CANCER THERAPY

Abstract

A method of treating haematological cancer with a therapy comprising a DHODH inhibitor. Also provided is a combination therapy comprising a pan-HER inhibitor and a DHODH inhibitor for treating a haematological cancer.

Claims

1. A method of treating a haematological cancer, comprising administering a DHODH inhibitor 2-(3,5-difluoro-3methoxybiphenyl-4-ylamino)nicotinic acid or a pharmaceutically acceptable salt thereof to a patient in need thereof.

2. A method according to claim 1, wherein the haematological cancer is selected from myeloma, lymphoma, Leukaemia, chronic myeloproliferative disease, monoclonal gammopathy of uncertain significance, myelodysplastic syndrome (MDS), amyloidosis and plasmacytoma.

3. A method of treatment according to claim 1, wherein the haematological cancer is selected from myelodysplastic syndrome (MDS), multiple myeloma (MM), acute myeloid leukaemia (AML) and chronic myeloid leukaemia (CML).

4. A method of treatment according to claim 1, wherein the haematological cancer is MDS.

5. A method of treatment according to claim 1, wherein the haematological cancer is multiple myeloma (MM).

6. A method of treatment according to claim 1, wherein the haematological cancer is lymphoma.

7. A method of treatment according to claim 6, wherein the lymphoma is selected from Hodgkin's lymphoma, and non-Hodgkin's lymphoma.

8. A method of treatment according to claim 6, wherein the lymphoma is independently selected from anaplastic large cell lymphoma, angioimmunoblastic lymphoma, Burkitt lymphoma, Burkitt-like lymphoma, blastic NK-cell lymphoma, cutaneous T-cell lymphoma, diffuse large B-cell lymphoma, diffuse large B-cell lymphoma, lymphoblastic lymphoma, MALT lymphoma, mantle cell lymphoma, mediastinal large B-cell lymphoma, nodal marginal zone B-cell lymphoma, small lymphocytic lymphoma, thyroid lymphoma, follicular lymphoma, Waldenstrom's macroglobulinaemia and combinations thereof.

9. A method of treatment according to claim 1, wherein the haematological cancer is a chronic myeloproliferative disease.

10. A method of treatment according to claim 9, wherein the chronic myeloproliferative disease is selected from essential thrombocythaemia, chronic idiopathic myelofibrosis, and polycythaemia rubra vera.

11. A method of treatment according to claim 1, wherein the haematological cancer is leukaemia.

12. A method of treatment according to claim 11, wherein the leukaemia is independently selected from AML (acute myeloid leukaemia), ALL (acute lymphoblastic leukaemia), CML (chronic myeloid leukaemia) and CLL (chronic lymphocytic leukaemia) and combinations thereof.

13. A method of treatment according to claim 11, wherein the leukaemia is selected from hairy cell leukaemia, acute lymphoblastic leukaemia, and chronic lymphoblastic leukaemia.

14. A method of treatment according to claim 11, wherein the leukaemia is independently selected from acute lymphoblastic leukaemia, chronic lymphoblastic leukaemia, acute myelogenous leukaemia (AML), chronic myelogenous leukaemia (CML), hairy cell leukaemia, T-cell prolymphocytic leukaemia, large granular lymphocytic leukaemia, adult T-cell leukaemia, clonal eosinophilias, T-cell granular leukaemia, NK-cell leukaemia, adult T-cell leukaemia and combinations thereof.

15. A method of treatment according to claim 1, wherein the haematological cancer is AML.

16. A method of treatment according to claim 1, wherein the haematological cancer is CML.

17. A method of treatment according to claim 1, wherein the DHODH inhibitor is employed in a combination therapy with a second therapy.

18. A method of treatment according to claim 17, wherein the second therapy is an inhibitor of DNA repair.

19. A method of treatment according to claim 18, wherein the inhibitor is selected from the group comprising: a TRC102, (2E)-2-[(4,5-Dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)methylene]-undecanoic acid [also known as E3330], NCS-666715 and NSC-124854, 8-oxoguamine, tanespirmycin, luminespib, alvespimycin, genetespib, retaspimycin, 6-Amino-8-[(6-iodo-1,3-benzodioxol-5-yl)thio]-N-(1-methylethyl)-9H-purine-9-propanamine (PU-H71), 4-[2-carbamoyl-5-[6,6-dimethyl-4-oxo-3-(trifluoromethyl)-5,7-dihydroindazol-1-yl]anilino]cyclohexyl] 2-aminoacetate (SNX-5422), luminespib (resorcyinylic), 2-(2-ethyl-3,5-dihydroxy-6-(3-methoxy-4-(2-morpholinoethoxy)benzoyl)phenyl)-N,N-bis(2-methoxyethyl)acetamide (KW-2478), AT13387, 5,6-bis((E)-benzylideneamino)-2-thioxo-2,3-dihydropyrimidin-4(1H)-one (SCR7) and combinations of two or more of the same; b olaparib, rucaparib, niraparib, iniparib, talazoparib, veliparib, CEP9722, E7016, BGB-290, AZD-2461, 3-aminobenzamide and combinations thereof; and c 7-hydroxystaurosporine [UCN-01], trabectedin, MCI13E, NERI01 and combinations of two or more of the same.

