WO2015057992A1 - Btk inhibitors for hematopoietic mobilization - Google Patents

Btk inhibitors for hematopoietic mobilization Download PDF

Info

Publication number
WO2015057992A1
WO2015057992A1 PCT/US2014/060953 US2014060953W WO2015057992A1 WO 2015057992 A1 WO2015057992 A1 WO 2015057992A1 US 2014060953 W US2014060953 W US 2014060953W WO 2015057992 A1 WO2015057992 A1 WO 2015057992A1
Authority
WO
WIPO (PCT)
Prior art keywords
pyrazin
benzamide
amino
imidazo
pyridin
Prior art date
Application number
PCT/US2014/060953
Other languages
French (fr)
Inventor
Raquel IZUMI
Francisco SALVA
Ahmed HAMDY
Original Assignee
Izumi Raquel
Salva Francisco
Hamdy Ahmed
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Izumi Raquel, Salva Francisco, Hamdy Ahmed filed Critical Izumi Raquel
Publication of WO2015057992A1 publication Critical patent/WO2015057992A1/en

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4985Pyrazines or piperazines ortho- or peri-condensed with heterocyclic ring systems
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • A61P35/02Antineoplastic agents specific for leukemia
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00Drugs for disorders of the blood or the extracellular fluid

Definitions

  • the invention is in the field of therapeutics.
  • the present invention relates generally to the administration of a Bruton's tyrosine kinase inhibitor to mobilize hematopoietic stem and progenitor cells from the bone marrow into the peripheral blood and the use of such hematopoietic cells to improve hematopoiesis and/or in the treatment of various disorders.
  • Bruton's tyrosine kinase (Btk) is a member of the Tec family of non-receptor tyrosine kinases and plays a role in several hematopoietic cell signaling pathways, e.g., Toll like receptor (TLR) and cytokine receptor-mediated TNF-a production in macrophages, IgE receptor (FcsRI) signaling in Mast cells, inhibition of Fas/APO-1 apoptotic signaling in B lineage lymphoid cells, and collagen- stimulated platelet aggregation.
  • TLR Toll like receptor
  • FcsRI IgE receptor
  • Btk Mutations in the Btk gene result in X-linked agammaglobulinemia, an immunodeficiency characterized by failure to produce mature B lymphocytes and associated with a failure of Ig heavy chain rearrangement.
  • point mutation or deletion of Btk causes X-linked immunodeficiency (xid), with about 50% fewer conventional B2 B cells, absent BI B cells, and reduced serum Ig levels. Khan et al (1995) Immunity 3:283-99; Rawlings et al (1993) Science 261 :358-61.
  • Btk is also expressed in specific cells of the myeloid lineage, and evidence suggests that it contributes to immune-complex mediated activation of the FcyR and FcsR signaling pathways in monocytes/macrophages, neutrophils, and mast cells. See, e.g., Jongstra-Bilen et al. (2008) J.
  • Btk inhibitors Due to the role of Btk in inhibiting Fas/APO-1 apoptotic signals in the B cell lineage, inhibitors of Btk, also referred to as Btk inhibitors, have been evaluated as agents for treating hematopoietic malignancies (e.g., B cell lymphoma). Additionally, due to the role of Btk in the signaling pathways of other immune cells, Btk inhibitors have also been evaluated as agents for suppressing the immune system, e.g., in patients with autoimmune disorders or organ transplants. See, e.g., Honinberg et al. (2010) Proc. Natl. Acad. Sci. USA 107:13075-80; Chang et al. (2011) Arthr. Res.
  • Btk-deficient mice evidence for the role of Btk in autoimmune and/or inflammatory disease has been established in Btk-deficient mouse models. For example, in standard murine preclinical models of systemic lupus erythematosus (SLE), Btk deficiency has been shown to result in a marked amelioration of disease progression. Moreover, Btk deficient mice are also resistant to developing collagen-induced arthritis and are less susceptible to Staphylococcus-induced arthritis.
  • SLE systemic lupus erythematosus
  • Inhibition of Btk activity is useful for the treatment of autoimmune and/or inflammatory diseases such as: SLE, rheumatoid arthritis, multiple vasculitides, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, and asthma. See, e.g., U.S. Patent No. 7,393,848.
  • Btk inhibitors have also been shown useful in preventing or reducing the risk of
  • Btk inhibitors can mobilize hematopoietic stem cells and progenitor cells to the peripheral blood of a subject, e.g., to increase the white blood cell count in the subject. Accordingly, provided herein are methods and compositions for improving hematopoiesis and increasing the white blood cell count in a subject in need thereof, including patients undergoing chemotherapy, radiation therapy and/or bone marrow transplantation.
  • the invention provides methods for mobilizing hematopoietic stem and/or progenitor cells in a subject in need thereof comprising administering to said subject a pharmaceutical composition comprising a mobilizing Bruton's Tyrosine Kinase (Btk) inhibitor in an amount effective to mobilize said cells into the peripheral blood of said subject.
  • Btk Bruton's Tyrosine Kinase
  • the inventive methods and uses can be advantageously employed in conjunction with bone marrow transplantation procedures, and/or subsequent to chemotherapy and/or radiation exposure to address leukopenia, neutropenia, granulocytopenia and/or thrombocytopenia in such patients.
  • the subject may be a bone marrow transplantation patient, and/or a leukopenic or neutropenic patient or a patient at risk of impaired hematopoiesis due to prior chemotherapy and/or radiation therapy.
  • the instant mobilizing Btk inhibitors may be administered as part of any therapeutic protocol aiming to restore or improve hematopoiesis in a patient in need thereof, e.g., to enhance the success of bone marrow
  • transplantation to reduce the extent or duration of leukopenia and neutropenia resulting from chemotherapy, radiation therapy or accidental radiation exposure, to enhance wound healing and burn treatment, and/or to aid in restoration of damaged organ tissue. They may also combat bacterial infections that are prevalent in leukemia.
  • the invention provides methods of obtaining mobilized hematopoietic stem and progenitor cells and uses thereof.
  • the subject methods comprise administering to a subject a mobilizing Btk inhibitor in an amount effective to increase the number of such cells in the subject, preferably in the peripheral blood of the subject.
  • the administering step comprises administration of a mobilizing Btk inhibitor alone.
  • the step comprises administration of a mobilizing Btk inhibitor in combination with other compounds, e.g., cytokines, that also increase the white blood cell count in the peripheral blood of a subject.
  • Suitable compounds may be selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM-CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor, thrombopoietin, growth related oncogene, and/or combinations thereof.
  • GM-CSF granulocyte-macrophage colony stimulating factor
  • IL-1 Interleukin-1
  • IL-3 Interleukin-3
  • IL-8 Interleukin-8
  • PIXY-321 GM-CSF/IL-3 fusion protein
  • the subject methods comprise administering to a subject a mobilizing Btk inhibitor (with or without other mobilizing factors) in an amount effective to increase the number of hematopoietic stem and progenitor and/or white blood cells in the peripheral blood of the subject, and obtaining the immune cells so mobilized, e.g., by apheresis.
  • a mobilizing Btk inhibitor with or without other mobilizing factors
  • the harvested cells may be used therapeutically, e.g., in hematopoietic stem and/or progenitor cell transplantation.
  • the invention provides methods of treating a patient in need of improved hematopoiesis comprising administering to a subject a mobilizing Btk inhibitor (with or without other mobilizing factors) in an amount effective to increase the number of hematopoietic stem, progenitor and/or white blood cells in the peripheral blood of the subject, obtaining the cells so mobilized, and introducing the cells into the patient.
  • the subject and the patient are histocompatible.
  • the histocompatible subject and the patient are syngeneic.
  • the histocompatible subject and the patient are allogeneic.
  • the harvested cells are enriched and/or cultured ex vivo prior to introduction into the patient.
  • ex vivo culture comprises differentiating the obtained cells into or enriching for myeloid cells, lymphoid cells, and common progenitors thereof etc.
  • a method of treating a patient in need thereof further comprises culturing the obtained hematopoietic cells in one or more differentiation factors prior and/or enriching the obtained hematopoietic cells for a common progenitor cell or cells prior to introducing the cells into the patient.
  • Mobilizing Btk inhibitors may be administered to any animal subject in order to mobilize hematopoietic stem and progenitor cells.
  • the mobilizing Btk inhibitor is administered to a mammal, and more preferably to a human.
  • the invention provides compounds for use in mobilizing hematopoietic stem and/or progenitor cells into the peripheral blood of a subject, wherein the compound is a Bruton's Tyrosine Kinase (Btk) inhibitor comprising a 6,5 fused heteroaryl ring system.
  • Btk Bruton's Tyrosine Kinase
  • the 6,5 fused heteroaryl ring system is selected from: imidazopyrazine and
  • imidazotriazine ring systems In some embodiments, the 6,5 fused heteroaryl ring system is an imidazotriazine ring system.
  • Btk inhibitors suitable for use in the subject invention comprise an imidazopyrazine ring system.
  • the subject has impaired hematopoiesis. In some embodiments, the subject has previously received a
  • the myelosuppressive or myeloablative treatment comprises chemotherapy.
  • the subject has previously been accidentally exposed to radiation.
  • the compound for use is for use in combination with an additional mobilizing agent selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM- CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor, thrombopoietin, growth related oncogene, and combinations thereof.
  • an additional mobilizing agent selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM- CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor, thrombopoietin, growth related oncogene, and combinations thereof.
  • GM-CSF granulocyte-
  • the compound for use is for use in harvesting the hematopoietic stem and/or progenitor cells from the subject. In some embodiments, the compound for use is for use in harvesting the hematopoietic stem and/or progenitor cells from the subject by apheresis. In some embodiments, the compound for use is for use in providing the hematopoietic stem and/or progenitor cells to a patient in need thereof. In some embodiments, the compound for use is for use in providing the hematopoietic stem and/or progenitor cells to a bone marrow transplantation patient. In some embodiments, the subject and the patient are histocompatible.
  • the subject and the patient are syngeneic. In some embodiments, the subject and the patient are allogeneic. In some embodiments, the hematopoietic stem and/or progenitor cells are cultured to enrich and/or differentiate the hematopoietic stem and/or progenitor cells prior to introducing the cells into the patient. In some embodiments, the hematopoietic stem and/or progenitor cells are enriched for or differentiated into myeloid cells. In some embodiments, the hematopoietic stem and/or progenitor cells are enriched for or differentiated into lymphoid cells.
  • the hematopoietic stem and/or progenitor cells are enriched for or differentiated into NK cells.
  • the Btk inhibitor is reversible. In some embodiments, the Btk inhibitor is irreversible.
  • the invention provides uses of a Bruton's Tyrosine Kinase (Btk) inhibitor comprising a 6,5 fused heteroaryl ring system in the manufacture of a medicament for mobilizing hematopoietic stem and/or progenitor cells into the peripheral blood of a subject.
  • the 6,5 fused heteroaryl ring system is selected from: imidazopyrazine and imidazotriazine ring systems.
  • the 6,5 fused heteroaryl ring system is an imidazotriazine ring system.
  • Btk inhibitors suitable for use in the subject invention comprise an imidazopyrazine ring system.
  • the subject has impaired hematopoiesis.
  • the subject has previously received a
  • the myelosuppressive or myeloablative treatment comprises chemotherapy.
  • the subject has previously been accidentally exposed to radiation.
  • the medicament is for use in combination with an additional mobilizing agent selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM- CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor, thrombopoietin, growth related oncogene, and combinations thereof.
  • an additional mobilizing agent selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM- CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor, thrombopoietin, growth related oncogene, and combinations thereof.
  • the medicament is for use for use in harvesting the hematopoietic stem and/or progenitor cells from the subject. In some embodiments, the medicament is for use in harvesting the hematopoietic stem and/or progenitor cells from the subject by apheresis. In some embodiments, the medicament is for use in providing the hematopoietic stem and/or progenitor cells to a patient in need thereof. In some embodiments, the medicament is for use in providing the hematopoietic stem and/or progenitor cells to a bone marrow transplantation patient. In some embodiments, the subject and the patient are histocompatible. In some embodiments, the subject and the patient are syngeneic.
  • the subject and the patient are allogeneic.
  • the hematopoietic stem and/or progenitor cells are cultured to enrich and/or differentiate the hematopoietic stem and/or progenitor cells prior to introducing the cells into the patient.
  • the hematopoietic stem and/or progenitor cells are enriched for or differentiated into myeloid cells.
  • the hematopoietic stem and/or progenitor cells are enriched for or differentiated into lymphoid cells.
  • the hematopoietic stem and/or progenitor cells are enriched for or differentiated into NK cells.
  • the Btk inhibitor is reversible. In some embodiments, the Btk inhibitor is irreversible.
  • Preferred mobilizing Btk inhibitors suitable for use in the subject invention comprise a 6,5 fused heteroaryl ring system.
  • the 6,5 fused heteroaryl ring system is selected from: imidazopyrazine and imidazotriazine ring systems.
  • the 6,5 fused heteroaryl ring system is an imidazotriazine ring system.
  • Btk inhibitors suitable for use in the subject invention comprise an imidazopyrazine ring system. These Btk inhibitors have higher aqueous solubility and a lower serum affinity for albumin.
  • these Btk inhibitors also have lower lipophilicity and basic pKa values which renders these Btk inhibitors amenable for salt formation. Accordingly, these Btk inhibitors are expected to have better pharmacokinetic properties and higher solubility. Higher exposure levels, and a higher free fraction in circulation will consequently result in higher uptake in target tissue.
  • the mobilizing Btk inhibitor is selected from a compound of structural Formula I:
  • X is CH, N, O or S
  • Y is C(R6), N, O or S;
  • Z is CH, N or bond; A is CH or N;
  • Bl isNorC(R7)
  • B2 is N or C(R8)
  • B3 is N or C(R9)
  • B4 isNorC(RlO);
  • Rl is Rl lC(O), R12S(0), R13S0 2 or ( 1 -6C)alkyl optionally substituted with R14;
  • R2 is H, (l-3C)alkyl or (3-7C)cycloalkyl;
  • R3 is H, (l-6C)alkyl or (3-7C)cycloalkyl); or
  • R2 and R3 form, together with the N and C atom they are attached to, a (3-7C)heterocycloalkyl optionally substituted with one or more fluorine, hydroxyl, (1 -3C)alkyl, (l-3C)alkoxy or oxo;
  • R5 is H, halogen, cyano, (l-4C)alkyl, (l-3C)alkoxy, (3-6C)cycloalkyl, any alkyl group of which is optionally substituted with one or more halogen; or
  • R5 is (6-10C)aryl or (2-6C)heterocycloalkyl
  • R6 isHor(l-3C)alkyl
  • R5 and R6 together may form a (3-7C)cycloalkenyl, or (2-6C)heterocycloalkenyl; each optionally substituted with (l-3C)alkyl, or one or more halogen;
  • R7 is H, halogen or (l-3C)alkoxy
  • R8 isHor(l-3C)alkyl
  • R7 and R8 form, together with the carbon atom they are attached to a (6-10C)aryl or (l-9C)heteroaryl;
  • R9 is H, halogen or (l-3C)alkoxy;
  • RIO is H, halogen or (l-3C)alkoxy
  • Rl 1 is independently selected from a group consisting of (l-6C)alkyl, (2-6C)alkenyl and (2- 6C)alkynyl each alkyl, alkenyl or alkynyl optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (1- 3C)alkoxy, (3-7C)cycloalkoxy, (6-10C)aryl or (3-7C)heterocycloalkyl; or
  • Rl l is (l-3C)alkyl-C(0)-S-(l-3C)alkyl; or
  • Rl 1 is (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen or cyano;
  • R12 and R13 are independently selected from a group consisting of (2-6C)alkenyl or (2-6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl,
  • R14 is independently selected from a group consisting of halogen, cyano or (2-6C)alkenyl or (2- 6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3-7C)cycloalkoxy, (6- 10C)aryl, (l-5C)heteroaryl or (3-7C)heterocycloalkyl;
  • X, Y, Z can simultaneously be a heteroatom
  • the mobilizing Btk inhibitor for use in the subject invention is a compound selected from the group consisting of:
  • the mobilizing Btk inhibitor for use in the subject invention is a compound selected from the group consisting of:
  • the compounds of Formula I may include an asymmetric center or centers, and may be in the form of a composition of a racemic mixture, a diastereoisomeric mixture, a single enantiomer, an enantiomeric diastereomer, a meso compound, a pure epimer, or a mixture of epimers thereof, etc.
  • the compounds of Formulas I or II may have one or more double bonds, and may be in a form of a cis/trans, E/Z mixture or an E or Z geometric isomer thereof.
  • the compounds of Formula I may also be prepared as a salt form, e.g., pharmaceutically acceptable salts, including suitable acid forms, e.g., salt forms selected from hydrochloride, hydrobromide, acetate, propionate, butyrate, sulphate, hydrogen sulphate, sulphite, carbonate, hydrogen carbonate, phosphate, phosphinate, oxalate, hemi-oxalate, malonate, hemi-malonate, fumarate, hemi-fumarate, maleate, hemi-maleate, citrate, hemi-citrate, tartrate, hemi-tartrate, aspartate, glutamate, etc.
  • suitable acid forms e.g., salt forms selected from hydrochloride, hydrobromide, acetate, propionate, butyrate, sulphate, hydrogen sulphate, sulphite, carbonate, hydrogen carbonate, phosphate, phosphinate, ox
  • Figure 1 shows the results of a flow cytometry analysis showing that CD34+ cells are mobilized to the peripheral blood in rats dosed with 300 mg/kg/day COMPOUND 1 dose.
  • Figure 2 shows a summary of CD34+ cell frequencies in rat PBMC samples (Day 7 Study).
  • Figure 3 shows the results from a CFC Assay from rats treated with vehicle or COMPOUND 1 (300 mg/kg/day) for 7 or 28 days.
  • White blood cells include neutrophils, macrophages, eosinophils, basophils, mast cells, and the B and T cells of the immune system. White blood cells are continuously replaced via the hematopoietic system, by the action of colony stimulating factors (CSF) and various cytokines on progenitor cells in hematopoietic tissues.
  • CSF colony stimulating factors
  • cytokines various cytokines on progenitor cells in hematopoietic tissues.
  • the nucleotide sequences encoding a number of these growth factors have been cloned and sequenced.
  • G-CSF granulocyte colony stimulating factor
  • GM-CSF granulocyte-macrophage colony stimulating factor
  • IL-1 Interleukin-1
  • IL-3 Interleukin-3
  • IL-8 Interleukin-8
  • ⁇ -321 GM- CSF/IL-3 fusion protein
  • macrophage inflammatory protein stem cell factor
  • thrombopoietin growth related oncogene
  • progenitor cell refers to a cell that, in response to certain stimuli, can form differentiated hematopoietic or myeloid cells.
  • the presence of progenitor cells can be assessed by the ability of the cells in a sample to form colony-forming units of various types, including, for example, CFU-GM (colony-forming units, granulocyte-macrophage); CFU-GEMM (colony-forming units, multipotential); BFU-E (burst-forming units, erythroid); HPP-CFC (high proliferative potential colony-forming cells); or other types of differentiated colonies which can be obtained in culture using known protocols.
  • CFU-GM colony-forming units, granulocyte-macrophage
  • CFU-GEMM colony-forming units, multipotential
  • BFU-E burst-forming units, erythroid
  • HPP-CFC high proliferative potential colony-forming cells
  • stem cells are less differentiated forms of progenitor cells. Typically, such cells are often positive for CD34. Some stem cells do not contain this marker, however. These CD34+ cells can be assayed using fluorescence activated cell sorting (FACS) and thus their presence can be assessed in a sample using this technique.
  • FACS fluorescence activated cell sorting
  • CD34+ cells are present only in low levels in the blood, but are present in large numbers in bone marrow. While other types of cells such as endothelial cells and mast cells also may exhibit this marker, CD34 is considered an index of stem cell presence.
  • Mature blood cells are derived from hematopoietic precursor cells (progenitor) cells and stem cells present in specific hematopoietic tissues including bone marrow. Within these environments hematopoietic cells proliferate and differentiate prior to entering the circulation.
  • the chemokine receptor CXCR4 and its natural ligand stromal cell derived factor-1 (SDF-1) appear to be important in this process. See, e.g., Maekawa et al (2000) Internal Med. 39:90-100; Nagasawa et al. (2000) Int. J. Hematol. 72:408 41 1).
  • CD34+ progenitor cells express CXCR4 and require SDF-1 produced by bone marrow stromal cells for chemoattraction and engraftment, Peled et al. (1999) Science 283 :845- 48, and that in vitro, SDF-1 is chemotactic for both CD34+ cells, Aiuti et al. (1997) J. Exp. Med. 185: 11 1-120; Viardot et al. (1998)Ann. Hematol. 77: 194 197, and for progenitor cells, Jo et al. (2000) J. Clin. Invest. 105: 101 -111.
  • SDF-1 is also an important chemoattractant, signaling via the CXCR4 receptor, for several other more committed hematopoietic progenitors and mature blood cells including T-lymphocytes and monocytes, Bleul et al. (1996) J. Exp. Med. 184: 1 101-1 109), pro-and pre-B lymphocytes, Fedyk et al. (1999) J. Leukoc. Biol. (1999) 66:667-673; Ma et al. (1999) Immunity 10:463-71 , and megakaryocytes. Hodohara et al. (2000) Blood 95:769-75; Riviere et al.
  • SDF-1 is a CXC chemokine and is considered as one of the most potent chemoattractants of hematopoietic stem cells (HSC) into the bone marrow.
  • HSC hematopoietic stem cells
  • SDF-1 binding to CXCR4 triggers G protein coupling (from inactive conformation to an active conformation) and subsequent dissociation of the heterotrimeric G protein into ⁇ and Gai subunits, which in turn bind to several downstream effectors, resulting in activation of PI3K, protein kinase C (PKC), and MAPK- mediated pathways.
  • Sharma et al. (201 1) Stem Cells Dev. 20 (6): 933-46.
  • Studies of the SDF 1/CXCR4 axis in B-cell migration have identified Bruton's tyrosine kinase (Btk) as a central kinase in the SDF-1/CXCR4 signaling cascade.
  • Btk Bruton's tyrosine kinase
  • Btk expression has been described in HSC, multipotent progenitor cells and cells of the myeloid lineage, Mohammed et al. (2009) Immunological Reviews 228: 58-73, including erythroid cells, platelets, monocytes, macrophages, granulocytes, and dendritic cells. In the lymphoid lineage, Btk is expressed in B cells, but not in T cells or natural killer cells (NK cells). Schmidt et al (2004) Int Arch Allergy Immunol. 134:65-78.
  • Mozobil® a small molecule inhibitor which binds to CXCR4 preventing binding of SDF-1 and subsequent downstream signaling, causes rapid and reversible mobilization of CD34+ HSC into the peripheral blood for harvesting by apheresis.
  • Blocking the SDF-1/CXCR4 signaling cascade with a Btk inhibitor (PCI-32765) in patients with B-cell malignancies produces rapid and clinically significant decreases in lymphadenopathy as the malignant B cells are mobilized out of the lymph nodes and into the peripheral blood producing a marked lymphocytosis.
  • the present invention elucidates the role of Btk in HSC homing and mobilization and provides compositions and methods of using Btk inhibitors for hematopoietic stem and/or progenitor cell mobilization.
  • Acyl means -C(0)-alkyl, where alkyl is as defined herein.
  • Acylthio means -SC(0)-alkyl, where alkyl is as defined herein.
  • alkenyl means a straight or branched chain, saturated monovalent hydrocarbon radical having one or more carbon-carbon double bonds.
  • the hydrocarbon chain may have from two to twenty carbons, two to sixteen carbons, two to fourteen carbons, two to twelve carbons, two to ten carbons, two to eight carbons, two to six carbons, two to four carbons, etc.
  • Lower alkenyl may refer to alkenyls having, e.g., two to six carbons, two to four carbons, etc.
  • a straight chain alkenyl may have from two to six carbon atoms and a branched alkenyl three to six carbon atoms, e.g., ethenyl, prop-l -en-l -yl, prop-2-en-2-yl, allyl, butenyl (including all isomeric forms), pentenyl (including all isomeric forms), and the like.
  • alkenylcarbonyl means -C(0)-alkenyl, where alkenyl is as defined herein.
  • alkenylsulfonyl means -S0 2 -alkenyl, where alkenyl is as defined herein.
  • Alkyl means a straight or branched chain, saturated monovalent hydrocarbon radical.
  • the hydrocarbon chain may have from one to twenty carbons, one to sixteen carbons, one to fourteen carbons, one to twelve carbons, one to ten carbons, one to eight carbons, one to six carbons, one to four carbons, etc.
  • “Lower alkyl” may refer to alkyls having, e.g., one to six carbons, one to four carbons, etc.
  • a straight chain alkyl may have from one to six carbon atoms and a branched alkyl three to six carbon atoms, e.g., methyl, ethyl, propyl, 2-propyl, butyl
  • Aryl means a monovalent monocyclic or bicyclic aromatic hydrocarbon radical, e.g., having from of 6 to 20 or 6 to 10 ring atoms e.g., phenyl or naphthyl.
  • Alkynyl means a straight or branched chain, saturated monovalent hydrocarbon radical having one or more carbon-carbon triple bonds.
  • the hydrocarbon chain may have from two to twenty carbons, two to sixteen carbons, two to fourteen carbons, two to twelve carbons, two to ten carbons, two to eight carbons, two to six carbons, two to four carbons, etc.
  • “Lower alkynyl” may refer to alkynyls having, e.g., two to six carbons, two to four carbons, etc.
  • a straight chain alkynyl may have from two to six carbon atoms and a branched alkynyl three to six carbon atoms, e.g., ethynyl, prop-l-yn-l -yl, prop-2-yn-2-yl, propargyl, butynyl (including all isomeric forms), pentynyl (including all isomeric forms), and the like.
  • Alkynylcarbonyl means -C(0)-alkynyl, where alkynyl is as defined herein.
  • Heteroarylcarbonyl means -C(0)-heteroaryl, where heteroaryl is as defined herein.
  • Alkylaryl means a -(alkylene)-R radical where R is aryl as defined above.
  • Cycloalkyl means a cyclic saturated or partially saturated monovalent hydrocarbon radical (or an alicyclic radical).
  • the cycloalkyl may have from three to twenty carbon atoms, from three to sixteen carbon atoms, from three to fourteen carbon atoms, from three to twelve carbon atoms, from three to ten carbon atoms, from three to eight carbon atoms, from three to six carbon atoms, etc., wherein one or two carbon atoms may be replaced by an oxo group, e.g., admantanyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, indanyl and the like.
  • Alkylcycloalkyl means a -(alkylene)-R radical where R is cycloalkyl as defined above; e.g., cyclopropylmethyl, cyclobutylmethyl, cyclopentylethyl, or cyclohexylmethyl, and the like.
  • Heterocyclyl or “heterocycloalkyl” means a saturated or unsaturated monovalent monocyclic group, in which one or two ring atoms are heteroatom selected from N, O, or S, the remaining ring atoms being C.
  • the heterocyclyl ring is optionally fused to a (one) aryl or heteroaryl ring as defined herein.
  • the heterocyclyl ring fused to monocyclic aryl or heteroaryl ring is also referred to in this Application as "bicyclic heterocyclyl” ring.
  • one or two ring carbon atoms in the heterocyclyl ring can optionally be replaced by a -CO- group.
  • heterocyclyl includes, but is not limited to, pyrrolidino, piperidino, homopiperidino, 2- oxopyrrolidinyl, 2-oxopiperidinyl, morpholino, piperazino, tetrahydropyranyl, thiomorpholino, and the like.
  • heterocyclyl ring is unsaturated it can contain one or two ring double bonds.
  • heterocyclyl group contains at least one nitrogen atom, it is also referred to herein as
  • heterocycloamino is a subset of the heterocyclyl group.
  • the heterocyclyl group is a saturated ring and is not fused to aryl or heteroaryl ring as stated above, it is also referred to herein as saturated monocyclic heterocyclyl.
  • Alkylheterocycloalkyl means a -(alkylene)-R radical where R is heterocyclyl ring as defined above e.g., tetraydrofuranylmethyl, piperazinylmethyl, morpholinylethyl, and the like.
  • Heteroaryl means a monovalent monocyclic or bicyclic aromatic radical, where one or more, preferably one, two, or three, ring atoms are heteroatom selected from N, O, or S, the remaining ring atoms being carbon.
  • Representative examples include, but are not limited to, pyrrolyl, thienyl, thiazolyl, imidazolyl, furanyl, indolyl, isoindolyl, oxazolyl, isoxazolyl, diazolyl, pyrazolyl, triazolyl, benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, tetrazolyl, and the like.
  • substituted means that the referenced group is substituted with one or more additional group(s) individually and independently selected from groups described herein.
  • an optional substituent is selected from oxo, halogen, -CN, -NH 2 , -OH, -NH(CH 3 ), -N(CH 3 ) 2 , alkyl (including straight chain, branched and/or unsaturated alkyl), substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, fluoroalkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted alkoxy, fluoroalkoxy, -S-alkyl, -S(0) 2 -alkyl, -CONH((substituted or unsubstituted alkyl) or (substituted or unsubstituted phenyl)), -CON(H
  • alkyl CO(substituted or unsubstituted alkyl), -NHCOO(substituted or unsubstituted alkyl), -C(OH)( substituted or unsubstituted alkyl) 2 , and -C(NH 2 )( substituted or unsubstituted alkyl) 2 .
  • an optional substituent is selected from oxo, fluorine, chlorine, bromine, iodine, -CN, -NH 2 , -OH, -NH(CH 3 ), -N(CH 3 ) 2 , -CH 3 , -CH 2 CH 3 , -CH(CH 3 ) 2 , -CF 3 , -CH 2 CF 3 , -OCH 3 , -OCH 2 CH 3 , -OCH(CH 3 ) 2 , -OCF 3 , -OCH 2 CF 3 , -S(0) 2 -CH 3 , -CONH 2 ,
  • substituted groups are substituted with one, two or three of the preceding groups. In some embodiments, substituted groups are substituted with one or two of the preceding groups. In some embodiments, substituted groups are substituted with one of the preceding groups. Further, unless stated to the contrary, a formula with chemical bonds shown only as solid lines and not as wedges or dashed lines contemplates each possible isomer, e.g., each enantiomer and diastereomer, and a mixture of isomers, such as racemic or scalemic mixtures.
  • amino acid includes any one of the twenty naturally-occurring amino acids or the D-form of any one of the naturally-occurring amino acids.
  • amino acid also includes other non-naturally occurring amino acids besides the D-amino acids, which are functional equivalents of the naturally-occurring amino acids.
  • non-naturally occurring amino acids include, for example, norleucine ("Nle”), norvaline (“Nva”), L- or D-naphthalanine, ornithine (“Orn”), homoarginine (homoArg) and others well known in the peptide art, such as those described in M.
  • Amino acids and amino acid analogs can be purchased commercially (Sigma Chemical Co.; Advanced Chemtech) or synthesized using methods known in the art.
  • “Therapeutically effective amount” or “effective amount” means the amount of a composition, compound, therapy, or course of treatment that, when administered to a subject for treating a disease, disorder, or condition, is sufficient to effect such treatment for the disease, disorder, or condition.
  • the “therapeutically effective amount” will vary depending on the composition, the compound, the therapy, the course of treatment, the disease, disorder, or condition, and its severity and the age, weight, etc., of the subject to be treated.
  • Bruton's tyrosine kinase is a member of the cytoplasmic Tec family of kinases. Like other Btk family members, it contains a pleckstrin homology (PH) domain, and Src homology SH3 and SH2 domains. Btk plays an important role in B cell development. Btk comprises of several domains from the N-terminus: the PH, Tec homology (TH), SH2, SH3, and kinase (SHI) domains. Each of these domains has the potential to interact with a plethora of proteins critical for intracellular signaling. Moreover, functional association of Btk with many of its partners is crucial for its activation and regulation. Btk is a metalloprotein enzyme requiring Zn2+ for optimal activity and stability.
  • Preferred mobilizing Btk inhibitors suitable for use in the subject invention comprise a 6,5 fused heteroaryl ring system.
  • Btk inhibitors suitable for use in the subject invention comprise an imidazopyrazine ring system.
  • the mobilizing Btk inhibitor is selected from a compound of structural Formula I:
  • Wis(l-3C)alkyl or NH 2 ;
  • X is CH, N, O or S
  • Y is C(R6), N, O or S;
  • Z is CH, N or bond; A is CH or N;
  • Bl isNorC(R7)
  • B2 is N or C(R8)
  • B3 is N or C(R9)
  • B4 isNorC(RlO);
  • Rl is Rl lC(O), R12S(0), R13S0 2 or (l-6C)alkyl optionally substituted with R14;
  • R2 is H, (l-3C)alkyl or (3-7C)cycloalkyl;
  • R3 is H, (l-6C)alkyl or (3-7C)cycloalkyl); or
  • R2 and R3 form, together with the N and C atom they are attached to, a (3-7C)heterocycloalkyl optionally substituted with one or more fluorine, hydroxyl, (l-3C)alkyl, (l-3C)alkoxy or oxo;
  • R5 is H, halogen, cyano, (l-4C)alkyl, (l-3C)alkoxy, (3-6C)cycloalkyl, any alkyl group of which is optionally substituted with one or more halogen; or
  • R5 is (6-10C)aryl or (2-6C)heterocycloalkyl
  • R6 isHor(l-3C)alkyl
  • R5 and R6 together may form a (3-7C)cycloalkenyl, or (2-6C)heterocycloalkenyl; each optionally substituted with (l-3C)alkyl, or one or more halogen;
  • R7 is H, halogen or (l-3C)alkoxy
  • R8 isHor(l-3C)alkyl
  • R7 and R8 form, together with the carbon atom they are attached to a (6-10C)aryl or ( 1 -9C)heteroaryl;
  • R9 is H, halogen or (l -3C)alkoxy;
  • RI O is H, halogen or (l-3C)alkoxy
  • Rl 1 is independently selected from a group consisting of (l-6C)alkyl, (2-6C)alkenyl and (2- 6C)alkynyl each alkyl, alkenyl or alkynyl optionally substituted with one or more groups selected from hydroxyl, (l -4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (1- 3C)alkoxy, (3-7C)cycloalkoxy, (6-10C)aryl or (3-7C)heterocycloalkyl; or
  • Rl l is (l-3C)alkyl-C(0)-S-(l-3C)alkyl; or
  • Rl 1 is (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen or cyano;
  • R12 and R13 are independently selected from a group consisting of (2-6C)alkenyl or (2-6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (l -4C)alkyl, (3- 7C)cycloalkyl, [(l -4C)alkyl]amino, di[(l -4C)alkyl]amino, (l-3C)alkoxy, (3-7C)cycloalkoxy, (6- 10C)aryl or (3-7C)heterocycloalkyl; or (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen, or cyano;
  • R14 is independently selected from a group consisting of halogen, cyano or (2-6C)alkenyl or (2- 6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3-7C)cycloalkoxy, (6- 10C)aryl, (l -5C)heteroaryl or (3-7C)heterocycloalkyl;
  • X, Y, Z can simultaneously be a heteroatom
  • the Btk inhibitor is selected from a compound of structural Formula II:
  • W is (l -3C)alkyl, or NH 2 ;
  • Rl is selected from: (l -6C)acyl, (2-6)alkenylcarbonyl, (2-6)alkenylsulfonyl, (l -6C)alkyl, (2- 6)alkynylcarbonyl, and (l-5C)heteroarylcarbonyl, each optionally substituted with one or more substituents selected from: (l-6C)acylthio, (l-3C)alkoxy, (6-10C)aryl, cyano, (3-7C)cycloalkyl, di[(l- 4C)alkyl]amino, (3-7C)heterocycloalkyl, halogen, and hydroxyl;
  • R2 is H or (l-3C)alkyl
  • R3 is H or (l-6C)alkyl
  • R2 and R3 form, together with the N and C atom they are attached to, a (5-7C)heterocycloalkyl;
  • R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2- yl, pyrimidin-4-yl, pyridazin-3-yl, and isoxazol-3-yl, each optionally substituted with one substituent selected from: (l -3C)alkoxy, (l -4C)alkyl, (6-10C)aryl, (3-6C)cycloalkyl, halogen, and (3- 6C)heterocycloalkyl, wherein (l -4C)alkyl is further optionally substituted with one or more halogens.
  • the Btk inhibitor is selected from a compound of structural Formula II:
  • W is methyl, or NH 2 ;
  • Rl is selected from: acetyl, acryloyl, but-2-enoyl, but-2-ynoyl, hex-2-ynoyl, methacryloyl, methyl, 4- methylpent-2-ynoyl, pent-2-ynoyl, pyrimidine-4-carbonyl, and vinylsulfonyl, each optionally substituted with one or more substituents selected from: chloro, cyano, dimethylamino, fluoro, hydroxyl, methoxy, phenyl, and pyrrolidinyl;
  • R2 is H or methyl
  • R3 is H or methyl
  • R2 and R3 form, together with the N and C atom they are attached to, pyrrolidin-2-yl, piperidin-2-yl, azepan-2-yl, or morpholin-3-yl;
  • R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2- yl, pyrimidin-4-yl, pyridazin-3-yl, and isoxazol-3-yl, each optionally substituted with one substituent selected from: cyano, ethyl, fluoro, isopropyl, methoxy, methyl, phenyl, n-propyl, pyrrolidinyl, trifluoromethyl.
  • the Btk inhibitor is selected from a compound of structural Formula II:
  • W is methyl, or NH 2 ;
  • Rl is selected from 2-(acetylthio)acetyl, acryloyl, but-2-enoyl, but-2-ynoyl, cinnamoyl, cyanomethyl,
  • R2 is H or methyl
  • R3 is H or methyl
  • R2 and R3 form, together with the N and C atom they are attached to, pyrrolidin-2-yl, piperidin-2-yl, azepan-2-yl, or morpholin-3-y;
  • R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2- yl, pyrimidin-4-yl, pyridazin-3-yl, isoxazol-3-yl, 4-fluoropyridin-2-yl, 4-methylpyridin-2-yl, 4- propylpyridin-2-yl, 4-(trifluoromethyl)pyridin-2-yl, 4-ethylpyridin-2-yl, 4-cyanopyridin-2-yl, 4- methylpyrimidin-2-yl, 5-ethylthiazol-2-yl, 4-(pyrrolidin-l-yl)pyridin-2-yl, 4-methoxypyridin-2-yl, 4- isopropylpyridin-2-yl, 4-phenylpyridin-2-yl.
  • the Btk inhibitor includes every combination of one or more compounds selected from the following group shown in Table A.
  • the Btk inhibitor includes every combination of one or more compounds selected from the following group shown in Table B.
  • the mobilizing Btk inhibitor for use in the subject invention is selected from the group consisting of:
  • the mobilizing Btk inhibitor for use in the subject invention is selected from the group consisting of:
  • the Btk inhibitors described herein include further forms of the compounds such as pharmaceutically acceptable salts, solvates (including hydrates), amorphous phases, partially crystalline and crystalline forms (including all polymorphs), prodrugs, metabolites, N-oxides, isotopically-labeled, epimers, pure epimers, epimer mixtures, enantiomers including but not limited to single enantiomers and enantiomeric diastereomers, meso compounds, stereoisomers, racemic mixtures and diastereoisomeric mixtures.
  • Btk inhibitors described herein having one or more double bonds include cis/trans isomers, E/Z isomers and geometric isomers.
  • Btk inhibitors described herein can be prepared as a pharmaceutically acceptable salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, for example an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base.
  • a metal ion for example an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base.
