1111111111111111 IIIIII IIIII 1111111111 11111 11111 1111111111 1111111111 111111111111111 11111111
`US 20150086507Al
`
`c19) United States
`c12) Patent Application Publication
`Izumi et al.
`
`c10) Pub. No.: US 2015/0086507 Al
`Mar. 26, 2015
`(43) Pub. Date:
`
`(54) BRUTON'S TYROSINE KINASE INHIBITORS
`FOR HEMATOPOIETIC MOBILZATION
`
`(71) Applicants:Raquel IZUMI, San Carlos, CA (US);
`Francisco SALVA, San Francisco, CA
`(US); Ahmed HAMDY, Soquel, CA
`(US)
`
`(72)
`
`Inventors: Raquel Izumi, San Carlos, CA (US);
`Francisco Salva, San Francisco, CA
`(US); Ahmed Hamdy, Soquel, CA (US)
`
`(21) Appl. No.:
`
`14/394,061
`
`(22) PCT Filed:
`
`Apr. 11, 2013
`
`(86) PCT No.:
`
`PCT/US13/36242
`
`§ 371 (c)(l),
`Oct. 10, 2014
`(2) Date:
`Related U.S. Application Data
`(60) Provisional application No. 61/622,843, filed on Apr.
`11, 2012.
`
`Publication Classification
`
`(51)
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`
`Int. Cl.
`C07D487/04
`A61K 45106
`C12N5/0789
`A61K 311519
`(52) U.S. Cl.
`CPC ............ C07D 487104 (2013.01); A61K 311519
`(2013.01); A61K 45106 (2013.01); C12N
`510647 (2013.01)
`USPC ...... 424/85.2; 514/265.1; 424/85.1; 435/325;
`424/93.7
`
`(57)
`
`ABSTRACT
`
`Methods to improve hematopoiesis and increase white blood
`cell counts in subjects and patients using pyrimidine-based
`inhibitors ofBruton's tyrosine kinase (Btk) are disclosed.
`
`

`

`Patent Application Publication
`
`Mar.26,2015
`
`US 2015/0086507 Al
`
`Figure 1
`
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`
`

`

`US 2015/0086507 Al
`
`Mar. 26, 2015
`
`1
`
`BRUTON'S TYROSINE KINASE INHIBITORS
`FOR HEMATOPOIETIC MOBILZATION
`
`FIELD OF INVENTION
`
`[0001] The invention is in the field of therapeutics and
`medicinal chemistry. 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
`Tee 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 pro(cid:173)
`duction in macrophages, IgE receptor (FcERI) 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 oflg heavy chain rearrangement. Rawlings and
`Witte (1994) Immun. Rev. 138:105-119. In the mouse, point
`mutation or deletion of Btk causes X-linked immunodefi(cid:173)
`ciency (xid), with about 50% fewer conventional B2 B cells,
`absent Bl B cells, and reduced serum lg 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 FcER
`signaling pathways in monocytes/macrophages, neutrophils,
`and mast cells. See, e.g., Jongstra-Bilen et al. (2008) J. Immu(cid:173)
`nol. 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 ofBtk in inhibiting Fas/ APO-1 apo(cid:173)
`ptotic 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 ofBtk in the signaling pathways
`of other immune cells, Btk inhibitors have also been evalu(cid:173)
`ated 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 :Rl 15. 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 defi(cid:173)
`ciency 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. Pat. No.
`7,393,848.
`[0004] Btk inhibitors have also been shown useful in pre(cid:173)
`venting or reducing the risk ofthromboembolism. See, e.g.,
`Uckun (2008) Int. Rev. Immunol. 27:43-69.
`
`SUMMARY OF INVENTION
`
`[0005]
`In contrast to the prior art uses ofBruton'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. Accord(cid:173)
`ingly, provided herein are methods and compositions for
`improving hematopoiesis and increasing the white blood cell
`count in a subject in need thereof, including patients under(cid:173)
`going chemotherapy, radiation therapy and/or bone marrow
`transplantation. Also provided herein are methods of deter(cid:173)
`mining 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 sub(cid:173)
`ject a pharmaceutical composition comprising a mobilizing
`Bruton' s Tyrosine Kinase (Btk) inhibitor in an amount effec(cid:173)
`tive to mobilize said cells into the peripheral blood of said
`subject. The inventive methods and uses can be advanta(cid:173)
`geously employed in conjunction with bone marrow trans(cid:173)
`plantation 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 neu(cid:173)
`tropenic 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 ofleukopenia and neutropenia
`resulting from chemotherapy, radiation therapy or accidental
`radiation exposure, to enhance wound healing and bum treat(cid:173)
`ment, 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 administer(cid:173)
`ing 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 administra(cid:173)
`tion of a mobilizing Btk inhibitor alone. In another embodi(cid:173)
`ment, the step comprises administration of a mobilizing Btk
`inhibitor in combination with other compounds, e.g., cytok(cid:173)
`ines, 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-macroph-
`
`

