NZ750789B2 - Imidazopyrazine syk inhibitors for the treatment of cancers and other hyperproliferative disorders - Google Patents

Abstract

The application provides methods of treating or reducing aplastic anemia in a patient in need thereof, comprising the step of administering an effective amount of a compound selected from Compounds 1-6, disclosed herein.

Description

IMIDAZOPYRAZINE SYK INHIBITORS FOR THE TREATMENT OF S AND OTHER HYPERPROLIFERATIVE DISORDERS CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. § 119(e) of United States Provisional Application Nos. 62/394,573, filed September 14, 2016, 62/416,047, filed er 1, 2016; and 62/429,209, filed December 2, 2016, which are hereby incorporated by reference in their entirety.

FIELD The present disclosure relates to methods of treatment of diseases and disorders using compounds and compositions that inhibit Spleen ne Kinase (SYK) activity.

BACKGROUND Protein kinases, the largest family of human s, encompass well over 500 proteins. Spleen Tyrosine Kinase (SYK) is a member of the family of tyrosine kinases, and is a regulator of early B-cell development as well as mature B-cell activation, signaling, and survival. SYK has roles in immunoreceptor- and in-mediated signaling in a variety of cell types, including B-cells, macrophages, monocytes, mast cells, eosinophils, basophils, neutrophils, tic cells, s, natural killer cells, platelets, and osteoclasts. The inhibition of Syk ty can be useful for the treatment cancers and inflammatory diseases.

U.S. Patents 8,455,493, 8,440,667, 9,376,441, 111, 9,353,066 and 9,376,418 disclose SYK inhibitors. Several SYK inhibitors are in advanced stages of clinical trials. Fostamatinib is a SYK inhibitor currently undergoing phase III clinical trials for chronic immune thrombocytopenic purpura (chronic ITP). (NCT02076412; Clinicaltrials.gov). Fostamatinib, however, has adverse side s including ension, penia and transaminitis.

(Nijjar J.S., et al., Rheumatology 2013, 1556-1562).

SUMMARY Some embodiments provide methods for treating or reducing thrombocytopenia, leukopenia, anemia, or neutropenia in a patient in need thereof, comprising the step of administering an effective amount of a compound selected from: [Annotation] r None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar ed set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar H W? HZN ,Nj/RNJ N\ IN\ 3 3 Compound-l Compound-2 N/ /N HZN ”YVNJ) 3 3 Compound-3 Compound-5 Compound-6 or a ceutically acceptable salt, ester or derivative thereof. Some embodiments provide methods of treatment for anemia, thrombocytopenia, leukopenia or penia wherein said patient is not undergoing chemotherapy.

Some embodiments provide methods for treating a disease or condition selected from the group ting of an inflammatory disorder, an allergic disorder, an autoimmune e, and a cancer in a subject in need thereof, the method comprising administering to the human in need thereof a therapeutically effective amount of a [Annotation] wilksar None set by wilksar [Annotation] wilksar ionNone set by wilksar [Annotation] wilksar Unmarked set by wilksar ation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar compound selected from Compounds 1-6, or a pharmaceutically acceptable salt, ester or derivative thereof, wherein said subject is additionally undergoing proton pump inhibitor therapy; and, wherein said compound selected from Compounds 1-6 is administered at a daily dosage of about 25% to about 75% higher than the daily dosage recommended for a subject not undergoing proton pump inhibitor therapy.

Some embodiments provide methods for treating a proliferative disease or disorder in a subject in need thereof, sing the step of administering a therapeutically effective amount of a compound selected from Compounds 1-6, or a pharmaceutically acceptable salt thereof, to said subject, wherein said subject has one or more abnormal core-binding karyotypes ed from )(q22,q22), inv(16)(p13.1q22) and t(16,16)(p13.1,q22), or wherein said subject has one or more chromosomal abnormalities selected from the deletion of 5, 5q, 7, 7q, 17p, inv(3)(q21,q26) and t(3,3)(q21,q26), or n said subject has a 11q23 rearrangement of MLL gene.

Some embodiments provide methods for treating a proliferative disease or disorder in a subject in need thereof, comprising the step of administering a eutically effective amount of a compound selected from Compounds 1-6, or a pharmaceutically able salt thereof, to said subject, wherein said subject has mixed e leukaemia gene-partial tandem ations (MLL-PTD). MLL-PTD confer a worse prognosis with shortened overall and event free survival in childhood and adult AML. (Basecke J., et al., British Journal ofHematology, 2006, 135(4):).

Some embodiments provide methods for treating a proliferative disease or er in a subject in need thereof, comprising the step of administering a therapeutically effective amount of a compound selected from Compounds 1-6, or a pharmaceutically acceptable salt thereof, to said t, wherein said subject has or ses a mixed lineage leukemia (MLL) fusion protein, a BCR-ABL fusion protein, FLT3 internal tandem duplications, NPMl mutation, IDH1 mutation, IDH2 mutation, a deleted or d p53, high or elevated levels of meningioma 1, and/or high or levels of lactate dehydrogenase. MLL fusion partners can vary and more than 51 fusion rs have been identified. MLL/AF9 often results in AML while MLL/AF4 oftent results in age ALL. (Basecke J. et al, supra). Some ments provide methods for identifying abnormal MLL fusion protein expression in a patient followed by the [Annotation] r None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar ation] wilksar Unmarked set by wilksar administration of a compound selected from Compounds 1-6, or a pharmaceutically acceptable salt thereof to a patient in need f.