  • the salt forms of the disclosed compounds can be prepared using salts of the starting materials or intermediates.
  • a Btk inhibitor of the disclosure is present in a composition as a salt.
  • salts are obtained by reacting a compound of the disclosure with acids.
  • pharmaceutically acceptable salts are obtained by reacting a compound of the disclosure with a base.
  • the compounds are used as free-acid or free-base form in the manufacture of the compositions described herein.
  • the type of salts include, but are not limited to: (1) acid addition salts, formed by reacting the free base form of the compound with a
  • inorganic acid such as, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, metaphosphoric acid, and the like; or with an organic acid, such as, for example, acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, trifluoroacetic acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2- hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo-[2.
  • organic acid
  • the Btk inhibitor described herein are reacted with an organic base, such as, but not limited to, ethanolamine, diethanolamine, triethanolamine, tromethamine, N— methylglucamine, dicyclohexylamine, tris(hydroxymethyl)methylamine.
  • an organic base such as, but not limited to, ethanolamine, diethanolamine, triethanolamine, tromethamine, N— methylglucamine, dicyclohexylamine, tris(hydroxymethyl)methylamine.
  • the compounds described herein form salts with amino acids such as, but not limited to, arginine, lysine, and the like.
  • Acceptable inorganic bases used to form salts with compounds that include an acidic proton include, but are not limited to, aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like.
  • stereochemistry of such chiral centers can independently be in the R or S configuration, or a mixture of the two.
  • the chiral centers can be further designated as R or S or R,S or d,D, 1,L or d,l, D,L.
  • the Btk inhibitors of the invention if they can be present in optically active form, can actually be present in the form of a racemic mixture of enantiomers, or in the form of either of the separate enantiomers in substantially isolated and purified form, or as a mixture comprising any relative proportions of the enantiomers.
  • diastereomers When the Btk inhibitors described herein contain two or more chiral centers then diastereomers are possible. Such diastereomers may be present as pure diastereomeric enantiomers, pure racemic mixtures of diastereomeric enantiomers, mixtures of diastereomers which may be racemic or may have optical activity in their own right due to complex permutations of enantiomeric diastereomers in the balance of the mixtures.
  • the Btk inhibitors of the invention can be present in geometrically isomeric forms, then they can actually be present in the form of a mixture of geometric isomers comprising any relative proportions of the isomers, or in some cases in the form of either of the separate geometric isomers in substantially isolated and purified form.
  • the geometry around such double bonds can be independently a cis/trans, E/Z mixture or an E or Z geometric isomer thereof.
  • the Btk inhibitors described herein include solvent addition forms or crystal forms thereof, particularly solvates or polymorphs.
  • Solvates contain either stoichiometric or non-stoichiometric amounts of a solvent, and may be formed during the process of crystallization with pharmaceutically acceptable solvents such as water, ethanol, and the like. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is alcohol.
  • the Btk inhibitors described herein possess one or more stereocenters and each center exists independently in either the R or S configuration.
  • the Btk inhibitors presented herein include all diastereomeric, enantiomeric, and epimeric forms as well as the appropriate mixtures thereof.
  • sites on the Btk inhibitors disclosed herein are susceptible to various metabolic reactions. Therefore incorporation of appropriate substituents at the places of metabolic reactions will reduce, minimize or eliminate the metabolic pathways.
  • the appropriate substituent to decrease or eliminate the susceptibility of the aromatic ring to metabolic reactions is, by way of example only, a halogen, deuterium or an alkyl group.
  • the Btk inhibitors described herein are isotopically-labeled, which are identical to those recited in the various formulae and structures presented herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature.
  • one or more hydrogen atoms are replaced with deuterium.
  • metabolic sites on the compounds described herein are deuterated.
  • substitution with deuterium affords certain therapeutic advantages resulting from greater metabolic stability, such as, for example, increased in vivo half-life or reduced dosage requirements.
  • groups and substituents thereof can be chosen by one skilled in the field to provide stable moieties and compounds.
  • Inhibitors of Btk kinase activity preferably inhibit Btk activity with an IC 50 of less than or equal to 10 micromolar, less than or equal to 1 micromolar, less than or equal to 500 nanomolar, less than or equal to 100 nanomolar, or less than or equal to 10 nanomolar in an ADP-GLOTM, bioluminescent, homogeneous assay or in a HTRF® (Homogeneous Time-Resolved Fluorescence) assay.
  • IC 50 of less than or equal to 10 micromolar, less than or equal to 1 micromolar, less than or equal to 500 nanomolar, less than or equal to 100 nanomolar, or less than or equal to 10 nanomolar in an ADP-GLOTM, bioluminescent, homogeneous assay or in a HTRF® (Homogeneous Time-Resolved Fluorescence) assay.
  • Inhibitors of Bruton's tyrosine kinase activity preferably inhibit phosphorylation of Y551 of Btk with an IC 50 of less than or equal to 10 micromolar, less than or equal to 1 micromolar, less than or equal to 500 nanomolar, less than or equal to 1 12 nanomolar, less than or equal to 100 nanomolar, or less than or equal to 10 nanomolar.
  • Inhibitors of Bruton's tyrosine kinase activity preferably inhibit phosphorylation of Y223 of Btk with an IC 50 of less than or equal to 10 micromolar, less than or equal to 1 micromolar, less than or equal to 500 nanomolar, less than or equal to 1 12 nanomolar, less than or equal to 100 nanomolar, or less than or equal to 10 nanomolar.
  • Inhibitors of Bruton's tyrosine kinase activity may inhibit phosphorylation of downstream PLCy2 with an IC 50 of less than or equal to 10 micromolar, less than or equal to 1 micromolar, less than or equal to 500 nanomolar, less than or equal to 100 nanomolar, or less than or equal to 10 nanomolar in, e.g., an IgM phospho-PLCg2 assay.
  • the pharmaceutical compositions will generally comprise a pharmaceutically acceptable carrier and a pharmacologically effective amount of the compounds, or mixture of thereof, or suitable salts thereof.
  • the pharmaceutical composition may be formulated as powders, granules, solutions, suspensions, aerosols, solids, pills, tablets, capsules, gels, topical creams, suppositories, transdermal patches, and other formulations known in the art.
  • pharmaceutically acceptable carrier comprises any of standard
  • diluents for example, saline, phosphate buffer saline (PBS), aqueous ethanol, or solutions of glucose, mannitol, dextran, propylene glycol, oils (e.g., vegetable oils, animal oils, synthetic oils, etc.), microcrystalline cellulose, carboxymethyl cellulose, hydroxylpropyl methyl cellulose, magnesium stearate, calcium phosphate, gelatin, polysorbate 80 or the like, or as solid formulations in appropriate excipients.
  • PBS phosphate buffer saline
  • aqueous ethanol or solutions of glucose, mannitol, dextran, propylene glycol, oils (e.g., vegetable oils, animal oils, synthetic oils, etc.), microcrystalline cellulose, carboxymethyl cellulose, hydroxylpropyl methyl cellulose, magnesium stearate, calcium phosphate, gelatin, polysorbate 80 or the like, or as solid formulations in appropriate excipients.
  • the pharmaceutical compositions will often further comprise one or more buffers (e.g., neutral buffered saline or phosphate buffered saline), carbohydrates (e.g., glucose, mannose, sucrose or dextrans), mannitol, proteins, polypeptides or amino acids such as glycine, antioxidants (e.g., ascorbic acid, sodium metabisulfite, butylated hydroxytoluene, butylated hydroxyanisole, etc.), bacteriostats, chelating agents such as EDTA or glutathione, solutes that render the formulation isotonic, hypotonic or weakly hypertonic with the blood of a recipient, suspending agents, thickening agents, preservatives, flavoring agents, sweetening agents, and coloring compounds as appropriate.
  • buffers e.g., neutral buffered saline or phosphate buffered saline
  • carbohydrates e.g., glucose, mannose, sucrose or dextrans
  • compositions may be formulated for any appropriate manner of administration, including for example, oral, nasal, mucosal, rectal, vaginal, topical, intravenous, intraperitoneal, intradermal, subcutaneous, and intramuscular administration.
  • compositions can be administered as injectable dosages of a solution or suspension of the substance in a physiologically acceptable diluent with a pharmaceutical carrier that can be a sterile liquid such as sterile pyrogen free water, oils, saline, glycerol, polyethylene glycol or ethanol.
  • a pharmaceutical carrier can be a sterile liquid such as sterile pyrogen free water, oils, saline, glycerol, polyethylene glycol or ethanol.
  • auxiliary substances such as wetting or emulsifying agents, surfactants, pH buffering substances and the like can be present in compositions.
  • Other components of pharmaceutical compositions are those of petroleum, animal, vegetable, or synthetic origin, for example, non-aqueous solutions of peanut oil, soybean oil, corn oil, cottonseed oil, ethyl oleate, and isopropyl myristate.
  • Sterile injectable solutions can be prepared by incorporating the mobilizing Btk inhibitor in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization.
  • dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above.
  • the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
  • an agent of the invention as described above may be formulated with one or more additional compounds that enhance the solubility of these agents. The invention also extends to such derivatives of such agents of the invention.
  • compositions described herein may be presented in unit-dose or multi- dose containers, such as sealed ampoules or vials. Such containers are typically sealed in such a way to preserve the sterility and stability of the formulation until use.
  • formulations may be stored as suspensions, solutions or emulsions in oily or aqueous vehicles, as indicated above.
  • a pharmaceutical composition may be stored in a freeze-dried condition requiring only the addition of a sterile liquid carrier immediately prior to use.
  • a pharmaceutical composition comprising expanded hematopoietic progenitor cells cryopreserved in a suitable cryopreservation medium, which can then be thawed and resuspended as needed for administration to a patient.
  • compositions described herein can also contain the mobilizing Btk inhibitor in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs.
  • Compositions intended for oral use can be prepared according to any method known to the art for the manufacture of pharmaceutical compositions, and such compositions can contain one or more agents selected from, by way of non-limiting example, sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations.
  • Formulations suitable for oral administration can be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion.
  • composition preparations which can be used orally include tablets, push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Tablets can be made by compression or molding, optionally with one or more accessory ingredients.
  • Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with binders (e.g., povidone, gelatin, hydroxypropylmethyl cellulose), inert diluents, preservative, disintegrant (e.g., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose) or lubricating, surface active or dispersing agents.
  • binders e.g., povidone, gelatin, hydroxypropylmethyl cellulose
  • inert diluents preservative
  • disintegrant e.g., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose
  • Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.
  • the tablets can optionally be coated or scored and can be formulated so as to provide slow or controlled release of the active ingredient therein. Tablets can optionally be provided with an enteric coating, to provide release in parts of the gut other than the stomach. All formulations for oral administration should be in dosages suitable for such administration.
  • the push- fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers.
  • the active compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • stabilizers can be added. Dragee cores are provided with suitable coatings.
  • concentrated sugar solutions can be used, which can optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.
  • Dyestuffs or pigments can be added to the tablets or Dragee coatings for identification or to characterize different combinations of active compound doses.
  • the compounds and compositions described herein can include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration can include flavoring agents.
  • one or more mobilizing Btk inhibitor may be formulated in pharmaceutical compositions with additional active ingredients, or administered in methods of treatment in conjunction with treatment with one or more additional medications, such as another agent as described herein that also increases mobilization of stem cells, for example, or one or more agents selected from the group consisting of: recombinant-methionyl human.
  • G-CSF G-CSF
  • GM-CSF G-CSF
  • LEUKINE® Sargramostim; Immunex
  • erythropoietin rhEPO, EPOGEN®; Amgen
  • thrombopoietin rhTPO; Genentech
  • interleukin-11 rhIL-1 1, NEUMEGA®; American Home Products
  • Flt3 ligand Flt3 ligand
  • Mobista Immunex
  • Mobista Immunex
  • Mobista multilineage hematopoietic factor
  • Melopoietin Leridistem; Searle
  • IL-3 myeloid progenitor inhibitory factor-1
  • Mirostipen Human Genome Sciences
  • stem cell factor rhSCF, STEMGEN®; Amgen
  • plerixafor Mozobil®; Genzyme-Sanofi.
  • the amount administered to the host will vary depending upon what is being administered, the purpose of the administration, such as prophylaxis or therapy, the state of the host, the manner of administration, the number of administrations, interval between administrations, and the like. These can be determined empirically by those skilled in the art and may be adjusted for the extent of the therapeutic response. Factors to consider in determining an appropriate dose include, but is not limited to, size and weight of the subject, the age and sex of the subject, the severity of the symptom, the stage of the disease, method of delivery of the agent, half-life of the agents, and efficacy of the agents. Stage of the disease to consider includes whether the disease is acute or chronic, relapsing or remitting phase, and the progressiveness of the disease.
  • an initial effective dose can be estimated from cell culture or other in vitro assays.
  • a dose can then be formulated in animal models to generate a circulating concentration or tissue concentration, including that of the IC 50 as determined by the cell culture assays.
  • toxicity and therapeutic efficacy are generally determined by cell culture assays and/or using experimental animals, typically by determining a LD 50 (lethal dose to 50% of the test population) and ED 50 (therapeutically effectiveness in 50% of the test population).
  • Guidance is found in standard reference works, for example, Goodman & Oilman's The Pharmacological Basis of Therapeutics, 10th Ed. (Hardman, J. G. et al., eds.) McGraw-Hill, New York, N.Y. (2001).
  • the methods of administration are chosen depending on the condition being treated, the form of the subject mobilizing Btk inhibitor(s), and the pharmaceutical composition.
  • Administration of the therapeutic compounds can be done in a variety of ways, including, but not limited to, orally subcutaneously, intravenously, intraperitoneally, intramuscularly, and possibly direct injection to specified organs such as e.g., spleen or bone marrow, although systemic and oral administration are preferred.
  • Administration of the pharmaceutical compositions may be through a single route or concurrently by several routes.
  • compositions may be administered once per day, a few or several times per day, or even multiple times per day, depending upon, among other things, the indication being treated and the judgment of the prescribing physician.
  • mobilizing Btk inhibitors and pharmaceutical compositions comprising the same.
  • the mobilizing Btk inhibitors and pharmaceutical compositions can be administered to a subject to mobilize the subject's hematopoietic stem and progenitor cells into the peripheral blood of the subject.
  • a mobilizing Btk inhibitor may be administered to a subject in need thereof, e.g., a patient having impaired hematopoiesis and/or deficient in white blood cells such as myeloid and lymphoid cells.
  • These therapies typically include high doses of irradiation and/or chemotherapeutic agents. While necessary to kill off the cancer cells, a significant side effect of these approaches is the pathological impact of such treatments on rapidly dividing normal cells, such as hair follicles, mucosal cells and the hematopoietic cells, including primitive hematopoietic progenitor cells and stem cells.
  • hematopoietic stem cells and progenitor/precursor cells leads to reduced mature blood cell counts, such as lymphocytes, neutrophils and platelets, eventually resulting in a loss of immune system function in these patients and a substantially increased risk of morbidity and mortality from opportunistic infections.
  • Leukopenia resulting from chemotherapy or irradiation therapy may occur within a few days following cytotoxic treatments, although the patient can be vulnerable to infection for up to one month until the white blood cell counts recover to within a normal range.
  • the therapy must be interrupted to allow for recovery of the white blood cells, which may in turn result in the survival of cancer cells, an increase drug resistance in the cancer cells, and may actually result in a relapse of the cancer.
  • the methods of mobilizing progenitor cells and/or providing hematopoietic cells to a patient disclosed herein may be used for the treatment of such impaired hematopoiesis.
  • a treatment modality that enhances the stem and/or progenitor cells in blood is helpful in treatments to ameliorate the effects of conditions that adversely affect the bone marrow, such as chemotherapy or irradiation (intentional or accidental) that results in leukopenia, including neutropenia, and thrombocytopenia.
  • the mobilizing Btk inhibitors disclosed herein may enhance the success of bone marrow transplantation, and may combat infections in the patient undergoing such therapies.
  • the compounds are also be used to mobilize and harvest hematopoietic stem cells or progenitor cells via apheresis and the harvested cells are used in treatments requiring stem cell transplantations.
  • the Btk inhibitors can be used both in vivo to promote mobilization of hematopoietic stem cells or progenitor cells from the bone marrow to the peripheral blood or can be used for ex vivo studies, whereby a patient's own stem cells are removed and expanded in culture for autologous transplants.
  • Also contemplated by the present invention are in vitro screens, whereby candidate or test compounds can be measured for their effects on mobilization before being administered in vivo.
  • the disorder relates to impairment of hematopoiesis caused by disease or myeloablative treatments.
  • treatment refers to therapeutic or prophylactic treatment.
  • Treatment encompasses administration of a Btk inhibitor capable of mobilizing hematopoietic stem and progenitor cells to the peripheral blood of a first subject, and optionally harvesting said mobilized progenitor cells and introducing them into a compatible subject.
  • the obtained progenitor cells are cultured ex vivo prior to reintroduction into the original subject.
  • the obtained progenitor cells are cultured ex vivo prior to introduction into a second subject.
  • the methods comprise administration of the obtained, and optionally cultured, cells in an appropriate form prior to the onset of disease symptoms and/or after clinical manifestations, or other manifestations of the disease or condition to reduce disease severity, halt disease progression, or eliminate the disease.
  • prophylactic or therapeutic treatment refers to administration to the subject of one or more of the Btk inhibitor compositions. If it is administered prior to clinical manifestation of the impaired hematopoiesis (e.g., leukopenia or neutropenia) then the treatment is prophylactic, i.e., it protects the host against developing the unwanted condition, whereas if administered after manifestation of impaired hematopoiesis, the treatment is therapeutic (i.e., it is intended to improve hematopoiesis and/or increase white blood cell count).
  • Subject refers to a mammal, preferably a human, in need of treatment for a condition, disorder or disease.
  • myelosuppressive chemotherapy regimens with cytotoxic agents such as doxorubicin, paclitaxel, cisplatin, carboplatin, etoposide, ifosfamide, daunorubicin, cytosine arabinoside, thioguanine, and the like typically induce a transient but profound myelosuppression in patients, at about seven to fourteen days after chemotherapy. This may be followed by rapid reappearance of leukocytes in the peripheral blood and a "rebound" increase of the circulating leukocytes above baseline levels. As the leukocyte count rises, hematopoietic progenitor cells also begin to appear in the peripheral blood and rapidly increase.
  • Protocols involving dose intensification (i.e., to increase the log-kill of the respective tumor therapy) or schedule compression may exacerbate the degree and duration of myelosuppression associated with the chemotherapy and/or radiation therapy.
  • dose intensification i.e., to increase the log-kill of the respective tumor therapy
  • schedule compression may exacerbate the degree and duration of myelosuppression associated with the chemotherapy and/or radiation therapy.
  • repeated cycles of doxorubicin-based treatment have been shown to produce cumulative and long-lasting damage in the bone marrow progenitor cell populations (Lorhrman et al., (1978) Br. J. Haematol. 40:369).
  • treatments using the mobilizing Btk inhibitors described may be provided to patients suffering from a cancerous condition or hyperproliferative disease, whereby the treatment of the disease with chemotherapy or irradiation therapy results in a decrease in bone marrow cellularity, thus making the patient more prone to acquiring infectious agents or diseases.
  • administration of the mobilizing Btk inhibitors of the present invention subsequent to the chemotherapy and/or radiation exposure allows for the mobilization of hematopoietic stem and/or progenitor cells from the bone marrow to the peripheral blood.
  • subsequent administration may be within 1 -24 hours, preferably within 12 to 24 or 24 to 36 or 48 hours, preferably within one to 30 days, more preferably within one to 14 or 21 days, more preferably within three or five to ten or fourteen days after the chemotherapy and/or radiation exposure. Improving hematopoiesis may advantageously allow for the use of accelerated doses of chemotherapy or irradiation therapy.
  • transplantation of hematopoietic stem and progenitor cells harvested from a subject may provide a more rapid and sustained hematopoietic recovery, e.g, after the administration of high-dose chemotherapy or radiation therapy in patients with hematological malignancies and solid tumours.
  • This has become the preferred source of hematopoietic stem cells for autologous transplantation because of the shorter time to engraftment and the lack of a need for surgical procedures such as are necessary for bone marrow harvesting (Demirer et al. (1996) Stem Cells 14: 106-1 16; Pettengel et al., (1992) Blood 82:2239-2248).
  • Hematopoietic stem cells and progenitors may be obtained from a subject according to the methods described herein, and enriched for and/or further differentiated into progenitors of different lineages according to well-known methods.
  • common lymphoid progenitor cells which give rise to B cells, T cells, and Natural Killer Cells
  • common myeloid progenitor cells which give rise to monocytes, granulocytes, megakaryocites, and erythrocytes may be enriched by sorting according to the expression levels of certain growth factor receptors. See, e.g., U.S. Patent Nos. 7,979,057; 7,235,623; 6,908,763; and 5,989,660, the disclosures of which are expressly incorporated by reference herein.
  • CLPs common lymphoid progenitors express low levels of c-kit (CD1 17) on their cell surface. Antibodies that specifically bind c-kit in humans, mice, rats, etc. are known in the art. Alternatively, the c-kit ligand, steel factor (Slf) may be used to identify cells expressing c-kit. The CLP cells express high levels of the IL-7 receptor alpha chain (CDwl27). Antibodies that bind to human or to mouse CDwl27 are known in the art. Alternatively, the cells are identified by binding of the ligand to the receptor, IL-7.
  • CDwl27 IL-7 receptor alpha chain
  • Murine CLPs express low levels of Sca-1 (Ly-6E and Ly-6A, see van de Rijn (1989) Proc Natl Acad Sci 86:4634-4638). Antibodies specific for Sca-1 are known in the art. The expression of high levels of Sca-1 on murine hematopoietic stem cells has been previously described (Spangrude et al. (1988) J Immunol 141 :3697-3707). A candidate human counterpart for Sca-1 is described in Hill et al. (1996) Exp Hematol 24:936-943, as high-level expression of a novel epitope of CD59 identifies a subset of CD34+ bone marrow cells that are highly enriched for pluripotent stem cells.
  • the CLP subset also has the phenotype of lacking expression of lineage specific markers, exemplified by B220, CD4, CD8, CD3, Gr-1 and Mac-1.
  • a cocktail of binding reagents herein designated “lin” may be used.
  • the lin panel will comprise binding reagents, e.g. antibodies and functional binding fragments thereof, ligands, peptidomimetics, etc., that recognize two or more of the lineage markers.
  • a lin panel will generally include at least one marker expressed on mature B cells, on mature T cells, on mature granulocytes and on mature macrophages.
  • Markers suitable for use in a lineage panel are typically expressed on these mature cells, but are not present on multiple lineages, or on stem and progenitor cells.
  • the subject cells are characterized as lacking expression of Thy-1 , a marker that is characteristic of hematopoietic stem cells.
  • the phenotype of the CLP may be further characterized as Mel-14 " , CD43 10 , HSA 10 , CD45 + and common cytokine receptor ⁇ chain
  • Myeloid progenitors may be subdivided into three distinct subsets: a common myeloid progenitor cell (CMP) which is characterized by progenitor cell activity for myeloid lineages, but lacking the potential to differentiate into lymphoid lineages; a granulocyte monocyte (GMP) committed progenitor cell; and an erythroid/megakaryocyte (MEP) committed progenitor cell.
  • CMP myeloid progenitor cell
  • GMP granulocyte monocyte
  • MEP erythroid/megakaryocyte
  • the CMP population is useful in transplantation to provide a recipient with myeloid cells, including megakaryocytes, platelets and erythroid cells, in addition to monocytes and granulocytes; for drug screening; experimental models of hematopoietic differentiation and interaction; screening in vitro assays to define growth and differentiation factors, and to characterize genes involved in myeloid development and regulation; and the like.
  • the native cells may be used for these purposes, or they may be genetically modified to provide altered capabilities.
  • Each of these progenitor subsets may be are separated from a complex mixture of cells by using reagents that specifically recognize markers on the cell surface.
  • all three of the myeloid lineage progenitors stain negatively for the markers Thy-1 (CD90), IL- 7R.alpha. (CD 127); and with a panel of lineage markers, which lineage markers may include CD2; CD3; CD4; CD7; CD8; CD10; CD1 lb; CD14; CD19; CD20; CD56; and glycophorin A (GPA) in humans and CD2; CD3; CD4; CD8; CD19; IgM; Terl 10; Gr-1 in mice.
  • all of the progenitor cells are CD34 positive.
  • all of the progenitor subsets may be further characterized as Sca-1 negative, (Ly-6E and Ly-6A), and c-kit high.
  • all three of the subsets are CD38+.
  • the progenitor subsets that is, the population of cells defined as Lin “ IL-7R " Thy- ⁇
  • the population can be divided into subsets for the CMP, GMP and MEP cells.
  • the markers IL-3Ra (CDwl27) and CD45RA are sufficient for separating the three subsets, where the CMP is IL- 3Ra 10 CD45RA " ; the GMP is IL-3Ra'° CD45Ra + ; and the MEP is IL-3Ra CD45RA " .
  • the CD34 and Fey receptor (FcyR) are useful in making these distinctions.
  • the CMP is characterized as FcyR 10 CD34 + population; the GMP is FcyR hi CD34 + ; and the MEP subset is FcyR 10 CD34 ⁇
  • CMP cells In the presence of steel factor (SLF), flt-3 ligand (FL), interleukin (IL)-3, IL-1 1, GM- CSF, thrombopoietin (Tpo) and erythropoietin (Epo), the CMP cells give rise to various types of myeloerythroid colonies, including CFU-GEMMeg, burst-forming unit-erythroid (BFU-E), CFU- megakaryocytes (CFU-Meg), CFU-granulocyte/macrophage (CFU-GM), CFU-granulocyte (CFU-G) and CFU-macrophage (CFU-M).
  • the GMP subset generates CFU-M, CFU-G, or CFU-GM colonies containing macrophages and/or granulocytes in response to the above growth factors.
  • the MEP subset gives rise to CFU-Meg, BFU-E, or CFU-MEP colonies containing only megakaryocytes and/or erythrocytes in response to IL-3, GM-CSF, Tpo and Epo, but do not form colonies in the absence of Tpo and Epo. All three myeloid progenitor populations do not require "early-acting cytokines" such as SLF, FL and IL- 11 to initiate colony formation.
  • CMP cells give rise to Gr-1+/Mac-1+ myelomonocytic cells and megakaryocyte colonies, as well as TER1 19+ erythroid cells in spleen and bone marrow.
  • the GMP progenitor population gives rise to Gr-1+/Mac- 1+ cells; and the MEP progenitor population to megakaryocytes and erythroid cells.
  • the subject subsets may be separated from a complex mixture of cells by techniques that enrich for cells having the above-described characteristics for the subset of interest.
  • An appropriate solution may be used for dispersion or suspension.
  • Such solution will generally be a balanced salt solution, e.g. normal saline, PBS, Hank's balanced salt solution, etc., conveniently supplemented with fetal calf serum or other naturally occurring factors, in conjunction with an acceptable buffer at low concentration, generally from 5-25 mM.
  • Convenient buffers include HEPES, phosphate buffers, lactate buffers, etc.
  • Separation of the subject cell populations may be attained by use of affinity separation to provide a substantially pure population.
  • Techniques for affinity separation may include magnetic separation, using antibody-coated magnetic beads, affinity chromatography, cytotoxic agents joined to a monoclonal antibody or used in conjunction with a monoclonal antibody, e.g. complement and cytotoxins, and "panning" with antibody attached to a solid matrix, e.g. plate, or other convenient technique.
  • Techniques providing accurate separation include fluorescence activated cell sorters, which can have varying degrees of sophistication, such as multiple color channels, low angle and obtuse light scattering detecting channels, impedance channels, etc.
  • the cells may be selected against dead cells by employing dyes associated with dead cells (e.g. propidium iodide). Any technique may be employed which is not unduly detrimental to the viability of the selected cells.
  • the affinity reagents may be specific receptors or ligands for the cell surface molecules indicated above.
  • peptide-MHC antigen and T cell receptor pairs may be used; peptide ligands and receptor; effector and receptor molecules, and the like.
  • Antibodies and T cell receptors may be monoclonal or polyclonal, and may be produced by transgenic animals, immunized animals, immortalized human or animal B-cells, cells transfected with DNA vectors encoding the antibody or T cell receptor, etc. The details of the preparation of antibodies and their suitability for use as specific binding members are well-known to those skilled in the art.
  • antibodies are conjugated with a label for use in separation.
  • Labels include magnetic beads, which allow for direct separation, biotin, which can be removed with avidin or streptavidin bound to a support, fluorochromes, which can be used with a fluorescence activated cell sorter, or the like, to allow for ease of separation of the particular cell type.
  • Fluorochromes that find use include phycobiliproteins, e.g. phycoerythrin and allophycocyanins, fluorescein and Texas red. Frequently each antibody is labeled with a different fluorochrome, to permit independent sorting for each marker.
  • the antibodies are added to a suspension of cells, and incubated for a period of time sufficient to bind the available cell surface antigens.
  • the incubation will usually be at least about 5 minutes and usually less than about 30 minutes. It is desirable to have a sufficient concentration of antibodies in the reaction mixture, such that the efficiency of the separation is not limited by lack of antibody.
  • the appropriate concentration is determined by titration.
  • the medium in which the cells are separated will be any medium which maintains the viability of the cells.
  • a preferred medium is phosphate buffered saline containing from 0.1 to 0.5% BSA.
  • Various media are commercially available and may be used according to the nature of the cells, including Dulbecco's Modified Eagle Medium (dMEM), Hank's Basic Salt Solution (HBSS), Dulbecco's phosphate buffered saline (dPBS), RPMI, Iscove's medium, PBS with 5 mM EDTA, etc., frequently supplemented with fetal calf serum, BSA, HSA, etc.
  • dMEM Dulbecco's Modified Eagle Medium
  • HBSS Hank's Basic Salt Solution
  • dPBS Dulbecco's phosphate buffered saline
  • RPMI Dulbecco's phosphate buffered saline
  • Iscove's medium PBS with 5 mM EDTA, etc., frequently supplemented with fetal calf serum, BSA, HSA, etc.
  • the labeled cells are then separated as to the expression of cell surface markers as previously described.
  • the separated cells may be collected in any appropriate medium that maintains the viability of the cells, usually having a cushion of serum at the bottom of the collection tube.
  • Various media are commercially available and may be used according to the nature of the cells, including dMEM, HBSS, dPBS, RPMI, Iscove's medium, etc., frequently supplemented with fetal calf serum.
  • the culture may contain growth factors to which the cells are responsive.
  • Growth factors as defined herein, are molecules capable of promoting survival, growth and/or differentiation of cells, either in culture or in the intact tissue, through specific effects on a transmembrane receptor.
  • Growth factors include polypeptides and non-polypeptide factors.
  • Specific growth factors that may be used in culturing the subject cells include steel factor (c-kit ligand), Flk-2 ligand, IL-1 1, IL-3, GM-CSF, erythropoietin and thrombopoietin.
  • the specific culture conditions are chosen to achieve a particular purpose, i.e. differentiation into erythroid of megakaryocyte populations, maintenance of progenitor cell activity, etc.
  • the subject cells may be grown in a co-culture with stromal or feeder layer cells.
  • Stromal cells suitable for use in the growth of hematopoietic cells are known in the art. These include bone marrow stroma as used in "Whitlock-Witte” (Whitlock et al.
  • the subject cultured cells may be used in a wide variety of ways.
  • the nutrient medium which is a conditioned medium, may be isolated at various stages and the components analyzed. Separation can be achieved with HPLC, reversed phase-HPLC, gel electrophoresis, isoelectric focusing, dialysis, or other non-degradative techniques, which allow for separation by molecular weight, molecular volume, charge, combinations thereof, or the like. One or more of these techniques may be combined to enrich further for specific fractions.
  • Example 1 Flow cytometry hierarchal gating scheme for PBMC samples.
  • FIG. 1 A 7-AAD negative "live” cells are gated, followed by forward and side scatter gating to identify cells from debris. Then CD45+ gate is applied to capture all hematopoietic, non- RBC precursors. Finally a CD34+, CD1 lb- gate identifies immature CD34+ cells.
  • Figure IB Individual rat data is shown for the No dose (vehicle control) and the 300 mg/kg/day dose. CD34+ percentages shown are of the total live cells.
  • Example 2 CD34 positive cell enumeration by flow cytometry.
  • CD45-FITC (BD Biosciences), CD34-PE (Santa Cruz Biotechnology), CD1 lb-APC (BD Biosciences)
  • CD45-FITC CD1 lb-APC, (no CD34 antibody)
  • CD45-FITC IgGl-PE (isotype control; BD Biosciences), CD1 lb Alexa 647
  • Hierarchical gating is used to discriminate discrete cell populations and the CD34+ gates are applied to the data using the no CD34+ (Stain #2) and the isotype control (Stain #3) samples as a guide to determine CD34+ from CD34- stained cells.
  • a rat bone marrow sample serves as a positive control for a sample containing CD34+ cells.
  • the CD34+ cells are plotted as a percentage live cells (see, figure 2). Student-T test is calculated using GraphPad Prism.
  • Example 3 Hematopoietic Colony Forming (CFC) assay.
  • Rat PBMC samples from the Day 7 and Day 29 studies are analyzed for the number of primitive hematopoietic cells that able to form colonies in semi-solid media.
  • MethoCultTM GF R3774 Stem Cell Technologies
  • rat IL-3, GM-CSF, and Stem Cell Factor growth factors
  • CFU-GM, CFU-M, CFU-G granulocyte-macrophage progenitors
  • Blocking the chemokine receptor, CXCR4 results in mobilization of hematopoietic progenitor cells from the bone marrow to the periphery as shown with the clinical use of AMD3100 (Mozobil), an antagonist of CXCR4.
  • This drug specifically increased CD34+ hematopoietic stem cell counts in the peripheral blood.
  • Stromal derived factor 1 (SDF-1) through its interaction with CXCR4, retains the stem cells in the bone marrow (a process referred to as "homing")and AMD3100 specifically antagonizes this interaction.
  • AMD3100 in combination with granulocyte colony- stimulating factor (G-CSF) results in the collection of more progenitor cells than G-CSF alone.
  • G-CSF granulocyte colony- stimulating factor
  • hematological malignancies such as non-Hodgkin's lymphoma or multiple myeloma.
  • BTK has been shown to play a role in signaling downstream of CXCR4 induced by the chemokine SDF-1.
  • blocking BTK with COMPOUND 1 shows similar biological effects as antagonizing CXCR4 on mobilizing hematopoietic progenitor cells to the periphery.
  • COMPOUND 1 treated rats at 300 mg/kg/day for 7 days results in an increase in circulating hematopoietic progenitor cells in the peripheral blood. These cells express CD34 on the cell surface and form CFC-GM colonies in semi-solid media supplemented with growth factors that induce proliferation in myeloid progenitor cells. Rats that are dosed for 28 days with COMPOUND 1 also showed an increased in CFC-GM colonies.