`

`US 2015/0086507 Al
`
`Mar. 26, 2015
`
`2
`
`[0015] Y is a 4-, 5-, 6-, or 7-membered cycloalkyl ring, or
`[0016] Yis a4-, 5-, 6-, or7-memberedmonocyclicnitrogen
`containing heterocycloalkyl ring; or
`[0017] Y is an optionally substituted group selected from
`among alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,
`and heteroaryl; or
`[0018] Y is selected form the group consisting of azetidi(cid:173)
`nyl, pyrrolidinyl, piperidinyl, and azepanyl;
`[0019] Z is C(=O), OC(=O), NHC(=O), NRC(=O),
`C(=S), S(=O)x, OS(=O)x, NHS(=O)x, where xis 1 or 2;
`[0020] R7 and Rg are independently selected from among
`H, unsubstituted cl -C4alkyl, substituted cl -C4 alkyl, cl -C6
`alkoxyalkyl, C 1 -Cg alkylamino alkyl, C 1 -C4 alkyl(phenyl),
`unsubstituted
`C 1 -C4heteroalkyl,
`substituted
`C 1-C4heteroalkyl, unsubstituted C3 -C6cycloalkyl, substi(cid:173)
`tuted C3 -C6cycloalkyl, unsubstituted C2 -C6heterocycloalkyl,
`and substituted C2-C6heterocycloalkyl; or
`[0021] R7 and Rg taken together form a bond;
`[0022] R6 is H, substituted or unsubstituted C 1 -C4alkyl,
`substituted
`or
`unsubstituted
`C1 -C4heteroalkyl,
`C 1 -C6alkoxyalkyl, C 1 -Cgalkylaminoalkyl, substituted or
`unsubstituted C3 -C6cycloalkyl, substituted or unsubstituted
`aryl, substituted or unsubstituted C2 -Cgheterocycloalkyl,
`substituted or unsubstituted heteroaryl, C 1-C4alkyl(aryl),
`C 1-C4alkyl(phenyl), C 1-C4alkyl(heteroaryl), C 1 -C4alkyl(C3 -
`Cgcycloalkyl), or C 1 -C4alkyl(C2 -Cgheterocycloalkyl), or
`C 1 -Cg alkylamino alkyl;
`[0023] R is H, or C 1 -C6alkyl; and
`pharmaceutically acceptable solvates or pharmaceutically
`acceptable salts thereof.
`[0024]
`In another embodiment, the mobilizing Btk inhibi(cid:173)
`tor for use in the subject invention is selected from a com(cid:173)
`pound of structural Formula II:
`
`age colony stimulating factor (GM-CSF), Interleukin-I (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 mobi(cid:173)
`lizing Btk inhibitor (with or without other mobilizing factors)
`in an amount effective to increase the number ofhematopoi(cid:173)
`etic 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 transplan(cid:173)
`tation. Accordingly, in another aspect the invention provides
`methods of treating a patient in need of improved hemato(cid:173)
`poiesis 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 hematopoi(cid:173)
`etic 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 mobi(cid:173)
`lizing Btk inhibitor is administered to a mammal, and more
`preferably to a human.
`[0012] Preferred mobilizing Btk inhibitors suitable for use
`in the subject invention comprise a pyrimidine ring, i.e., a 1,3
`diazine. In one embodiment, the mobilizing Btk inhibitor is
`selected from a compound of structural Formula I:
`
`_,,Ar
`La
`
`wherein:
`[0025] Ra, Rb, Re, Rd, and Re, are each independently
`selected from H, F, Cl, Br, I, -CN, -SR2 , ---OR3 , C02R3 ; or
`[0026] Ra, or Rb together with one of Re, Rd and Re, and
`the carbon atoms to which they are attached form an epoxide;
`[0027] wherein Ra, Rb, Re, Rd, and Re, cannot all be H;
`[0028] R2 is selected from H, methyl, ethyl, n-propyl, iso(cid:173)
`propyl, n-butyl, iso-butyl, tert-butyl, a cysteinyl, a glutathio(cid:173)
`nyl, C 1 -C4alkyl, a cysteinyl, or a glutathionyl;
`[0029] R3 is selected from H, C 1 -C4alkyl, phenyl, or ben(cid:173)
`zyl; and
`pharmaceutically acceptable solvates or pharmaceutically
`acceptable salts thereof.
`
`wherein:
`[0013] La is CH2 , 0, NH or S;
`[0014] Ar is a substituted or unsubstituted aryl, unsubsti(cid:173)
`tuted phenyl, or a substituted or unsubstituted heteroaryl;
`
`