Homeobox (HOX) proteins are homeo-domain DNA binding transcription factors that play an important role in regulation of hematopoiesis (Sitwala, Int. J. Clin.

Exp. Pathol. 2008, Mar 30, 1(6):461-74). Myeloid Ecotropic Viral Integration Site 1 (MEISl) is a co-factor of ox A9 ) that recruits additional DNA binding proteins to HOXA9 binding sites in order to maintain the Mixed Lineage Leukemia (MLL) stem cell riptional program ns, Curr. Opin. Hematol. 2016, Jul, 23(4):354-61). Overexpression of these two ns in hematopoietic stem cells (HSCs) results in a rapid and fatal leukemia when transduced HSCs are transplanted in mice (Thorsteinsdottir, EMBO J. 2001, Feb 1, 20(3):350-61). A subset of acute lymphoid leukemia (ALL), pro-B-acute lymphocytic leukemia (pro-B ALL), myelodysplastic syndrome (MDS) and T-ALL patients can be characterized by tumor cells collected from bone marrow showing high co-eXpression of HOXA9 and MEISl that are associated with increased SYK protein levels. Some embodiments provide a method for treating ts with ALL, pro-B ALL, MDS, AML, MDS and T-ALL where HOXA9 and MEISl are overeXpressed by the administration of a therapeutically ive amount of a SYK inhibitor selected from Compounds 1-6, or a pharmaceutically acceptable salt, ester or derivative thereof Some embodiments provide methods for ng a proliferative disease or disorder in a subject in need thereof, comprising the step of administering a therapeutically effective amount of a compound selected from Compounds 1-6, or a pharmaceutically acceptable salt f, to said t, wherein said subject is administered one or more anti-cancer agents selected from the group consisting of AMG— 330, anti-miR155, AntiJL-1+, CD117-MTX aptamer, CD47mAb, CD-98mAb, c, hBF4-mAb, LAIRl-mAb, IL-IRAP-mAb, MG006, miR-150, MRX-6313, MSK-777, SP-2065, SPM-2, SIRPOLFc, TTT-3002, MLL/Menin inhibitor, AML-IL2/CD80, EM, AG—221, , AKN-028, ALT 801, Anti-CD44 mAb, 15, AT- 109, AT-406, AT-9283, AZ-1208, BGB-324, BI 836858, EMS-936564, BP01, Carbozatinib, CB-839, CART-CD123, CART-CD33, CPI-613, CPI-0610, CSL-362, 2291, Dacogen, Dinacilib, DC/AML fusion, Elesclomol, EPZ-5676, GSK2879552, IGN 523, INNO-305, Iomab-B, KX-2391, LeY, LOR-253, MK-8242, [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar MGD006, Oxi-4503, PF-449913, Plerixafor, PRI-724, Rebistanib, Rigosertib, RO- 5503781, PF-04449913, SG—2000, SL-401, SGN-CD33a, TCN-P, Tiecycline, Triciribine, Vismodegib, idar, Actimab-A, AEG 35156, Belinostat, B1811283, Bismab-A, Birinapant, BL-8040, , CNDO-109, Crenolanib, Deformolus, DFP- 10917, Flavopiridol, GVAX, KB-004, KPT-330, Lestartinib, Lirilumab, LO-02040, LY209314, MK8776, Omacetaxine, Pacritinib, nostat, PD-616, umab, PR- 104, ostat, PLX-3397, R—115777, Ribovarin, tinib, SGI-110, nib, Tresosulfan, Temodar, Tosedostat, stat, WT-1, Arsenix trioxide, Clofarabine, Decitabine, Laromustine, Tipfarnib, Palbocyclib, Quizartinib and Mylotarg.

DESCRIPTION OF THE FIGURES Neutrophil counts in patients receiving chemotherapy and 200 or 400 mg twice daily dose of entospletinib (ENTO or Compound-1). The figure shows daily levels for individual patients and neutrophil recovery through induction treatment.

Platelet counts in patients receiving herapy and 200 or 400 mg doses of entospletinib (ENTO or Compound-1). The figure shows daily levels for individual patients and platelet recovery h induction treatment.

Platelet recovery during the entospletinib (ENTO or Compound-1) monotherapy lead-in window.

Scatter Plot of MEIS1 and a HOXA10 Expression in Leukemia Subtypes. Leukemia subtypes with high co-expression 10 and MEISl are boxed in red. Healthy bone marrow is boxed in blue. Data source: Microarray Innovations in Leukemia (MILE) study (GSE13159, PMID: 18573112 and 20406941).