Abstract

Methods to improve hematopoiesis and increase white blood cell counts in subjects and patients using 6,5 fused heteroaryl ring system-based inhibitors of Bruton's tyrosine kinase (Btk) are disclosed.

Description

BTK INHIBITORS FOR HEMATOPOIETIC MOBILIZATION
FIELD OF INVENTION
[0001] The invention is in the field of therapeutics. The present invention relates generally to the administration of a Bruton's tyrosine kinase inhibitor to mobilize hematopoietic stem and progenitor cells from the bone marrow into the peripheral blood and the use of such hematopoietic cells to improve hematopoiesis and/or in the treatment of various disorders.
BACKGROUND OF THE INVENTION
[0002] Bruton's tyrosine kinase (Btk) is a member of the Tec family of non-receptor tyrosine kinases and plays a role in several hematopoietic cell signaling pathways, e.g., Toll like receptor (TLR) and cytokine receptor-mediated TNF-a production in macrophages, IgE receptor (FcsRI) signaling in Mast cells, inhibition of Fas/APO-1 apoptotic signaling in B lineage lymphoid cells, and collagen- stimulated platelet aggregation. See, e.g., Jeffries, et al. (2003) J. Biol. Chem. 278:26258-26264; Horwood et al. (2003) J. Exp. Med. 197: 1603-1611; Iwaki et al. (2005) J. Biol. Chem. 280(48):40261- 40270; Vassilev et al. (1999) J. Biol. Chem. 274(3): 1646-1656, and Quek et al. (1998), Curr. Bio. 8(20): 1137-1140. It is a particularly important in the signaling pathway initiated upon stimulation of the B cell receptor and during B cell development. Mutations in the Btk gene result in X-linked agammaglobulinemia, an immunodeficiency characterized by failure to produce mature B lymphocytes and associated with a failure of Ig heavy chain rearrangement. Rawlings and Witte (1994) Immun. Rev. 138:105-119. In the mouse, point mutation or deletion of Btk causes X-linked immunodeficiency (xid), with about 50% fewer conventional B2 B cells, absent BI B cells, and reduced serum Ig levels. Khan et al (1995) Immunity 3:283-99; Rawlings et al (1993) Science 261 :358-61. Btk is also expressed in specific cells of the myeloid lineage, and evidence suggests that it contributes to immune-complex mediated activation of the FcyR and FcsR signaling pathways in monocytes/macrophages, neutrophils, and mast cells. See, e.g., Jongstra-Bilen et al. (2008) J.
Immunol. 181 :288-298; Wang et al. (2007) Int. Immunopharmacol. 7:541-546; Hata et al. (1998) J Exp Med. 187:1235-1247.
[0003] Due to the role of Btk in inhibiting Fas/APO-1 apoptotic signals in the B cell lineage, inhibitors of Btk, also referred to as Btk inhibitors, have been evaluated as agents for treating hematopoietic malignancies (e.g., B cell lymphoma). Additionally, due to the role of Btk in the signaling pathways of other immune cells, Btk inhibitors have also been evaluated as agents for suppressing the immune system, e.g., in patients with autoimmune disorders or organ transplants. See, e.g., Honinberg et al. (2010) Proc. Natl. Acad. Sci. USA 107:13075-80; Chang et al. (2011) Arthr. Res. & Ther. 13:R115. Evidence for the role of Btk in autoimmune and/or inflammatory disease has been established in Btk-deficient mouse models. For example, in standard murine preclinical models of systemic lupus erythematosus (SLE), Btk deficiency has been shown to result in a marked amelioration of disease progression. Moreover, Btk deficient mice are also resistant to developing collagen-induced arthritis and are less susceptible to Staphylococcus-induced arthritis. Inhibition of Btk activity is useful for the treatment of autoimmune and/or inflammatory diseases such as: SLE, rheumatoid arthritis, multiple vasculitides, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, and asthma. See, e.g., U.S. Patent No. 7,393,848.
[0004] Btk inhibitors have also been shown useful in preventing or reducing the risk of
thromboembolism. See, e.g., Uckun (2008) Int. Rev. Immunol. 27:43-69.
SUMMARY OF INVENTION
[0005] In contrast to the prior art uses of Bruton's Tyrosine Kinase inhibitors to suppress immune cells and/or the immune system, disclosed herein is the surprising discovery that Btk inhibitors can mobilize hematopoietic stem cells and progenitor cells to the peripheral blood of a subject, e.g., to increase the white blood cell count in the subject. Accordingly, provided herein are methods and compositions for improving hematopoiesis and increasing the white blood cell count in a subject in need thereof, including patients undergoing chemotherapy, radiation therapy and/or bone marrow transplantation. Also provided herein are methods of determining whether a Btk inhibitor is a "mobilizing Btk inhibitor" capable of mobilizing hematopoietic stem and/or progenitor cells to the peripheral blood of a subject; and methods of using a mobilizing Btk inhibitor to mobilize such cells, including harvesting such cells for subsequent reinfusion into the same or a different subject.
[0006] In one aspect, the invention provides methods for mobilizing hematopoietic stem and/or progenitor cells in a subject in need thereof comprising administering to said subject a pharmaceutical composition comprising a mobilizing Bruton's Tyrosine Kinase (Btk) inhibitor in an amount effective to mobilize said cells into the peripheral blood of said subject. The inventive methods and uses can be advantageously employed in conjunction with bone marrow transplantation procedures, and/or subsequent to chemotherapy and/or radiation exposure to address leukopenia, neutropenia, granulocytopenia and/or thrombocytopenia in such patients. Accordingly, in some embodiments the subject may be a bone marrow transplantation patient, and/or a leukopenic or neutropenic patient or a patient at risk of impaired hematopoiesis due to prior chemotherapy and/or radiation therapy.
[0007] As compounds that increase the white blood cell count in a subject, the instant mobilizing Btk inhibitors may be administered as part of any therapeutic protocol aiming to restore or improve hematopoiesis in a patient in need thereof, e.g., to enhance the success of bone marrow
transplantation, to reduce the extent or duration of leukopenia and neutropenia resulting from chemotherapy, radiation therapy or accidental radiation exposure, to enhance wound healing and burn treatment, and/or to aid in restoration of damaged organ tissue. They may also combat bacterial infections that are prevalent in leukemia.
[0008] In another aspect, the invention provides methods of obtaining mobilized hematopoietic stem and progenitor cells and uses thereof. The subject methods comprise administering to a subject a mobilizing Btk inhibitor in an amount effective to increase the number of such cells in the subject, preferably in the peripheral blood of the subject. In one embodiment, the administering step comprises administration of a mobilizing Btk inhibitor alone. In another embodiment, the step comprises administration of a mobilizing Btk inhibitor in combination with other compounds, e.g., cytokines, that also increase the white blood cell count in the peripheral blood of a subject. Suitable compounds may be selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM-CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor, thrombopoietin, growth related oncogene, and/or combinations thereof. Thus, the subject methods comprise administering to a subject a mobilizing Btk inhibitor (with or without other mobilizing factors) in an amount effective to increase the number of hematopoietic stem and progenitor and/or white blood cells in the peripheral blood of the subject, and obtaining the immune cells so mobilized, e.g., by apheresis.
[0009] The harvested cells may be used therapeutically, e.g., in hematopoietic stem and/or progenitor cell transplantation. Accordingly, in another aspect the invention provides methods of treating a patient in need of improved hematopoiesis comprising administering to a subject a mobilizing Btk inhibitor (with or without other mobilizing factors) in an amount effective to increase the number of hematopoietic stem, progenitor and/or white blood cells in the peripheral blood of the subject, obtaining the cells so mobilized, and introducing the cells into the patient. Preferably, the subject and the patient are histocompatible. In one embodiment, the histocompatible subject and the patient are syngeneic. In another embodiment, the histocompatible subject and the patient are allogeneic.
[0010] In another embodiment, the harvested cells are enriched and/or cultured ex vivo prior to introduction into the patient. Such ex vivo culture comprises differentiating the obtained cells into or enriching for myeloid cells, lymphoid cells, and common progenitors thereof etc. Accordingly, in one embodiment, a method of treating a patient in need thereof further comprises culturing the obtained hematopoietic cells in one or more differentiation factors prior and/or enriching the obtained hematopoietic cells for a common progenitor cell or cells prior to introducing the cells into the patient.
[0011] Mobilizing Btk inhibitors may be administered to any animal subject in order to mobilize hematopoietic stem and progenitor cells. In a preferred embodiment, the mobilizing Btk inhibitor is administered to a mammal, and more preferably to a human.
[0012] In another aspect, the invention provides compounds for use in mobilizing hematopoietic stem and/or progenitor cells into the peripheral blood of a subject, wherein the compound is a Bruton's Tyrosine Kinase (Btk) inhibitor comprising a 6,5 fused heteroaryl ring system. In some embodiments, the 6,5 fused heteroaryl ring system is selected from: imidazopyrazine and
imidazotriazine ring systems. In some embodiments, the 6,5 fused heteroaryl ring system is an imidazotriazine ring system. In a particularly preferred aspect, Btk inhibitors suitable for use in the subject invention comprise an imidazopyrazine ring system. In some embodiments, the subject has impaired hematopoiesis. In some embodiments, the subject has previously received a
myelosuppressive or myeloablative treatment. In some embodiments, the myelosuppressive or myeloablative treatment comprises chemotherapy. In some embodiments, the subject has previously been accidentally exposed to radiation.
[0013] In some embodiments, the compound for use is for use in combination with an additional mobilizing agent selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM- CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor, thrombopoietin, growth related oncogene, and combinations thereof.
[0014] In some embodiments, the compound for use is for use in harvesting the hematopoietic stem and/or progenitor cells from the subject. In some embodiments, the compound for use is for use in harvesting the hematopoietic stem and/or progenitor cells from the subject by apheresis. In some embodiments, the compound for use is for use in providing the hematopoietic stem and/or progenitor cells to a patient in need thereof. In some embodiments, the compound for use is for use in providing the hematopoietic stem and/or progenitor cells to a bone marrow transplantation patient. In some embodiments, the subject and the patient are histocompatible. In some embodiments, the subject and the patient are syngeneic. In some embodiments, the subject and the patient are allogeneic. In some embodiments, the hematopoietic stem and/or progenitor cells are cultured to enrich and/or differentiate the hematopoietic stem and/or progenitor cells prior to introducing the cells into the patient. In some embodiments, the hematopoietic stem and/or progenitor cells are enriched for or differentiated into myeloid cells. In some embodiments, the hematopoietic stem and/or progenitor cells are enriched for or differentiated into lymphoid cells. In some embodiments, the hematopoietic stem and/or progenitor cells are enriched for or differentiated into NK cells. In some embodiments, the Btk inhibitor is reversible. In some embodiments, the Btk inhibitor is irreversible.
[0015] In another aspect, the invention provides uses of a Bruton's Tyrosine Kinase (Btk) inhibitor comprising a 6,5 fused heteroaryl ring system in the manufacture of a medicament for mobilizing hematopoietic stem and/or progenitor cells into the peripheral blood of a subject. In some embodiments, the 6,5 fused heteroaryl ring system is selected from: imidazopyrazine and imidazotriazine ring systems. In some embodiments, the 6,5 fused heteroaryl ring system is an imidazotriazine ring system. In a particularly preferred aspect, Btk inhibitors suitable for use in the subject invention comprise an imidazopyrazine ring system. In some embodiments, the subject has impaired hematopoiesis. In some embodiments, the subject has previously received a
myelosuppressive or myeloablative treatment. In some embodiments, the myelosuppressive or myeloablative treatment comprises chemotherapy. In some embodiments, the subject has previously been accidentally exposed to radiation.
[0016] In some embodiments, the medicament is for use in combination with an additional mobilizing agent selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM- CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor, thrombopoietin, growth related oncogene, and combinations thereof.
[0017] In some embodiments, the medicament is for use for use in harvesting the hematopoietic stem and/or progenitor cells from the subject. In some embodiments, the medicament is for use in harvesting the hematopoietic stem and/or progenitor cells from the subject by apheresis. In some embodiments, the medicament is for use in providing the hematopoietic stem and/or progenitor cells to a patient in need thereof. In some embodiments, the medicament is for use in providing the hematopoietic stem and/or progenitor cells to a bone marrow transplantation patient. In some embodiments, the subject and the patient are histocompatible. In some embodiments, the subject and the patient are syngeneic. In some embodiments, the subject and the patient are allogeneic. In some embodiments, the hematopoietic stem and/or progenitor cells are cultured to enrich and/or differentiate the hematopoietic stem and/or progenitor cells prior to introducing the cells into the patient. In some embodiments, the hematopoietic stem and/or progenitor cells are enriched for or differentiated into myeloid cells. In some embodiments, the hematopoietic stem and/or progenitor cells are enriched for or differentiated into lymphoid cells. In some embodiments, the hematopoietic stem and/or progenitor cells are enriched for or differentiated into NK cells. In some embodiments, the Btk inhibitor is reversible. In some embodiments, the Btk inhibitor is irreversible.
[0018] Preferred mobilizing Btk inhibitors suitable for use in the subject invention comprise a 6,5 fused heteroaryl ring system. In some embodiments, the 6,5 fused heteroaryl ring system is selected from: imidazopyrazine and imidazotriazine ring systems. In some embodiments, the 6,5 fused heteroaryl ring system is an imidazotriazine ring system. In a particularly preferred aspect, Btk inhibitors suitable for use in the subject invention comprise an imidazopyrazine ring system. These Btk inhibitors have higher aqueous solubility and a lower serum affinity for albumin. In one aspect, these Btk inhibitors also have lower lipophilicity and basic pKa values which renders these Btk inhibitors amenable for salt formation. Accordingly, these Btk inhibitors are expected to have better pharmacokinetic properties and higher solubility. Higher exposure levels, and a higher free fraction in circulation will consequently result in higher uptake in target tissue.
[0019] In one embodiment, the mobilizing Btk inhibitor is selected from a compound of structural Formula I:
Figure imgf000007_0001
Formula I
wherein,
Wis (l-3C)alkyl, or NH2;
X is CH, N, O or S;
Y is C(R6), N, O or S;
Z is CH, N or bond; A is CH or N;
Bl isNorC(R7);
B2 is N or C(R8);
B3 is N or C(R9);
B4 isNorC(RlO);
Rl is Rl lC(O), R12S(0), R13S02 or ( 1 -6C)alkyl optionally substituted with R14;
R2 is H, (l-3C)alkyl or (3-7C)cycloalkyl;
R3 is H, (l-6C)alkyl or (3-7C)cycloalkyl); or
R2 and R3 form, together with the N and C atom they are attached to, a (3-7C)heterocycloalkyl optionally substituted with one or more fluorine, hydroxyl, (1 -3C)alkyl, (l-3C)alkoxy or oxo;
R4isHor(l-3C)alkyl;
R5 is H, halogen, cyano, (l-4C)alkyl, (l-3C)alkoxy, (3-6C)cycloalkyl, any alkyl group of which is optionally substituted with one or more halogen; or
R5 is (6-10C)aryl or (2-6C)heterocycloalkyl;
R6 isHor(l-3C)alkyl;or
R5 and R6 together may form a (3-7C)cycloalkenyl, or (2-6C)heterocycloalkenyl; each optionally substituted with (l-3C)alkyl, or one or more halogen;
R7 is H, halogen or (l-3C)alkoxy;
R8 isHor(l-3C)alkyl;or
R7 and R8 form, together with the carbon atom they are attached to a (6-10C)aryl or (l-9C)heteroaryl; R9 is H, halogen or (l-3C)alkoxy;
RIO is H, halogen or (l-3C)alkoxy;
Rl 1 is independently selected from a group consisting of (l-6C)alkyl, (2-6C)alkenyl and (2- 6C)alkynyl each alkyl, alkenyl or alkynyl optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (1- 3C)alkoxy, (3-7C)cycloalkoxy, (6-10C)aryl or (3-7C)heterocycloalkyl; or
Rl l is (l-3C)alkyl-C(0)-S-(l-3C)alkyl; or
Rl 1 is (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen or cyano;
R12 and R13 are independently selected from a group consisting of (2-6C)alkenyl or (2-6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl,
(l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3-
7C)cycloalkoxy, (6-10C)aryl or (3-7C)heterocycloalkyl; or (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen, or
cyano;
R14 is independently selected from a group consisting of halogen, cyano or (2-6C)alkenyl or (2- 6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3-7C)cycloalkoxy, (6- 10C)aryl, (l-5C)heteroaryl or (3-7C)heterocycloalkyl;
with the proviso that
- 0 to 2 atoms of X, Y, Z can simultaneously be a heteroatom;
- when one atom selected from X, Y is O or S, then Z is a bond and the other atom selected from X, Y cannot be O or S;
- when Z is C or N, then Y is C(R6) or N and X is C or N;
- 0 to 2 atoms of Bl, B2, B3 and B4 are N; or
pharmaceutically acceptable solvates or pharmaceutically acceptable salts thereof.
[0020] In another embodiment, the mobilizing Btk inhibitor for use in the subject invention is a compound selected from the group consisting of:
(S)-4 (3-(l-Acryloylpyrrolidin-2-yl)-8-aminoimidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2-yl)benzamide; (S,E)-4-(8-amino-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide; (S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(4-fluoropyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin-2- yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- propylpyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- ethylpyridin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4,5,6,7- tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-fluoro-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-methoxy-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(thiazol-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-fluoropyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin-2- yl)benzamide;
(S)-4-(8-Amino-3-(l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyrimidin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyrimidin-2- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyrimidin-4- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridazin-3- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(isoxazol-3- yl)benzamide;
(S,E)-4-(8-Amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(5- ethylthiazol-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-fluoro-N-(4-propylpyridin- 2-yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo [ 1 ,5-a]pyrazin-l -yl)-2- methoxy-N-(4-propylpyridin-2-yl)benzamide;
4-(8-Amino-3-((S)-l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l-yl)-3-methyl-N-(pyridin yl)benzamide;
4-(3-(Acrylamidomethyl)-8-aminoimidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynamidoethyl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide; (S)-S-2-(2-(8-Amino- 1 -(4-(pyridin-2-ylcarbamoyl)phenyl)imidazo [ 1 ,5-a]pyrazin-3 -yl)pyrrolidin- 1 - yl)-2-oxoethyl ethanethioate;
(S)-4-(8-Amino-3 -( 1 -(4-hydroxy-4-methylpent-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(pyridin-2-yl)benz amide;
(S)-4-(8-Amino-3 -( 1 -(6-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin-l -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l -pent-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-Amino-3-(l -(3-cyclopropylpropioloyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -y^
(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -hex-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
4-(3-(l -Acryloylazepan-2-yl)-8-aminoimidazo[l,5-a]pyrazin-l -yl)-N-(pyridin-2-yl)benzamide;
(R)-4-(8-Amino-3-(4-but-2-ynoylmoφholin-3-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-( 1 -(N-methylbut-2-ynamido)ethyl)imidazo[ 1 ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(4-methoxybut-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-fluoropyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-(pyrrolidin- 1 - yl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4-fluoropyridin2- yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4-propylpyridin2- yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxy-N-methylbut-2-enamido)ethyl)imidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4-propylpyridm yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2-fluoro-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methoxypyridin-2 -yl)b enzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2-fluoro- N-(4-methoxypyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (isoxazol-3-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (pyrimidin-2-yl)benz amide;
4-(8-amino-3-((S)-l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazi^ methyl-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin-2 -yl)b enzamide ;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- isopropylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (4-methylpyridin-2-yl)benz amide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(thiazol-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4-propylpyridin2- yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)^
(4-isopropylpyridin-2-yl)benzamide;
4-(8-amino-3-((S)-l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-3-methyl-N-(pyri 2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2-fluoro-N-(4- propylpyridin-2-yl)benzamide;
4-(3 -((S)- 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-3-methyl-N-(pyridin-2- yl)benzamide;
(E)-4-(8-amino-3-((4-(dimethyl amino)but-2-enamido)methyl) imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloro pyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4-isopropylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloro pyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (pyridazin-3 -yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l-yl)-N- (pyridazin-3 -yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-^ (pyridazin-3 -yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxy-N-methylbut-2-enamido)ethyl)imidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-(dimethylamino)-N-methylbut-2-enamido)ethyl)imidazo [ 1 ,5-a]pyrazin- 1 - yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l ^ (4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-^ (4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- fluoropyridin-2-yl)benzamide; (S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-
(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S)-4-(8-amino-3-( 1 -(2-chloropyrimidine-4-carbony^
methoxy-N-(pyridin-2-yl)benz amide;
(S)-4-(8-amino-3-( 1 -(2-chloropyrimidine-4-carbo
fluoro -N-(pyridin-2 -yl)b enzamide ;
4-(8-amino-3-((S)-l -((E)-4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-3-^ N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (pyrimidin-4-yl)benz amide;
4-(8-amino-3-((S)-l -((E)-4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-3- methyl-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- methylpyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4-methylpyrimidin-2- yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (pyrimidin-2-yl)benz amide;
(S)-4-(8-amino-3-(l -methacryloylpyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l -(2-(trifluoromethyl)acryloyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -but-2-enoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(pyridin-^ yl)benzamide;
(S)-4-(8-amino-3-(l -(cyanomethyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(E)-4-(8-amino-3-((4-methoxybut-2-enamido)methyl)imidazo[l,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4-(pyrrolidm^ yl)pyridin-2-yl)benzamide;
(E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)azepan-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- cyanopyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2-methoxy-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acrylamidoethyl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide; (S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(thiazol-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-isopropylpyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2- methoxy-N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l-cinnamoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyri
yl)benzamide;
(S)-N-(l-(8-amino-l-(4-(pyridin-2-ylcarbamoyl)phenyl)imidazo[l,5-a]pyrazin-3-yl)ethyl)-2- chloropyrimidine-4-carboxamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-fluoropyridin2- yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbon^^
(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-methoxy-N- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-methoxy- N-(4-propylpyridin-2-yl)benzamide;
4-(8-amino-3-(but-2-ynamidomethyl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-( 1 -(N-methylbut-2-ynamido)ethyl)imidazo[ 1 ,5-a]pyrazin-l -yl)-N-(4-propylpyridin- 2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-fl^ (4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-( 1 -(2-chloropyrimidine-4-carbo
(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(5-ethylthiazol-2- yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l,5-a]pyrazin-l-yl)-N-(5-ethylthiazol-2- yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbon^^
(5-ethylthiazol-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l^ (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(R,E)-4-(8-amino-3-(4-(4-methoxybut-2-enoyl)moφholin-3-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-methoxypyridin yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-y^ (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l^ (4-methylpyridin-2-yl)benz amide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-cyanopyridin2- yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-ethylpyridin2- yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-phenylpyridin-2- yl)benzamide; and
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-phenylpyridin-2- yl)benzamide.
[0021] In another embodiment, the mobilizing Btk inhibitor for use in the subject invention is a compound selected from the group consisting of:
(S)-4-(3 -( 1 -But-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-2- yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(Dimethylamino)but-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-Methyl-3-(l-(vinylsulfonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-propylpyridin- 2-yl)benzamide; (S)-4-(3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)-8-methyHmidazo[l,5-a]pyrazin-l-y (4-propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-Methoxybut-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -But-2-ynamidoethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-(trifluoromethyl)pyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -Acrylamidoethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-(trifluoromethyl)pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methoxypyridin2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methoxypyridin-2- yl)benzamide;
(S)-4-(3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)-8-methyHmidazo[l,5-a]pyrazin-l-yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)-N-methylbut-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 - yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide; (S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin2- yl)benzamide;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-but-2-ynoylpiperidin-2-yl)-8-methyHmidazo[l,5-a]pyrazin-l-yl)-N-(4-propylpyridin2- yl)benzamide;
(S)-4-(3-(l-acrylamidoethyl)-8-methylimidazo[l,5-a]pyrazin-l-yl)-N-(4-propylpyridin-2- yl)benzamide; (S)-4-(8-methyl-3 -( 1 -(N-methylb^
2-yl)benzamide;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(pyridin-2-yl)benz amide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpiperidin-2-yl)-8-methylimidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-methoxybut-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(pyridin-2-yl)benz amide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin2- yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-2- yl)benzamide;
(S,E)-4-(3 -( 1 -(4-methoxybut-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4-propylpyridin-2-yl)benzamide;
(S,E)-4-(3-(l-(4-methoxybut-2-enamido)ethyl)-8-methylimidazo[l ,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-but-2-ynamidoethyl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4-propylpyridin-2- yl)benzamide; and
(S,E)-4-(3-(l-(4-methoxy-N-methylbut-2-enamido)ethyl)-8-methylimidazo[l ,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide.
[0022] In certain embodiments, the compounds of Formula I may include an asymmetric center or centers, and may be in the form of a composition of a racemic mixture, a diastereoisomeric mixture, a single enantiomer, an enantiomeric diastereomer, a meso compound, a pure epimer, or a mixture of epimers thereof, etc. Further, the compounds of Formulas I or II may have one or more double bonds, and may be in a form of a cis/trans, E/Z mixture or an E or Z geometric isomer thereof. [0023] The compounds of Formula I may also be prepared as a salt form, e.g., pharmaceutically acceptable salts, including suitable acid forms, e.g., salt forms selected from hydrochloride, hydrobromide, acetate, propionate, butyrate, sulphate, hydrogen sulphate, sulphite, carbonate, hydrogen carbonate, phosphate, phosphinate, oxalate, hemi-oxalate, malonate, hemi-malonate, fumarate, hemi-fumarate, maleate, hemi-maleate, citrate, hemi-citrate, tartrate, hemi-tartrate, aspartate, glutamate, etc.
BRIEF DESCRIPTION OF THE FIGURES
[0024] Figure 1 shows the results of a flow cytometry analysis showing that CD34+ cells are mobilized to the peripheral blood in rats dosed with 300 mg/kg/day COMPOUND 1 dose.
[0025] Figure 2 shows a summary of CD34+ cell frequencies in rat PBMC samples (Day 7 Study).
[0026] Figure 3 shows the results from a CFC Assay from rats treated with vehicle or COMPOUND 1 (300 mg/kg/day) for 7 or 28 days.
DETAILED DESCRIPTION
[0027] Blood cells play a crucial part in maintaining the health and viability of animals, including humans. White blood cells include neutrophils, macrophages, eosinophils, basophils, mast cells, and the B and T cells of the immune system. White blood cells are continuously replaced via the hematopoietic system, by the action of colony stimulating factors (CSF) and various cytokines on progenitor cells in hematopoietic tissues. The nucleotide sequences encoding a number of these growth factors have been cloned and sequenced. Perhaps the most widely known of these is granulocyte colony stimulating factor (G-CSF) which has been approved for use in counteracting the negative effects of chemotherapy by stimulating the production of white blood cells and progenitor cells (peripheral blood stem cell mobilization). A discussion of the hematopoietic effects of this factor can be found, for example, in U.S. Pat. No. 5,582,823, incorporated herein by reference.
[0028] Several other factors have been reported to increase white blood cells and progenitor cells in both human and animal subjects. These agents include granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), ΡΓΧΥ-321 (GM- CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, thrombopoietin and growth related oncogene, as single agents or in combination. Dale et al. (1998) Am. J. of Hematol. 57:7-15; Rosenfeld et al. (1997) Bone Marrow Transplantation 17:179-183; Pruijt et al.,(1999) Cur. Op. in Hematol. 6:152-158; Broxmeyer et al. (1995) Exp. Hematol. 23:335-340; Broxmeyer et al. (1998) Blood Cells, Molecules and Diseases 24:14-30; Glaspy et al. (1996) Cancer Chemother. Pharmacol. 38 (suppl): S53-S57; Vadhan-Raj et al. (1997) Ann. Intern. Med.l26:673-81 ; King et al. (2001) Blood 97:1534-1542; Glaspy et al. (1997) Blood 90:2939-2951.
[0029] While endogenous growth factors are pharmacologically effective, the well-known disadvantages of employing proteins and peptides as pharmaceuticals underlies the need to add to the repertoire of such growth factors with agents that are small molecules. In another aspect, such small molecules are advantageous over proteins and peptides where production in large quantities are desired.
[0030] As used herein, the term "progenitor cell" refers to a cell that, in response to certain stimuli, can form differentiated hematopoietic or myeloid cells. The presence of progenitor cells can be assessed by the ability of the cells in a sample to form colony-forming units of various types, including, for example, CFU-GM (colony-forming units, granulocyte-macrophage); CFU-GEMM (colony-forming units, multipotential); BFU-E (burst-forming units, erythroid); HPP-CFC (high proliferative potential colony-forming cells); or other types of differentiated colonies which can be obtained in culture using known protocols.
[0031] As used herein, "stem cells" are less differentiated forms of progenitor cells. Typically, such cells are often positive for CD34. Some stem cells do not contain this marker, however. These CD34+ cells can be assayed using fluorescence activated cell sorting (FACS) and thus their presence can be assessed in a sample using this technique.
[0032] In general, CD34+ cells are present only in low levels in the blood, but are present in large numbers in bone marrow. While other types of cells such as endothelial cells and mast cells also may exhibit this marker, CD34 is considered an index of stem cell presence.
[0033] The development and maturation of blood cells is a complex process. Mature blood cells are derived from hematopoietic precursor cells (progenitor) cells and stem cells present in specific hematopoietic tissues including bone marrow. Within these environments hematopoietic cells proliferate and differentiate prior to entering the circulation. The chemokine receptor CXCR4 and its natural ligand stromal cell derived factor-1 (SDF-1) appear to be important in this process. See, e.g., Maekawa et al (2000) Internal Med. 39:90-100; Nagasawa et al. (2000) Int. J. Hematol. 72:408 41 1). This is demonstrated by reports that CXCR4 or SDF-1 knock-out mice exhibit hematopoietic defects. Ma et al. (1998) Proc. Natl. Acad. Sci USA 95:9448 9453; Tachibana et al. (1998) Nature 393 :591 594; Zou et al. (1998) Nature 393 :595-599.
[0034] It is also known that CD34+ progenitor cells express CXCR4 and require SDF-1 produced by bone marrow stromal cells for chemoattraction and engraftment, Peled et al. (1999) Science 283 :845- 48, and that in vitro, SDF-1 is chemotactic for both CD34+ cells, Aiuti et al. (1997) J. Exp. Med. 185: 11 1-120; Viardot et al. (1998)Ann. Hematol. 77: 194 197, and for progenitor cells, Jo et al. (2000) J. Clin. Invest. 105: 101 -111. SDF-1 is also an important chemoattractant, signaling via the CXCR4 receptor, for several other more committed hematopoietic progenitors and mature blood cells including T-lymphocytes and monocytes, Bleul et al. (1996) J. Exp. Med. 184: 1 101-1 109), pro-and pre-B lymphocytes, Fedyk et al. (1999) J. Leukoc. Biol. (1999) 66:667-673; Ma et al. (1999) Immunity 10:463-71 , and megakaryocytes. Hodohara et al. (2000) Blood 95:769-75; Riviere et al. (1999) Blood 95: 151 1-23; Majk et al. (2000) Blood 96:4142-51 ; Gear et al. (2001) Blood 97:937-45; Abi-Younes et al. (2000) Circ. Res. 86: 131 -38
[0035] The SDF-1/CXCR4 signaling axis is known to direct homing and engraftment of hematopoietic stem cells to the bone marrow. Kucia et al. (2005) Stem Cells 23 :879-94. SDF-1 is a CXC chemokine and is considered as one of the most potent chemoattractants of hematopoietic stem cells (HSC) into the bone marrow. Lapid et al (2009) "Egress and Mobilization of Hematopoietic Stem and Progenitor Cells" in StemBook (available at www.stembook.org/node/558). SDF-1 binding to CXCR4 triggers G protein coupling (from inactive conformation to an active conformation) and subsequent dissociation of the heterotrimeric G protein into Οβγ and Gai subunits, which in turn bind to several downstream effectors, resulting in activation of PI3K, protein kinase C (PKC), and MAPK- mediated pathways. Sharma et al. (201 1) Stem Cells Dev. 20 (6): 933-46. Studies of the SDF 1/CXCR4 axis in B-cell migration have identified Bruton's tyrosine kinase (Btk) as a central kinase in the SDF-1/CXCR4 signaling cascade. De Gorter et al. (2007) Immunity 26: 93-104. Btk expression has been described in HSC, multipotent progenitor cells and cells of the myeloid lineage, Mohammed et al. (2009) Immunological Reviews 228: 58-73, including erythroid cells, platelets, monocytes, macrophages, granulocytes, and dendritic cells. In the lymphoid lineage, Btk is expressed in B cells, but not in T cells or natural killer cells (NK cells). Schmidt et al (2004) Int Arch Allergy Immunol. 134:65-78.
[0036] Notably, blocking of the SDF-1/CXCR4 signaling axis by AMD3100 (plerixafor or
Mozobil®), a small molecule inhibitor which binds to CXCR4 preventing binding of SDF-1 and subsequent downstream signaling, causes rapid and reversible mobilization of CD34+ HSC into the peripheral blood for harvesting by apheresis. Blocking the SDF-1/CXCR4 signaling cascade with a Btk inhibitor (PCI-32765) in patients with B-cell malignancies produces rapid and clinically significant decreases in lymphadenopathy as the malignant B cells are mobilized out of the lymph nodes and into the peripheral blood producing a marked lymphocytosis. The present invention elucidates the role of Btk in HSC homing and mobilization and provides compositions and methods of using Btk inhibitors for hematopoietic stem and/or progenitor cell mobilization.