`

`US 2015/0086507 Al
`
`Mar. 26, 2015
`
`3
`
`[0030]
`In another embodiment, the mobilizing Btk inhibi(cid:173)
`tor for use in the subject invention is selected from a com(cid:173)
`pound of structural Formula III:
`
`,..,.Ar
`La
`
`wherein;
`[0031] La is O or S;
`[0032] Ar is an unsubstituted phenyl;
`[0033] Y is a 4-, 5-, 6-, or 7-membered cycloalkyl ring, or
`[0034] Y is a 4-, 5-, 6-, or7-memberedmonocyclic nitrogen
`containing heterocyclic ring;
`[0035] Z is C(=O), OC(=O), NHC(=O), S(=O)x, or
`NHS(=O)x, where xis 2;
`[0036] R 8 is H; R7 is H, unsubstituted C 1 -C4 alkyl, C 1 -C6
`alkoxyalkyl, C1 -C8 alkylaminoalkyl, or C 1 -C4 alkyl(phenyl);
`or
`[0037] R7 and R 8 taken together form a bond;
`[0038] R 6 is H, unsubstituted C 1 -C4 alkyl, C 1 -C6 alkoxy(cid:173)
`alkyl, C 1 -C8 alkylaminoalkyl, or C 1 -C4 alkyl(phenyl); and
`pharmaceutically acceptable solvates or pharmaceutically
`acceptable salts thereof.
`[0039]
`In certain embodiments, the compounds of Formu(cid:173)
`las I-III 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 enantio(cid:173)
`meric diastereomer, a mesa 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.
`[0040] The compounds of Formulas I, II and III 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(cid:173)
`oxalate, malonate, hemi-malonate, fumarate, hemi-fumarate,
`maleate, hemi-maleate, citrate, hemi-citrate, tartrate, hemi(cid:173)
`tartrate, aspartate, glutamate, etc.
`
`BRIEF DESCRIPTION OF THE FIGURES
`
`[0041] FIG. 1 shows the results of an in vitro transwell
`assay using human CD34+ cells and migration towards an
`SDF-1 gradient with varying concentrations of a pyrimidine(cid:173)
`based Btk inhibitor.
`
`DETAILED DESCRIPTION
`
`[0042] 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 granu(cid:173)
`locyte 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 mobili(cid:173)
`zation). 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.
`[0043] Several other factors have been reported to increase
`white blood cells and progenitor cells in both human and
`animal subjects. These agents include granulocyte-macroph(cid:173)
`age colony stimulating factor (GM-CSF), Interleukin-I (IL-
`1), Interleukin-3 (IL-3), Interleukin-8 (IL-8), PIXY-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; Broxmeye 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. (l997)Ann. Intern. Med. 126:673-81; King
`et al. (2001) Blood 97:1534-1542; Glaspy et al. (1997) Blood
`90:2939-2951.
`[0044] While endogenous growth factors are pharmaco(cid:173)
`logically effective, the well-known disadvantages of employ(cid:173)
`ing 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.
`[0045] As used herein, the term "progenitor cell" refers to a
`cell that, in response to certain stimuli, can form differenti(cid:173)
`ated hematopoietic or myeloid cells. The presence of progeni(cid:173)
`tor 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-mac(cid:173)
`rophage ); CFU-GEMM (colony-forming units, multipoten(cid:173)
`tial); 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.
`[0046] As used herein, "stem cells" are less differentiated
`forms of progenitor cells. Typically, such cells are often posi(cid:173)
`tive for CD34. Some stem cells do not contain this marker,
`however. These CD34+ cells can be assayed using fluores(cid:173)
`cence activated cell sorting (FACS) and thus their presence
`can be assessed in a sample using this technique.
`[0047]
`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 consid(cid:173)
`ered an index of stem cell presence.
`
`

