustments / modifications are achieved using groups that adjust the Michael acceptor chemical reactivity, acidity, basicity, lipophilicity, solubility, and other physical properties of a molecule. The physical and biological properties adjusted by such modifications to G include, by way of example only, enhancing the chemical reactivity of Michael acceptor groups, solubility, in vivo absorption, and in vivo metabolism. In addition, in vivo metabolism may include, by way of example only, controlling in vivo PK properties, off-target activity, potential toxicity associated with CYP P450 interactions, drug-drug interactions, and the like. Furthermore, modifications to G allow for the adjustment of the in vivo efficacy of a compound by, by way of example only, adjusting specific and nonspecific protein binding to plasma proteins and lipids, and in vivo tissue distribution.

[0202] In some embodiments, the BTK inhibitor has a structure of formula (D):

[0203] [ka] During the ceremony, L a is CH, O, NH, or S; Ar is an optionally substituted aromatic carbocycle or aromatic heterocycle; Y is an optionally substituted alkyl, heteroalkyl, carbocyclyl, heterocyclyl, or combinations thereof; Z is C(O), OC(O), NHC(O), C(S), S(O) x , OS(O) x , NHS(O) x where x is 1 or 2; and R6, R7, and R8 are independently selected from H, alkyl, heteroalkyl, carbocycle, heterocycle, or combinations thereof.

[0204] In some embodiments, L a is O.

[0205] In some embodiments, Ar is phenyl.

[0206] In some embodiments, Z is C(O).

[0207] In some embodiments, each of R1, R2, and R3 is H.

[0208] In some embodiments, provided herein are compounds of Formula (D) below, and pharmaceutically active metabolites, pharmaceutically acceptable solvates, pharmaceutically acceptable salts, or pharmaceutically acceptable prodrugs thereof:

[0209] [ka] During the ceremony: L a is CH, O, NH, or S; Ar is a substituted or unsubstituted aryl or a substituted or unsubstituted heteroaryl; Y is an optionally substituted group selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl; Z is C(=O), OC(=O), NHC(=O), C(=S), S(=O) x , OS(=O) x , NHS(=O) x where x is 1 or 2; R7 and R8 are independently selected from H, unsubstituted C1-C4 alkyl, substituted C1-C4 alkyl, unsubstituted C1-C4 heteroalkyl, substituted C1-C4 heteroalkyl, unsubstituted C3-C6 cycloalkyl, substituted C3-C6 cycloalkyl, unsubstituted C2-C6 heterocycloalkyl, substituted C2-C6 heterocycloalkyl; or R7 and R8 taken together form a single bond; R6 is H, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 heteroalkyl, C1-C6 alkoxyalkyl, C1-C8 alkylaminoalkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted C2-C8 heterocycloalkyl, substituted or unsubstituted heteroaryl, C1-C4 alkyl(aryl), C1-C4 alkyl(heteroaryl), C1-C4 alkyl(C3-C8 cycloalkyl), or C1-C4 alkyl(C2-C8 heterocycloalkyl).

[0210] For any and all of the embodiments, the substituents may be selected from among the subgroups of the described alternatives. For example, in some embodiments, L a is CH, O, or NH. In other embodiments, L a is O or NH. In yet another embodiment, L a is O.

[0211] In some embodiments, Ar is substituted or unsubstituted aryl. In still other embodiments, Ar is 6-membered aryl. In some other embodiments, Ar is phenyl.

[0212] In some embodiments, x is 2. In still other embodiments, Z is C(=O), OC(=O), NHC(=O), S(=O) x , OS(=O) x , or NHS(=O) x In some other embodiments, Z is C(=O), NHC(=O), or S(=O)2.

[0213] In some embodiments, R7 and R8 are independently selected from H, unsubstituted C1-C4 alkyl, substituted C1-C4 alkyl, unsubstituted C1-C4 heteroalkyl, and substituted C1-C4 heteroalkyl; or R7 and R8 taken together form a single bond. In still other embodiments, each of R7 and R8 is H; or R7 and R8 taken together form a single bond.