Definitions
[0037] "Acyl" means -C(0)-alkyl, where alkyl is as defined herein.
[0038] "Acylthio" means -SC(0)-alkyl, where alkyl is as defined herein.
[0039] "Alkenyl" means a straight or branched chain, saturated monovalent hydrocarbon radical having one or more carbon-carbon double bonds. By way of example, the hydrocarbon chain may have from two to twenty carbons, two to sixteen carbons, two to fourteen carbons, two to twelve carbons, two to ten carbons, two to eight carbons, two to six carbons, two to four carbons, etc. "Lower alkenyl" may refer to alkenyls having, e.g., two to six carbons, two to four carbons, etc. In certain examples, a straight chain alkenyl may have from two to six carbon atoms and a branched alkenyl three to six carbon atoms, e.g., ethenyl, prop-l -en-l -yl, prop-2-en-2-yl, allyl, butenyl (including all isomeric forms), pentenyl (including all isomeric forms), and the like.
[0040] "Alkenylcarbonyl" means -C(0)-alkenyl, where alkenyl is as defined herein.
[0041] "Alkenylsulfonyl" means -S02-alkenyl, where alkenyl is as defined herein.
[0042] "Alkyl" means a straight or branched chain, saturated monovalent hydrocarbon radical. By way of example, the hydrocarbon chain may have from one to twenty carbons, one to sixteen carbons, one to fourteen carbons, one to twelve carbons, one to ten carbons, one to eight carbons, one to six carbons, one to four carbons, etc. "Lower alkyl" may refer to alkyls having, e.g., one to six carbons, one to four carbons, etc. In certain examples, a straight chain alkyl may have from one to six carbon atoms and a branched alkyl three to six carbon atoms, e.g., methyl, ethyl, propyl, 2-propyl, butyl
(including all isomeric forms), pentyl (including all isomeric forms), and the like. "Me" means methyl, "Et" means ethyl, and "iPr" means isopropyl.
[0043] "Aryl" means a monovalent monocyclic or bicyclic aromatic hydrocarbon radical, e.g., having from of 6 to 20 or 6 to 10 ring atoms e.g., phenyl or naphthyl.
[0044] "Alkynyl" means a straight or branched chain, saturated monovalent hydrocarbon radical having one or more carbon-carbon triple bonds. By way of example, the hydrocarbon chain may have from two to twenty carbons, two to sixteen carbons, two to fourteen carbons, two to twelve carbons, two to ten carbons, two to eight carbons, two to six carbons, two to four carbons, etc. "Lower alkynyl" may refer to alkynyls having, e.g., two to six carbons, two to four carbons, etc. In certain examples, a straight chain alkynyl may have from two to six carbon atoms and a branched alkynyl three to six carbon atoms, e.g., ethynyl, prop-l-yn-l -yl, prop-2-yn-2-yl, propargyl, butynyl (including all isomeric forms), pentynyl (including all isomeric forms), and the like.
[0045] "Alkynylcarbonyl" means -C(0)-alkynyl, where alkynyl is as defined herein.
[0046] "Heteroarylcarbonyl" means -C(0)-heteroaryl, where heteroaryl is as defined herein.
[0047] "Alkylaryl" means a -(alkylene)-R radical where R is aryl as defined above.
[0048] "Cycloalkyl" means a cyclic saturated or partially saturated monovalent hydrocarbon radical (or an alicyclic radical). By way of example, the cycloalkyl may have from three to twenty carbon atoms, from three to sixteen carbon atoms, from three to fourteen carbon atoms, from three to twelve carbon atoms, from three to ten carbon atoms, from three to eight carbon atoms, from three to six carbon atoms, etc., wherein one or two carbon atoms may be replaced by an oxo group, e.g., admantanyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, indanyl and the like.
[0049] "Alkylcycloalkyl" means a -(alkylene)-R radical where R is cycloalkyl as defined above; e.g., cyclopropylmethyl, cyclobutylmethyl, cyclopentylethyl, or cyclohexylmethyl, and the like.
[0050] "Heterocyclyl" or "heterocycloalkyl" means a saturated or unsaturated monovalent monocyclic group, in which one or two ring atoms are heteroatom selected from N, O, or S, the remaining ring atoms being C. The heterocyclyl ring is optionally fused to a (one) aryl or heteroaryl ring as defined herein. The heterocyclyl ring fused to monocyclic aryl or heteroaryl ring is also referred to in this Application as "bicyclic heterocyclyl" ring. Additionally, one or two ring carbon atoms in the heterocyclyl ring can optionally be replaced by a -CO- group. More specifically the term heterocyclyl includes, but is not limited to, pyrrolidino, piperidino, homopiperidino, 2- oxopyrrolidinyl, 2-oxopiperidinyl, morpholino, piperazino, tetrahydropyranyl, thiomorpholino, and the like. When the heterocyclyl ring is unsaturated it can contain one or two ring double bonds. When the heterocyclyl group contains at least one nitrogen atom, it is also referred to herein as
heterocycloamino and is a subset of the heterocyclyl group. When the heterocyclyl group is a saturated ring and is not fused to aryl or heteroaryl ring as stated above, it is also referred to herein as saturated monocyclic heterocyclyl.
[0051] "Alkylheterocycloalkyl" means a -(alkylene)-R radical where R is heterocyclyl ring as defined above e.g., tetraydrofuranylmethyl, piperazinylmethyl, morpholinylethyl, and the like.
[0052] "Heteroaryl" means a monovalent monocyclic or bicyclic aromatic radical, where one or more, preferably one, two, or three, ring atoms are heteroatom selected from N, O, or S, the remaining ring atoms being carbon. Representative examples include, but are not limited to, pyrrolyl, thienyl, thiazolyl, imidazolyl, furanyl, indolyl, isoindolyl, oxazolyl, isoxazolyl, diazolyl, pyrazolyl, triazolyl, benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, tetrazolyl, and the like.
[0053] "Oxo" or "carbonyl" means =(0) group or C=0 group, respectively.
[0054] The term "substituted" means that the referenced group is substituted with one or more additional group(s) individually and independently selected from groups described herein. In some embodiments, an optional substituent is selected from oxo, halogen, -CN, -NH2, -OH, -NH(CH3), -N(CH3)2, alkyl (including straight chain, branched and/or unsaturated alkyl), substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, fluoroalkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted alkoxy, fluoroalkoxy, -S-alkyl, -S(0)2-alkyl, -CONH((substituted or unsubstituted alkyl) or (substituted or unsubstituted phenyl)), -CON(H or alkyl)2, -OCON(substituted or unsubstituted alkyl)2, -NHCONH((substituted or unsubstituted alkyl) or (substituted or unsubstituted phenyl)), -NHCOalkyl, -N(substituted or unsubstituted
alkyl)CO(substituted or unsubstituted alkyl), -NHCOO(substituted or unsubstituted alkyl), -C(OH)( substituted or unsubstituted alkyl)2, and -C(NH2)( substituted or unsubstituted alkyl)2. In some embodiments, by way of example, an optional substituent is selected from oxo, fluorine, chlorine, bromine, iodine, -CN, -NH2, -OH, -NH(CH3), -N(CH3)2, -CH3, -CH2CH3, -CH(CH3)2, -CF3, -CH2CF3, -OCH3, -OCH2CH3, -OCH(CH3)2, -OCF3, -OCH2CF3, -S(0)2-CH3, -CONH2,
-CONHCH3, -NHCONHCH3, -COCH3, -COOH and the like. In some embodiments, substituted groups are substituted with one, two or three of the preceding groups. In some embodiments, substituted groups are substituted with one or two of the preceding groups. In some embodiments, substituted groups are substituted with one of the preceding groups. Further, unless stated to the contrary, a formula with chemical bonds shown only as solid lines and not as wedges or dashed lines contemplates each possible isomer, e.g., each enantiomer and diastereomer, and a mixture of isomers, such as racemic or scalemic mixtures.
[0055] The term "amino acid" includes any one of the twenty naturally-occurring amino acids or the D-form of any one of the naturally-occurring amino acids. In addition, the term "amino acid" also includes other non-naturally occurring amino acids besides the D-amino acids, which are functional equivalents of the naturally-occurring amino acids. Such non-naturally occurring amino acids include, for example, norleucine ("Nle"), norvaline ("Nva"), L- or D-naphthalanine, ornithine ("Orn"), homoarginine (homoArg) and others well known in the peptide art, such as those described in M. Bodanzsky, "Principles of Peptide Synthesis," 1st and 2nd Revised Ed., Springer- Verlag, New York, N.Y., 1984 and 1993, and Stewart and Young, "Solid Phase Peptide Synthesis," 2nd Ed., Pierce Chemical Co., Rockford, 111., 1984, both of which are incorporated herein by reference.
[0056] Amino acids and amino acid analogs can be purchased commercially (Sigma Chemical Co.; Advanced Chemtech) or synthesized using methods known in the art.
[0057] "Therapeutically effective amount" or "effective amount" means the amount of a composition, compound, therapy, or course of treatment that, when administered to a subject for treating a disease, disorder, or condition, is sufficient to effect such treatment for the disease, disorder, or condition. The "therapeutically effective amount" will vary depending on the composition, the compound, the therapy, the course of treatment, the disease, disorder, or condition, and its severity and the age, weight, etc., of the subject to be treated.
Btk Inhibitors
[0058] Bruton's tyrosine kinase is a member of the cytoplasmic Tec family of kinases. Like other Btk family members, it contains a pleckstrin homology (PH) domain, and Src homology SH3 and SH2 domains. Btk plays an important role in B cell development. Btk comprises of several domains from the N-terminus: the PH, Tec homology (TH), SH2, SH3, and kinase (SHI) domains. Each of these domains has the potential to interact with a plethora of proteins critical for intracellular signaling. Moreover, functional association of Btk with many of its partners is crucial for its activation and regulation. Btk is a metalloprotein enzyme requiring Zn2+ for optimal activity and stability. Mohammed (2009) Immuno. Rev. 228:58-73.
[0059] Preferred mobilizing Btk inhibitors suitable for use in the subject invention comprise a 6,5 fused heteroaryl ring system. In a particularly preferred aspect, Btk inhibitors suitable for use in the subject invention comprise an imidazopyrazine ring system. In one embodiment, the mobilizing Btk inhibitor is selected from a compound of structural Formula I:
Figure imgf000024_0001
Formula I
wherein:
Wis(l-3C)alkyl, or NH2;
X is CH, N, O or S;
Y is C(R6), N, O or S;
Z is CH, N or bond; A is CH or N;
Bl isNorC(R7);
B2 is N or C(R8);
B3 is N or C(R9);
B4 isNorC(RlO);
Rl is Rl lC(O), R12S(0), R13S02 or (l-6C)alkyl optionally substituted with R14;
R2 is H, (l-3C)alkyl or (3-7C)cycloalkyl;
R3 is H, (l-6C)alkyl or (3-7C)cycloalkyl); or
R2 and R3 form, together with the N and C atom they are attached to, a (3-7C)heterocycloalkyl optionally substituted with one or more fluorine, hydroxyl, (l-3C)alkyl, (l-3C)alkoxy or oxo;
R4isHor(l-3C)alkyl;
R5 is H, halogen, cyano, (l-4C)alkyl, (l-3C)alkoxy, (3-6C)cycloalkyl, any alkyl group of which is optionally substituted with one or more halogen; or
R5 is (6-10C)aryl or (2-6C)heterocycloalkyl;
R6 isHor(l-3C)alkyl;or
R5 and R6 together may form a (3-7C)cycloalkenyl, or (2-6C)heterocycloalkenyl; each optionally substituted with (l-3C)alkyl, or one or more halogen;
R7 is H, halogen or (l-3C)alkoxy;
R8 isHor(l-3C)alkyl;or
R7 and R8 form, together with the carbon atom they are attached to a (6-10C)aryl or ( 1 -9C)heteroaryl; R9 is H, halogen or (l -3C)alkoxy;
RI O is H, halogen or (l-3C)alkoxy;
Rl 1 is independently selected from a group consisting of (l-6C)alkyl, (2-6C)alkenyl and (2- 6C)alkynyl each alkyl, alkenyl or alkynyl optionally substituted with one or more groups selected from hydroxyl, (l -4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (1- 3C)alkoxy, (3-7C)cycloalkoxy, (6-10C)aryl or (3-7C)heterocycloalkyl; or
Rl l is (l-3C)alkyl-C(0)-S-(l-3C)alkyl; or
Rl 1 is (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen or cyano;
R12 and R13 are independently selected from a group consisting of (2-6C)alkenyl or (2-6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (l -4C)alkyl, (3- 7C)cycloalkyl, [(l -4C)alkyl]amino, di[(l -4C)alkyl]amino, (l-3C)alkoxy, (3-7C)cycloalkoxy, (6- 10C)aryl or (3-7C)heterocycloalkyl; or (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen, or cyano;
R14 is independently selected from a group consisting of halogen, cyano or (2-6C)alkenyl or (2- 6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3-7C)cycloalkoxy, (6- 10C)aryl, (l -5C)heteroaryl or (3-7C)heterocycloalkyl;
with the proviso that
- 0 to 2 atoms of X, Y, Z can simultaneously be a heteroatom;
- when one atom selected from X, Y is O or S, then Z is a bond and the other atom selected from X, Y cannot be O or S;
- when Z is C or N, then Y is C(R6) or N and X is C or N;
- 0 to 2 atoms of Bl , B2, B3 and B4 are N; and
pharmaceutically acceptable solvates or pharmaceutically acceptable salts thereof.
[0060] In one embodiment, the Btk inhibitor is selected from a compound of structural Formula II:
Figure imgf000025_0001
Formula II
or a pharmaceutically acceptable salt thereof, wherein:
W is (l -3C)alkyl, or NH2; Rl is selected from: (l -6C)acyl, (2-6)alkenylcarbonyl, (2-6)alkenylsulfonyl, (l -6C)alkyl, (2- 6)alkynylcarbonyl, and (l-5C)heteroarylcarbonyl, each optionally substituted with one or more substituents selected from: (l-6C)acylthio, (l-3C)alkoxy, (6-10C)aryl, cyano, (3-7C)cycloalkyl, di[(l- 4C)alkyl]amino, (3-7C)heterocycloalkyl, halogen, and hydroxyl;
R2 is H or (l-3C)alkyl;
R3 is H or (l-6C)alkyl; or
R2 and R3 form, together with the N and C atom they are attached to, a (5-7C)heterocycloalkyl; R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2- yl, pyrimidin-4-yl, pyridazin-3-yl, and isoxazol-3-yl, each optionally substituted with one substituent selected from: (l -3C)alkoxy, (l -4C)alkyl, (6-10C)aryl, (3-6C)cycloalkyl, halogen, and (3- 6C)heterocycloalkyl, wherein (l -4C)alkyl is further optionally substituted with one or more halogens.
[0061] In one embodiment, the Btk inhibitor is selected from a compound of structural Formula II:
Figure imgf000026_0001
Formula II
or a pharmaceutically acceptable salt thereof, wherein:
W is methyl, or NH2;
Rl is selected from: acetyl, acryloyl, but-2-enoyl, but-2-ynoyl, hex-2-ynoyl, methacryloyl, methyl, 4- methylpent-2-ynoyl, pent-2-ynoyl, pyrimidine-4-carbonyl, and vinylsulfonyl, each optionally substituted with one or more substituents selected from: chloro, cyano, dimethylamino, fluoro, hydroxyl, methoxy, phenyl, and pyrrolidinyl;
R2 is H or methyl;
R3 is H or methyl; or
R2 and R3 form, together with the N and C atom they are attached to, pyrrolidin-2-yl, piperidin-2-yl, azepan-2-yl, or morpholin-3-yl; and
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2- yl, pyrimidin-4-yl, pyridazin-3-yl, and isoxazol-3-yl, each optionally substituted with one substituent selected from: cyano, ethyl, fluoro, isopropyl, methoxy, methyl, phenyl, n-propyl, pyrrolidinyl, trifluoromethyl.
[0062] In one embodiment, the Btk inhibitor is selected from a compound of structural Formula II:
Figure imgf000027_0001
Formula
or a pharmaceutically acceptable salt thereof, wherein:
W is methyl, or NH2;
Rl is selected from 2-(acetylthio)acetyl, acryloyl, but-2-enoyl, but-2-ynoyl, cinnamoyl, cyanomethyl,
3- cyclopropylpropioloyl, 4-(dimethylamino)but-2-enoyl, 4-(dimethylamino)but-2-ynoyl, hex-2-ynoyl,
4- hydroxy-4-methylpent-2-ynoyl, methacryloyl, 4-methoxybut-2-enoyl, 4-methoxybut-2-ynoyl, 4- methylpent-2-ynoyl, pent-2-ynoyl, 2-chloropyrimidine-4-carbonyl, 4-(pyrrolidin-l-yl)but-2-enoyl, 2- (trifluoromethyl)acryloyl, and vinylsulfonyl;
R2 is H or methyl;
R3 is H or methyl; or
R2 and R3 form, together with the N and C atom they are attached to, pyrrolidin-2-yl, piperidin-2-yl, azepan-2-yl, or morpholin-3-y; and
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2- yl, pyrimidin-4-yl, pyridazin-3-yl, isoxazol-3-yl, 4-fluoropyridin-2-yl, 4-methylpyridin-2-yl, 4- propylpyridin-2-yl, 4-(trifluoromethyl)pyridin-2-yl, 4-ethylpyridin-2-yl, 4-cyanopyridin-2-yl, 4- methylpyrimidin-2-yl, 5-ethylthiazol-2-yl, 4-(pyrrolidin-l-yl)pyridin-2-yl, 4-methoxypyridin-2-yl, 4- isopropylpyridin-2-yl, 4-phenylpyridin-2-yl.
[0063] Suitable embodiments of such 6,5 fused heteroaryl ring system-based Btk inhibitor compounds are described in more detail in U.S. Patent Nos. 7,071,199, 7,459,554, as well as PCT Appln. Nos. PCT EP2012 063552 and PCT EP2012 063556, and related pending applications, the disclosures of which are expressly incorporated by reference herein.
[0064] In one embodiment, the Btk inhibitor includes every combination of one or more compounds selected from the following group shown in Table A.
Table A
No. Chemical Structure Chemical Name
Figure imgf000028_0001
yl)benzamide
Figure imgf000029_0001
yl)benzamide
Figure imgf000030_0001
Figure imgf000031_0001
Figure imgf000032_0001
Figure imgf000033_0001
Figure imgf000034_0001
Figure imgf000035_0001
Figure imgf000036_0001
Figure imgf000037_0001
0 yl)benzamide
Figure imgf000038_0001
Figure imgf000039_0001
Figure imgf000040_0001
Figure imgf000041_0001
yl)benzamide
Figure imgf000042_0001
Figure imgf000043_0001
Figure imgf000044_0001
Figure imgf000045_0001
Figure imgf000046_0001
yl)benzamide
Figure imgf000047_0001
Figure imgf000048_0001
Figure imgf000049_0001
yl)benzamide
Figure imgf000050_0001
[0065] In one embodiment, the Btk inhibitor includes every combination of one or more compounds selected from the following group shown in Table B.
Table B
Figure imgf000050_0002
Figure imgf000051_0001
Figure imgf000052_0001
Figure imgf000053_0001
Figure imgf000054_0001
Figure imgf000055_0001
Figure imgf000056_0001
[0066] In another embodiment, the mobilizing Btk inhibitor for use in the subject invention is selected from the group consisting of:
(S)-4 (3-(l-Acryloylpyrrolidin-2-yl)-8-aminoimidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2-yl)benzamide; (S,E)-4-(8-amino-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(4-fluoropyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin-2- yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- propylpyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- ethylpyridin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4,5,6,7- tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-fluoro-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-methoxy-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(thiazol-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-fluoropyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin-2- yl)benzamide;
(S)-4-(8-Amino-3-(l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyrimidin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyrimidin-2- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyrimidin-4- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridazin-3- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(isoxazol-3- yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(5- ethylthiazol-2-yl)benzamide; (S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-fluoro-N-(4-propylpyridin- 2-yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo [ 1 ,5-a]pyrazin-l -yl)-2- methoxy-N-(4-propylpyridin-2-yl)benzamide;
4-(8-Amino-3-((S)-l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l-yl)-3-methyl-N-(pyridin yl)benzamide;
4-(3-(Acrylamidomethyl)-8-aminoimidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynamidoethyl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide; (S)-S-2-(2-(8-Amino- 1 -(4-(pyridin-2-ylcarbamoyl)phenyl)imidazo [ 1 ,5-a]pyrazin-3 -yl)pyrrolidin- 1 - yl)-2-oxoethyl ethanethioate;
(S)-4-(8-Amino-3 -( 1 -(4-hydroxy-4-methylpent-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(pyridin-2-yl)benz amide;
(S)-4-(8-Amino-3 -( 1 -(6-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin-l -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l -pent-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-Amino-3-(l -(3-cyclopropylpropioloyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -y^
(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -hex-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
4-(3-(l -Acryloylazepan-2-yl)-8-aminoimidazo[l,5-a]pyrazin-l -yl)-N-(pyridin-2-yl)benzamide;
(R)-4-(8-Amino-3-(4-but-2-ynoylmoφholin-3-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-( 1 -(N-methylbut-2-ynamido)ethyl)imidazo[ 1 ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(4-methoxybut-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-fluoropyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-(pyrrolidin- 1 - yl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4-fluoropyridin2- yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide; (S)-4-(3 -( 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-propylpyridin2- yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxy-N-methylbut-2-enamido)ethyl)imidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(vinylsulfonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-propylpyridm yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-fluoro-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methoxypyridin-2 -yl)b enzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2-fluoro- N-(4-methoxypyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (isoxazol-3-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyrimidin-2-yl)benz amide;
4-(8-amino-3-((S)-l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazi^ methyl-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin-2 -yl)b enzamide ;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- isopropylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (4-methylpyridin-2-yl)benz amide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(thiazol-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-propylpyridin2- yl)benzamide; (S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyraz
(4-isopropylpyridin-2-yl)benzamide;
4-(8-amino-3-((S)-l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-3-methyl-N-(pyri 2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2-fluoro-N-(4- propylpyridin-2-yl)benzamide;
4-(3 -((S)- 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-3-methyl-N-(pyridin-2- yl)benzamide;
(E)-4-(8-amino-3-((4-(dimethyl amino)but-2-enamido)methyl) imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloro pyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4-isopropylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloro pyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (pyridazin-3 -yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l-yl)-N- (pyridazin-3 -yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-^ (pyridazin-3 -yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxy-N-methylbut-2-enamido)ethyl)imidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-(dimethylamino)-N-methylbut-2-enamido)ethyl)imidazo [ 1 ,5-a]pyrazin- 1 - yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l ^
(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-( 1 -(2-chloropyrimidine-4-carbony^
(4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l ^ methoxy-N-(pyridin-2-yl)benz amide;
(S)-4-(8-amino-3-( 1 -(2-chloropyrimidine-4-ca^
fliioro -N-(pyridin-2 -yl)b enzamide ;
4-(8-amino-3-((S)-l-((E)-4-methoxybut-2-enoy
N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyrimidin-4-yl)benz amide;
4-(8-amino-3-((S)-l -((E)-4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-3- methyl-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- methylpyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-methylpyrimidin-2- yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyrimidin-2-yl)benz amide;
(S)-4-(8-amino-3-(l-methacryloylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l-(2-(trifluoromethyl)acryloyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-but-2-enoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-^ yl)benzamide;
(S)-4-(8-amino-3-(l-(cyanomethyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(E)-4-(8-amino-3-((4-methoxybut-2-enamido)methyl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-(pyrrolidm^ yl)pyridin-2-yl)benzamide;
(E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)azepan-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- cyanopyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-methoxy-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acrylamidoethyl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(thiazol-2-yl)benzamide; (S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4-isopropylpyridin-2- yl)benzamide; (S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2- methoxy-N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l -cinnamoylpyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l-yl)-N-(pyri
yl)benzamide;
(S)-N-(l-(8-amino-l -(4-(pyridin-2-ylcarbamoyl)phenyl)imidazo[l ,5-a]pyrazin-3-yl)ethyl)-2- chloropyrimidine-4-carboxamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4-fluoropyridin2- yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbon^^
(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2-methoxy-N- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2-methoxy- N-(4-propylpyridin-2-yl)benzamide;
4-(8-amino-3-(but-2-ynamidomethyl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-( 1 -(N-methylbut-2-ynamido)ethyl)imidazo[ 1 ,5-a]pyrazin-l -yl)-N-(4-propylpyridin- 2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2-fl^
(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbon^^
(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(5-ethylthiazol-2- yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(5-ethylthiazol-2- yl)benzamide;
(S)-4-(8-amino-3-( 1 -(2-chloropyrimidine-4-carbo
(5-ethylthiazol-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-^ (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(R,E)-4-(8-amino-3-(4-(4-methoxybut-2-enoyl)moφholin-3-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide; (S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4-methoxypyridin yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-en^
(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-( 1 -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l ^ (4-methylpyridin-2-yl)benz amide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4-cyanopyridin2- yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4-ethylpyridin2- yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4-phenylpyridin-2- yl)benzamide; and
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-phenylpyridin-2- yl)benzamide.
[0067] In another embodiment, the mobilizing Btk inhibitor for use in the subject invention is selected from the group consisting of:
(S)-4-(3 -( 1 -But-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-2- yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(Dimethylamino)but-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-Methyl-3-(l -(vinylsulfonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4-propylpyridin- 2-yl)benzamide;
(S)-4-(3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N- (4-propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-Methoxybut-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -But-2-ynamidoethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-(trifluoromethyl)pyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -Acrylamidoethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-(trifluoromethyl)pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methoxypyridin2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methoxypyridin-2- yl)benzamide;
(S)-4-(3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)-8-methyHmidazo[l ,5-a]pyrazin-l-yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)-N-methylbut-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 - yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide; (S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin2- yl)benzamide;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-but-2-ynoylpiperidin-2-yl)-8-methyHmidazo[l ,5-a]pyrazin-l-yl)-N-(4-propylpyridin2- yl)benzamide;
(S)-4-(3-(l-acrylamidoethyl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4-propylpyridin-2- yl)benzamide;
(S)-4-(8-methyl-3-(l-(N-methylbut-2-ynamido)ethyl)imidazo[l,5-a]pyrazin-l -yl)-N-(4-propylpyridm 2-yl)benzamide;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(pyridin-2-yl)benz amide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpiperidin-2-yl)-8-methylimidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-methoxybut-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(pyridin-2-yl)benz amide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin2- yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-2- yl)benzamide;
(S,E)-4-(3 -( 1 -(4-methoxybut-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N- (4-propylpyridin-2-yl)benzamide;
(S,E)-4-(3-(l-(4-methoxybut-2-enamido)ethyl)-8-methylimidazo[l ,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-but-2-ynamidoethyl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4-propylpyridin-2- yl)benzamide; and
(S,E)-4-(3-(l-(4-methoxy-N-methylbut-2-enamido)ethyl)-8-methylimidazo[l ,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide.
[0068] In the scope of the embodiments, the Btk inhibitors described herein include further forms of the compounds such as pharmaceutically acceptable salts, solvates (including hydrates), amorphous phases, partially crystalline and crystalline forms (including all polymorphs), prodrugs, metabolites, N-oxides, isotopically-labeled, epimers, pure epimers, epimer mixtures, enantiomers including but not limited to single enantiomers and enantiomeric diastereomers, meso compounds, stereoisomers, racemic mixtures and diastereoisomeric mixtures. Btk inhibitors described herein having one or more double bonds include cis/trans isomers, E/Z isomers and geometric isomers. Btk inhibitors described herein can be prepared as a pharmaceutically acceptable salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, for example an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base. In addition, the salt forms of the disclosed compounds can be prepared using salts of the starting materials or intermediates.
[0069] In some embodiments, a Btk inhibitor of the disclosure is present in a composition as a salt. In some embodiments, salts are obtained by reacting a compound of the disclosure with acids. In some other embodiments, pharmaceutically acceptable salts are obtained by reacting a compound of the disclosure with a base. In other embodiments, the compounds are used as free-acid or free-base form in the manufacture of the compositions described herein. The type of salts, include, but are not limited to: (1) acid addition salts, formed by reacting the free base form of the compound with a
pharmaceutically acceptable: inorganic acid, such as, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, metaphosphoric acid, and the like; or with an organic acid, such as, for example, acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, trifluoroacetic acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2- hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo-[2.2.2]oct-2-ene-l -carboxylic acid, glucoheptonic acid, 4,4 '-methyl en ebis-(3- hydroxy-2-ene-l-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, butyric acid, phenylacetic acid, phenylbutyric acid, valproic acid, and the like; (2) salts formed when an acidic proton present in the parent compound is replaced by a metal ion, e.g., an alkali metal ion (e.g. lithium, sodium, potassium), an alkaline earth ion (e.g. magnesium, or calcium), or an aluminum ion. In some cases, the Btk inhibitor described herein are reacted with an organic base, such as, but not limited to, ethanolamine, diethanolamine, triethanolamine, tromethamine, N— methylglucamine, dicyclohexylamine, tris(hydroxymethyl)methylamine. In other cases, the compounds described herein form salts with amino acids such as, but not limited to, arginine, lysine, and the like. Acceptable inorganic bases used to form salts with compounds that include an acidic proton, include, but are not limited to, aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like.
[0070] When the Btk inhibitors described herein include one or more chiral centers, the
stereochemistry of such chiral centers can independently be in the R or S configuration, or a mixture of the two. The chiral centers can be further designated as R or S or R,S or d,D, 1,L or d,l, D,L.
Correspondingly, the Btk inhibitors of the invention, if they can be present in optically active form, can actually be present in the form of a racemic mixture of enantiomers, or in the form of either of the separate enantiomers in substantially isolated and purified form, or as a mixture comprising any relative proportions of the enantiomers.
[0071] When the Btk inhibitors described herein contain two or more chiral centers then diastereomers are possible. Such diastereomers may be present as pure diastereomeric enantiomers, pure racemic mixtures of diastereomeric enantiomers, mixtures of diastereomers which may be racemic or may have optical activity in their own right due to complex permutations of enantiomeric diastereomers in the balance of the mixtures.
[0072] When the Btk inhibitors of the invention can be present in geometrically isomeric forms, then they can actually be present in the form of a mixture of geometric isomers comprising any relative proportions of the isomers, or in some cases in the form of either of the separate geometric isomers in substantially isolated and purified form.
[0073] When the Btk inhibitors described herein include one or more isolated or linearly conjugated double bonds, the geometry around such double bonds can be independently a cis/trans, E/Z mixture or an E or Z geometric isomer thereof.
[0074] In some embodiments, the Btk inhibitors described herein include solvent addition forms or crystal forms thereof, particularly solvates or polymorphs. Solvates contain either stoichiometric or non-stoichiometric amounts of a solvent, and may be formed during the process of crystallization with pharmaceutically acceptable solvents such as water, ethanol, and the like. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is alcohol.
[0075] As noted above, in some embodiments the Btk inhibitors described herein possess one or more stereocenters and each center exists independently in either the R or S configuration. The Btk inhibitors presented herein include all diastereomeric, enantiomeric, and epimeric forms as well as the appropriate mixtures thereof.
[0076] In some embodiments, sites on the Btk inhibitors disclosed herein are susceptible to various metabolic reactions. Therefore incorporation of appropriate substituents at the places of metabolic reactions will reduce, minimize or eliminate the metabolic pathways. In specific embodiments, the appropriate substituent to decrease or eliminate the susceptibility of the aromatic ring to metabolic reactions is, by way of example only, a halogen, deuterium or an alkyl group.
[0077] In some embodiments, the Btk inhibitors described herein are isotopically-labeled, which are identical to those recited in the various formulae and structures presented herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. In some embodiments, one or more hydrogen atoms are replaced with deuterium. In some embodiments, metabolic sites on the compounds described herein are deuterated. In some embodiments, substitution with deuterium affords certain therapeutic advantages resulting from greater metabolic stability, such as, for example, increased in vivo half-life or reduced dosage requirements. Throughout the specification, groups and substituents thereof can be chosen by one skilled in the field to provide stable moieties and compounds.
[0078] Inhibitors of Btk kinase activity preferably inhibit Btk activity with an IC50 of less than or equal to 10 micromolar, less than or equal to 1 micromolar, less than or equal to 500 nanomolar, less than or equal to 100 nanomolar, or less than or equal to 10 nanomolar in an ADP-GLO™, bioluminescent, homogeneous assay or in a HTRF® (Homogeneous Time-Resolved Fluorescence) assay.
[0079] Inhibitors of Bruton's tyrosine kinase activity preferably inhibit phosphorylation of Y551 of Btk with an IC50 of less than or equal to 10 micromolar, less than or equal to 1 micromolar, less than or equal to 500 nanomolar, less than or equal to 1 12 nanomolar, less than or equal to 100 nanomolar, or less than or equal to 10 nanomolar.
[0080] Inhibitors of Bruton's tyrosine kinase activity preferably inhibit phosphorylation of Y223 of Btk with an IC50 of less than or equal to 10 micromolar, less than or equal to 1 micromolar, less than or equal to 500 nanomolar, less than or equal to 1 12 nanomolar, less than or equal to 100 nanomolar, or less than or equal to 10 nanomolar. Inhibitors of Bruton's tyrosine kinase activity may inhibit phosphorylation of downstream PLCy2 with an IC50 of less than or equal to 10 micromolar, less than or equal to 1 micromolar, less than or equal to 500 nanomolar, less than or equal to 100 nanomolar, or less than or equal to 10 nanomolar in, e.g., an IgM phospho-PLCg2 assay.
[0081] The compounds described herein, and other related compounds having different substituents can be synthesized using techniques and materials known to those of skill in the art, such as described, for example, in March, ADVANCED ORGANIC CHEMISTRY 4th Ed., (Wiley 1992); Carey and Sundberg, ADVANCED ORGANIC CHEMISTRY 4th Ed., Vols. A and B (Plenum 2000, 2001), and Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3rd Ed., (Wiley 1999) (all of which are incorporated by reference in their entirety). General methods for the preparation of the compounds as disclosed herein may be derived from known reactions in the field, and the reactions may be modified by the use of appropriate reagents and conditions, as would be recognized by the skilled person, for the introduction of the various moieties found in the formulae as provided herein. Appropriate synthesis methods for the subject compounds are described in more detail in U.S. Patent Nos. 8,088,781, 8,008,309, 7,718,662, 7,514,444 and 7,960,396, as well as PCT Appln. Nos. PCT EP2012 063552 and PCT EP2012 063556. Also disclosed herein are pharmaceutical compositions. The pharmaceutical compositions will generally comprise a pharmaceutically acceptable carrier and a pharmacologically effective amount of the compounds, or mixture of thereof, or suitable salts thereof. The pharmaceutical composition may be formulated as powders, granules, solutions, suspensions, aerosols, solids, pills, tablets, capsules, gels, topical creams, suppositories, transdermal patches, and other formulations known in the art.