`

`US 2015/0086507 Al
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`Mar. 26, 2015
`
`4
`
`[0048] 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 pro(cid:173)
`liferate and differentiate prior to entering the circulation. The
`chemokine receptor CXCR4 and its natural ligand stromal
`cell derived factor-I (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. 72:408 411).
`This is demonstrated by reports that CXCR4 or SDF-1 knock(cid:173)
`out mice exhibithematopoietic defects. Ma eta!. (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.
`It is also known that CD34+ progenitor cells express
`[0049]
`CXCR4 and require SDF-1 produced by bone marrow stro(cid:173)
`mal 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. 185:111-120; 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 animportantchemoattrac(cid:173)
`tant, signaling via the CXCR 4 receptor, for several other more
`committed hematopoietic progenitors and mature blood cells
`including T-lymphocytes and monocytes, Bleul et al. (1996)
`J. Exp. Med. 184:1101-1109), pro- and pre-B lymphocytes,
`Fedyket al. (1999)J. Leukoc. Biol. (1999) 66:667-673; Ma et
`al.
`(1999) Immunity 10:463-71, and megakaryocytes.
`Hodohara et al. (2000)Blood95:769-75; Riviere et al. (1999)
`Blood 95:1511-23; 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.
`[0050] The SDF-1/CXCR4 signaling axis is known to
`direct homing and engraftment ofhematopoietic 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 ofHematopoietic 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 subse(cid:173)
`quent dissociation of the heterotrimeric G protein into G~y
`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. (2011)
`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 signal(cid:173)
`ing cascade. De Gorter et al. (2007)Immunity 26: 93-104. Btk
`expression has been described in HSC, multipotent progeni(cid:173)
`tor cells and cells of the myeloid lineage, Mohammed et al.
`(2009) Immunological Reviews 228: 58-73, including eryth(cid:173)
`roid 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.
`[0051] Notably, blocking of the SDF-1/CXCR4 signaling
`axis by AMD3100 (plerixafor or Mozobil®), a small mol(cid:173)
`ecule inhibitor which binds to CXCR4 preventing binding of
`SDF-1 and subsequent downstream signaling, causes rapid
`and reversible mobilization of CD34+ HSC into the periph(cid:173)
`eral 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 clini(cid:173)
`cally significant decreases in lymphadenopathy as the malig(cid:173)
`nant 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
`
`"Alkyl" means a straight or branched chain, satu(cid:173)
`[0052]
`rated 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, an straight chain alkyl may have
`from one to six carbon atoms and a branched alky I 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.
`"Ary!" means a monovalent monocyclic or bicyclic
`[0053]
`aromatic hydrocarbon radical, e.g., having from of 6 to 20 or
`6 to 10 ring atoms e.g., phenyl or naphthyl.
`"Alkylaryl" means a (alkylene)-Rradical where Ris
`[0054]
`aryl as defined above.
`"Cycloalkyl" means a cyclic saturated or partially
`[0055]
`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, cyclo(cid:173)
`pentyl, cyclohexyl, cyclohexenyl, indanyl and the like.
`"Alkylcycloalkyl" means a (alkylene)-R radical
`[0056]
`where R is cycloalkyl as defined above; e.g., cyclopropylm(cid:173)
`ethyl, cyclobutylmethyl, cyclopentylethyl, or cyclohexylm(cid:173)
`ethyl, and the like.
`"Heterocyclyl" or "heterocycloalkyl" means a satu(cid:173)
`[0057]
`rated or unsaturated monovalent monocyclic group, in which
`one or two ring atoms are heteroatom selected from N, 0, 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 "bicy(cid:173)
`clic 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, mor(cid:173)
`pholino, 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.
`
`