HOXA10 and HOXA9 RNA sion is Highly Correlated in AML. Data Source: The Cancer Genome Atlas , RNAseq data, release 2016_01_28.

Expression of HOXA9 and MEIS1 RNA in Cancer Types from TCGA. MEISl and HOXA9 RNA levels were plotted for cancers in the TCGA. Blue lines in each panel represent the linear regression line. The red box signifies the high HOXA9 and MEISl population in AML. Data Source: TCGA, RNAseq data, release 2016_01_28.

[Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar ed set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] r ed set by wilksar Expression of HOXA9 and MEISl RNA in Tumor and Normal from Bladder, Breast Invasive Carcinoma, Prostate and Uterine s from TCGA Data.

Data Source: TCGA, RNAseq data, release 2016_01_28.

DETAILED DESCRIPTION Some embodiments provide methods for treating or reducing thrombocytopenia, enia, anemia, or neutropenia in a patient in need thereof, comprising the step of administering an effective amount of a compound selected from: 0/} “/1 K/NONH K/ 3 3 Compound-1 Compound-2 C C 9 9 Mk”\ N&N H2N /N \N/ H2N /N \N\/) I I l I N and Compound-5 Compound-6 [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar ation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar or a pharmaceutically able salt, ester or derivative thereof. Some embodiments provide methods of treatment for anemia, thrombocytopenia, enia or neutropenia wherein said patient is not undergoing chemotherapy.

Without being bound to a mechanism or theory, the disclosure herein provides methods to increase neutrophil and platelet levels in patients in need thereof. It is the expectation that the compounds herein will be useful in the treatment of, for example, conditions that cause or exacerbate neutropenia, anemia, leukopenia and thrombocytopenia.

Some embodiments provide methods for treating anemia comprising the step of stering a therapeutically effective amount of a compound ed from Compounds 1-6 to a subject in need thereof. In some embodiments, anemia is selected from the group consisting of hemolytic , microangiopathic hemolytic anemia, hypersplenism, thropoietic anemia, spherocytosis, sideroblastic , autoimmune hemolytic anemia, sickle cell anemia, thalassemia, Glucosephosphate dehydrogenase —def1cient anemia, ious , aplastic anemia, anemia caused by liver disease or renal disease, and anemia caused by the deficiency of one or more vitamins or nutrients. Some embodiments e a method of treatment where anemia is caused by deficiency of one or more nutrients or vitamins, for example, a vitamin B12, B2, B6, C, A1, D, E or K, iron, folic acid, zinc, copper, calcium or protein.

Some embodiments provide methods for treating thrombocytopenia comprising the step of administering a therapeutically effective amount of a compound selected from Compounds 1-6 to a subject in need thereof In some embodiments, thrombocytopenia is selected from idiopathic thrombocytopenia, alcohol-induced thrombocytopenia, drug-induced immune ocytopenia, thrombotic thrombocytopenic purpura, transfusion-induced thrombocytopenia, primary thrombocythemia, disseminated ascular coagulation, hypersplenism, hemolytic uremic syndrome, paroxysmal nocturnal hemoglobinuria and immune thrombocytopenia.

In some embodiments, thrombocytopenia is due to low production of platelets in bone marrow, while in some embodiments, it is due to increased own of platelets in the bloodstream, spleen or liver, or due to lution.

Some embodiments provide methods for treating neutropenia comprising the step of administering a therapeutically ive amount of a compound selected from [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by r [Annotation] wilksar ed set by wilksar Compounds 1-6 to a t in need thereof. In some embodiments, neutropenia is selected from primary neutropenia, acute neutropenia, severe chronic neutropenia, severe congenital neutropenia (Kostmann's syndrome), severe infantile genetic agranulocytosis, benign neutropenia, cyclic neutropenia, chronic idiopathic neutropenia, secondary neutropenia, syndrome ated penia, and immune-mediated neutropenia.

Some embodiments provide methods for treating neutropenia comprising the step of administering a therapeutically effective amount of a compound selected from Compounds 1-6 to a subject in need thereof, wherein neutropenia is caused or ated with ion, alcoholism, allergic disorders, aplastic anemia, autoimmune disease, myelodysplasia, myelofibrosis, dysgammaglobulinemia, smal nocturnal hemoglobinuria, vitamin B12 deficiency, folate deficiency, viral ion, bacterial infection, spleen disorder, hemodialysis or transplantation, myeloma, lymphoma, metastatic solid tumors which infiltrate and replace the bone marrow, toxins, bone marrow failure, Schwachman-Diamond syndrome, cartilage-hair hypoplasia, dyskeratosis congenita, glycogen storage disease type IB, splenomegaly, or sic defects in myeloid cells.