[0214] In some embodiments, R6 is H, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 heteroalkyl, C1-C6 alkoxyalkyl, C1-C2 alkyl-N(C1-C3 alkyl)2, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, C1-C4 alkyl(aryl), C1-C4 alkyl(heteroaryl), C1-C4 alkyl(C3-C8 cycloalkyl), or C1-C4 alkyl(C2-C8 heterocycloalkyl). In some other embodiments, R6 is H, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 heteroalkyl, C1-C6 alkoxyalkyl, C1-C2 alkyl-N(C1-C3 alkyl), C1-C4 alkyl(aryl), C1-C4 alkyl(heteroaryl), C1-C4 alkyl(C3-C8 cycloalkyl), or C1-C4 alkyl(C2-C8 heterocycloalkyl). In still other embodiments, R6 is H, substituted or unsubstituted C1-C4 alkyl, —CH2—O—(C1-C3 alkyl), —CH2—N(C1-C3 alkyl), C1-C4 alkyl(phenyl), or C1-C4 alkyl(5- or 6-membered heteroaryl). In some embodiments, R6 is H, substituted or unsubstituted C1-C4 alkyl, -CH2-O-(C1-C3 alkyl), -CH2-N(C1-C3 alkyl)2, C1-C4 alkyl(phenyl), or C1-C4 alkyl(5- or 6-membered heteroaryl containing 1 or 2 N atoms), or C1-C4 alkyl(5- or 6-membered heterocycloalkyl containing 1 or 2 N atoms).

[0215] In some embodiments, Y is an optional substituent selected from alkyl, heteroalkyl, cycloalkyl, and heterocycloalkyl. In other embodiments, Y is an optional substituent selected from C1-C6 alkyl, C1-C6 heteroalkyl, 4-, 5-, 6-, or 7-membered cycloalkyl, and 4-, 5-, 6-, or 7-membered heterocycloalkyl. In still other embodiments, Y is C1-C6 alkyl, C1-C6 heteroalkyl, 5- or 6-membered cycloalkyl, and 5- or 6-membered heterocycloalkyl containing 1 or 2 N atoms. In some other embodiments, Y is 5- or 6-membered cycloalkylene or 5- or 6-membered heterocycloalkyl containing 1 or 2 N atoms.

[0216] Any combination of the groups described above for various variations is contemplated herein. It is understood that substituents and substitution patterns for the compounds provided herein can be selected by one skilled in the art to provide compounds that are chemically stable and that can be synthesized by techniques described herein as well as those known in the art.

[0217] In some embodiments, the BTK inhibitor compound of Formula (A), Formula (B), Formula (C), or Formula (D) includes, but is not limited to, a compound selected from the group consisting of:

[0218] [ka]

[0219] [ka]

[0220] In some embodiments, the Btk inhibitor is selected from the following:

[0221] [ka]

[0222] In some embodiments, the Btk inhibitor is selected from the following: 1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)prop-2-en-1-one (Compound 4); (E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)but-2-en-1-one (Compound 5); 1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)but-2-en-1-one (Compound 6); N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)sulfonylethylene (Compound 6); 1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)prop-2-yn-1-one (Compound 8); 1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (Compound 9); N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1 1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 11); 1-((S)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 12); 1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 13); 1-((S)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (Compound 13); 1-((S)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (Compound 14); and (E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound 15).

[0223] Throughout the specification, groups and substituents thereof may be chosen by one skilled in the art to provide stable moieties and compounds.

[0224] Compounds of any of Formula (A), Formula (B), Formula (C), or Formula (D) irreversibly inhibit Btk and may be used to treat patients afflicted with Bruton's tyrosine kinase-dependent or Bruton's tyrosine kinase-mediated diseases or disorders, including, but not limited to, cancer, autoimmune and other inflammatory diseases.

[0225] "Ibrutinib" or "1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one" or "1-{(3R)-3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl}prop-2-en-1-one" or "2-propen-1-one, 1-[(3R)-3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl" or ibrutinib or other suitable name refers to a compound having the structure:

[0226] [ka]

[0227] A wide variety of pharmaceutically acceptable salts may be formed from ibrutinib, including:

[0228] Acid addition salts formed by reacting ibrutinib with organic acids (including aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxylalkanoic acids, alkanediol (alkanediol) acids, aromatic acids, aliphatic and aromatic sulfonic acids, amino acids, etc.), including, for example, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

[0229] Acid addition salts formed by reacting ibrutinib with inorganic acids (including hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, etc.).

[0230] The term "pharmaceutically acceptable salt," in reference to ibrutinib, refers to a salt of ibrutinib that does not cause significant irritation to the mammal to which it is administered and that does not substantially abrogate the biological activity and properties of the compound.