[0082] As used herein, "pharmaceutically acceptable carrier" comprises any of standard
pharmaceutically accepted carriers known to those of ordinary skill in the art in formulating pharmaceutical compositions. Thus, the compounds, by themselves, such as being present as pharmaceutically acceptable salts, or as conjugates, may be prepared as formulations in
pharmaceutically acceptable diluents; for example, saline, phosphate buffer saline (PBS), aqueous ethanol, or solutions of glucose, mannitol, dextran, propylene glycol, oils (e.g., vegetable oils, animal oils, synthetic oils, etc.), microcrystalline cellulose, carboxymethyl cellulose, hydroxylpropyl methyl cellulose, magnesium stearate, calcium phosphate, gelatin, polysorbate 80 or the like, or as solid formulations in appropriate excipients.
[0083] The pharmaceutical compositions will often further comprise one or more buffers (e.g., neutral buffered saline or phosphate buffered saline), carbohydrates (e.g., glucose, mannose, sucrose or dextrans), mannitol, proteins, polypeptides or amino acids such as glycine, antioxidants (e.g., ascorbic acid, sodium metabisulfite, butylated hydroxytoluene, butylated hydroxyanisole, etc.), bacteriostats, chelating agents such as EDTA or glutathione, solutes that render the formulation isotonic, hypotonic or weakly hypertonic with the blood of a recipient, suspending agents, thickening agents, preservatives, flavoring agents, sweetening agents, and coloring compounds as appropriate.
[0084] While any suitable carrier known to those of ordinary skill in the art may be employed in the compositions, the type of carrier will typically vary depending on the mode of administration. The therapeutic compositions may be formulated for any appropriate manner of administration, including for example, oral, nasal, mucosal, rectal, vaginal, topical, intravenous, intraperitoneal, intradermal, subcutaneous, and intramuscular administration.
[0085] For parenteral administration, the compositions can be administered as injectable dosages of a solution or suspension of the substance in a physiologically acceptable diluent with a pharmaceutical carrier that can be a sterile liquid such as sterile pyrogen free water, oils, saline, glycerol, polyethylene glycol or ethanol. Additionally, auxiliary substances, such as wetting or emulsifying agents, surfactants, pH buffering substances and the like can be present in compositions. Other components of pharmaceutical compositions are those of petroleum, animal, vegetable, or synthetic origin, for example, non-aqueous solutions of peanut oil, soybean oil, corn oil, cottonseed oil, ethyl oleate, and isopropyl myristate.
[0086] Sterile injectable solutions can be prepared by incorporating the mobilizing Btk inhibitor in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. In accordance with an alternative aspect of the invention, an agent of the invention as described above may be formulated with one or more additional compounds that enhance the solubility of these agents. The invention also extends to such derivatives of such agents of the invention.
[0087] The pharmaceutical compositions described herein may be presented in unit-dose or multi- dose containers, such as sealed ampoules or vials. Such containers are typically sealed in such a way to preserve the sterility and stability of the formulation until use. In general, formulations may be stored as suspensions, solutions or emulsions in oily or aqueous vehicles, as indicated above. Alternatively, a pharmaceutical composition may be stored in a freeze-dried condition requiring only the addition of a sterile liquid carrier immediately prior to use. In one embodiment, a pharmaceutical composition is provided comprising expanded hematopoietic progenitor cells cryopreserved in a suitable cryopreservation medium, which can then be thawed and resuspended as needed for administration to a patient.
[0088] The pharmaceutical compositions described herein can also contain the mobilizing Btk inhibitor in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use can be prepared according to any method known to the art for the manufacture of pharmaceutical compositions, and such compositions can contain one or more agents selected from, by way of non-limiting example, sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations.
[0089] Formulations suitable for oral administration can be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion.
[0090] Composition preparations which can be used orally include tablets, push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Tablets can be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with binders (e.g., povidone, gelatin, hydroxypropylmethyl cellulose), inert diluents, preservative, disintegrant (e.g., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose) or lubricating, surface active or dispersing agents. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets can optionally be coated or scored and can be formulated so as to provide slow or controlled release of the active ingredient therein. Tablets can optionally be provided with an enteric coating, to provide release in parts of the gut other than the stomach. All formulations for oral administration should be in dosages suitable for such administration. The push- fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers can be added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions can be used, which can optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments can be added to the tablets or Dragee coatings for identification or to characterize different combinations of active compound doses. [0091] It should be understood that in addition to the ingredients particularly mentioned above, the compounds and compositions described herein can include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration can include flavoring agents.
[0092] In some embodiments, one or more mobilizing Btk inhibitor may be formulated in pharmaceutical compositions with additional active ingredients, or administered in methods of treatment in conjunction with treatment with one or more additional medications, such as another agent as described herein that also increases mobilization of stem cells, for example, or one or more agents selected from the group consisting of: recombinant-methionyl human. G-CSF (NEUPOGEN®, Filgastim; Amgen), GM-CSF (LEUKINE®, Sargramostim; Immunex), erythropoietin (rhEPO, EPOGEN®; Amgen), thrombopoietin (rhTPO; Genentech), interleukin-11 (rhIL-1 1, NEUMEGA®; American Home Products), Flt3 ligand (Mobista; Immunex), multilineage hematopoietic factor (MARSTEM™; Maret Pharm.), myelopoietin (Leridistem; Searle), IL-3, myeloid progenitor inhibitory factor-1 (Mirostipen; Human Genome Sciences), stem cell factor (rhSCF, STEMGEN®; Amgen), and plerixafor (Mozobil®; Genzyme-Sanofi).
[0093] The amount administered to the host will vary depending upon what is being administered, the purpose of the administration, such as prophylaxis or therapy, the state of the host, the manner of administration, the number of administrations, interval between administrations, and the like. These can be determined empirically by those skilled in the art and may be adjusted for the extent of the therapeutic response. Factors to consider in determining an appropriate dose include, but is not limited to, size and weight of the subject, the age and sex of the subject, the severity of the symptom, the stage of the disease, method of delivery of the agent, half-life of the agents, and efficacy of the agents. Stage of the disease to consider includes whether the disease is acute or chronic, relapsing or remitting phase, and the progressiveness of the disease.
[0094] Determining the dosages and times of administration for a therapeutically effective amount are well within the skill of the ordinary person in the art. For example, an initial effective dose can be estimated from cell culture or other in vitro assays. A dose can then be formulated in animal models to generate a circulating concentration or tissue concentration, including that of the IC50 as determined by the cell culture assays.
[0095] In addition, toxicity and therapeutic efficacy are generally determined by cell culture assays and/or using experimental animals, typically by determining a LD50 (lethal dose to 50% of the test population) and ED50 (therapeutically effectiveness in 50% of the test population). Guidance is found in standard reference works, for example, Goodman & Oilman's The Pharmacological Basis of Therapeutics, 10th Ed. (Hardman, J. G. et al., eds.) McGraw-Hill, New York, N.Y. (2001).
[0096] For the purposes of this invention, the methods of administration are chosen depending on the condition being treated, the form of the subject mobilizing Btk inhibitor(s), and the pharmaceutical composition. Administration of the therapeutic compounds can be done in a variety of ways, including, but not limited to, orally subcutaneously, intravenously, intraperitoneally, intramuscularly, and possibly direct injection to specified organs such as e.g., spleen or bone marrow, although systemic and oral administration are preferred. Administration of the pharmaceutical compositions may be through a single route or concurrently by several routes.
[0097] The compositions may be administered once per day, a few or several times per day, or even multiple times per day, depending upon, among other things, the indication being treated and the judgment of the prescribing physician.
[0098] As disclosed above, disclosed herein are mobilizing Btk inhibitors and pharmaceutical compositions comprising the same. The mobilizing Btk inhibitors and pharmaceutical compositions can be administered to a subject to mobilize the subject's hematopoietic stem and progenitor cells into the peripheral blood of the subject. Accordingly, a mobilizing Btk inhibitor may be administered to a subject in need thereof, e.g., a patient having impaired hematopoiesis and/or deficient in white blood cells such as myeloid and lymphoid cells.
Impaired Hematopoiesis
[0099] A variety of diseases, in particular cancers and hyperproliferative disorders, require treatment with agents that are preferentially cytotoxic to dividing cells. These therapies typically include high doses of irradiation and/or chemotherapeutic agents. While necessary to kill off the cancer cells, a significant side effect of these approaches is the pathological impact of such treatments on rapidly dividing normal cells, such as hair follicles, mucosal cells and the hematopoietic cells, including primitive hematopoietic progenitor cells and stem cells. This indiscriminate destruction of hematopoietic stem cells and progenitor/precursor cells leads to reduced mature blood cell counts, such as lymphocytes, neutrophils and platelets, eventually resulting in a loss of immune system function in these patients and a substantially increased risk of morbidity and mortality from opportunistic infections. Leukopenia resulting from chemotherapy or irradiation therapy may occur within a few days following cytotoxic treatments, although the patient can be vulnerable to infection for up to one month until the white blood cell counts recover to within a normal range. If the reduced leukocyte count (leukopenia), neutrophil count (neutropenia), granulocyte count (granulocytopenia) and/or platelet count (thrombocytopenia) become sufficiently serious, the therapy must be interrupted to allow for recovery of the white blood cells, which may in turn result in the survival of cancer cells, an increase drug resistance in the cancer cells, and may actually result in a relapse of the cancer.
[00100] The methods of mobilizing progenitor cells and/or providing hematopoietic cells to a patient disclosed herein may be used for the treatment of such impaired hematopoiesis. A treatment modality that enhances the stem and/or progenitor cells in blood is helpful in treatments to ameliorate the effects of conditions that adversely affect the bone marrow, such as chemotherapy or irradiation (intentional or accidental) that results in leukopenia, including neutropenia, and thrombocytopenia. The mobilizing Btk inhibitors disclosed herein may enhance the success of bone marrow transplantation, and may combat infections in the patient undergoing such therapies. In this context the compounds are also be used to mobilize and harvest hematopoietic stem cells or progenitor cells via apheresis and the harvested cells are used in treatments requiring stem cell transplantations. Furthermore, the Btk inhibitors can be used both in vivo to promote mobilization of hematopoietic stem cells or progenitor cells from the bone marrow to the peripheral blood or can be used for ex vivo studies, whereby a patient's own stem cells are removed and expanded in culture for autologous transplants. Also contemplated by the present invention are in vitro screens, whereby candidate or test compounds can be measured for their effects on mobilization before being administered in vivo.
[00101] In some embodiments, the disorder relates to impairment of hematopoiesis caused by disease or myeloablative treatments. As used herein, "treatment" refers to therapeutic or prophylactic treatment. Treatment encompasses administration of a Btk inhibitor capable of mobilizing hematopoietic stem and progenitor cells to the peripheral blood of a first subject, and optionally harvesting said mobilized progenitor cells and introducing them into a compatible subject. In one embodiment, the obtained progenitor cells are cultured ex vivo prior to reintroduction into the original subject. In one embodiment, the obtained progenitor cells are cultured ex vivo prior to introduction into a second subject. In one embodiment, the methods comprise administration of the obtained, and optionally cultured, cells in an appropriate form prior to the onset of disease symptoms and/or after clinical manifestations, or other manifestations of the disease or condition to reduce disease severity, halt disease progression, or eliminate the disease.
[00102] "Prophylactic" or "therapeutic" treatment refers to administration to the subject of one or more of the Btk inhibitor compositions. If it is administered prior to clinical manifestation of the impaired hematopoiesis (e.g., leukopenia or neutropenia) then the treatment is prophylactic, i.e., it protects the host against developing the unwanted condition, whereas if administered after manifestation of impaired hematopoiesis, the treatment is therapeutic (i.e., it is intended to improve hematopoiesis and/or increase white blood cell count). "Subject" refers to a mammal, preferably a human, in need of treatment for a condition, disorder or disease.
[00103] For example, myelosuppressive chemotherapy regimens with cytotoxic agents such as doxorubicin, paclitaxel, cisplatin, carboplatin, etoposide, ifosfamide, daunorubicin, cytosine arabinoside, thioguanine, and the like typically induce a transient but profound myelosuppression in patients, at about seven to fourteen days after chemotherapy. This may be followed by rapid reappearance of leukocytes in the peripheral blood and a "rebound" increase of the circulating leukocytes above baseline levels. As the leukocyte count rises, hematopoietic progenitor cells also begin to appear in the peripheral blood and rapidly increase. Protocols involving dose intensification (i.e., to increase the log-kill of the respective tumor therapy) or schedule compression may exacerbate the degree and duration of myelosuppression associated with the chemotherapy and/or radiation therapy. For instance, in the adjuvant setting, repeated cycles of doxorubicin-based treatment have been shown to produce cumulative and long-lasting damage in the bone marrow progenitor cell populations (Lorhrman et al., (1978) Br. J. Haematol. 40:369).
[00104] The effect of this short-term hematopoietic cell damage resulting from chemotherapy has been addressed to some extent by the concurrent use of G-CSF (Neupogen®), which is used to accelerate the regeneration of neutrophils (Le Chevalier (1994) Eur. J. Cancer 30A:410). This approach has met with limitations, however, as it may be accompanied by progressive
thrombocytopenia and cumulative bone marrow damage as reflected by a reduction in the quality of mobilized progenitor cells over successive cycles of treatment. Because of the current interest in chemotherapy dose intensification as a means of improving tumor response rates and patient survival, the necessity for alternative therapies to either improve or replace current treatments to rescue the myelosuppressive effects of chemotherapy and/or radiation therapy has escalated, and is currently one of the major rate limiting factors for tumor therapy dose escalations.
[00105] In the present invention, treatments using the mobilizing Btk inhibitors described may be provided to patients suffering from a cancerous condition or hyperproliferative disease, whereby the treatment of the disease with chemotherapy or irradiation therapy results in a decrease in bone marrow cellularity, thus making the patient more prone to acquiring infectious agents or diseases. Thus, administration of the mobilizing Btk inhibitors of the present invention subsequent to the chemotherapy and/or radiation exposure allows for the mobilization of hematopoietic stem and/or progenitor cells from the bone marrow to the peripheral blood. As used herein "subsequent" administration may be within 1 -24 hours, preferably within 12 to 24 or 24 to 36 or 48 hours, preferably within one to 30 days, more preferably within one to 14 or 21 days, more preferably within three or five to ten or fourteen days after the chemotherapy and/or radiation exposure. Improving hematopoiesis may advantageously allow for the use of accelerated doses of chemotherapy or irradiation therapy.
[00106] Additionally, transplantation of hematopoietic stem and progenitor cells harvested from a subject may provide a more rapid and sustained hematopoietic recovery, e.g, after the administration of high-dose chemotherapy or radiation therapy in patients with hematological malignancies and solid tumours. This has become the preferred source of hematopoietic stem cells for autologous transplantation because of the shorter time to engraftment and the lack of a need for surgical procedures such as are necessary for bone marrow harvesting (Demirer et al. (1996) Stem Cells 14: 106-1 16; Pettengel et al., (1992) Blood 82:2239-2248). Moreover, this approach has been successfully used in the allogeneic setting as well as in the syngeneic setting, including in the context of more differentiated hematopoietic progenitor cells, e.g., myeloid progenitor cells. See, e.g., U.S. Patent Nos. 8,252,587 and 8,383,095, the disclosures of which are expressly incorporated by reference herein.
[00107] It is possible to expand hematopoietic stem and progenitor cells in stroma-containing or nonstromal systems. Expansion systems have reportedly shown increases in CFU_GM of more than 100-fold. Enrichment of CD34+ cells may be required before expansion in nonstromal culture but may not be necessary in stroma-containing systems. Early results of clinical trials are encouraging and have been taken to demonstrate that the engraftment potential of the expanded hematopoietic cells is not compromised by culture. Other possible applications of stem cell expansion include purging of tumor cells; production of immune-competent cells, such as dendritic cells and NK cells, and gene therapy.
Enrichment of Common Lymphoid Progenitors
[00108] Hematopoietic stem cells and progenitors may be obtained from a subject according to the methods described herein, and enriched for and/or further differentiated into progenitors of different lineages according to well-known methods. For example, common lymphoid progenitor cells, which give rise to B cells, T cells, and Natural Killer Cells, common myeloid progenitor cells which give rise to monocytes, granulocytes, megakaryocites, and erythrocytes may be enriched by sorting according to the expression levels of certain growth factor receptors. See, e.g., U.S. Patent Nos. 7,979,057; 7,235,623; 6,908,763; and 5,989,660, the disclosures of which are expressly incorporated by reference herein.
[00109] For example, common lymphoid progenitors (CLPs) express low levels of c-kit (CD1 17) on their cell surface. Antibodies that specifically bind c-kit in humans, mice, rats, etc. are known in the art. Alternatively, the c-kit ligand, steel factor (Slf) may be used to identify cells expressing c-kit. The CLP cells express high levels of the IL-7 receptor alpha chain (CDwl27). Antibodies that bind to human or to mouse CDwl27 are known in the art. Alternatively, the cells are identified by binding of the ligand to the receptor, IL-7.
[00110] Murine CLPs express low levels of Sca-1 (Ly-6E and Ly-6A, see van de Rijn (1989) Proc Natl Acad Sci 86:4634-4638). Antibodies specific for Sca-1 are known in the art. The expression of high levels of Sca-1 on murine hematopoietic stem cells has been previously described (Spangrude et al. (1988) J Immunol 141 :3697-3707). A candidate human counterpart for Sca-1 is described in Hill et al. (1996) Exp Hematol 24:936-943, as high-level expression of a novel epitope of CD59 identifies a subset of CD34+ bone marrow cells that are highly enriched for pluripotent stem cells.
[00111] The CLP subset also has the phenotype of lacking expression of lineage specific markers, exemplified by B220, CD4, CD8, CD3, Gr-1 and Mac-1. For staining purposes a cocktail of binding reagents, herein designated "lin", may be used. The lin panel will comprise binding reagents, e.g. antibodies and functional binding fragments thereof, ligands, peptidomimetics, etc., that recognize two or more of the lineage markers. A lin panel will generally include at least one marker expressed on mature B cells, on mature T cells, on mature granulocytes and on mature macrophages. Markers suitable for use in a lineage panel are typically expressed on these mature cells, but are not present on multiple lineages, or on stem and progenitor cells. The subject cells are characterized as lacking expression of Thy-1 , a marker that is characteristic of hematopoietic stem cells. The phenotype of the CLP may be further characterized as Mel-14", CD4310, HSA10, CD45+ and common cytokine receptor γ chain
Enrichment of myeloid progenitors
[00112] Myeloid progenitors may be subdivided into three distinct subsets: a common myeloid progenitor cell (CMP) which is characterized by progenitor cell activity for myeloid lineages, but lacking the potential to differentiate into lymphoid lineages; a granulocyte monocyte (GMP) committed progenitor cell; and an erythroid/megakaryocyte (MEP) committed progenitor cell. The CMP gives rise to the other two subsets.
[00113] The CMP population is useful in transplantation to provide a recipient with myeloid cells, including megakaryocytes, platelets and erythroid cells, in addition to monocytes and granulocytes; for drug screening; experimental models of hematopoietic differentiation and interaction; screening in vitro assays to define growth and differentiation factors, and to characterize genes involved in myeloid development and regulation; and the like. The native cells may be used for these purposes, or they may be genetically modified to provide altered capabilities.
[00114] Each of these progenitor subsets may be are separated from a complex mixture of cells by using reagents that specifically recognize markers on the cell surface. In both human and mouse cells, all three of the myeloid lineage progenitors stain negatively for the markers Thy-1 (CD90), IL- 7R.alpha. (CD 127); and with a panel of lineage markers, which lineage markers may include CD2; CD3; CD4; CD7; CD8; CD10; CD1 lb; CD14; CD19; CD20; CD56; and glycophorin A (GPA) in humans and CD2; CD3; CD4; CD8; CD19; IgM; Terl 10; Gr-1 in mice. With the exception of the mouse MEP subset, all of the progenitor cells are CD34 positive. In the mouse all of the progenitor subsets may be further characterized as Sca-1 negative, (Ly-6E and Ly-6A), and c-kit high. In the human, all three of the subsets are CD38+.
[00115] Among the progenitor subsets, that is, the population of cells defined as Lin" IL-7R" Thy-Γ, the population can be divided into subsets for the CMP, GMP and MEP cells. In humans, the markers IL-3Ra (CDwl27) and CD45RA are sufficient for separating the three subsets, where the CMP is IL- 3Ra10 CD45RA"; the GMP is IL-3Ra'° CD45Ra+; and the MEP is IL-3Ra CD45RA". In the mouse, the CD34 and Fey receptor (FcyR) are useful in making these distinctions. The CMP is characterized as FcyR10 CD34+ population; the GMP is FcyRhi CD34+; and the MEP subset is FcyR10 CD34\
[00116] In the presence of steel factor (SLF), flt-3 ligand (FL), interleukin (IL)-3, IL-1 1, GM- CSF, thrombopoietin (Tpo) and erythropoietin (Epo), the CMP cells give rise to various types of myeloerythroid colonies, including CFU-GEMMeg, burst-forming unit-erythroid (BFU-E), CFU- megakaryocytes (CFU-Meg), CFU-granulocyte/macrophage (CFU-GM), CFU-granulocyte (CFU-G) and CFU-macrophage (CFU-M).
[00117] The GMP subset generates CFU-M, CFU-G, or CFU-GM colonies containing macrophages and/or granulocytes in response to the above growth factors. In contrast, the MEP subset gives rise to CFU-Meg, BFU-E, or CFU-MEP colonies containing only megakaryocytes and/or erythrocytes in response to IL-3, GM-CSF, Tpo and Epo, but do not form colonies in the absence of Tpo and Epo. All three myeloid progenitor populations do not require "early-acting cytokines" such as SLF, FL and IL- 11 to initiate colony formation.
[00118] All of these progenitors are capable of rapid differentiation activity in vivo. CMP cells give rise to Gr-1+/Mac-1+ myelomonocytic cells and megakaryocyte colonies, as well as TER1 19+ erythroid cells in spleen and bone marrow. The GMP progenitor population gives rise to Gr-1+/Mac- 1+ cells; and the MEP progenitor population to megakaryocytes and erythroid cells.
[00119] The subject subsets may be separated from a complex mixture of cells by techniques that enrich for cells having the above-described characteristics for the subset of interest. An appropriate solution may be used for dispersion or suspension. Such solution will generally be a balanced salt solution, e.g. normal saline, PBS, Hank's balanced salt solution, etc., conveniently supplemented with fetal calf serum or other naturally occurring factors, in conjunction with an acceptable buffer at low concentration, generally from 5-25 mM. Convenient buffers include HEPES, phosphate buffers, lactate buffers, etc
[00120] Separation of the subject cell populations may be attained by use of affinity separation to provide a substantially pure population. Techniques for affinity separation may include magnetic separation, using antibody-coated magnetic beads, affinity chromatography, cytotoxic agents joined to a monoclonal antibody or used in conjunction with a monoclonal antibody, e.g. complement and cytotoxins, and "panning" with antibody attached to a solid matrix, e.g. plate, or other convenient technique. Techniques providing accurate separation include fluorescence activated cell sorters, which can have varying degrees of sophistication, such as multiple color channels, low angle and obtuse light scattering detecting channels, impedance channels, etc. The cells may be selected against dead cells by employing dyes associated with dead cells (e.g. propidium iodide). Any technique may be employed which is not unduly detrimental to the viability of the selected cells.
[00121] The affinity reagents may be specific receptors or ligands for the cell surface molecules indicated above. In addition to antibody reagents, peptide-MHC antigen and T cell receptor pairs may be used; peptide ligands and receptor; effector and receptor molecules, and the like. Antibodies and T cell receptors may be monoclonal or polyclonal, and may be produced by transgenic animals, immunized animals, immortalized human or animal B-cells, cells transfected with DNA vectors encoding the antibody or T cell receptor, etc. The details of the preparation of antibodies and their suitability for use as specific binding members are well-known to those skilled in the art.
[00122] Of particular interest is the use of antibodies as affinity reagents. Conveniently, these antibodies are conjugated with a label for use in separation. Labels include magnetic beads, which allow for direct separation, biotin, which can be removed with avidin or streptavidin bound to a support, fluorochromes, which can be used with a fluorescence activated cell sorter, or the like, to allow for ease of separation of the particular cell type. Fluorochromes that find use include phycobiliproteins, e.g. phycoerythrin and allophycocyanins, fluorescein and Texas red. Frequently each antibody is labeled with a different fluorochrome, to permit independent sorting for each marker.
[00123] The antibodies are added to a suspension of cells, and incubated for a period of time sufficient to bind the available cell surface antigens. The incubation will usually be at least about 5 minutes and usually less than about 30 minutes. It is desirable to have a sufficient concentration of antibodies in the reaction mixture, such that the efficiency of the separation is not limited by lack of antibody. The appropriate concentration is determined by titration. The medium in which the cells are separated will be any medium which maintains the viability of the cells. A preferred medium is phosphate buffered saline containing from 0.1 to 0.5% BSA. Various media are commercially available and may be used according to the nature of the cells, including Dulbecco's Modified Eagle Medium (dMEM), Hank's Basic Salt Solution (HBSS), Dulbecco's phosphate buffered saline (dPBS), RPMI, Iscove's medium, PBS with 5 mM EDTA, etc., frequently supplemented with fetal calf serum, BSA, HSA, etc.
[00124] The labeled cells are then separated as to the expression of cell surface markers as previously described. The separated cells may be collected in any appropriate medium that maintains the viability of the cells, usually having a cushion of serum at the bottom of the collection tube. Various media are commercially available and may be used according to the nature of the cells, including dMEM, HBSS, dPBS, RPMI, Iscove's medium, etc., frequently supplemented with fetal calf serum.
[00125] The culture may contain growth factors to which the cells are responsive. Growth factors, as defined herein, are molecules capable of promoting survival, growth and/or differentiation of cells, either in culture or in the intact tissue, through specific effects on a transmembrane receptor. Growth factors include polypeptides and non-polypeptide factors. Specific growth factors that may be used in culturing the subject cells include steel factor (c-kit ligand), Flk-2 ligand, IL-1 1, IL-3, GM-CSF, erythropoietin and thrombopoietin. The specific culture conditions are chosen to achieve a particular purpose, i.e. differentiation into erythroid of megakaryocyte populations, maintenance of progenitor cell activity, etc.
[00126] In addition to, or instead of growth factors, the subject cells may be grown in a co-culture with stromal or feeder layer cells. Stromal cells suitable for use in the growth of hematopoietic cells are known in the art. These include bone marrow stroma as used in "Whitlock-Witte" (Whitlock et al.
[1985] Annu Rev Immunol 3 :213-235) or "Dexter" culture conditions (Dexter et al. J Exp Med 145: 1612-1616); and heterogeneous thymic stromal cells (Small and Weissman [1996] Scand J Immunol 44: 1 15-121).
[00127] The subject cultured cells may be used in a wide variety of ways. The nutrient medium, which is a conditioned medium, may be isolated at various stages and the components analyzed. Separation can be achieved with HPLC, reversed phase-HPLC, gel electrophoresis, isoelectric focusing, dialysis, or other non-degradative techniques, which allow for separation by molecular weight, molecular volume, charge, combinations thereof, or the like. One or more of these techniques may be combined to enrich further for specific fractions. EXAMPLES
Example 1: Flow cytometry hierarchal gating scheme for PBMC samples.
[00128] (Figure 1 A) 7-AAD negative "live" cells are gated, followed by forward and side scatter gating to identify cells from debris. Then CD45+ gate is applied to capture all hematopoietic, non- RBC precursors. Finally a CD34+, CD1 lb- gate identifies immature CD34+ cells. (Figure IB) Individual rat data is shown for the No dose (vehicle control) and the 300 mg/kg/day dose. CD34+ percentages shown are of the total live cells.
Example 2: CD34 positive cell enumeration by flow cytometry.
[00129] Cells are thawed and washed with RPMI 1% FBS two times. Fc receptors are blocked with mouse anti-rat CD32 (BD Biosciences) at 1 μg per million cells for 10 minutes on ice. Cells are then stained for 1 hour at 4°C in three antibody cocktails recognizing the following rat surface proteins:
1. CD45-FITC (BD Biosciences), CD34-PE (Santa Cruz Biotechnology), CD1 lb-APC (BD Biosciences)
2. CD45-FITC, CD1 lb-APC, (no CD34 antibody)
3. CD45-FITC, IgGl-PE (isotype control; BD Biosciences), CD1 lb Alexa 647
[00130] The manufacturer's recommended antibody concentrations are used. Samples are washed via centrifugation two times in PBS+0.5% BSA. The dead cell discriminator, 7-Aminoactinomycin D (7- ADD; Calbiochem), is added at 1 μg/ml to stained cells and incubated for 30 minutes on ice. Samples are acquired on a FacsVerse™ flow cytometer (BD Biosciences) and FCS express 4 (DeNovo Software) is used to analyze the fluorescent data. Hierarchical gating is used to discriminate discrete cell populations and the CD34+ gates are applied to the data using the no CD34+ (Stain #2) and the isotype control (Stain #3) samples as a guide to determine CD34+ from CD34- stained cells. A rat bone marrow sample serves as a positive control for a sample containing CD34+ cells. The CD34+ cells are plotted as a percentage live cells (see, figure 2). Student-T test is calculated using GraphPad Prism.
Example 3: Hematopoietic Colony Forming (CFC) assay.
[00131] Rat PBMC samples from the Day 7 and Day 29 studies are analyzed for the number of primitive hematopoietic cells that able to form colonies in semi-solid media. MethoCult™ GF R3774 (Stem Cell Technologies) is optimized for detection and quantification of rat hematopoietic progenitors using CFC assays and has been formulated with growth factors (rat IL-3, GM-CSF, and Stem Cell Factor) to support optimal growth of granulocyte-macrophage progenitors (CFU-GM, CFU-M, CFU-G). For each rat sample, 3 Petri dishes are prepared containing 75,000 cells and 1 mL of MethoCult per dish. Cultures are incubated in a 37°C, 5% C02 incubator and colonies are counted after 14 days in a blinded fashion. Each point is the average count of 3 replicate cultures for each rat PBMC sample. P values are calculated by Student-T test. The vehicle group shows one rat from day 7 (value = 0 colonies) and 4 rats from the day 29 rat study (see, figure 3). Example 4: Results.
[00132] Flow cytometry phenotyping using antibodies directed to CD45, CD1 lb, and CD34 provided a clear way to discriminate the CD34+ population from more mature populations, and can be measured as a percentage of live cells (Figure 1). Rats that receive COMPOUND 1 for 7 days show an increase in CD34+ percentage compared to rats that received vehicle (p-value = 0.0448) (Figure 2)
[00133] After conducting the CD34+ phenotyping, four day 7 rat samples (3 from the COMPOUND 1 group and 1 from the vehicle group) contained adequate numbers of cells to test in a CFC assay. Also included in this experiment are samples from the day 29 rat study. A statically relevant increase in colonies numbers are counted in cultures from the COMPOUND 1 treated animals compared to vehicle (p=.0142 for the 7-day study and p=.0105 for the 29-day study).
Example 5: Conclusion.
[00134] Blocking the chemokine receptor, CXCR4, results in mobilization of hematopoietic progenitor cells from the bone marrow to the periphery as shown with the clinical use of AMD3100 (Mozobil), an antagonist of CXCR4. This drug specifically increased CD34+ hematopoietic stem cell counts in the peripheral blood. Stromal derived factor 1 (SDF-1), through its interaction with CXCR4, retains the stem cells in the bone marrow (a process referred to as "homing")and AMD3100 specifically antagonizes this interaction. AMD3100 in combination with granulocyte colony- stimulating factor (G-CSF) results in the collection of more progenitor cells than G-CSF alone. At present, the major indication for clinical use of AMD3100 is the mobilization of hematopoietic stem cells from the bone marrow into the circulating blood for transplantation in patients with
hematological malignancies such as non-Hodgkin's lymphoma or multiple myeloma.
[00135] BTK has been shown to play a role in signaling downstream of CXCR4 induced by the chemokine SDF-1. As provided for the first time herein, blocking BTK with COMPOUND 1, a BTK- specific covalent small molecule inhibitor, shows similar biological effects as antagonizing CXCR4 on mobilizing hematopoietic progenitor cells to the periphery.
[00136] COMPOUND 1 treated rats at 300 mg/kg/day for 7 days results in an increase in circulating hematopoietic progenitor cells in the peripheral blood. These cells express CD34 on the cell surface and form CFC-GM colonies in semi-solid media supplemented with growth factors that induce proliferation in myeloid progenitor cells. Rats that are dosed for 28 days with COMPOUND 1 also showed an increased in CFC-GM colonies.
[00137] Certain modifications and improvements will occur to those skilled in the art upon a reading of the foregoing description. It should be understood that all such modifications and improvements have been omitted herein for the sake of conciseness and readability but are properly within the scope of the following claims.