Some embodiments provide methods for treating leukopenia comprising the step of stering a therapeutically effective amount of a compound selected from Compounds 1-6 to a subject in need thereof. In some embodiments, leukopenia is caused or associated with one or more diseases or ers selected from aplastic anemia, mune diseases, herapy, ion, hyperthyroidism, rheumatoid arthritis, liver disorders, spleen disorders, congenital disorders, viral infections, bacterial infections and parasitic infections. In some embodiments, leukopenia is caused or associated with parasitic infection, for example, infection selected from Acanthamoeba keratitism, amoebiasis, ascariasis, babesiosis, balantidiasis, baylisascariasis, Chagas disease, clonorchiasis, cochliomyia, cryptosporidiosis, diphyllobothriasis, dracunculiasis, coccosis, elephantiasis, enterobiasis, fascioliasis, fasciolopsiasis, filariasis, giardiasis, gnathostomiasis, hymenolepiasis, isosporiasis, Katayama fever, aniasis, Lyme disease, malaria, metagonimiasis, myiasis, onchocerciasis, pediculosis, scabies, schistosomiasis, sleeping ss, strongyloidiasis, sis, toxocariasis, toxoplasmosis, nosis, and trichuriasis. In some embodiments, leukopenia is caused by or associated with viral ion, for example, infection selected from HCV, HIV, influenza, Ebola virus, Marburg virus, flavivirus, Venezuelean equine encephalitis, ation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by r [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar ation] wilksar Unmarked set by wilksar Chikungunya virus, and West Nile virus. In some embodiments, flavivirus is selected from the group consisting of Absettarov, Alfuy, Apoi, Aroa, Bagaza, Banzi, Bouboui, Bussuquara, Cacipacore, Carey , Dakar bat, Dengue l, Dengue 2, Dengue 3, Dengue 4, Edge Hill, Entebbe bat, Gadgets Gully, Hanzalova, Hypr, Ilheus, Israel turkey meningoencephalitis, Japanese encephalitis, Jugra, Jutiapa, Kadam, Karshi, Kedougou, Kokobera, Koutango, Kumlinge, Kunjin, Kyasanur Forest e, Langat, Louping ill, Meaban, Modoc, Montana myotis leukoencephalitis, Murray valley encephalitis, Naranjal, Negishi, Ntaya, Omsk hemorrhagic fever, Phnom—Penh bat, Powassan, Rio Bravo, Rocio, royal farm, Russian spring-summer encephalitis, Saboya, St. Louis encephalitis, Sal Vieja, San Perlita, Saumarez Reef, Sepik, Sokuluk, Spondweni, Stratford, Tembusu, Tyuleniy, Uganda S, Usutu, Wesselsbron, west Nile, Yaounde, yellow fever, and Zika. In some embodiments, leukopenia is caused or ated with bacterial infection, for example, infection ing from Acidobacteria, Actinobacteria, Aquificae, Bacteroidetes, Caldiserica, Chlamydiae, Chlorobi, Chloroflexi, Chrysiogenetes, Cyanobacteria, Deferribacteres, Deinococcus-Thermus, Dictyoglomi, Elusimicrobia, Fibrobacteres, Firmicutes, Fusobacteria, Gemmatimonadetes, Lentisphaerae, Nitrospira, Planctomycetes, bacteria, Spirochaetes, istetes, Tenericutes, utes, Thermodesulfobacteria, Thermomicrobia, Thermotogae, or Verrucomicrobia.

Some embodiments provide methods for treating a disease or condition selected from the group consisting of an inflammatory disorder, an allergic disorder, an autoimmune disease, and a cancer in a subject in need thereof, the method comprising administering to the human in need thereof a therapeutically effective amount of a compound ed from Compounds 1-6, or a pharmaceutically acceptable salt, ester or derivative thereof, n said subject is onally undergoing proton pump inhibitor therapy, and, n said compound selected from Compounds 1-6 is administered at an increased daily dosage compared to the daily dosage recommended for a t not undergoing proton pump inhibitor therapy. In some embodiments, proton pump inhibitor is selected selected from omeprazole, hydroxyomeprazole, esomeprazole, prazole, lansoprazole, pantoprazole, rabeprazole, dontoprazole, habeprazole, perprazole, ransoprazole, pariprazole, prazole, tenatoprazole, S-tenatoprazole-Na, and deXlansoprazole. For example, a patient oing proton pump therapy is administered a daily dosage of a compound selected from Compounds 1-6 that is increased by [Annotation] r None set by wilksar [Annotation] wilksar ionNone set by wilksar [Annotation] wilksar Unmarked set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar ionNone set by wilksar [Annotation] wilksar Unmarked set by wilksar between 25-75% of the ended daily dosage. In one embodiment, the daily dosage of compound 1 is increased by about 200%, 100%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30% or 25%. For example, a daily dosage of recommendation of about 800 mg of compound-1 can be increased by about 200%, 100%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30% or 25% for patient undergoing proton pump inhibitor therapy. For example, the increased dosage can be about 2400 mg or 1600 mg or 1400 mg or about 1360 mg or about 1320 mg or about 1280 mg or about 1240 mg or about 1200 mg or about 1160 mg or about 1140 mg or about 1100 mg or about 1060 mg or about 1020 mg or about 980 mg or about 940 mg or about 900 mg. In one embodiment, the ation provides methods for treating a proliferative disease or disorder in a subject in need thereof, comprising the step of administering a therapeutically effective amount of a compound selected from Compounds 1-6, or a pharmaceutically acceptable salt thereof, to said subject, wherein said subject has one or more al inding ypes selected from t(8,21)(q22,q22), inv(16)(p13.1q22) and t(16,16)(p13.1,q22), or wherein said subject has one or more chromosomal abnormalities selected from the deletion of 5, 5q, 7, 7q, 17p, inv(3)(q21,q26) and t(3,3)(q21,q26), or wherein said subject has a 11q23 rearrangement of MLL gene.