[0231] It should be understood that a reference to a pharmaceutically acceptable salt includes its solvent addition forms (solvates). Solvates contain either stoichiometric or non-stoichiometric amounts of the solvate and are formed during the process of product formation or isolation with pharmaceutically acceptable solvents (such as water, ethanol, methanol, methyl tert-butyl ether (MTBE), diisopropyl ether (DIPE), ethyl acetate, isopropyl acetate, isopropyl alcohol, methyl isobutyl ketone (MIBK), methyl ethyl ketone (MEK), acetone, nitromethane, tetrahydrofuran (THF), dichloromethane (DCM), dioxane, heptane, toluene, anisole, acetonitrile, etc.). In one embodiment, solvates are formed using, but not limited to, Class 3 solvents. Categories of solvents are defined, for example, in the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH), "Impurities: Guidelines for Residual Solvents, Q3C(R3), (November 2005). Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is alcohol. In some embodiments, solvates of ibrutinib, or a pharmaceutically acceptable salt thereof, are advantageously prepared or formed during the processes described herein. In some embodiments, solvates of ibrutinib are anhydrous. In some embodiments, ibrutinib, or a pharmaceutically acceptable salt thereof, exists in an unsolvated form. In some embodiments, ibrutinib, or a pharmaceutically acceptable salt thereof, exists in an unsolvated form and is anhydrous.

[0232] In yet other embodiments, ibrutinib, or a pharmaceutically acceptable salt thereof, is prepared in various forms, including, but not limited to, amorphous, crystalline, milled, and nanoparticulate forms. In some embodiments, ibrutinib or a pharmaceutically acceptable salt thereof is amorphous. In some embodiments, ibrutinib or a pharmaceutically acceptable salt thereof is amorphous and anhydrous. In some embodiments, ibrutinib or a pharmaceutically acceptable salt thereof is crystalline. In some embodiments, ibrutinib or a pharmaceutically acceptable salt thereof is crystalline and anhydrous.

[0233] In some embodiments, ibrutinib is prepared as outlined in U.S. Patent No. 7,514,444.

[0234] Additionally, Btk blocks are PCI-45292, PCI-45466, AVL-101 / CC-101(Avila Therapeutics / Celgene). Corporation)、AVL-263 / CC-263(Avila Therapeutics / Celgene Corporation)、AVL-292 / CC-292(Avila Therapeutics / Celgene Corporation)、AVL-291 / CC-291(Avila Therapeutics / Celgene Corporation)、CNX774(Avila Therapeutics)、BMS-488516(Bristol-Myers Squibb)、BMS-509744(Bristol-Myers Squibb)、CGI-1746(CGI Pharma / Gilead Sciences)、CGI-560(CGI Pharma / Gilead). Sciences), CTA-056, GDC-0834 (Genentech), HY-11066 (CTK4I7891), HMS3265G21, HM S3265G22, HMS3265H21, HMS3265H22, 439574-61-5, AG-F-54930, ONO-4059(Ono Pharmaceutical Co., Ltd., ONO-WG37 (Ono Pharmaceutical Co., Ltd.), PLS-123 (Peking University), RN486 (Hoffmann-La Roche), HM71224 (Hanmi Pharmaceutical Company). Limited, LFM-A13, BGB-3111 (Beigene), KBP-7536 (KBP BioSciences), ACP-196 (Acerta Pharma), and JTE-051 (Japan Tobacco Inc).

[0235] In some embodiments, the Btk inhibitor is 4-(tert-butyl)-N-(2-methyl-3-(4-methyl-6-((4-(morpholine-4-carbonyl)phenyl)amino)-5-oxo-4,5-dihydropyrazin-2-yl)phenyl)benzamide (CGI-1746); 7-benzyl-1-(3-(piperidin-1-yl)propyl)-2-(4-(pyridin-4-yl)phenyl)-1H-imidazoline Dazo[4,5-g]quinoxalin-6(5H)-one (CTA-056); (R)-N-(3-(6-(4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenylamino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide (GDC-0834); 6-cyclopropyl-8-fluorobenzo[b]thiophene-2-carboxamide N-(2-hydroxymethyl-3-{1-methyl-5-[5-(4-methyl-piperazin-1-yl)-pyridin-2-ylamino]-6-oxo-1,6-dihydro-pyridin-3-yl}-phenyl)-2H-isoquinolin-1-one (RN-486); N-(5-[5-(4-acetylpiperazine-1-carbonyl)-4-methoxy-2-methylphenyl]sulfanyl-1,3-thiazol-2-yl) ]-4-[(3,3-dimethylbutan-2-ylamino)methyl]benzamide (BMS-509744, HY-11092); or N-(5-((5-(4-acetylpiperazine-1-carbonyl)-4-methoxy-2-methylphenyl)thio)thiazol-2-yl)-4-(((3-methylbutan-2-yl)amino)methyl)benzamide (HY11066); or a pharmaceutically acceptable salt thereof.