`

`US 2015/0086507 Al
`
`Mar. 26, 2015
`
`5
`
`a-(alkylene)-R
`"Alkylheterocycloalkyl" means
`[0058]
`radical where R is heterocyclyl ring as defined above e.g.,
`tetraydrofuranylmethyl, piperazinylmethyl, morpholinyl(cid:173)
`ethyl, and the like.
`"Heteroaryl" means a monovalent monocyclic or
`[0059]
`bicyclic aromatic radical, where one or more, preferably one,
`two, or three, ring atoms are heteroatom selected from N, 0,
`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, pyrimidi(cid:173)
`nyl, pyrazinyl, pyridazinyl, tetrazolyl, and the like.
`"Oxo" or "carbonyl" means =(0) group or C=O
`[0060]
`group, respectively.
`[0061] 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 substitu(cid:173)
`ent is selected from oxo, halogen, ----CN, -NH2 , -OH,
`-NH(CH3), -N(CH3)2 , alkyl (including straight chain,
`branched and/or unsaturated alkyl), substituted or unsubsti(cid:173)
`tuted cycloalkyl, substituted or unsubstituted heterocy(cid:173)
`cloalkyl, fluoroalkyl, substituted or unsubstituted het(cid:173)
`eroalkyl, substituted or unsubstituted alkoxy, fluoroalkoxy,
`-S-alkyl, -S(O)2 -alkyl, ----CONH ((substituted or unsub(cid:173)
`stituted 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 unsubsti(cid:173)
`tuted 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,
`-OH,
`-NH2 ,
`-NH(CH3),
`-N(CH3)2 ,
`----CH3,
`-CH2CH3, ----CH(CH3)2 , ----CF3, -CH2CF3, ---OCH3,
`-OCH2CH3,
`---OCH(CH3)2 ,
`---OCF3,
`---OCH2CF3,
`-S(O)2----CH3, -CONH2 ,
`----CONHCH3, -NHCON-
`HCH3, ----COCH3, --COOR and the like. In some embodi(cid:173)
`ments, substituted groups are substituted with one, two or
`three of the preceding groups. In some embodiments, substi(cid:173)
`tuted groups are substituted with one or two of the preceding
`groups. In some embodiments, substituted groups are substi(cid:173)
`tuted 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 mix(cid:173)
`tures.
`[0062] 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 occur(cid:173)
`ring amino acids besides the D-amino acids, which are func(cid:173)
`tional equivalents of the naturally-occurring amino acids.
`Such non-naturally-occurring amino acids include, for
`example, norleucine ("Nie"), norvaline ("Nva"), L- or
`D-naphthalanine, omithine ("Om"), homoarginine (ho(cid:173)
`moArg) and others well known in the peptide art, such as
`those described in M. Bodanzsky, "Principles of Peptide Syn(cid:173)
`thesis," 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,
`Ill., 1984, both of which are incorporated herein by reference.
`[0063] Amino acids and amino acid analogs can be pur(cid:173)
`chased commercially (Sigma Chemical Co.; Advanced
`Chemtech) or synthesized using methods known in the art.
`"Therapeutically effective amount" or "effective
`[0064]
`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 suffi(cid:173)
`cient to effect such treatment for the disease, disorder, or
`condition. The "therapeutically effective amount" will