Claims

Claims:
1. A method for mobilizing hematopoietic stem and/or progenitor cells in a subject, comprising administering to said subject a pharmaceutical composition comprising a Bruton's Tyrosine Kinase (Btk) inhibitor comprising a 6,5 fused heteroaryl ring system in an amount effective to mobilize said hematopoietic stem and/or progenitor cells into the peripheral blood of said subject.
2. The method of claim 1, wherein said Btk inhibitor is a compound selected from Formula I:
Figure imgf000081_0001
Formula I
or a pharmaceutically acceptable salt thereof, wherein:
W is (l-3C)alkyl, or NH2;
X is CH, N, O or S;
Y is C(R6), N, O or S;
Z is CH, N or bond;
A is CH or N;
Bl is N or C(R7);
B2 is N or C(R8);
B3 is N or C(R9);
B4 is N or C(RlO);
Rl is Rl lC(O), R12S(0), R13S02 or (l-6C)alkyl optionally substituted with R14;
R2 is H, (l-3C)alkyl or (3-7C)cycloalkyl;
R3 is H, (l-6C)alkyl or (3-7C)cycloalkyl); or
R2 and R3 form, together with the N and C atom they are attached to, a (3- 7C)heterocycloalkyl optionally substituted with one or more fluorine, hydroxyl, (1 -3C)alkyl, (l-3C)alkoxy or oxo;
R4 is H or (l-3C)alkyl;
R5 is H, halogen, cyano, (l-4C)alkyl, (l-3C)alkoxy, (3-6C)cycloalkyl, any alkyl group of which is optionally substituted with one or more halogen; or
R5 is (6-10C)aryl or (2-6C)heterocycloalkyl;
R6 is H or (l-3C)alkyl; or
R5 and R6 together may form a (3-7C)cycloalkenyl, or (2-6C)heterocycloalkenyl; each optionally substituted with (l-3C)alkyl, or one or more halogen;
R7 is H, halogen or (l-3C)alkoxy;
R8 is H or (l-3C)alkyl; or
R7 and R8 form, together with the carbon atom they are attached to a (6-10C)aryl or (1- 9C)heteroaryl;
R9 is H, halogen or (l-3C)alkoxy;
RIO is H, halogen or (l-3C)alkoxy;
Rl 1 is independently selected from a group consisting of (l-6C)alkyl, (2-6C)alkenyl and (2- 6C)alkynyl each alkyl, alkenyl or alkynyl optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino,
di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3-7C)cycloalkoxy, (6-10C)aryl or (3- 7C)heterocycloalkyl; or
Rl l is (l-3C)alkyl-C(0)-S-(l-3C)alkyl; or
Rl 1 is (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen or cyano;
R12 and R13 are independently selected from a group consisting of (2-6C)alkenyl or (2- 6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (1- 4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3- 7C)cycloalkoxy, (6-10C)aryl or (3-7C)heterocycloalkyl; or (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen, or
cyano;
R14 is independently selected from a group consisting of halogen, cyano or (2-6C)alkenyl or (2-6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3- 7C)cycloalkoxy, (6-10C)aryl, (l-5C)heteroaryl or (3-7C)heterocycloalkyl;
with the proviso that
0 to 2 atoms of X, Y, Z can simultaneously be a heteroatom;
when one atom selected from X, Y is O or S, then Z is a bond and the other atom selected from X, Y cannot be O or S;
- when Z is C or N, then Y is C(R6) or N and X is C or N; 0 to 2 atoms of Bl, B2, B3 and B4 are N.
3. The method according to claim 2, wherein W is NH2.
4. The method according to claim 2, wherein W is CH3.
5. The method according to claim 2, wherein Bl is C(R7); B2 is C(R8); B3 is C(R9); B4 is C(R10);
R7, R9, and RI O each are H; and
R8 is selected from a group consisting of hydrogen and methyl.
6. The method according to any one of claims 2 to 5, wherein R4 is selected from a group consisting of hydrogen and methyl.
7. The method according to any one of claims 2 to 6, wherein the ring containing X, Y, and Z is selected from a group consisting of pyridyl, pyrimidyl, pyridazyl, triazinyl, thiazolyl, oxazolyl, and isoxazolyl.
8. The method according to any one of claims 2 to 7, wherein R5 is selected from a group consisting of hydrogen, fluorine, chlorine, (l -3C)alkyl and (1-2C) alkoxy; the (l -3C)alkyl group of which is optionally substituted with one or more halogen.
9. The method according to any one of claims 2 to 8, wherein R5 is selected from a group consisting of hydrogen, fluorine, methyl, ethyl, propyl, methoxy and trifluoromethyl.
10. The method according to any one of claims 2 to 9, wherein R2 is hydrogen or (l-3C)alkyl; and R3 is (l-6C)alkyl.
11. The method according to any one of claims 2 to 10, wherein R2 and R3 together form a heterocycloalkyl ring selected from azetidinyl, pyrrolidinyl, piperidinyl, homopiperidinyl or morpholinyl, optionally substituted with one or more fluoro, hydroxyl, (1 -3C) alkyl, (1 - 3C)alkoxy, or oxo.
12. The method according to any one of claims 2 to 1 1 , wherein Rl is Rl lC(O) and Rl 1 is independently selected from a group consisting of (l -6C)alkyl, (2-6C)alkenyl or (2- 6C)alkynyl each optionally substituted with one or more groups selected from hydroxyl, (1 - 4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l -4C)alkyl]amino, (l -3C)alkoxy, (3- 7C)cycloalkoxy, (6-10C)aryl or (3-7C)heterocycloalkyl; or Rl l is (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen or cyano.
13. The method according to any one of claims 2 to 12, wherein Rl is Rl lC(O) and Rl 1 is selected from a group consisting of (2-6C)alkenyl or (2-6C)alkynyl each optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3-7C)cycloalkoxy or (3-7C)heterocycloalkyl.
14. The method according to claim 1 , wherein said Btk inhibitor is a compound selected from Formula II:
Figure imgf000084_0001
Formula
or a pharmaceutically acceptable salt thereof, wherein:
W is (l-3C)alkyl, or NH2;
Rl is selected from: (l-6C)acyl, (2-6)alkenylcarbonyl, (2-6)alkenylsulfonyl, (l-6C)alkyl, (2- 6)alkynylcarbonyl, and (l-5C)heteroarylcarbonyl, each optionally substituted with one or more substituents selected from: (l-6C)acylthio, (l-3C)alkoxy, (6-10C)aryl, cyano, (3- 7C)cycloalkyl, di[(l-4C)alkyl]amino, (3-7C)heterocycloalkyl, halogen, and hydroxyl;
R2 is H or (l-3C)alkyl;
R3 is H or (l-6C)alkyl; or
R2 and R3 form, together with the N and C atom they are attached to, a (5- 7C)heterocycloalkyl;
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazin-3-yl, and isoxazol-3-yl, each optionally substituted with one substituent selected from: (l-3C)alkoxy, (l-4C)alkyl, (6-10C)aryl, (3-6C)cycloalkyl, halogen, and (3-6C)heterocycloalkyl, wherein (l-4C)alkyl is further optionally substituted with one or more halogens.
The method according to claim 1 , wherein said Btk inhibitor is a compound selected from Formula II:
Figure imgf000085_0001
Formula II
or a pharmaceutically acceptable salt thereof, wherein:
W is methyl, or NH2;
Rl is selected from: acetyl, acryloyl, but-2-enoyl, but-2-ynoyl, hex-2-ynoyl, methacryloyl, methyl, 4-methylpent-2-ynoyl, pent-2-ynoyl, pyrimidine-4-carbonyl, and vinylsulfonyl, each optionally substituted with one or more substituents selected from: chloro, cyano, dimethylamino, fluoro, hydroxyl, methoxy, phenyl, and pyrrolidinyl;
R2 is H or methyl;
R3 is H or methyl; or
R2 and R3 form, together with the N and C atom they are attached to, pyrrolidin-2-yl, piperidin-2-yl, azepan-2-yl, or morpholin-3-yl; and
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazin-3-yl, and isoxazol-3-yl, each optionally substituted with one substituent selected from: cyano, ethyl, fluoro, isopropyl, methoxy, methyl, phenyl, n-propyl, pyrrolidinyl, trifluoromethyl.
The method according to claim 1 , wherein said Btk inhibitor is a compound selected from Formula II:
Figure imgf000085_0002
Formula II
or a pharmaceutically acceptable salt thereof, wherein:
W is methyl, or NH2; Rl is selected from 2-(acetylthio)acetyl, acryloyl, but-2-enoyl, but-2-ynoyl, cinnamoyl, cyanomethyl, 3-cyclopropylpropioloyl, 4-(dimethylamino)but-2-enoyl, 4- (dimethylamino)but-2-ynoyl, hex-2-ynoyl, 4-hydroxy-4-methylpent-2-ynoyl, methacryloyl, 4- methoxybut-2-enoyl, 4-methoxybut-2-ynoyl, 4-methylpent-2-ynoyl, pent-2-ynoyl, 2- chloropyrimidine-4-carbonyl, 4-(pyrrolidin-l-yl)but-2-enoyl, 2-(trifluoromethyl)acryloyl, and vinylsulfonyl;
R2 is H or methyl;
R3 is H or methyl; or
R2 and R3 form, together with the N and C atom they are attached to, pyrrolidin-2-yl, piperidin-2-yl, azepan-2-yl, or morpholin-3-y; and
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazin-3-yl, isoxazol-3-yl, 4-fluoropyridin-2-yl, 4- methylpyridin-2-yl, 4-propylpyridin-2-yl, 4-(trifluoromethyl)pyridin-2-yl, 4-ethylpyridin-2-yl, 4-cyanopyridin-2-yl, 4-methylpyrimidin-2-yl, 5-ethylthiazol-2-yl, 4-(pyrrolidin-l-yl)pyridin- 2-yl, 4-methoxypyridin-2-yl, 4-isopropylpyridin-2-yl, 4-phenylpyridin-2-yl.
17. The method according to claim 1, wherein said Btk inhibitor is a compound selected from the group consisting of:
(S)-4 (3 -( 1 -Acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(pyridin-2-yl)benz amide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(4-fluoropyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin-2 -yl)b enzamide ;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)- N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)- N-(4-ethylpyridin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4,5,6,7- tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-fluoro-N-(pyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-methoxy-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(thiazol-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-fluoropyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin- 2-yl)benzamide;
(S)-4-(8-Amino-3-(l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyrimidin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyrimidin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyrimidin-4- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridazin-3- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(isoxazol-3- yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (5-ethylthiazol-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-fluoro-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo [ 1 ,5-a]pyrazin- l-yl)-2-methoxy-N-(4-propylpyridin-2-yl)benzamide;
4-(8-Amino-3-((S)-l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-3-methyl-N- (pyridin-2-yl)benzamide; 4-(3-(Acrylamidomethyl)-8-aminoimidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2-yl)benzam (S)-4-(8-Amino-3 -( 1 -but-2-ynamidoethyl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-S-2-(2-(8-Amino-l -(4-(pyridin-2-ylcarbamoyl)phenyl)imidazo[l ,5-a]pyrazin-3- yl)pyrrolidin-l -yl)-2-oxoethyl ethanethioate;
(S)-4-(8-Amino-3 -( 1 -(4-hydroxy-4-methylpent-2-ynoyl)pyrrolidin-2-yl)imidazo [1 ,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(6-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l -pent-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-Amino-3-(l -(3-cyclopropylpropioloyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -y^ N-(pyridin-2-yl)benz amide;
(S)-4-(8-Amino-3 -( 1 -hex-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
4-(3 -( 1 -Acryloylazepan-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(R)-4-(8-Amino-3-(4-but-2-ynoylmoφholin-3-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-( 1 -(N-methylbut-2-ynamido)ethyl)imidazo[ 1 ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(4-methoxybut-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-fluoropyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-(pyrrolidin- 1 - yl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- fluoropyridin2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- propylpyridin2-yl)benzamide; (S,E)-4-(8-amino-3-(l -(4-methoxy-N-methylbut-2-enamido)ethyl)imidazo[ 1 ,5-a]pyrazin- 1 - yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2-fluoro-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-methoxypyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2- fluoro-N-(4-methoxypyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (isoxazol-3-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (pyrimidin-2-yl)benz amide;
4-(8-amino-3-((S)-l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l- yl)-3-methyl-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-methylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-isopropylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-
1- yl)-N-(4-methylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(thiazol-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-
2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- propylpyridin2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-isopropylpyridin-2-yl)benzamide;
4-(8-amino-3-((S)-l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-3 -methyl -N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-fluoro-N-(4- propylpyridin-2-yl)benzamide;
4-(3 -((S)- 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-3-methyl-N-(pyridin- 2-yl)benzamide;
(E)-4-(8-amino-3-((4-(dimethylamino)but-2-enamido)methyl)imidazo[ 1 ,5-a]pyrazin- 1 -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloro pyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-isopropylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloro pyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridazin-3 -yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin- 1 -yl)-N-(pyridazin-3 -yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l- yl)-N-(pyridazin-3-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxy-N-methylbut-2-enamido)ethyl)imidazo[ 1 ,5-a]pyrazin- 1 - yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)-N-methylbut-2-enamido)ethyl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin- l-yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l- yl)-N-(4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-2-methoxy-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-2-fluoro-N-(pyridin-2-yl)benzamide; 4-(8-amino-3-((S) -((E)-4-methoxybut-2-enoyl)piperi^
3 -m ethyl -N- (pyridin-2 -yl)b enzamide ;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyrimidin-4-yl)benz amide;
4-(8-amino-3-((S)-l -((E)-4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- 3 -m ethyl -N- (4 -propylpyridin-2 -yl)b enzamide ;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (4-methylpyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- methylpyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l- yl)-N-(pyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-methacryloylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l-(2-(trifluoromethyl)acryloyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)- N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l-but-2-enoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin^ yl)benzamide;
(S)-4-(8-amino-3-(l-(cyanomethyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(E)-4-(8-amino-3-((4-methoxybut-2-enamido)methyl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin- 2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- (pyrrolidin- 1 -yl)pyridin-2-yl)benzamide;
(E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)azepan-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-cyanopyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-methoxy-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acrylamidoethyl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(thiazol-2- yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- isopropylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2- methoxy-N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l -cinnamoylpyrrolidm
yl)benzamide;
(S)-N-(l-(8-amino-l -(4-(pyridin-2-ylcarbamoyl)phenyl)imidazo[l ,5-a]pyrazin-3-yl)ethyl)-2- chloropyrimidine-4-carboxamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- fluoropyridin2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2-methoxy-N- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2- methoxy-N-(4-propylpyridin-2-yl)benzamide;
4-(8-amino-3-(but-2-ynamidomethyl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2-yl)benzamide; (S)-4-(8-amino-3-( 1 -(N-methylbut-2-ynamido)ethyl)imidazo[ 1 ,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2- fluoro-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(5- ethylthiazol-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(5-ethylthiazol-2- yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl) -N-(5 - ethyl thiazol-2 -yl)b enzamide ;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(R,E)-4-(8-amino-3-(4-(4-methoxybut-2-enoyl)moφholin-3-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin- 2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- methoxypyridin-2 -yl)b enzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyre^
1 -yl)-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-(trifluoromethyl)pyridin-2-yl)benz amide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-
1- yl)-N-(4-methylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- cyanopyridin2 -yl)b enzamide ;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- ethy lpyridin2 -yl)b enzamide ;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- phenylpyridin-2-yl)benzamide; and
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-phenylpyridin-
2- yl)benzamide.
18. The method according to claim 1 , wherein said Btk inhibitor is a compound selected from the group consisting of:
(S)-4-(3 -( 1 -But-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin- 2-yl)benzamide;
(S,E)-4-(3-(l-(4-(Dimethylamino)but-2-enoyl)piperidin-2-yl)-8-methylimidazo[l,5- a]pyrazin- 1 -yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-Methyl-3-(l-(vinylsulfonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)-8-methylimidazo[l,5-a]pyrazin-l- yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-Methoxybut-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -But-2-ynamidoethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acrylamidoethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methoxypyridin2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- cyanopyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin-2 -yl)b enzamide ;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methoxypyridin-2 -yl)b enzamide;
(S)-4-(3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)-8-methylimidazo[l ,5-a]pyrazin-l- yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)-N-methylbut-2-enamido)ethyl)-8-methylimidazo [1 ,5- a]pyrazin- 1 -yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- ethy lpyridin2 -yl)b enzamide ;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin- l-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpiperidin-2-yl)-8-methylimidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin2-yl)benzamide;
(S)-4-(3-(l-acrylamidoethyl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4-propylpyridin-2- yl)benzamide;
(S)-4-(8-methyl-3-(l-(N-methylbut-2-ynamido)ethyl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide; (S,E)-4-(8-methyl-3-(l -(4-(pyrrolidm^
a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpiperidin-2-yl)-8-methylimidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-methoxybut-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)-8-methylimidazo [1 ,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- cyanopyridin2 -yl)b enzamide ;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-methoxybut-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)piperidin-2-yl)imidazo[l ,^ l-yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(3-(l-(4-methoxybut-2-enamido)ethyl)-8-methylimidazo[l ,5-a]pyrazin-l -yl)-N-(4 propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- l-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-but-2-ynamidoethyl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4-propylpyridin-2 yl)benzamide; and
(S,E)-4-(3-(l-(4-methoxy-N-methylbut-2-enamido)ethyl)-8-methylimidazo[l ,5-a]pyrazin-l - yl)-N-(4-propylpyridin-2-yl)benzamide.
19. The method according to claim 1 , wherein said subject has impaired hematopoiesis.
20. The method according to claim 19, wherein said administering step is performed subsequent to a myelosuppressive or myeloablative treatment of said subject.
21. The method according to claim 20, wherein said myelosuppressive or myeloablative treatment comprises chemotherapy.
22. The method according to claim 19, wherein said administering step is performed after
accidental radiation exposure to said subject.
23. The method according to any one of claims 1-22, said method further comprising the
simultaneous or sequential administration of an additional mobilizing agent selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF),
Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM-CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor,
thrombopoietin, growth related oncogene, and combinations thereof.
24. A method of obtaining hematopoietic stem and/or progenitor cells from a subject comprising (a) mobilizing hematopoietic stem and/or progenitor cells according to the method of any one of claims 1-18; and (b) harvesting said cells.
25. The method according to claim 24, wherein said cells are harvested by apheresis.
26. A method of providing hematopoietic cells to a patient in need thereof comprising introducing cells obtained by a method according to any one of claims 24 or 25 to said patient.
27. The method according to claim 26, wherein said patient is undergoing bone marrow
transplantation.
28. The method according to claim 27, wherein said subject and said patient are histocompatible.
29. The method according to claim 28, wherein said subject and said patient are syngeneic.
30. The method according to claim 28, wherein said subject and said patient are allogeneic.
31. The method according to any one of claims 26-30, wherein said method comprises culturing the obtained cells to enrich and/or differentiate the hematopoietic stem and/or progenitor cells prior to introducing the cells into the patient.
32. The method of claim 31 , wherein the cells are enriched for or differentiated into myeloid cells.
33. The method of claim 26, wherein the cells are enriched for or differentiated into lymphoid cells.
34. The method of claim 26, wherein the cells are enriched for or differentiated into NK cells. 35. The method of claim 1, wherein the mobilizing Btk inhibitor is either reversible or
irreversible.
36. A compound for use in mobilizing hematopoietic stem and/or progenitor cells into the peripheral blood of a subject, wherein said compound is a Bruton's Tyrosine Kinase (Btk) inhibitor comprising a 6,5 fused heteroaryl ring system. 37. The compound for use according to claim 36, wherein said Btk inhibitor is a compound selected from Formula I:
Figure imgf000097_0001
Formula I
or a pharmaceutically acceptable salt thereof, wherein:
W is (l-3C)alkyl, or NH2;
X is CH, N, O or S;
Y is C(R6), N, O or S;
Z is CH, N or bond;
A is CH or N;
Bl is N or C(R7);
B2 is N or C(R8);
B3 is N or C(R9); B4 is N or C(RlO);
Rl is Rl lC(O), R12S(0), R13S02 or (l-6C)alkyl optionally substituted with R14;
R2 is H, (l-3C)alkyl or (3-7C)cycloalkyl;
R3 is H, (l-6C)alkyl or (3-7C)cycloalkyl); or
R2 and R3 form, together with the N and C atom they are attached to, a (3- 7C)heterocycloalkyl optionally substituted with one or more fluorine, hydroxyl, (l-3C)alkyl, (l-3C)alkoxy or oxo;
R4 is H or (l-3C)alkyl;
R5 is H, halogen, cyano, (l-4C)alkyl, (l-3C)alkoxy, (3-6C)cycloalkyl, any alkyl group of which is optionally substituted with one or more halogen; or
R5 is (6-10C)aryl or (2-6C)heterocycloalkyl;
R6 is H or (l-3C)alkyl; or
R5 and R6 together may form a (3-7C)cycloalkenyl, or (2-6C)heterocycloalkenyl; each optionally substituted with (l-3C)alkyl, or one or more halogen;
R7 is H, halogen or (l-3C)alkoxy;
R8 is H or (l-3C)alkyl; or
R7 and R8 form, together with the carbon atom they are attached to a (6-10C)aryl or (1- 9C)heteroaryl;
R9 is H, halogen or (l-3C)alkoxy;
RIO is H, halogen or (l-3C)alkoxy;
Rl 1 is independently selected from a group consisting of (l-6C)alkyl, (2-6C)alkenyl and (2- 6C)alkynyl each alkyl, alkenyl or alkynyl optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino,
di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3-7C)cycloalkoxy, (6-10C)aryl or (3- 7C)heterocycloalkyl; or
Rl l is (l-3C)alkyl-C(0)-S-(l-3C)alkyl; or
Rl 1 is (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen or cyano;
R12 and R13 are independently selected from a group consisting of (2-6C)alkenyl or (2- 6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (1- 4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3- 7C)cycloalkoxy, (6-10C)aryl or (3-7C)heterocycloalkyl; or (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen, or
cyano;
R14 is independently selected from a group consisting of halogen, cyano or (2-6C)alkenyl or (2-6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3- 7C)cycloalkoxy, (6-10C)aryl, (l-5C)heteroaryl or (3-7C)heterocycloalkyl;
with the proviso that
0 to 2 atoms of X, Y, Z can simultaneously be a heteroatom;
when one atom selected from X, Y is O or S, then Z is a bond and the other atom selected from X, Y cannot be O or S;
- when Z is C or N, then Y is C(R6) or N and X is C or N;
- 0 to 2 atoms of B 1 , B2, B3 and B4 are N.
The compound for use according to claim 36, wherein said Btk inhibitor is a compound selected from Formula II:
Figure imgf000099_0001
Formula
or a pharmaceutically acceptable salt thereof, wherein:
W is (l -3C)alkyl, or NH2;
Rl is selected from: (l -6C)acyl, (2-6)alkenylcarbonyl, (2-6)alkenylsulfonyl, (l -6C)alkyl, (2- 6)alkynylcarbonyl, and (l-5C)heteroarylcarbonyl, each optionally substituted with one or more substituents selected from: (l -6C)acylthio, (l -3C)alkoxy, (6-10C)aryl, cyano, (3- 7C)cycloalkyl, di[(l-4C)alkyl]amino, (3-7C)heterocycloalkyl, halogen, and hydroxyl;
R2 is H or (l-3C)alkyl;
R3 is H or (l-6C)alkyl; or
R2 and R3 form, together with the N and C atom they are attached to, a (5- 7C)heterocycloalkyl;
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazin-3-yl, and isoxazol-3-yl, each optionally substituted with one substituent selected from: (l -3C)alkoxy, (l -4C)alkyl, (6-10C)aryl, (3-6C)cycloalkyl, halogen, and (3-6C)heterocycloalkyl, wherein (l-4C)alkyl is further optionally substituted with one or more halogens.
The compound for use according to claim 36, wherein said Btk inhibitor is a compound selected from Formula II:
Figure imgf000100_0001
Formula II
or a pharmaceutically acceptable salt thereof, wherein:
W is methyl, or NH2;
Rl is selected from: acetyl, acryloyl, but-2-enoyl, but-2-ynoyl, hex-2-ynoyl, methacryloyl, methyl, 4-methylpent-2-ynoyl, pent-2-ynoyl, pyrimidine-4-carbonyl, and vinylsulfonyl, each optionally substituted with one or more substituents selected from: chloro, cyano, dimethylamino, fluoro, hydroxyl, methoxy, phenyl, and pyrrolidinyl;
R2 is H or methyl;
R3 is H or methyl; or
R2 and R3 form, together with the N and C atom they are attached to, pyrrolidin-2-yl, piperidin-2-yl, azepan-2-yl, or morpholin-3-yl; and
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazin-3-yl, and isoxazol-3-yl, each optionally substituted with one substituent selected from: cyano, ethyl, fluoro, isopropyl, methoxy, methyl, phenyl, n-propyl, pyrrolidinyl, trifluoromethyl.
The compound for use according to claim 36, wherein said Btk inhibitor is a compound selected from Formula II:
Figure imgf000100_0002
Formula II
or a pharmaceutically acceptable salt thereof, wherein:
W is methyl, or NH2; Rl is selected from 2-(acetylthio)acetyl, acryloyl, but-2-enoyl, but-2-ynoyl, cinnamoyl, cyanomethyl, 3-cyclopropylpropioloyl, 4-(dimethylamino)but-2-enoyl, 4- (dimethylamino)but-2-ynoyl, hex-2-ynoyl, 4-hydroxy-4-methylpent-2-ynoyl, methacryloyl, 4- methoxybut-2-enoyl, 4-methoxybut-2-ynoyl, 4-methylpent-2-ynoyl, pent-2-ynoyl, 2- chloropyrimidine-4-carbonyl, 4-(pyrrolidin-l-yl)but-2-enoyl, 2-(trifluoromethyl)acryloyl, and vinylsulfonyl;
R2 is H or methyl;
R3 is H or methyl; or
R2 and R3 form, together with the N and C atom they are attached to, pyrrolidin-2-yl, piperidin-2-yl, azepan-2-yl, or morpholin-3-y; and
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazin-3-yl, isoxazol-3-yl, 4-fluoropyridin-2-yl, 4- methylpyridin-2-yl, 4-propylpyridin-2-yl, 4-(trifluoromethyl)pyridin-2-yl, 4-ethylpyridin-2-yl, 4-cyanopyridin-2-yl, 4-methylpyrimidin-2-yl, 5-ethylthiazol-2-yl, 4-(pyrrolidin-l-yl)pyridin- 2-yl, 4-methoxypyridin-2-yl, 4-isopropylpyridin-2-yl, 4-phenylpyridin-2-yl.
41. The compound for use according to claim 36, wherein said Btk inhibitor is a compound
selected from the group consisting of:
(S)-4 (3 -( 1 -Acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(pyridin-2-yl)benz amide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(4-fluoropyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin-2 -yl)b enzamide ;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)- N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)- N-(4-ethylpyridin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4,5,6,7- tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-fluoro-N-(pyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-methoxy-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(thiazol-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-fluoropyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin- 2-yl)benzamide;
(S)-4-(8-Amino-3-(l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyrimidin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyrimidin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyrimidin-4- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridazin-3- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(isoxazol-3- yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (5-ethylthiazol-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-fluoro-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo [ 1 ,5-a]pyrazin- l-yl)-2-methoxy-N-(4-propylpyridin-2-yl)benzamide;
4-(8-Amino-3-((S)-l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-3-methyl-N- (pyridin-2-yl)benzamide; 4-(3-(Acrylamidomethyl)-8-aminoimidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2-yl)benzam (S)-4-(8-Amino-3 -( 1 -but-2-ynamidoethyl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-S-2-(2-(8-Amino-l -(4-(pyridin-2-ylcarbamoyl)phenyl)imidazo[l ,5-a]pyrazin-3- yl)pyrrolidin-l -yl)-2-oxoethyl ethanethioate;
(S)-4-(8-Amino-3 -( 1 -(4-hydroxy-4-methylpent-2-ynoyl)pyrrolidin-2-yl)imidazo [1 ,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(6-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l -pent-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-Amino-3-(l -(3-cyclopropylpropioloyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -y^ N-(pyridin-2-yl)benz amide;
(S)-4-(8-Amino-3 -( 1 -hex-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
4-(3 -( 1 -Acryloylazepan-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(R)-4-(8-Amino-3-(4-but-2-ynoylmoφholin-3-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-( 1 -(N-methylbut-2-ynamido)ethyl)imidazo[ 1 ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(4-methoxybut-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-fluoropyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-(pyrrolidin- 1 - yl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- fluoropyridin2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- propylpyridin2-yl)benzamide; (S,E)-4-(8-amino-3-(l -(4-methoxy-N-methylbut-2-enamido)ethyl)imidazo[ 1 ,5-a]pyrazin- 1 - yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(vinylsulfonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-fluoro-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-methoxypyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2- fluoro-N-(4-methoxypyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (isoxazol-3-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyrimidin-2-yl)benz amide;
4-(8-amino-3-((S)-l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l- yl)-3-methyl-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-methylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-isopropylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-
1- yl)-N-(4-methylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(thiazol-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-