Cytogenetic data help stratify patients in terms of diagnosis and evaluating prognosis for survival and risk of transformation to AML (US. Patent Publication No. 20100009364, Hofmann, W.K., et al., Myelodysplastic syndrome. Annual Review of Medicine. 2005, vol. 56, pp. 1-16). Characteristic chromosomal deletions involve chromosome 5 [del(5q), -5], chromosome 11 [del(11q)], chromosome 12 [del(12q)], chromosome 20 [del(20q)], chromosome 7 [del(7q), -7], chromosome 17 [del(17p)], and chromosome 13 [del(13q)]. Other frequent structural and/or numerical chromosomal aberrations include trisomy 8, trisomy 21, and ion 26). Rare reciprocal ocations include t(1, 7)(q10, p10), t(l, 3)(p36, q21), t(3, 3)(q21, q26), t(6, 9)(p23, q34), and t(5, 12)-fusion between PDGFRB and TEL(ETV-6), (q33, p13), t(5, , 11.2). Some embodiments provide methods for ng proliferative diseases including AML and MDS by the administration of a compound selected from Compounds 1-6 or a pharmaceutically acceptable salt thereof to a t in need thereof.

Some embodiments provide methods for treating a proliferative disease or disorder in a subject in need thereof, comprising the step of administering a therapeutically effective amount of a nd selected from Compounds 1-6, or a ation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar pharmaceutically acceptable salt thereof, to said subject, wherein said t has or expresses a mixed lineage ia (MLL) fusion n, a L fusion protein, FLT3 internal tandem duplications, a deleted or mutated p53, high or elevated levels of meningioma 1, and/or high or levels of lactate dehydrogenase.

Homeobox (HOX) proteins are homeo-domain DNA binding transcription factors that play an important role in regulation of hematopoiesis (Sitwala, Int. J. Clin.

Exp. Pathol. 2008, Mar 30, 1(6):461-74). Myeloid pic Viral Integration Site 1 ) is a co-factor of Homeobox A9 (HOXA9) that recruits additional DNA binding proteins to HOXA9 binding sites in order to maintain the Mixed Lineage Leukemia (MLL) stem cell transcriptional m (Collins, Curr. Opin. Hematol. 2016, Jul, 23(4):354-61). Over expression of these two proteins in hematopoietic stem cells (HSCs) results in a rapid and fatal leukemia when transduced HSCs are transplanted in mice (Thorsteinsdottir, EMBO J. 2001, Feb 1, 20(3):350-61). A subset of acute lymphoid leukemia (ALL), acute myeloid leukemia (AML), pro-B-acute lymphocytic leukemia (pro-B ALL), myelodysplastic syndrome (MDS) and T-cell acute lymphoblastic leukaemia ) patients can be characterized by tumor cells collected from bone marrow showing high co-expression of HOXA9 and MEISl that may be associated with increased SYK protein levels. Increased SYK protein expression can be measured by evaluating pSYK . Some embodiments provide a method for treating patients with ALL, pro-B ALL, AML, MDS and T-ALL, where HOXA9 and MEISl are overexpressed, by the administration of a therapeutically effective amount of a SYK tor selected from Compounds 1-6, or a pharmaceutically acceptable salt, ester or derivative thereof.

Some embodiments provide methods for treating a proliferative disease or disorder in a subject in need thereof, comprising the step of stering a therapeutically ive amount of a compound selected from nds 1-6, or a pharmaceutically able salt thereof, to said subject, wherein said subject is administered one or more anti-cancer agents selected from the group consisting of AMG— 330, anti-miR155, AntiJL-l+, CDl l7-MTX aptamer, CD47mAb, CD-98mAb, GO-203c, hBF4-mAb, LAIR1-mAb, IL-IRAP-mAb, MG006, miR-150, MRX-6313, MSK-777, SP-2065, SPM-2, FC, TTT-3002, MLL/Menin inhibitor, AML-IL2/CD80, 4GSSDIEM, , AG—l20, AKN-028, ALT 801, Anti-CD44 mAb, ASP-2215, AT- E AT-406, AT-9283, AZ-1208, BGB-324, BI 836858, EMS-936564, BP01, [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar ed set by wilksar Carbozatinib, CB-839, CART-CD123, CART-CD33, CPI-613, CPI-0610, CSL-362, CWP-232291, n, Dinacilib, DC/AML fusion, Elesclomol, EPZ-5676, 9552, IGN 523, INNO-305, Iomab-B, KX-2391, LeY, LOR-253, MK-8242, MGD006, OXi-4503, PF-449913, Plerixafor, FRI-724, anib, Rigosertib, RO- 5503781, PF-04449913, SG—2000, SL-401, SGN—CD33a, TCN—P, Tiecycline, Triciribine, Vismodegib, Zosuquidar, Actimab-A, AEG 35156, Belinostat, B1811283, -A, Birinapant, BL-8040, CC-486, CNDO-109, Crenolanib, Deformolus, DFP- 10917, Flavopiridol, GVAX, KB-004, KPT-330, Lestartinib, Lirilumab, LO-02040, LY209314, MK8776, OmacetaXine, Pacritinib, Panobinostat, , Pdilizumab, PR- 104, Pracinostat, PLX-3397, R-115777, Ribovarin, Selumetinib, SGI-110, Sorafenib, Tresosulfan, Temodar, Tosedostat, Vorinostat, WT-1, ArseniX trioxide, Clofarabine, Decitabine, Laromustine, Tipfamib and Mylotarg.