[0236] In some embodiments, the Btk inhibitor is:

[0237] [ka]

[0238] [ka]

[0239]

Chem.

[0240] <ITK inhibitor> In some embodiments, the ACK inhibitor is an ITK inhibitor. In some embodiments, the ITK inhibitor covalently binds to cysteine 442 of ITK. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2002 / 0500071, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2005 / 070420, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2005 / 079791, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2007 / 076228, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2007 / 058832, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2004 / 016610, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2004 / 016611, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2004 / 016600, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2004 / 016615, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2005 / 026175, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2006 / 065946, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2007 / 027594, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2007 / 017455, which is incorporated by reference in its entirety.In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2008 / 025820, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2008 / 025821, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2008 / 025822, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2011 / 017219, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2011 / 090760, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2009 / 158571, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2009 / 051822, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in US20110281850, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2014 / 082085, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2014 / 093383, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in US8759358, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2014 / 105958, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in US2014 / 0256704, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in US20140315909, which is incorporated by reference in its entirety.In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in US20140303161, which is incorporated by reference in its entirety. In some embodiments, the ITK inhibitor is an ITK inhibitor compound described in WO2014 / 145403, which is incorporated by reference in its entirety.

[0241] In some embodiments, the ITK inhibitor has a structure selected from the following:

[0242] [ka]

[0243] <Pharmaceutical Compositions / Preparations> In certain embodiments, disclosed herein are compositions comprising a therapeutically effective amount of an ACK inhibitory compound and a pharmaceutically acceptable excipient. In some embodiments, the ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is a compound of Formula (A). In some embodiments, the ACK inhibitory compound is (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (i.e., PCI-32765 / ibrutinib).

[0244] Pharmaceutical compositions of ACK inhibitor compounds (e.g., ITK or BTK inhibitors, such as ibrutinib) are formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliary agents, which facilitate processing of the active compound into pharmaceutically usable preparations. Appropriate formulations depend on the route of administration selected. Summaries of the pharmaceutical compositions described herein can be found, for example, in Remington's The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999).

[0245] Pharmaceutical composition, as used herein, refers to a mixture of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients.

[0246] The pharmaceutical compositions are optionally manufactured in a conventional manner, such as by means of, but not limited to, conventional mixing, dissolving, granulating, dragee-making, pulverizing, emulsifying, encapsulating, entrapping, or compressing processes.

[0247] The pharmaceutical formulations described herein may be administered by any suitable route of administration, including, but not limited to, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular), intranasal, buccal, topical, rectal, or transdermal routes of administration.

[0248] The pharmaceutical compositions described herein are formulated into dosage forms suitable for oral ingestion by a patient to be treated, including, but not limited to, aqueous oral dispersions, liquids, gels, syrups, elixirs, slurries, suspensions, and the like, solid oral dosage forms, aerosols, controlled-release formulations, fast-dissolve formulations, effervescent formulations, lyophilized formulations, tablets, powders, pills, dragees, capsules, delayed-release formulations, extended-release formulations, pulsed-release formulations, multiparticulate formulations, and immediate mixed release and controlled-release formulations. In some embodiments, the compositions are formulated into capsules. In some embodiments, the compositions are formulated into solutions (e.g., for IV administration).

[0249] The pharmaceutical solid dosage forms described herein optionally comprise a compound described herein and one or more pharmaceutically acceptable excipients such as a compatible carrier, binder, filler, suspending agent, flavoring agent, sweetener, disintegration agent, dispersing agent, surfactant, lubricant, coloring agent, diluent, solubilizer, moistening agent, plasticizer, stabilizer, transdermal absorption enhancer, wetting agent, antifoaming agent, antioxidant, preservative, or one or more combinations thereof.

[0250] In some embodiments, a film coating is provided around the composition using standard coating procedures, such as those described in Remington's Pharmaceutical Sciences, 20th Edition (2000). In some embodiments, the composition is formulated into particles (e.g., for administration via capsules), and some or all of the particles are coated. In some embodiments, the composition is formulated into particles (e.g., for administration via capsules), and some or all of the particles are microencapsulated. In some embodiments, the composition is formulated into particles (e.g., for administration via capsules), and some or all of the particles are not microencapsulated or coated.