2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- propylpyridin2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-isopropylpyridin-2-yl)benzamide;
4-(8-amino-3-((S)-l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-3 -methyl -N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-fluoro-N-(4- propylpyridin-2-yl)benzamide;
4-(3 -((S)- 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-3-methyl-N-(pyridin- 2-yl)benzamide;
(E)-4-(8-amino-3-((4-(dimethylamino)but-2-enamido)methyl)imidazo[ 1 ,5-a]pyrazin- 1 -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloro pyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-isopropylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloro pyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridazin-3 -yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin- 1 -yl)-N-(pyridazin-3 -yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l- yl)-N-(pyridazin-3-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxy-N-methylbut-2-enamido)ethyl)imidazo[ 1 ,5-a]pyrazin- 1 - yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)-N-methylbut-2-enamido)ethyl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin- l-yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l- yl)-N-(4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-2-methoxy-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-2-fluoro-N-(pyridin-2-yl)benzamide; 4-(8-amino-3-((S) -((E)-4-methoxybut-2-enoyl)piperi^
3 -m ethyl -N- (pyridin-2 -yl)b enzamide ;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N^ (pyrimidin-4-yl)benz amide;
4-(8-amino-3-((S)-l -((E)-4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- 3 -m ethyl -N- (4 -propylpyridin-2 -yl)b enzamide ;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (4-methylpyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- methylpyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l- yl)-N-(pyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-methacryloylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l-(2-(trifluoromethyl)acryloyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)- N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l-but-2-enoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin^ yl)benzamide;
(S)-4-(8-amino-3-(l-(cyanomethyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(E)-4-(8-amino-3-((4-methoxybut-2-enamido)methyl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin- 2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- (pyrrolidin- 1 -yl)pyridin-2-yl)benzamide;
(E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)azepan-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-cyanopyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-methoxy-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acrylamidoethyl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(thiazol-2- yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- isopropylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2- methoxy-N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l -cinnamoylpyrrolidm
yl)benzamide;
(S)-N-(l-(8-amino-l -(4-(pyridin-2-ylcarbamoyl)phenyl)imidazo[l ,5-a]pyrazin-3-yl)ethyl)-2- chloropyrimidine-4-carboxamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- fluoropyridin2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2-methoxy-N- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2- methoxy-N-(4-propylpyridin-2-yl)benzamide;
4-(8-amino-3-(but-2-ynamidomethyl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2-yl)benzamide; (S)-4-(8-amino-3-( 1 -(N-methylbut-2-ynamido)ethyl)imidazo[ 1 ,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2- fluoro-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(5- ethylthiazol-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(5-ethylthiazol-2- yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl) -N-(5 - ethyl thiazol-2 -yl)b enzamide ;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(R,E)-4-(8-amino-3-(4-(4-methoxybut-2-enoyl)moφholin-3-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin- 2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- methoxypyridin-2 -yl)b enzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-(trifluoromethyl)pyridin-2-yl)benz amide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-
1- yl)-N-(4-methylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- cyanopyridin2 -yl)b enzamide ;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- ethy lpyridin2 -yl)b enzamide ;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- phenylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-phenylpyridin-
2- yl)benzamide;
(S)-4-(3 -( 1 -But-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin- 2-yl)benzamide;
(S,E)-4-(3-(l-(4-(Dimethylamino)but-2-enoyl)piperidin-2-yl)-8-methylimidazo[l,5- a]pyrazin- 1 -yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-Methyl-3-(l-(vinylsulfonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)-8-methylimidazo[l,5-a]pyrazin-l- yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-Methoxybut-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -But-2-ynamidoethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide; (S)-4-(3 -( 1 -Acrylamidoethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methoxypyridin2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- cyanopyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin-2 -yl)b enzamide ;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methoxypyridin-2 -yl)b enzamide;
(S)-4-(3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)-8-methylimidazo[l,5-a]pyrazin-l- yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)-N-methylbut-2-enamido)ethyl)-8-methylimidazo [1,5- a]pyrazin- 1 -yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- ethy lpyridin2 -yl)b enzamide ;
(S,E)-4-(8-methyl-3-(l-(4-(pyrrolidin-l-yl)but-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpiperidin-2-yl)-8-methylimidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin2-yl)benzamide;
(S)-4-(3-(l-acrylamidoethyl)-8-methylimidazo[l,5-a]pyrazin-l-yl)-N-(4-propylpyridin-2- yl)benzamide;
(S)-4-(8-methyl-3-(l-(N-methylbut-2-ynamido)ethyl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-methyl-3-(l-(4-(pyrrolidin-l-yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpiperidin-2-yl)-8-methylimidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-methoxybut-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)-8-methylimidazo [1 ,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- cyanopyridin2 -yl)b enzamide ;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-methoxybut-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)piperidin-2-yl)imidazo[l ,^ l-yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(3-(l-(4-methoxybut-2-enamido)ethyl)-8-methylimidazo[l ,5-a]pyrazin-l -yl)-N-(4 propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- l-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-but-2-ynamidoethyl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4-propylpyridin- yl)benzamide; and
(S,E)-4-(3-(l-(4-methoxy-N-methylbut-2-enamido)ethyl)-8-methylimidazo[l ,5-a]pyrazin-l - yl)-N-(4-propylpyridin-2-yl)benzamide.
42. The compound for use according to any one of claims 36 to 41 , wherein said subject has impaired hematopoiesis.
43. The compound for use according to claim 42, wherein said subject has previously received a myelosuppressive or myeloablative treatment.
44. The compound for use according to claim 43, wherein said myelosuppressive or
myeloablative treatment comprises chemotherapy.
45. The compound for use according to claim 42, wherein said subject has previously been accidentally exposed to radiation.
46. The compound for use according to any one of claims 36 to 45, in combination with an additional mobilizing agent selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM-CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor, thrombopoietin, growth related oncogene, and combinations thereof.
47. The compound for use according to any one of claims 36 to 41 , for use in harvesting said hematopoietic stem and/or progenitor cells from said subject.
48. The compound for use according to claim 47, for use in harvesting said hematopoietic stem and/or progenitor cells from said subject by apheresis.
49. The compound for use according to claim 47 or 48, for use in providing said hematopoietic stem and/or progenitor cells to a patient in need thereof.
50. The compound for use according to claim 49, for use in providing said hematopoietic stem and/or progenitor cells to a bone marrow transplantation patient.
51. The compound for use according to claim 50, wherein said subject and said patient are histocompatible.
52. The compound for use according to claim 51, wherein said subject and said patient are syngeneic.
53. The compound for use according to claim 51, wherein said subject and said patient are allogeneic.
54. The compound for use according to any one of claims 49 to 53, wherein said hematopoietic stem and/or progenitor cells are cultured to enrich and/or differentiate said hematopoietic stem and/or progenitor cells prior to introducing the cells into the patient.
55. The compound for use according to claim 54, wherein said hematopoietic stem and/or progenitor cells are enriched for or differentiated into cells selected from: myeloid cells, lymphoid cells, and NK cells. The compound for use according to any one of claims 36 to 55, wherein said Btk inhibitor is reversible.
The compound for use according to any one of claims 36 to 55, wherein said Btk inhibitor is irreversible.
Use of a Bruton's Tyrosine Kinase (Btk) inhibitor comprising a 6,5 fused heteroaryl ring system in the manufacture of a medicament for mobilizing hematopoietic stem and/or progenitor cells into the peripheral blood of a subject.
The use according to claim 58, wherein said Btk inhibitor is a compound selected from Formula I:
Figure imgf000112_0001
Formula I
or a pharmaceutically acceptable salt thereof, wherein:
W is (l-3C)alkyl, or NH2;
X is CH, N, O or S;
Y is C(R6), N, O or S;
Z is CH, N or bond;
A is CH or N;
Bl is N or C(R7);
B2 is N or C(R8);
B3 is N or C(R9);
B4 is N or C(RlO);
Rl is Rl lC(O), R12S(0), R13S02 or (l-6C)alkyl optionally substituted with R14; R2 is H, (l-3C)alkyl or (3-7C)cycloalkyl;
R3 is H, (l-6C)alkyl or (3-7C)cycloalkyl); or
R2 and R3 form, together with the N and C atom they are attached to, a (3- 7C)heterocycloalkyl optionally substituted with one or more fluorine, hydroxyl, (l-3C)alkyl, (l-3C)alkoxy or oxo;
R4 is H or (l-3C)alkyl;
R5 is H, halogen, cyano, (l-4C)alkyl, (l-3C)alkoxy, (3-6C)cycloalkyl, any alkyl group of which is optionally substituted with one or more halogen; or
R5 is (6-10C)aryl or (2-6C)heterocycloalkyl;
R6 is H or (l-3C)alkyl; or
R5 and R6 together may form a (3-7C)cycloalkenyl, or (2-6C)heterocycloalkenyl; each optionally substituted with (l-3C)alkyl, or one or more halogen;
R7 is H, halogen or (l-3C)alkoxy;
R8 is H or (l-3C)alkyl; or
R7 and R8 form, together with the carbon atom they are attached to a (6-10C)aryl or (1- 9C)heteroaryl;
R9 is H, halogen or (l-3C)alkoxy;
RIO is H, halogen or (l-3C)alkoxy;
Rl 1 is independently selected from a group consisting of (l-6C)alkyl, (2-6C)alkenyl and (2- 6C)alkynyl each alkyl, alkenyl or alkynyl optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino,
di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3-7C)cycloalkoxy, (6-10C)aryl or (3- 7C)heterocycloalkyl; or
Rl l is (l-3C)alkyl-C(0)-S-(l-3C)alkyl; or
Rl 1 is (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen or cyano;
R12 and R13 are independently selected from a group consisting of (2-6C)alkenyl or (2- 6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (1- 4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3- 7C)cycloalkoxy, (6-10C)aryl or (3-7C)heterocycloalkyl; or (l-5C)heteroaryl optionally substituted with one or more groups selected from halogen, or
cyano;
R14 is independently selected from a group consisting of halogen, cyano or (2-6C)alkenyl or (2-6C)alkynyl both optionally substituted with one or more groups selected from hydroxyl, (l-4C)alkyl, (3-7C)cycloalkyl, [(l-4C)alkyl]amino, di[(l-4C)alkyl]amino, (l-3C)alkoxy, (3- 7C)cycloalkoxy, (6-10C)aryl, (l-5C)heteroaryl or (3-7C)heterocycloalkyl;
with the proviso that 0 to 2 atoms of X, Y, Z can simultaneously be a heteroatom;
when one atom selected from X, Y is O or S, then Z is a bond and the other atom selected from X, Y cannot be O or S;
- when Z is C or N, then Y is C(R6) or N and X is C or N;
- 0 to 2 atoms of B 1 , B2, B3 and B4 are N.
The use according to claim 58, wherein said Btk inhibitor is a compound selected from Formula II:
Figure imgf000114_0001
Formula
or a pharmaceutically acceptable salt thereof, wherein:
W is (l-3C)alkyl, or NH2;
Rl is selected from: (l-6C)acyl, (2-6)alkenylcarbonyl, (2-6)alkenylsulfonyl, (l-6C)alkyl, (2- 6)alkynylcarbonyl, and (l-5C)heteroarylcarbonyl, each optionally substituted with one or more substituents selected from: (l-6C)acylthio, (l-3C)alkoxy, (6-10C)aryl, cyano, (3- 7C)cycloalkyl, di[(l-4C)alkyl]amino, (3-7C)heterocycloalkyl, halogen, and hydroxyl;
R2 is H or (l-3C)alkyl;
R3 is H or (l-6C)alkyl; or
R2 and R3 form, together with the N and C atom they are attached to, a (5- 7C)heterocycloalkyl;
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazin-3-yl, and isoxazol-3-yl, each optionally substituted with one substituent selected from: (l-3C)alkoxy, (l-4C)alkyl, (6-10C)aryl, (3-6C)cycloalkyl, halogen, and (3-6C)heterocycloalkyl, wherein (l-4C)alkyl is further optionally substituted with one or more halogens.
The use according to claim 58, wherein said Btk inhibitor is a compound selected from Formula II:
Figure imgf000115_0001
Formula II
or a pharmaceutically acceptable salt thereof, wherein:
W is methyl, or NH2;
Rl is selected from: acetyl, acryloyl, but-2-enoyl, but-2-ynoyl, hex-2-ynoyl, methacryloyl, methyl, 4-methylpent-2-ynoyl, pent-2-ynoyl, pyrimidine-4-carbonyl, and vinylsulfonyl, each optionally substituted with one or more substituents selected from: chloro, cyano, dimethylamino, fluoro, hydroxyl, methoxy, phenyl, and pyrrolidinyl;
R2 is H or methyl;
R3 is H or methyl; or
R2 and R3 form, together with the N and C atom they are attached to, pyrrolidin-2-yl, piperidin-2-yl, azepan-2-yl, or morpholin-3-yl; and
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazin-3-yl, and isoxazol-3-yl, each optionally substituted with one substituent selected from: cyano, ethyl, fluoro, isopropyl, methoxy, methyl, phenyl, n-propyl, pyrrolidinyl, trifluoromethyl.
The use according to claim 58, wherein said Btk inhibitor is a compound selected from Formula II:
Figure imgf000115_0002
Formula II
or a pharmaceutically acceptable salt thereof, wherein:
W is methyl, or NH2; Rl is selected from 2-(acetylthio)acetyl, acryloyl, but-2-enoyl, but-2-ynoyl, cinnamoyl, cyanomethyl, 3-cyclopropylpropioloyl, 4-(dimethylamino)but-2-enoyl, 4- (dimethylamino)but-2-ynoyl, hex-2-ynoyl, 4-hydroxy-4-methylpent-2-ynoyl, methacryloyl, 4- methoxybut-2-enoyl, 4-methoxybut-2-ynoyl, 4-methylpent-2-ynoyl, pent-2-ynoyl, 2- chloropyrimidine-4-carbonyl, 4-(pyrrolidin-l-yl)but-2-enoyl, 2-(trifluoromethyl)acryloyl, and vinylsulfonyl;
R2 is H or methyl;
R3 is H or methyl; or
R2 and R3 form, together with the N and C atom they are attached to, pyrrolidin-2-yl, piperidin-2-yl, azepan-2-yl, or morpholin-3-y; and
R15 is selected from: pyridin-2-yl, 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl, thiazol-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazin-3-yl, isoxazol-3-yl, 4-fluoropyridin-2-yl, 4- methylpyridin-2-yl, 4-propylpyridin-2-yl, 4-(trifluoromethyl)pyridin-2-yl, 4-ethylpyridin-2-yl, 4-cyanopyridin-2-yl, 4-methylpyrimidin-2-yl, 5-ethylthiazol-2-yl, 4-(pyrrolidin-l-yl)pyridin- 2-yl, 4-methoxypyridin-2-yl, 4-isopropylpyridin-2-yl, 4-phenylpyridin-2-yl.
63. The use according to claim 58, wherein said Btk inhibitor is a compound selected from the group consisting of:
(S)-4 (3 -( 1 -Acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(pyridin-2-yl)benz amide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(4-fluoropyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin-2 -yl)b enzamide ;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)- N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -but-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)- N-(4-ethylpyridin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4,5,6,7- tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-fluoro-N-(pyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-methoxy-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5- a]pyrazin- 1 -yl)-N-(thiazol-2-yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-fluoropyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin- 2-yl)benzamide;
(S)-4-(8-Amino-3-(l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyrimidin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyrimidin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyrimidin-4- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridazin-3- yl)benzamide;
(S)-4-(8-Amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(isoxazol-3- yl)benzamide;
(S,E)-4-(8-Amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (5-ethylthiazol-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-2-fluoro-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo [ 1 ,5-a]pyrazin- l-yl)-2-methoxy-N-(4-propylpyridin-2-yl)benzamide;
4-(8-Amino-3-((S)-l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-3-methyl-N- (pyridin-2-yl)benzamide; 4-(3-(Acrylamidomethyl)-8-aminoimidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2-yl)benzam (S)-4-(8-Amino-3 -( 1 -but-2-ynamidoethyl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-S-2-(2-(8-Amino-l -(4-(pyridin-2-ylcarbamoyl)phenyl)imidazo[l ,5-a]pyrazin-3- yl)pyrrolidin-l -yl)-2-oxoethyl ethanethioate;
(S)-4-(8-Amino-3 -( 1 -(4-hydroxy-4-methylpent-2-ynoyl)pyrrolidin-2-yl)imidazo [1 ,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(6-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3-(l -pent-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-Amino-3-(l -(3-cyclopropylpropioloyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -y^ N-(pyridin-2-yl)benz amide;
(S)-4-(8-Amino-3 -( 1 -hex-2-ynoylpyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
4-(3 -( 1 -Acryloylazepan-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(R)-4-(8-Amino-3-(4-but-2-ynoylmoφholin-3-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-( 1 -(N-methylbut-2-ynamido)ethyl)imidazo[ 1 ,5-a]pyrazin-l -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(4-(dimethylamino)but-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-Amino-3 -( 1 -(4-methoxybut-2-ynoyl)pyrrolidin-2-yl)imidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-fluoropyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-(pyrrolidin- 1 - yl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(4- fluoropyridin2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- propylpyridin2-yl)benzamide; (S,E)-4-(8-amino-3-(l -(4-methoxy-N-methylbut-2-enamido)ethyl)imidazo[ 1 ,5-a]pyrazin- 1 - yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(vinylsulfonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-fluoro-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-methoxypyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2- fluoro-N-(4-methoxypyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (isoxazol-3-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyrimidin-2-yl)benz amide;
4-(8-amino-3-((S)-l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l- yl)-3-methyl-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-methylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-isopropylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-
1- yl)-N-(4-methylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(thiazol-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin-
2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- propylpyridin2-yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-isopropylpyridin-2-yl)benzamide;
4-(8-amino-3-((S)-l -(vinylsulfonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-3 -methyl -N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2-fluoro-N-(4- propylpyridin-2-yl)benzamide;
4-(3 -((S)- 1 -acryloylpiperidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-3-methyl-N-(pyridin- 2-yl)benzamide;
(E)-4-(8-amino-3-((4-(dimethylamino)but-2-enamido)methyl)imidazo[ 1 ,5-a]pyrazin- 1 -yl)-N- (pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloro pyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin- l-yl)-N-(4-isopropylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloro pyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin- l-yl)-N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (pyridazin-3 -yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin- 1 -yl)-N-(pyridazin-3 -yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl)-N-(pyridazin-3-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxy-N-methylbut-2-enamido)ethyl)imidazo[ 1 ,5-a]pyrazin- 1 - yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)-N-methylbut-2-enamido)ethyl)imidazo[l ,5- a]pyrazin- 1 -yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin- l-yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl)-N-(4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (4-fluoropyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-2-methoxy-N-(pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-2-fluoro-N-(pyridin-2-yl)benzamide; 4-(8-amino-3-((S) -((E)-4-methoxybut-2-enoyl)piperi^
3 -m ethyl -N- (pyridin-2 -yl)b enzamide ;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyrimidin-4-yl)benz amide;
4-(8-amino-3-((S)-l -((E)-4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- 3 -m ethyl -N- (4 -propylpyridin-2 -yl)b enzamide ;
(S,E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (4-methylpyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpiperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- methylpyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l- yl)-N-(pyrimidin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-methacryloylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(8-amino-3-(l-(2-(trifluoromethyl)acryloyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)- N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l-but-2-enoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin^ yl)benzamide;
(S)-4-(8-amino-3-(l-(cyanomethyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin-2- yl)benzamide;
(E)-4-(8-amino-3-((4-methoxybut-2-enamido)methyl)imidazo[l,5-a]pyrazin-l-yl)-N-(pyridin- 2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- (pyrrolidin- 1 -yl)pyridin-2-yl)benzamide;
(E)-4-(8-amino-3-(l-(4-methoxybut-2-enoyl)azepan-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N- (pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-cyanopyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-2-methoxy-N- (pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acrylamidoethyl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(thiazol-2- yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- isopropylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2- methoxy-N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l -cinnamoylpyrrolidm
yl)benzamide;
(S)-N-(l-(8-amino-l -(4-(pyridin-2-ylcarbamoyl)phenyl)imidazo[l ,5-a]pyrazin-3-yl)ethyl)-2- chloropyrimidine-4-carboxamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- fluoropyridin2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-2-methoxy-N- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2- methoxy-N-(4-propylpyridin-2-yl)benzamide;
4-(8-amino-3-(but-2-ynamidomethyl)imidazo[l ,5-a]pyrazin-l -yl)-N-(pyridin-2-yl)benzamide; (S)-4-(8-amino-3-( 1 -(N-methylbut-2-ynamido)ethyl)imidazo[ 1 ,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-2- fluoro-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l -but-2-ynoylpiperidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)-N-(5- ethylthiazol-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-aminoimidazo[l ,5-a]pyrazin-l-yl)-N-(5-ethylthiazol-2- yl)benzamide;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin-l - yl) -N-(5 - ethyl thiazol-2 -yl)b enzamide ;
(S)-4-(8-amino-3-(l -(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- l-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(R,E)-4-(8-amino-3-(4-(4-methoxybut-2-enoyl)moφholin-3-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(pyridin-2-yl)benz amide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l -yl)-N- (4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-cyanopyridin- 2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- methoxypyridin-2 -yl)b enzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-methylpyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin-2- yl)benzamide;
(S,E)-4-(8-amino-3-(l-(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S,E)-4-(8-amino-3-(l -(4-methoxybut-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5-a]pyrazin-l -yl)- N-(4-(trifluoromethyl)pyridin-2-yl)benz amide;
(S)-4-(8-amino-3-(l-(2-chloropyrimidine-4-carbonyl)pyrrolidin-2-yl)imidazo[l,5-a]pyrazin-
1- yl)-N-(4-methylpyridin-2-yl)benzamide;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- cyanopyridin2 -yl)b enzamide ;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- ethy lpyridin2 -yl)b enzamide ;
(S)-4-(8-amino-3-(l-but-2-ynoylpyrrolidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- phenylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-aminoimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-phenylpyridin-
2- yl)benzamide;
(S)-4-(3 -( 1 -But-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpiperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-propylpyridin- 2-yl)benzamide;
(S,E)-4-(3-(l-(4-(Dimethylamino)but-2-enoyl)piperidin-2-yl)-8-methylimidazo[l,5- a]pyrazin- 1 -yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(8-Methyl-3-(l-(vinylsulfonyl)piperidin-2-yl)imidazo[l,5-a]pyrazin-l-yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)-8-methylimidazo[l,5-a]pyrazin-l- yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-Methoxybut-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -But-2-ynamidoethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide; (S)-4-(3 -( 1 -Acrylamidoethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methoxypyridin2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- cyanopyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -Acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4-ethylpyridin- 2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methylpyridin-2 -yl)b enzamide ;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- methoxypyridin-2 -yl)b enzamide;
(S)-4-(3-(l-(2-chloropyrimidine-4-carbonyl)piperidin-2-yl)-8-methylimidazo[l ,5-a]pyrazin-l- yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)-N-methylbut-2-enamido)ethyl)-8-methylimidazo [1 ,5- a]pyrazin- 1 -yl)-N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -acryloylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(pyridin-2- yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- ethy lpyridin2 -yl)b enzamide ;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)piperidin-2-yl)imidazo[l ,5-a]pyrazin- l-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpiperidin-2-yl)-8-methylimidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin2-yl)benzamide;
(S)-4-(3-(l-acrylamidoethyl)-8-methylimidazo[l ,5-a]pyrazin-l-yl)-N-(4-propylpyridin-2- yl)benzamide;
(S)-4-(8-methyl-3-(l-(N-methylbut-2-ynamido)ethyl)imidazo[l,5-a]pyrazin-l -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(8-methyl-3-(l -(4-(pyrrolidin-l -yl)but-2-enoyl)pyrrolidin-2-yl)imidazo[l ,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-acryloylpiperidin-2-yl)-8-methylimidazo[l,5-a]pyrazin-l-yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enamido)ethyl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)- N-(4-propylpyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpiperidin-2-yl)-8-methylimidazo[ 1 ,5-a]pyrazin- 1 -yl)-N-(4- (trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-methoxybut-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enoyl)pyrrolidin-2-yl)-8-methylimidazo [1,5- a]pyrazin- 1 -yl)-N-(pyridin-2-yl)benzamide;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- cyanopyridin2 -yl)b enzamide ;
(S)-4-(3 -( 1 -but-2-ynoylpyrrolidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-methoxybut-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- 1 -yl)-N- (4-propylpyridin-2-yl)benzamide;
(S,E)-4-(8-methyl-3-(l-(4-(pyrrolidin-l-yl)but-2-enoyl)piperidin-2-yl)imidazo[l,5-a]pyra l-yl)-N-(4-propylpyridin-2-yl)benzamide;
(S,E)-4-(3-(l-(4-methoxybut-2-enamido)ethyl)-8-methylimidazo[l,5-a]pyrazin-l-yl)-N-(4- propylpyridin-2-yl)benzamide;
(S,E)-4-(3 -( 1 -(4-(dimethylamino)but-2-enoyl)piperidin-2-yl)-8-methylimidazo [ 1 ,5-a]pyrazin- l-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;
(S)-4-(3-(l-but-2-ynamidoethyl)-8-methylimidazo[l,5-a]pyrazin-l-yl)-N-(4-propylpyridin-2- yl)benzamide; and
(S,E)-4-(3-(l-(4-methoxy-N-methylbut-2-enamido)ethyl)-8-methylimidazo[l,5-a]pyrazin-l- yl)-N-(4-propylpyridin-2-yl)benzamide.
64. The use according to any one of claims 58 to 63, wherein said subject has impaired
hematopoiesis.
65. The use according to claim 64, wherein said subject has previously received a
myelosuppressive or myeloablative treatment.
66. The use according to claim 65, wherein said myelosuppressive or myeloablative treatment comprises chemotherapy.
67. The use according to claim 64, wherein said subject has previously been accidentally exposed to radiation.
68. The use according to any one of claims 58 to 67, wherein said medicament is for use in
combination with an additional mobilizing agent selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-1 (IL-1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-321 (GM-CSF/IL-3 fusion protein), macrophage inflammatory protein, stem cell factor, plerixafor, thrombopoietin, growth related oncogene, and combinations thereof.
69. The use according to any one of claims 58 to 63, wherein said medicament is for use in
harvesting said hematopoietic stem and/or progenitor cells from said subject.
70. The use according to claim 69, wherein said medicament is for use in harvesting said
hematopoietic stem and/or progenitor cells from said subject by apheresis.
71. The use according to claim 69 or 70, wherein said medicament is for use in providing said hematopoietic stem and/or progenitor cells to a patient in need thereof.
72. The use according to claim 71 , wherein said medicament is for use in providing said
hematopoietic stem and/or progenitor cells to a bone marrow transplantation patient.
73. The use according to claim 72, wherein said subject and said patient are histocompatible.
74. The use according to claim 73, wherein said subject and said patient are syngeneic.
75. The use according to claim 73, wherein said subject and said patient are allogeneic.
76. The use according to any one of claims 71 to 75, wherein said hematopoietic stem and/or progenitor cells are cultured to enrich and/or differentiate said hematopoietic stem and/or progenitor cells prior to introducing the cells into the patient.
77. The use according to claim 76, wherein said hematopoietic stem and/or progenitor cells are enriched for or differentiated into cells selected from: myeloid cells, lymphoid cells, and NK cells.
78. The use according to any one of claims 58 to 77, wherein said Btk inhibitor is reversible.
79. The use according to any one of claims 58 to 77, wherein said Btk inhibitor is irreversible.
PCT/US2014/060953 2013-10-16 2014-10-16 Btk inhibitors for hematopoietic mobilization WO2015057992A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201361891641P 2013-10-16 2013-10-16
US61/891,641 2013-10-16