Some embodiments provide s of treatment for anemia, thrombocytopenia, leukopenia or neutropenia wherein said patient is not undergoing chemotherapy. For example, the subject is treated with a compound selected from Compounds 1-6 while the subject is not undergoing chemotherapy with agents such as DNA damaging agents, antibiotic agents, antimitotic agents, steroids or glucocorticoids.

DNA ting agents can be selected from, for example, mycin, amsacrine, busulfan, carboplatin, chlorambucil, tin, cyclophosphamide, Cytoxan, dactinomycin, daunorubicin, doxorubicin, epirubicin, iphosphamide, melphalan, merchlorethamine, mitomycin, mitoxantrone, nitrosourea, procarbazine, Taxol, Taxotere, teniposide, etoposide and triethylenethiophosphoramide. The antibiotic chemotherapy agents can be selected from dactinomycin (actinomycin D), daunorubicin, doxorubicin (adriamycin), idarubicin, cyclines, mitoxantrone, bleomycins, plicamycin (mithramycin) and mitomycin. The antimitotic agents can be selected from vinca alkaloids, a taxane, nocodazole, epothilones, navelbine and epidipodophyllotoxin. Vinca alkaloids can be selected from vinblastine and vincristine, or derivatives thereof. s can be selected from paclitaxel and xel, or derivatives thereof. ceutical itions andModes ofAdministration Compounds ed herein are usually stered in the form of pharmaceutical compositions. Thus, provided herein are also pharmaceutical compositions that contain one or more of the compounds of any of the formulae Elosed herein or a pharmaceutically acceptable salt, isomer, g, or solvate [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar ation] wilksar None set by wilksar ation] wilksar MigrationNone set by wilksar [Annotation] r Unmarked set by r thereof, and one or more pharmaceutically acceptable vehicles ed from carriers, adjuvants and excipients. Suitable pharmaceutically acceptable vehicles may e, for example, inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants. Such compositions are ed in a manner well known in the pharmaceutical art. See, 6. g. , Remington’s Pharmaceutical es, Mace Publishing Co., Philadelphia, Pa. 17th Ed. (1985), and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd Ed. (GS. Banker & C.T.

Rhodes, Eds).

The pharmaceutical compositions may be administered in either single or multiple doses. The pharmaceutical composition may be administered by various s including, for example, rectal, buccal, intranasal and transdermal routes. In certain ments, the pharmaceutical composition may be administered by intra- arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, or as an inhalant. In some embodiments, the pharmaceutical composition is administered orally.

One mode for administration is eral, for example, by inj ection. The forms in which the ceutical compositions described herein may be orated for administration by injection include, for example, aqueous or oil suspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, se, or a sterile aqueous solution, and similar pharmaceutical vehicles.

Oral administration may be another route for administration of the compounds described herein. Administration may be via, for example, capsule or enteric coated tablets. In making the pharmaceutical compositions that include at least one compound of any of the formulae described herein or a pharmaceutically acceptable salt, g, or solvate thereof, the active ingredient is usually diluted by an ent and/or enclosed within such a carrier that can be in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it can be in the form of a solid, semi- solid, or liquid al, which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for e, up to 10% by weight of the active compound, soft and hard gelatin capsules, sterile inj ectable solutions, and sterile [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar [Annotation] r None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar packaged s. In certain embodiments, the pharmaceutical ition is in the form of tablets.

As used herein, “pharmaceutically acceptable carrier” or aceutically able ent” includes any and all solvents, dispersion media, coatings, antibacterial and ngal agents, ic and absorption delaying agents and the like.

The use of such media and agents for pharmaceutically active substances is well known in the art. Except r as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated.

Supplementary active ingredients can also be incorporated into the compositions.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose. The formulations can onally include ating agents such as talc, magnesium stearate, and mineral oil, wetting agents, emulsifying and suspending agents, preserving agents such as methyl and propylhydroxy-benzoates, sweetening agents, and flavoring agents.