[0251] In some embodiments, the pharmaceutical compositions are formulated so that the amount of ACK inhibitor (e.g., an ITK or BTK inhibitor, such as ibrutinib) in each unit dosage form is about 140 mg per unit dosage form.

[0252] <Kit / manufactured product> Described herein are kits for treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient in need thereof, the kits comprising a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib).

[0253] Further described herein is a kit for preventing the onset of alloantibody-driven chronic graft-versus-host disease (cGVHD) or reducing the severity of an alloantibody-driven cGVHD onset in a patient requiring cell transplantation, the kit comprising a therapeutically effective amount of an ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib), wherein the therapeutically effective amount of the ACK inhibitory compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) is administered prior to or simultaneously with allogeneic hematopoietic stem cells and / or allogeneic T cells.

[0254] Kits and articles of manufacture are also described herein for use in the therapeutic applications described herein. In some embodiments, the kits include carriers, packages, or containers that are compartmentalized to accommodate one or more containers, such as vials, tubes, etc., each containing one of the distinct components used in the methods described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers can be formed from a variety of materials, such as glass or plastic.

[0255] The articles of manufacture provided herein include packaging materials. Pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubing, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging material appropriate for the selected formulation and intended mode of administration and treatment. Numerous formulations of the compounds and compositions provided herein are contemplated, as are various treatments for any disease that would benefit from inhibition of BTK or for which BTK is a mediator or contributor to the symptoms or causation of the disease.

[0256] The container optionally has a sterile access port (for example, the container is an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). Such kits optionally contain the compound along with an identifying instruction, or label, or instructions for use in the methods described herein.

[0257] Kits typically include one or more additional containers, each containing one or more of a variety of materials (e.g., reagents, optionally in concentrated form, and / or devices) desirable from a commercial and user standpoint for use of the compounds described herein. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, filters, needles, syringes, carriers, packaging, containers, vial and / or tube labels listing the contents and / or instructions for use, and inserts with instructions for use. A set of instructions is also typically included.

[0258] In some embodiments, a label is on or associated with a packaging container. A label can be attached to a container when letters, numbers, or other indicia forming the label are affixed, molded, or engraved into the container itself. A label can be associated with a container when present in a receptacle or carrier that also holds the container, for example, as a package insert. A label can be used to indicate that the contents are to be used for a specific therapeutic application. A label can also indicate instructions for using the contents, such as by the methods described herein.

[0259] In certain embodiments, pharmaceutical compositions comprising an ACK inhibitor compound (e.g., an ITK or BTK inhibitor, such as ibrutinib) are provided in a pack or dispenser device that may contain one or more unit dosage forms. The pack may, for example, comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied by a notice attached to the container in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, the notice reflecting approval by the agency of the drug form for human or animal administration. Such notice may, for example, be labeling approved by the U.S. Food and Drug Administration for prescription drugs or an approved product insert. Compositions containing a compound provided herein formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated disease. [Example]

[0260] Example 1 To determine whether ibrutinib could reverse established cGVHD, we used a murine model (MHC-distinct, C57BL / 6→B10.BR) of alloantibody-driven multi-organ system cGVHD, including bronchiolar obliterans (BO).

[0261] Materials and Methods Mice: C57BL / 6 (H2b) mice were purchased from the National Cancer Institute or The Jackson Laboratory. B10.BR (H2k) mice were purchased from The Jackson Laboratory. C57BL / 6 XID mice (in which BTK kinase activity is genetically abrogated) were obtained commercially from The Jackson Laboratory, and ITK- / - mice were a gift. Both strains are maintained on a defined C57BL / 6 genetic background. All mice were housed in a pathogen-free facility and used with approval from the respective institutional animal care committees.

[0262] Therapeutic allo-HSCT model: The C57BL / 6 → B10.BR model was previously described (Srinivasan, M. et al. Blood 119, 1570-1580 (2012)). Briefly, IP cyclophosphamide (Cy) at 120 mg / kg / day on days -3 and -2 and 8.3 Gy TBI ( 137 Recipients of conditioned B10.BR (using a Cesium irradiator) were given 1 × 10 6 1 × 10 with or without allogeneic splenocytes 7 Thy1.2-depleted C57BL / 6-derived bone marrow (BM) cells were transplanted.