20. A method of treatment according to claim 18, wherein the second therapy is a topoisomerase inhibitor selected from the group comprising: a irinotecan, topotecan, camptothecin lamellarin D and combinations thereof; and b etoposide (VP-16), teniposide, doxorubicin, daunorubicin, mitoxantrone, amsacrine, ellipticines, aurintricarboxylic acid, 3-Hydroxy-2-[(1R)-6-isopropenyl-3-methyl-cyclohex-2-en-1-yl]-5-pentyl-1,4-benzoquinone (HU-331) and combinations thereof.

Description

BRIEF DESCRIPTION OF FIGURES

[0262] FIGS. 1A&B Shows ASLAN003 induction of CD11b and CD14 in various AML cell lines (A) MOLM-14 (M5), THP-1 (M5) and NB-4 (M3) (B) KG-1 (M0/1) and HL-60 (M2)

[0263] FIG. 2 Shows NBT reduction activity for ASLAN003

[0264] FIG. 3 Shows Giemsa Staining for MOLM-14 cells after treatment with ASLAN003 or a control for 96 hours

[0265] FIG. 4 Shows NBT assay results for MOLM-14 cells after treatment for with ASLAN003 or a control for 96 hours

[0266] FIG. 5 Shows Giemsa Staining for THP-1 (M5) cells after treatment with ASLAN003 or a control for 96 hours

[0267] FIG. 6 Shows NBT assay results for THP-1 (M5) cells after treatment for with ASLAN003 or a control for 96 hours

[0268] FIG. 7 Shows Giemsa Staining for NB-4 cells after treatment with ASLAN003 or a control for 96 hours

[0269] FIG. 8 Shows NBT assay results for NB-4 cells after treatment for with ASLAN003 or a control for 96 hours

[0270] FIG. 9 Shows survival of MOLM-14 xenograft mice following administration of ASLAN003. Median survival: Vehicle group 24 days, ASLAN003 group 27 days

[0271] FIG. 10 Shows percentage of CD45+ cells in bone marrow, peripheral blood, spleen and liver of MOLM-14 xenograft following administration of ASLAN003

[0272] FIG. 11 Shows percentage of hCD11b+ cells in bone marrow of MOLM-14 xenograft mice following administration of ASLAN003

[0273] FIG. 12 Shows flow cytometry results for primary AD345 MDS cells after 96 hours treatment with ASLAN003.

[0274] FIG. 13 Shows flow cytometry results for primary AD537 MDS cells after 96 hours treatment with ASLAN003.

[0275] FIG. 14 Shows Wright-Giemsa Staining for AD537 cells.

[0276] FIG. 15 Shows NBT reduction staining for AD537 cells.

[0277] FIG. 16 Shows cell viability and % inhibition for JJN3 human multiple myeloma (MM) cell line following administration of ASLAN003.

[0278] FIG. 17 Shows cell viability and % inhibition for KMS12 human multiple myeloma (MM) cell line following administration of ASLAN003.

[0279] FIG. 18 Shows cell viability and % inhibition for U266 human multiple myeloma (MM) cell line following administration of ASLAN003.

[0280] FIG. 19 Shows cell viability for KMS28BM human multiple myeloma (MM) cell line following administration of ASLAN003.

[0281] FIG. 20 Shows cell viability and % inhibition for H929 human multiple myeloma (MM) cell line following administration of ASLAN003.

[0282] FIG. 21 Shows cell viability and % inhibition for MM1.S human multiple myeloma (MM) cell line following administration of ASLAN003.

[0283] FIG. 22 Shows cell viability for OPM2 human multiple myeloma (MM) cell line following administration of ASLAN003.

[0284] FIG. 23 Shows cell viability for RPMI8226 human multiple myeloma (MM) cell line following administration of ASLAN003.

[0285] FIG. 24 Shows cell viability and % inhibition for K562 human chronic myeloid leukaemia (CML) cell line following administration of ASLAN003.

[0286] FIG. 25 Shows cell viability and % inhibition for KCL-22 human chronic myeloid leukaemia (CML) cell line following administration of ASLAN003.