The compositions that include at least one compound of any of the formulae described herein or a. pharmaceutical ly acceptable salt, isomer, prodrug, or solvate thereof, can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the t by employing procedures known in the art. Controlled e drug delivery systems for oral stration include osmotic pump systems and dissolution systems containing polymer-coated oirs or drug- polymer matrix formulations. Examples of controlled release systems are given in US.

Patent Nos. 3,845,770, 4,326,525, 4,902,514 and 5,616,345. Another formulation for use in the methods of the present application employs transdermal delivery s (“patches”). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds described herein in controlled amounts. The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, e.g., US. Patent Nos. 5,023,252, 4,992,445 and 5,001,139.

Such patches may be ucted for continuous, pulsatile, or on demand delivery of pharmaceutical agents.

[Annotation] wilksar None set by wilksar ation] wilksar ionNone set by wilksar [Annotation] wilksar Unmarked set by wilksar [Annotation] r None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar For preparing solid compositions such as tablets, the principal active ingredient may be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound of any of the above formulae or a pharmaceutically acceptable salt, prodrug, or e thereof. When referring to these preformulation compositions as neous, the active ingredient may be dispersed evenly throughout the ition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.

The tablets or pills of the compounds described herein may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged , or to protect from the acid conditions of the stomach. For example, the tablet or pill can include an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.

Compositions for inhalation or insufflation may e ons and suspensions in ceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described supra. In some embodiments, the compositions are administered by the oral or nasal atory route for local or systemic . In other embodiments, compositions in pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized ons may be inhaled directly from the nebulizing device or the nebulizing device may be attached to a facemask tent, or ittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, ably orally or nasally, from devices that deliver the formulation in an appropriate manner.

Dosing The specific dose level of compounds described herein for any particular subject will depend upon a variety of factors including the ty of the specific [Annotation] wilksar None set by wilksar [Annotation] r MigrationNone set by r [Annotation] wilksar Unmarked set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by r [Annotation] wilksar Unmarked set by wilksar compound employed, the age, body weight, l health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination and the severity of the particular disease in the subject undergoing y. For example, a dosage may be expressed as a number of milligrams of a compound of the formula per kilogram of the subj ect’s body weight (mg/kg). Dosages of between about 0.01 and 200 mg/kg may be appropriate. In some embodiments, about 0.01 and 150 mg/kg may be riate. In other embodiments a dosage of between 0.05 and 100 mg/kg may be appropriate. Normalizing according to the subj ect’s body weight is particularly useful when adjusting dosages between subjects of widely disparate size, such as occurs when using the drug in both children and adult humans or when converting an effective dosage in a non-human subject such as dog to a dosage suitable for a human subject.

The daily dosage may also be described as a total amount of a compound of the formulae administered per dose or per day. Daily dosage of a compound may be n about 1 mg and 2,000 mg, between about 1,000 to 2,000 , between about 1 to 1,000 mg/day, between about 1 to 500 mg/day, between about 100 to 150 mg/day, n about 1 to 100 mg/day, between about 1 to 50 mg/day, between about 50 to 100 mg/day, between about 100 to 125 mg/day, n about 100 to 150 mg/day, between about 100 to 175 mg/day, between about 100 to 200 mg/day, between about 100 to 225 mg/day, between about 100 to 250 mg/day, between about 100 to 350 mg/day, between about 100 to 400 mg/day, n about 100 to 450 mg/day, or between about 100 to 500 mg/day.

When stered orally, the total daily dosage for a human subject may be between 1 mg and 1,000 mg/day, between about 1 to 100 mg/day, between about 1 to 50 mg/day, between about 50 to 100 mg/day, n 50 to 300 mg/day, between 50 to 200 mg/day, between 75 to 200 mg/day, between 75 to 150 mg/day, between 100 to 200 mg/day, between about 200 to 300 mg/day, between about 300 to 400 mg/day, between about 400 to 500 mg/day, between about 100 to 150 mg/day, between about 150 to 200 mg/day, between about 200 to 250 mg/day, between about 75 to 150 mg/day, or between about 150 to 300 .

The compounds of the present application or the compositions thereof may be administered once, twice, three, or four times daily, using any suitable mode described above.

[Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by r [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar ation] wilksar Unmarked set by wilksar Example Methods: In this phase 1b/2 study (NCT02343939), patients age 18 to 70 years with previously untreated AML, preserved organ on, and ECOG S 2 were eligible to receive dose escalated letinib (ENTO or Compound-1) for 14 days as monotherapy (days -14 to 0) followed by combination with daunorubicin 60 mg/m2/d, cycle 1 day 1 to 3, and cytarabine 100 mg/m2/d, cycle 1 day 1 to 7. All patients received entospletinib (ENTO or Compound-1) monotherapy for up to 14 days prior to starting induction. Chemotherapy could be initiated after 5 days of monotherapy (and entospletinib continued for 4+ weeks) in patients with leukemia-related cations necessitating chemotherapy. Patients enrolled to dose level (DL) 0 and DL 1 received entospletinib 200 mg po BID and 400 mg po BID, respectively. Patients with residual disease two weeks after chemotherapy received a second induction cycle identical to the first. Entospletinib was continued without interruption until remission was assessed at count recovery.