[0263] Therapeutic administration of ibrutinib via drinking water was performed as previously described (Dubovsky 2013). Mice received the equivalent dose of 15 mg / kg / day in 0.4% methylcellulose by intraperitoneal injection starting 28 days after transplantation for the C57BL / 6→B10.BR model. Cyclosporine A was administered IP in 0.2% CMC at 10 mg / kg / day for 2 weeks, then three times a week (3X) starting on day 25 (Blazar, BR et al. Blood 92, 3949-3959 (1998)).

[0264] Pulmonary function tests: Pulmonary function tests (PFTs) were performed in anesthetized mice using whole-body plethysmography with the Flexivent system (SCIREQ).

[0265] GC detection: GC detection was performed using 6 μm spleen cryosections stained with rhodamine-peanut agglutinin as previously described.

[0266] Masson trichrome staining: 6 μm frozen sections were fixed in acetone for 5 minutes, stained with hematoxylin-eosin, and stained with Masson's trichrome staining kit (Sigma) for the detection of collagen deposition to determine pathology. Histopathological scores were assigned as described (Blazar, BR et al. Blood 92, 3949-3959 (1998)). Collagen deposition was quantified for trichrome-stained sections as the ratio of blue-stained area to total contact area using Adobe Photoshop CS3 analysis tools.

[0267] Histopathological Scoring: Coded pathological analysis of H&E-stained sections was performed by trained veterinary pathologists in an unbiased manner. Scores ranged from 0 to 4 and indicated the maximum number of lymphoplasmacytic and histiocytic cell cuffs infiltrating the surrounding airways or vasculature and the number of infiltrating aggregates in two different 4X microscopic fields. 0 cuffs = 0, 1 to 5 cuffs = 1, 6 to 10 cuffs and <6 aggregates = 2, 11 to 15 cuffs and <15 aggregates = 4, and >16 cuffs = 4. Limited foci of alveolar histiocytosis present with a 0 cuff were considered concomitant. For H&E-stained sections of the kidneys, both perivascular lymphoplasmacytic infiltrates and intraductal protein were quantified by trained veterinary pathologists on coded samples. Scoring ranged from 0 to 4 according to the following guidelines: no inflammatory infiltrate and no hyaline eosinophilic material present in the tubular lumen = 0; ducts characteristic of scattered focal lymphocytes and plasma cells around the renal vessels or containing <6 hyaline eosinophilic material = 1; 6 to 10 ducts containing between 1 and 2 aggregates of inflammatory cells or hyaline eosinophilic material with a diameter of <10 = 3; 3 to 4 foci of inflammatory cells up to 20 in diameter or between 11 and 15 ducts containing hyaline eosinophilic material = 3; 5 inflammatory cell foci or more or less than 5 cells with a diameter of >20 or >15 ducts containing hyaline eosinophilic material = 4.

[0268] Statistical analysis: Unless otherwise stated, a two-tailed Student's T-test was used for normal data with equal variance. Significance was considered at p<0.05.

[0269] <Result> Therapeutic administration of ibrutinib improved the progression of pulmonary fibrosis and bronchiolitis obliterans.

[0270] cGVHD is characterized by a variety of autoimmune phenomena that are incompletely mimicked by any single in vivo animal model. The consensus criteria recently published by the National Institutes of Health consider BO to be the sole pathognomonic manifestation of cGVHD in the lung. The C57BL / 6→B10.BR model has been shown to develop multi-organ system disease, including BO, beginning 28 days after HSCT. Therapeutic administration of ibrutinib, beginning on day 28 and continuing indefinitely, reduced the progression of BO in vivo, as measured by pulmonary circulatory resistance (p=0.0090), elasticity (p=0.0019), and compliance (p=0.0071) (Figure 1A, B, and C).

[0271] BO is causally associated with pulmonary collagen deposition and tissue fibrosis. Masson trichrome staining of expanded lung tissue from four mice from three experiments demonstrated less peribronchiolar collagen fibrillation in ibrutinib-treated animals (Figure 1D). Quantified trichrome staining data confirmed that ibrutinib treatment ameliorated cGVHD-induced pulmonary fibrosis (p<0.0001) (Figure 1E). Death due to cGVHD was rare in this model, with virtually 100% survival observed in the ibrutinib cohort (Figure 2). Weekly assessment of mouse body weight revealed little variation between groups (Figure 3). These functional data demonstrate that ibrutinib therapeutically combats the underlying fibrotic pathogenesis of BO in the C57BL / 6→B10.BR cGVHD model.