Publications (1)

Publication Number Publication Date
WO2015057992A1 true WO2015057992A1 (en) 2015-04-23

Family

ID=51844872

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2014/060953 WO2015057992A1 (en) 2013-10-16 2014-10-16 Btk inhibitors for hematopoietic mobilization

Country Status (1)

Country Link
WO (1) WO2015057992A1 (en)

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015110923A3 (en) * 2014-01-21 2015-11-26 Acerta Pharma B.V. Btk inhibitor for the treatment of chronic lymphocytic and small lymphocytic leukemia
WO2016116777A1 (en) * 2014-08-11 2016-07-28 Acerta Pharma B.V. Compositions and methods for treatment of chronic lymphocytic leukemia and small lymphocytic leukemia using a btk inhibitor
WO2016192074A1 (en) * 2015-06-04 2016-12-08 Merck Sharp & Dohme Corp. Btk inhibitors
WO2018001331A1 (en) * 2016-06-30 2018-01-04 杭州华东医药集团新药研究院有限公司 Imidazopyridinamine phenyl derivative and use thereof
CN108129483A (en) * 2018-01-26 2018-06-08 成都施贝康生物医药科技有限公司 A kind of BTK inhibitor and its application
EP2836214B1 (en) * 2012-04-11 2018-06-27 Acerta Pharma B.V. Bruton's tyrosine kinase inhibitors for hematopoietic mobilization
WO2018130213A1 (en) * 2017-01-16 2018-07-19 东莞市真兴贝特医药技术有限公司 Imidazopyrazine compound, preparation method therefor and use thereof
JP2018522877A (en) * 2015-07-02 2018-08-16 アセルタ ファーマ ビー.ブイ. (S) -4- (8-amino-3- (1- (but-2-inoyl) pyrrolidin-2-yl) imidazo [1,5-a] pyrazin-1-yl) -N- (pyridine-2) -Yl) benzamide solid form and formulation
CN108602827A (en) * 2015-11-06 2018-09-28 安塞塔制药公司 The Imidazopyrazines inhibitor of Bruton tyrosine kinase
US10272083B2 (en) 2014-01-21 2019-04-30 Acerta Pharma B.V. Methods of treating chronic lymphocytic leukemia and small lymphocytic leukemia using a BTK inhibitor
CN110494433A (en) * 2017-03-22 2019-11-22 廖细斌 Bruton's tyrosine kinase inhibitor
CN110545826A (en) * 2016-12-03 2019-12-06 朱诺治疗学股份有限公司 Methods and compositions for using therapeutic T cells in combination with kinase inhibitors
JP2020503266A (en) * 2016-11-24 2020-01-30 中国科学院上海薬物研究所Shanghai Institute Of Materia Medica, Chinese Academy Of Sciences Pyrimido [5,4-b] indolizine or pyrimido [5,4-b] pyrrolidine compound, production method and use thereof
WO2021142257A1 (en) * 2020-01-08 2021-07-15 Telios Pharma, Inc. Methods of treating splenomegaly
JP2022533818A (en) * 2019-05-10 2022-07-26 河南知微生物医薬有限公司 Substituted 1-amino-1H-imidazole-5-carboxamides as Bruton's Tyrosine Kinase Inhibitors
US11642343B2 (en) 2020-01-08 2023-05-09 Telios Pharma, Inc. Methods of treating splenomegaly

Citations (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5582823A (en) 1985-08-23 1996-12-10 Amgen Inc. Methods of treating bacterial inflammation and granulocytopoiesis by administering human pluripotent granulocyte colony-stimulating factor
US5989660A (en) 1995-09-18 1999-11-23 Toray Industries, Inc. Base fabric for air bags, a process for producing it and an air bag comprising it
US6908763B1 (en) 1997-08-22 2005-06-21 The Board Of Trustees Of The Leland Stanford Junior University Mammalian common lymphoid progenitor cell
US7071199B1 (en) 1999-09-17 2006-07-04 Abbott Gmbh & Cco. Kg Kinase inhibitors as therapeutic agents
US7235623B2 (en) 2003-11-26 2007-06-26 Eastman Chemical Company Polyester compositions for calendering
US7393848B2 (en) 2003-06-30 2008-07-01 Cgi Pharmaceuticals, Inc. Certain heterocyclic substituted imidazo[1,2-A]pyrazin-8-ylamines and methods of inhibition of Bruton's tyrosine kinase by such compounds
US7459554B2 (en) 2003-10-15 2008-12-02 Osi Pharmaceuticals, Inc. Imidazopyrazine tyrosine kinase inhibitors
US7514444B2 (en) 2006-09-22 2009-04-07 Pharmacyclics, Inc. Inhibitors of bruton's tyrosine kinase
US7718662B1 (en) 2009-10-12 2010-05-18 Pharmacyclics, Inc. Pyrazolo-pyrimidine inhibitors of bruton's tyrosine kinase
US7979057B2 (en) 2000-10-06 2011-07-12 S.F. Ip Properties 62 Llc Third-party provider method and system
US8252587B2 (en) 2004-10-25 2012-08-28 Celerant Therapeutics, Inc. Methods of expanding myeloid cell populations and uses thereof
WO2013010868A1 (en) * 2011-07-19 2013-01-24 Msd Oss B.V. 4 - imidazopyridazin- 1 -yl-benzamides and 4 - imidazotriazin- 1 - yl - benzamides as btk- inhibitors
US8383095B2 (en) 2006-02-14 2013-02-26 Cellerant Therapeutics, Inc. Methods and compositions for enhancing engraftment of hematopoietic stem cells
WO2013155347A1 (en) * 2012-04-11 2013-10-17 Izumi Raquel Bruton's tyrosine kinase inhibitors for hematopoietic mobilization

Patent Citations (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5582823A (en) 1985-08-23 1996-12-10 Amgen Inc. Methods of treating bacterial inflammation and granulocytopoiesis by administering human pluripotent granulocyte colony-stimulating factor
US5989660A (en) 1995-09-18 1999-11-23 Toray Industries, Inc. Base fabric for air bags, a process for producing it and an air bag comprising it
US6908763B1 (en) 1997-08-22 2005-06-21 The Board Of Trustees Of The Leland Stanford Junior University Mammalian common lymphoid progenitor cell
US7071199B1 (en) 1999-09-17 2006-07-04 Abbott Gmbh & Cco. Kg Kinase inhibitors as therapeutic agents
US7979057B2 (en) 2000-10-06 2011-07-12 S.F. Ip Properties 62 Llc Third-party provider method and system
US7393848B2 (en) 2003-06-30 2008-07-01 Cgi Pharmaceuticals, Inc. Certain heterocyclic substituted imidazo[1,2-A]pyrazin-8-ylamines and methods of inhibition of Bruton's tyrosine kinase by such compounds
US7459554B2 (en) 2003-10-15 2008-12-02 Osi Pharmaceuticals, Inc. Imidazopyrazine tyrosine kinase inhibitors
US7235623B2 (en) 2003-11-26 2007-06-26 Eastman Chemical Company Polyester compositions for calendering
US8252587B2 (en) 2004-10-25 2012-08-28 Celerant Therapeutics, Inc. Methods of expanding myeloid cell populations and uses thereof
US8383095B2 (en) 2006-02-14 2013-02-26 Cellerant Therapeutics, Inc. Methods and compositions for enhancing engraftment of hematopoietic stem cells
US8088781B2 (en) 2006-09-22 2012-01-03 Pharmacyclics, Inc. Inhibitors of brutons tyrosine kinase
US8008309B2 (en) 2006-09-22 2011-08-30 Pharmacyclics, Inc. Inhibitors of bruton's tyrosine kinase
US7960396B2 (en) 2006-09-22 2011-06-14 Pharmacyclics, Inc. Inhibitors of Bruton's tyrosine kinase
US7514444B2 (en) 2006-09-22 2009-04-07 Pharmacyclics, Inc. Inhibitors of bruton's tyrosine kinase
US7718662B1 (en) 2009-10-12 2010-05-18 Pharmacyclics, Inc. Pyrazolo-pyrimidine inhibitors of bruton's tyrosine kinase
WO2013010868A1 (en) * 2011-07-19 2013-01-24 Msd Oss B.V. 4 - imidazopyridazin- 1 -yl-benzamides and 4 - imidazotriazin- 1 - yl - benzamides as btk- inhibitors
WO2013155347A1 (en) * 2012-04-11 2013-10-17 Izumi Raquel Bruton's tyrosine kinase inhibitors for hematopoietic mobilization

Non-Patent Citations (62)

* Cited by examiner, † Cited by third party
Title
"Goodman & Gilman's The Pharmacological Basis of Therapeutics", 2001, MCGRAW-HILL
ABI-YOUNES ET AL., CIRC. RES., vol. 86, 2000, pages 131 - 38
AIUTI ET AL., J. EXP. MED., vol. 185, 1997, pages 111 - 120
BLEUL ET AL., J. EXP. MED., vol. 184, 1996, pages 1101 - 1109
BROXMEYER ET AL., BLOOD CELLS, MOLECULES AND DISEASES, vol. 24, 1998, pages 14 - 30
BROXMEYER ET AL., EXP. HEMATOL., vol. 23, 1995, pages 335 - 340
CAREY; SUNDBERG: "ADVANCED ORGANIC CHEMISTRY", vol. A, B, PLENUM
CHANG ET AL., ARTHR. RES. & THER., vol. 13, 2011, pages R115
DALE ET AL., AM. J. OF HEMATOL., vol. 57, 1998, pages 7 - 15
DE GORTER ET AL., IMMUNITY, vol. 26, 2007, pages 93 - 104
DEMIRER ET AL., STEM CELLS, vol. 14, 1996, pages 106 - 116
DEXTER ET AL., J EXP MED, vol. 145, pages 1612 - 1616
FEDYK ET AL., J. LEUKOC. BIOL., vol. 66, 1999, pages 667 - 673
GEAR ET AL., BLOOD, vol. 97, 2001, pages 937 - 45
GLASPY ET AL., BLOOD, vol. 90, 1997, pages 2939 - 2951
GLASPY ET AL., CANCER CHEMOTHER. PHARMACOL., vol. 38, 1996, pages S53 - S57
GREEN; WUTS: "PROTECTIVE GROUPS IN ORGANIC SYNTHESIS", 1999, WILEY
HATA ET AL., J EXP MED., vol. 187, 1998, pages 1235 - 1247
HILL ET AL., EXP HEMATOL, vol. 24, 1996, pages 936 - 943
HODOHARA ET AL., BLOOD, vol. 95, 2000, pages 769 - 75
HONINBERG ET AL., PROC. NATL. ACAD. SCI. USA, vol. 107, 2010, pages 13075 - 80
HORWOOD ET AL., J. EXP. MED., vol. 197, 2003, pages 1603 - 1611
IWAKI ET AL., J. BIOL. CHEM., vol. 280, no. 48, 2005, pages 40261 - 40270
JEFFRIES ET AL., J. BIOL. CHEM., vol. 278, 2003, pages 26258 - 26264
JO ET AL., J. CLIN. INVEST., vol. 105, 2000, pages 101 - 111
JONGSTRA-BILEN ET AL., J. IMMUNOL., vol. 181, 2008, pages 288 - 298
KHAN ET AL., IMMUNITY, vol. 3, 1995, pages 283 - 99
KING; 2001 ET AL., BLOOD, vol. 97, pages 1534 - 1542
KUCIA ET AL., STEM CELLS, vol. 23, 2005, pages 879 - 94
LAPID ET AL.: "Egress and Mobilization of Hematopoietic Stem and Progenitor Cells", STEMBOOK, 2009
LORHRMAN ET AL., BR. J. HAEMATOL., vol. 40, 1978, pages 369
M. BODANZSKY: "Principles of Peptide Synthesis", 1984, SPRINGER-VERLAG
MA ET AL., IMMUNITY, vol. 10, 1999, pages 463 - 71
MA ET AL., PROC. NATL. ACAD. SCI USA, vol. 95, 1998, pages 9448 - 9453
MAEKAWA ET AL., INTERNAL MED., vol. 39, 2000, pages 90 - 100
MAJK ET AL., BLOOD, vol. 96, 2000, pages 4142 - 51
MARCH: "ADVANCED ORGANIC CHEMISTRY", 1992, WILEY
MOHAMMED ET AL., IMMUNOLOGICAL REVIEWS, vol. 228, 2009, pages 58 - 73
MOHAMMED, IMMUNO. REV., vol. 228, 2009, pages 58 - 73
NAGASAWA ET AL., INT. J. HEMATOL., vol. 72, 2000, pages 408 - 411
PELED ET AL., SCIENCE, vol. 283, 1999, pages 845 - 48
PETTENGEL ET AL., BLOOD, vol. 82, 1992, pages 2239 - 2248
PRUIJT ET AL., CUR. OP. IN HEMATOL., vol. 6, 1999, pages 152 - 158
QUEK ET AL., CURR. BIO., vol. 8, no. 20, 1998, pages 1137 - 1140
RAWLINGS ET AL., SCIENCE, vol. 261, 1993, pages 358 - 61
RAWLINGS; WITTE, IMMUN. REV., vol. 138, 1994, pages 105 - 119
RIVIERE ET AL., BLOOD, vol. 95, 1999, pages 1511 - 23
ROSENFELD ET AL., BONE MARROW TRANSPLANTATION, vol. 17, 1997, pages 179 - 183
SCHMIDT ET AL., INT ARCH ALLERGY IMMUNOL., vol. 134, 2004, pages 65 - 78
SHARMA ET AL., STEM CELLS DEV., vol. 20, no. 6, 2011, pages 933 - 46
SMALL; WEISSMAN, SCAND J IMMUNOL, vol. 44, 1996, pages 115 - 121
SPANGRUDE ET AL., J IMMUNOL, vol. 141, 1988, pages 3697 - 3707
STEWART; YOUNG: "Solid Phase Peptide Synthesis", 1984, PIERCE CHEMICAL CO.
TACHIBANA ET AL., NATURE, vol. 393, 1998, pages 591
UCKUN, INT. REV. IMMUNOL., vol. 27, 2008, pages 43 - 69
VADHAN-RAJ ET AL., ANN. INTERN. MED, vol. 126, 1997, pages 673 - 81
VAN DE RIJN, PROC NATL ACAD SCI, vol. 86, 1989, pages 4634 - 4638
VASSILEV ET AL., J. BIOL. CHEM., vol. 274, no. 3, 1999, pages 1646 - 1656
VIARDOT ET AL., ANN. HEMATOL., vol. 77, 1998, pages 194 - 197
WANG ET AL., INT. IMMUNOPHARMACOL., vol. 7, 2007, pages 541 - 546
WHITLOCK ET AL., ANNU REV IMMUNOL, vol. 3, 1985, pages 213 - 235
ZOU ET AL., NATURE, vol. 393, 1998, pages 595 - 599

Cited By (48)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2836214B1 (en) * 2012-04-11 2018-06-27 Acerta Pharma B.V. Bruton's tyrosine kinase inhibitors for hematopoietic mobilization
WO2015110923A3 (en) * 2014-01-21 2015-11-26 Acerta Pharma B.V. Btk inhibitor for the treatment of chronic lymphocytic and small lymphocytic leukemia
US10272083B2 (en) 2014-01-21 2019-04-30 Acerta Pharma B.V. Methods of treating chronic lymphocytic leukemia and small lymphocytic leukemia using a BTK inhibitor
US11090302B2 (en) 2014-01-21 2021-08-17 Acerta Pharma B.V. Methods of treating chronic lymphocytic leukemia and small lymphocytic leukemia using a BTK inhibitor
US11771696B2 (en) 2014-01-21 2023-10-03 Acerta Pharma B.V. Methods of treating chronic lymphocytic leukemia and small lymphocytic leukemia using a BTK inhibitor
WO2016116777A1 (en) * 2014-08-11 2016-07-28 Acerta Pharma B.V. Compositions and methods for treatment of chronic lymphocytic leukemia and small lymphocytic leukemia using a btk inhibitor
WO2016192074A1 (en) * 2015-06-04 2016-12-08 Merck Sharp & Dohme Corp. Btk inhibitors
US11820777B2 (en) 2015-07-02 2023-11-21 Acerta Pharma B.V. Crystal forms of (s)-4-(8-amino-3-(1-(but-2-ynoyl)pyrrolidin-2-yl)imidazo[1,5-a]pyrazin-1-yl)-n-(pyridin-2-yl)benzamide
JP7091494B2 (en) 2015-07-02 2022-06-27 アセルタ ファーマ ビー.ブイ. (S) -4- (8-Amino-3- (1- (Gnat-2-inoyl) pyrrolidine-2-yl) imidazole [1,5-a] pyrazine-1-yl) -N- (pyridine-2) -Il) Solid form and formulation of benzamide
JP2018522877A (en) * 2015-07-02 2018-08-16 アセルタ ファーマ ビー.ブイ. (S) -4- (8-amino-3- (1- (but-2-inoyl) pyrrolidin-2-yl) imidazo [1,5-a] pyrazin-1-yl) -N- (pyridine-2) -Yl) benzamide solid form and formulation
US10640509B2 (en) 2015-07-02 2020-05-05 Acerta Pharma B.V. Pharmaceutical composition comprising a crystal form of (S)-4-(8-amino-3-(1-(but-2-ynoyl)pyrrolidin-2-yl)imidazo[1,5-a]pyrazin-1-yl)-N-(pyridin-2-yl)benzamide for treating diffuse large B-cell lymphoma
US11059829B2 (en) 2015-07-02 2021-07-13 Acerta Pharma B.V. Crystal forms of (S)-4-(8-amino-3-(1-(but-2-ynoyl)pyrrolidin-2-yl)imidazo[1,5-a]pyrazin-1-yl)-N-(pyridin-2-yl)benzamide maleate
JP2021073235A (en) * 2015-07-02 2021-05-13 アセルタ ファーマ ビー.ブイ. Solid forms and formulations of (s)-4-(8-amino-3-(1-(but-2-ynoyl)pyrrolidin-2-yl)imidazo[1,5-a]pyrazin-1-yl)-n-(pyridin-2-yl)benzamide
US11110088B2 (en) 2015-11-06 2021-09-07 Acerta Pharma B.V. Imidazopyrazine inhibitors of Bruton's tyrosine kinase
JP2018536705A (en) * 2015-11-06 2018-12-13 アセルタ ファーマ ビー.ブイ. An imidazopyrazine inhibitor of breton-type tyrosine kinase
US20180318297A1 (en) * 2015-11-06 2018-11-08 Acerta Pharma B.V. Imidazopyrazine Inhibitors of Bruton's Tyrosine Kinase
CN108602827A (en) * 2015-11-06 2018-09-28 安塞塔制药公司 The Imidazopyrazines inhibitor of Bruton tyrosine kinase
US10736893B2 (en) * 2015-11-06 2020-08-11 Acerta Pharma B.V. Imidazopyrazine inhibitors of Bruton's tyrosine kinase
KR20190033538A (en) * 2016-06-30 2019-03-29 항저우 산잉타이 파마슈티컬 테크놀로지 컴퍼니, 리미티드 Imidazopyridine amine phenyl derivatives and uses thereof
CN107556317A (en) * 2016-06-30 2018-01-09 杭州华东医药集团新药研究院有限公司 A kind of Imidazopyridine amine phenyl derivatives and application thereof
EP3480199A4 (en) * 2016-06-30 2020-02-26 Hangzhou Sanyintai Pharmaceutical Technology Co., Ltd. Imidazopyridinamine phenyl derivative and use thereof
WO2018001331A1 (en) * 2016-06-30 2018-01-04 杭州华东医药集团新药研究院有限公司 Imidazopyridinamine phenyl derivative and use thereof
AU2017287553B2 (en) * 2016-06-30 2020-02-06 Hangzhou BANGSHUN Pharmaceutical Co., Ltd Imidazopyrazinamine phenyl derivative and use thereof
US10640512B2 (en) 2016-06-30 2020-05-05 Hangzhou Sanyintai Pharmaceutical Technology Co., Ltd. Imidazopyrazinamine phenyl derivative and use thereof
CN107556317B (en) * 2016-06-30 2020-06-09 杭州三因泰医药科技有限公司 Imidazole pyrazinamine phenyl derivative and application thereof
KR102215172B1 (en) * 2016-06-30 2021-02-16 항저우 방순 파마슈티컬 컴퍼니, 리미티드 Imidazopyrazineamine phenyl derivatives and uses thereof
JP2020503266A (en) * 2016-11-24 2020-01-30 中国科学院上海薬物研究所Shanghai Institute Of Materia Medica, Chinese Academy Of Sciences Pyrimido [5,4-b] indolizine or pyrimido [5,4-b] pyrrolidine compound, production method and use thereof
US11590167B2 (en) 2016-12-03 2023-02-28 Juno Therapeutic, Inc. Methods and compositions for use of therapeutic T cells in combination with kinase inhibitors
CN110545826A (en) * 2016-12-03 2019-12-06 朱诺治疗学股份有限公司 Methods and compositions for using therapeutic T cells in combination with kinase inhibitors
US10919899B2 (en) 2017-01-16 2021-02-16 Dongguan Zhenxing-Beite Medicine Technology Co., Ltd. Imidazopyrazine compounds, preparation methods and uses thereof
WO2018130213A1 (en) * 2017-01-16 2018-07-19 东莞市真兴贝特医药技术有限公司 Imidazopyrazine compound, preparation method therefor and use thereof
JP2020506957A (en) * 2017-01-16 2020-03-05 ▲東▼莞市真▲興▼▲貝▼特医▲薬▼技▲術▼有限公司Dongguan Zhenxing−Beite Medicine Technology Co., Ltd. Imidazopyrazine compounds, their production methods and applications
US11554118B2 (en) 2017-03-22 2023-01-17 Xibin Liao Bruton's tyrosine kinase inhibitors
JP7219902B2 (en) 2017-03-22 2023-02-09 スジョウ・バイジブゴン・ファーマスーティカル・テクノロジー・カンパニー・リミテッド Bruton's Tyrosine Kinase Inhibitor
JP2020511547A (en) * 2017-03-22 2020-04-16 スジョウ・バイジブゴン・ファーマスーティカル・テクノロジー・カンパニー・リミテッドSuzhou Baijibugong Pharmaceutical Technology Co. Ltd. Bruton's tyrosine kinase inhibitor
EP3601264A4 (en) * 2017-03-22 2021-03-24 Xibin Liao Bruton's tyrosine kinase inhibitors
IL269152B1 (en) * 2017-03-22 2023-07-01 Liao Xibin Bruton's tyrosine kinase inhibitors
CN110494433B (en) * 2017-03-22 2023-03-17 廖细斌 Bruton's tyrosine kinase inhibitors
AU2018237123B2 (en) * 2017-03-22 2022-08-04 Xibin Liao Bruton's tyrosine kinase inhibitors
US20210121458A1 (en) * 2017-03-22 2021-04-29 Xibin Liao Bruton's tyrosine kinase inhibitors
CN110494433A (en) * 2017-03-22 2019-11-22 廖细斌 Bruton's tyrosine kinase inhibitor
CN108129483A (en) * 2018-01-26 2018-06-08 成都施贝康生物医药科技有限公司 A kind of BTK inhibitor and its application
CN108129483B (en) * 2018-01-26 2019-06-04 成都施贝康生物医药科技有限公司 A kind of BTK inhibitor and its application
JP2022533818A (en) * 2019-05-10 2022-07-26 河南知微生物医薬有限公司 Substituted 1-amino-1H-imidazole-5-carboxamides as Bruton's Tyrosine Kinase Inhibitors
EP3980069A4 (en) * 2020-01-08 2022-08-17 Telios Pharma, Inc. Methods of treating splenomegaly
US11642343B2 (en) 2020-01-08 2023-05-09 Telios Pharma, Inc. Methods of treating splenomegaly
EP4000624A1 (en) * 2020-01-08 2022-05-25 Telios Pharma, Inc. Btk inhibitors for treating splenomegaly
WO2021142257A1 (en) * 2020-01-08 2021-07-15 Telios Pharma, Inc. Methods of treating splenomegaly

Similar Documents

Publication Publication Date Title
WO2015057992A1 (en) Btk inhibitors for hematopoietic mobilization
EP2836214B1 (en) Bruton's tyrosine kinase inhibitors for hematopoietic mobilization
US10260044B1 (en) Methods of expanding myeloid cell populations and uses thereof
Morabito et al. Expression of myelomonocytic antigens on chronic lymphocytic leukemia B cells correlates with their ability to produce interleukin 1
US8241903B2 (en) Catecholamine receptor modulation
JP6110309B2 (en) Methods for promoting hematopoietic stem cell engraftment
CA3091865A1 (en) Expansion of nk and dc cells in vivo mediating immune response
JP2019513836A (en) Method of Depleting Hematopoietic Stem / Hematopoietic Progenitor Cells (HSC / HP) in a Patient Using a Bispecific Antibody
CA2307624A1 (en) Hematopoietic stem cells
EP2946008A1 (en) Methods for mobilizing hematopoietic stem cells
US20020098521A1 (en) Method and marker for the isolation of human multipotent hematopoietic stem cells
Ho et al. Hematopoietic stem cell transplantation
Perry et al. Direct effects of cyclosporin A on proliferation of hematopoietic stem and progenitor cells
WO2008070861A2 (en) Methods and compositions for treating systemic lupus erythematosus
Carlson et al. Stem Cell Biology in Bone Marrow Transplantation
Hennessy A study of the haematological and immunological effects of GroβT in normal healthy volunteers
Pfeilstöcker et al. Hematopoietic recovery after IEV chemotherapy for malignant lymphoma followed by different cytokines can be monitored by analysis of Gα 16 and CD34
Morabito et al. Expression of myelomonocytic antigens on chronic lymphocytic leukemia
Hofmann G-CSF and Thrombopoietin: hematopoietic growth factors to the rescue
JP2014062067A (en) Proliferation acceleration and/or mobilization agent for hematopoietic stem cell and/or hemopoietic precursor cell
Civin STEM CELLS EXPRESSING FAS-LIGAND RESIST IMMUNE REJECTION

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 14792684

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 14792684

Country of ref document: EP

Kind code of ref document: A1