Results: Twelve patients enrolled with a median age of 54 (range, 18—69) years. Patients were in the following an LeukemiaNet genetic risk : favorable (n=1), intermediate I (n=3), intermediate II (n=2), and adverse (n=4), respectively. Three patients were not evaluable for dose limiting toxicity (DLT) assessment and were replaced (due to detection of CNS disease requiring non-study therapy (n=1), and withdrawal of consent unrelated to drug ty (n=2)). Single-agent entospletinib during the window period was well tolerated, ties after combination with intensive herapy were common and typical. Among three patients treated at 200 mg BID, no DLT was observed. Of three patients treated at 400 mg BID, a patient with documented fungal pneumonia ped grade 3 pneumonitis that was possibly related to entospletinib. Although this did not meet DLT criteria, DL 1 was expanded with 3 additional ts, none of whom experienced DLT. Overall, the most common non hematologic adverse events (inclusive of intensive chemotherapy periods) were febrile neutropenia, nausea, and diarrhea. Based on this clinical experience and compiled cokinetic data demonstrating lack of benefit to r dose escalation, 400 mg BID was selected as the recommended phase 2 dose. Responses were seen at both levels.

Among the 3 patients treated at 200 mg BID, two required a second ion but each ed a complete remission (CR) (3/3, 100%). Of the 6 patients treated at 400 mg BID, none required a second induction and the CR rate was also 100%. Remarkably, an [Annotation] wilksar None set by wilksar ation] wilksar MigrationNone set by wilksar [Annotation] wilksar ed set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by r [Annotation] wilksar Unmarked set by wilksar 18 year old male with 11q23-rearranged AML achieved morphologic and cytogenetic CR after only the 14 day entospletinib monotherapy window (prior to chemotherapy).

Another patient with 11q23-rearranged AML had significant platelet response during the window period (this patient refused disease evaluation by marrow aspiration prior to chemotherapy).

A neutrophil recovery plot for all enrolled subjects in the above study is provided in Figure 1. During the letinib monotherapy lead-in period, te neutrophil counts (ANC) remained y stable. All three of the ble subjects at the entospletinib 400 mg dose level who were under the severe neutropenia cutoff of 200/uL ANC at baseline recovered to above 200/uL during treatment. Severe neutropenia in these subjects was short lived, with a median (Q1, Q3) time to recovery of 8 (8, 14) days, individual values were 8, 8, and 14 days. The only evaluable subject with severe neutropenia at baseline at the entospletinib 200 mg dose level also recovered during treatment (recovery time of 66 days). Among the five subjects (treated at both entospletinib dose levels) who did not have severe neutropenia at baseline and had at least one post-baseline ANC assessment, none experienced severe neutropenia at any time during induction treatment, however, two of the five ts, both treated at the entospletinib 400 mg dose level, did ence severe neutropenia during the entospletinib monotherapy lead-in period, which recovered to above 200/uL by the end of the first induction cycle in both subjects (Figure 1).

A plot of et counts through induction treatment for all ed subjects in the above study is provided in Figure 2. During the two week entospletinib monotherapy window, platelet count remained stable, or in some cases, improved from baseline. One subject received platelet transfusions at the entospletinib 200 mg dose level and no transfusions were required at the entospletinib 400 mg dose level. During induction ent, the mean number of transfusions ed per subject was 4.1 during Cycle 1 and 5.5 during Cycle 2. Across the monotherapy lead-in and induction treatment cycles, the mean number of platelet transfusions ed per subject was 8.7 at the ENTO 200 mg dose level, 4.1 at the entospletinib 400 mg dose level, and 5.5 overall (Figure 3).

Throughout this specification, various patents, patent applications and other types of publications (e.g., journal articles) are referenced. The disclosure of all patents, [Annotation] wilksar None set by wilksar ation] wilksar MigrationNone set by r [Annotation] wilksar Unmarked set by wilksar [Annotation] wilksar None set by wilksar [Annotation] wilksar MigrationNone set by wilksar [Annotation] wilksar Unmarked set by wilksar patent applications, and publications cited herein are hereby incorporated by reference in their entirety for all purposes.

Claims (6)

CLAIMS What is d is:

1. Use of a therapeutically effective amount of a compound selected from , , Compound-1 Compound-2 , , and Compound-3 Compound-4 Compound-5 or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating or reducing aplastic anemia in a subject.

2. Use according to claim 1 n the compound is or a pharmaceutically acceptable salt thereof.

3. Use according to claim 1 wherein the compound is or a ceutically acceptable salt thereof.

4. Use according to claim 1 wherein the compound is or a pharmaceutically acceptable salt thereof.

5. Use according to claim 1 wherein the compound is or a pharmaceutically acceptable salt thereof.

6. Use according to claim 1 wherein the compound is or a pharmaceutically able salt thereof.