[0272] Ibrutinib limited in vivo germinal center reactions and Ig deposition in lung tissue.

[0273] While the ability of ibrutinib to block BCR-induced activation of BTK is well defined, it remains unclear whether alloreactive B cells are effectively inhibited in the context of GC. To investigate this, we used a C57BL / 6→B10.BR mouse model in which a robust GC response persists pathogenic alloreactive B lymphocytes, leading to Ig deposition in the liver and lungs and the progression of BO. Compared to vehicle-treated mice with active cGVHD, GC responses, as revealed by peanut agglutinin staining in the spleen, and ibrutinib treatment reduced the overall size, cellularity, and number of GC responses (Figure 4A). Sixty days after HSCT, splenocytes isolated from eight mice per group were analyzed for CD19+GL7+CD38lo germinal center B cells by flow cytometry. The data revealed that ibrutinib significantly inhibited cGVHD-induced germinal center B cell formation in the spleen (p=0.0222) (Figure 4B). These results indicated a significant reduction in the response of alloreactive GCs, potentially related to TEC kinase blockade caused by ibrutinib.

[0274] The functional product of alloreactive GC B cells is soluble Ig, which is deposited in healthy tissue. In the C57BL / 6→B10.BR cGVHD model, BO is closely associated with the deposition of soluble Ig in lung tissue and the resulting fibrosis cascade. By blocking B cell reactivity, ibrutinib limited lung deposition of allo-Ig, as quantified 60 days after HSCT using immunofluorescence microscopy (Figure 4C). As expected, quantified immunofluorescence signals revealed significant and complete ablation of lung Ig deposits after therapeutic ibrutinib treatment (p<0.001) (Figure 4D). These data confirmed that a clinically relevant downstream effect of ibrutinib treatment in the setting of cGVHD is the blockade of Ig deposition in healthy tissue.

[0275] Genetic ablation of BTK or ITK activity in allogeneic donor cell transplants confirmed that both TEC kinases are required for the development of cGVHD.

[0276] XID mice, in which the kinase activity of BTK is genetically abrogated, and ITK- / - mice have been thoroughly characterized on the C57BL / 6 genetic background (Numata et al., Int Immunol 9(1):139-46, 1997; and Liu et al., J Exp Med 187(10):1721-7, 1998). Given the ability of ibrutinib to inhibit both ITK and BTK, the relative independent contributions of ITK and BTK to the progression of cGVHD were examined. To answer this question, pulmonary function was examined 60 days after HSCT, as it represents the primary functional measure of cGVHD-induced lung injury and fibrosis in the C57BL / 6→B10.BR model.

[0277] The T cells that sustain cGVHD in this model arise from mature lymphocytes incorporated into the donor cell transplant. To characterize the effects of ITK inhibition within these cGVHD-causing T lymphocytes, ITK- / - splenic T cells were transplanted into allogeneic recipients with wild-type BM. Pulmonary function tests at day 60, including tolerance, resilience, and adaptation, were uniformly and significantly restored to healthy levels in mice receiving ITK- / - splenic T cells as part of the transplant compared with mice receiving wild-type splenic T cells (p=0.0014; p=0.0028; p=0.0003) (Figure 5). These data demonstrate that T cell ITK activity is required for the progression of cGVHD.

[0278] Pathogenic B cells in cGVHD arise from the ontogeny of donor hematopoietic stem cells; therefore, we transplanted XID BM with wild-type splenic T cells to demonstrate BTK inhibition in allogeneic B cells. Pulmonary function tests performed 60 days after HSCT revealed that BTK activity is essential for the progression of BO (Figure 6). Lung metrics of resistance, elasticity, and adaptability were significantly improved in mice receiving XID BM compared to mice receiving wild-type bone marrow (p=0.0025; p=0.0025; p=0.0496).

[0279] In summary, in a C57BL / 6→B10.BR cGVHD model, ibrutinib restored lung function, attenuated germinal center reactions and tissue immunoglobulin deposition, and reversed lung and liver fibrosis. Our analysis revealed that ibrutinib therapeutically blocked alloreactive germinal center (GC) B cells, immunoglobulin (Ig) deposition, and pulmonary fibrosis associated with the progression of cGVHD.

[0280] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous modifications, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It will be understood that various alternatives to the embodiments of the invention described herein may be utilized in practicing the invention. It is the following claims that define the scope of the invention, and it is intended that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

1. 1. Use of a compound of formula (A), or a pharmaceutically acceptable salt thereof, for treating alloantibody-driven chronic graft-versus-host disease (cGVHD) in a patient, wherein formula (A) has the following structure: 【Chemical 1】 During the ceremony: A is N; R １ is phenyl-O-phenyl or phenyl-S-phenyl; R ２ and R ３ are independently H; R ４ Is, L ３ -XL ４ -G, where: L ３ is optional and, if present, is a single bond, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl; X is optional and, if present, is a single bond, —O—, —C(═O)—, —S—, —S(═O)—, or —S(═O) ２ -, -NH-, -NR ９ -, -NHC(O)-, -C(O)NH-, -NR ９ C(O)-, -C(O)NR ９ -, -S(=O) ２ NH-, -NHS (=O) ２ -, -S(=O) ２ NR ９ -, -NR ９ S (= O) ２ -, -OC(O)NH-, -NHC(O)O-, -OC(O)NR ９ -, -NR ９ C(O)O-, -CH=NO-, -ON=CH-, -NR １０ C(O)NR １０ -, heteroaryl-, aryl-, -NR １０ C (=NR １１ ) NR １０ -, -NR １０ C (=NR １１ ) -, -C(=NR １１ ) NR １０ -, -OC(=NR １１ ) - or -C(=NR １１ ) O—; L ４ is optional and, if present, is a single bond, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle; Or, the L obtained together ３ , X, and L ４ forms a nitrogen-containing heterocycle; G is 【Chemistry 2】 where: R ６ , R ７ , and R ８ is independently selected from among H, halogen, CN, OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; Each R ９ is independently selected from among H, substituted or unsubstituted lower alkyl, and substituted or unsubstituted lower cycloalkyl; Each R １０ are independently H, substituted or unsubstituted lower alkyl, or substituted or unsubstituted lower cycloalkyl; or Two R's １０ the groups together can form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or R １０ and R １１ can together form a 5-, 6-, 7-, or 8-membered heterocyclic ring; or Each R １１ are independently selected from H or substituted or unsubstituted alkyl Use characterized by:

2. The L obtained together ３ , X, and L ４ The use according to claim 1, characterized in that:

3. 3. The use according to claim 1 or 2, characterized in that the nitrogen-containing heterocycle is a piperidine group.

4. G is 【Chemistry 3】 4. The use according to any one of claims 1 to 3, characterized in that:

5. 5. The use according to any one of claims 1 to 4, wherein the compound of formula (A) is (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (ibrutinib), or a pharmaceutically acceptable salt thereof. 【Chemistry 4】

6. 6. The use according to any one of claims 1 to 5, characterized in that the cGVHD is non-sclerodermic cGVHD.

7. The use according to any one of claims 1 to 5, characterized in that the cGVHD is multi-organ cGVHD.

8. 6. The use according to any one of claims 1 to 5, wherein the cGVHD is bronchiolitis obliterans syndrome.

9. 6. The use according to any one of claims 1 to 5, wherein the cGVHD is pulmonary cGVHD.

10. 10. The use according to any one of claims 1 to 9, characterized in that fibrosis is reduced.

11. 11. The use according to any one of claims 1 to 10, wherein the patient has undergone a cell transplant.

12. The use according to claim 11, wherein the cell transplant is a hematopoietic cell transplant.

13. 12. The use according to claim 11, wherein the cell transplant is an allogeneic bone marrow or hematopoietic stem cell transplant.

14. 12. The use according to claim 11, wherein the compound of formula (A) is administered simultaneously with allogeneic bone marrow or hematopoietic stem cell transplantation.

15. 15. The use according to any one of claims 1 to 14, wherein the patient suffers from relapsed or refractory CLL.

16. 16. The use according to any one of claims 1 to 15, wherein the compound of formula (A) is administered in an amount corresponding to a dose of between about 0.1 mg / kg per day and about 100 mg / kg per day.

17. 16. The use according to any one of claims 1 to 15, wherein the compound of formula (A) is administered in an amount of about 40 mg / day, about 140 mg / day, about 420 mg / day, about 560 mg / day, or about 840 mg / day.

18. 18. Use according to any one of claims 1 to 17, characterized in that the compound of formula (A) is suitable for oral administration.

19. 19. The use according to any one of claims 1 to 18, wherein the compound of formula (A) is administered in combination with one or more additional therapeutic agents.

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