EXAMPLE 1IN VITRO ANALYSIS OF ASLAN003 AGAINST AML CELL LINES FROM DIFFERENT FAB CLASSIFICATION SUBTYPES AND THAT ARE IN DIFFERENT STAGES OF BLAST CELL DIFFERENTIATION

[0287] The AML cell lines were dosed with the varying concentrations of ASLAN003 and differentiation of the cell lines were observed via upregulation of CD11b and CD14 on the cell surface via flow cytometry. Differentiation was also determined using the NBT assay and from morphological observations from Wright-Giemsa staining.

TABLE-US-00001 AML Cell Line FAB Classification ASLAN003 Differentiation KG-1 M0/M1 Positive MOLM-14 M5 Positive THP-1 M5 Positive HL-60 M2 Negative NB-4 M3 Negative

[0288] The results are shown in FIGS. 1 to 8 and summarised in the table above.

[0289] The myeloid differentiation effects of ASLAN003 were observed in KG-1, MOLM-14 and THP-1 cell lines. However, dosing of HL-60 and NB-4 AML cell lines with ASLAN003 did not induce observed differentiation effects.

EXAMPLE 2AN IN VIVO ANALYSIS OF ASLAN003 CARRIED OUT IN MOLM-14 XENOGRAFT MODEL

[0290] NSG mice were inoculated with MOLM-14 cells via tail vein injection. Treatment by oral gavage with either ASLAN003 (50 mg/kg QD) or vehicle control was started 3 days past inoculation. 11 mice were treated in the ASLAN003 group and 12 mice were treated in the vehicle control group. The following data is collected at the end of the experiment:

[0291] The results are shown in FIGS. 9 to 11.

[0292] FIG. 9 demonstrates that ASLAN003 significantly (p=0.031) prolonged survival of MOLM-14 xenograft micethe median survival for the vehicle group was 24 days vs 27 days for the group treated with ASLAN003.

[0293] FIG. 10 shows the results of the flow cytometry analysis of CD45+ cells in the bone marrow, peripheral blood, spleen and liver of MOLM-14 xenograft mice. The percentage of CD45+ cells is an indication of leukaemic burden. Thus, FIG. 10 indicates that ASLAN003 significantly decreased leukaemic burden in the bone marrow, peripheral blood, spleen and liver of MOLM-14 xenograft mice.

[0294] FIG. 11 shows the results of the flow cytometry analysis of hCD11b+ and hCD14+ cells in the bone marrow of MOLM-14 xenograft mice. CD11b and CD14 are markers of cell differentiation. Thus, the results in FIG. 11 shows that ASLAN003 significantly induced differentiation of leukaemic cells in the bone marrow of MOLM-14 xenograft mice.

[0295] Taken together, the above studies clearly demonstrate that ASLAN003 was able to significantly prolong the survival, reduce the leukaemic burden and induce differentiation of leukaemic cells in an AML animal model.

EXAMPLE 3AN IN VITRO ANALYSIS OF ASLAN003 CARRIED OUT ON AD345 AND AD537 MDS CELLS

[0296] AD345 primary MDS cells (refractory cytopenia with multilineage dysplasia, normal karyotype) and AD537 primary MDS bone marrow cells (refractory cytopenia with multilineage dysplasia, karyotype 46, XY, 43.1% myeloid) were treated with ASLAN003 (4000 nM) or DMSO control for 96 hours. A FACs analysis was then performed using Pacific blue dye. The AD537 cells were also stained with Wright-Giemsa and NBT reduction.

[0297] FIG. 12 shows the results of the FACs analysis for the AD345 MDS cells. A reduction in % viable cells from 93% to 86.5% was observed between the AD345 cells treated with ASLAN003 vs DMSO control. We also observed an about 11% increase in CD11b+CD14+ cells in 4 M ASLAN003 treated samples. CD13+CD33+ cells were increased about 12.3% in 4 M ASLAN003 treated samples.

[0298] FIG. 13 shows the results of the FACs analysis for the AD537 MDS cells. A reduction in % viable cells from 84.5% to 73% was observed between the AD537 cells treated with ASLAN003 vs DMSO control. We also observed an about 20% increase in CD14+ cells and 30% increase of CD11b+ cells in 4 M ASLAN003 treated samples. CD13+CD33+ cells were increased about 12.3% in 4 M ASLAN003 treated samples.

[0299] These results thus suggest that ASLAN003 was able to reduce the viability of MDS cells, indicating the potential for ASLAN003 as a treatment for MDS.

EXAMPLE 4IN VITRO ANALYSIS OF ASLAN003 CARRIED OUT ON CHRONIC MYELOID LEUKEMIA (CML) AND MULTIPLE MYELOMA (MM) CELLS

[0300] Multiple myeloma (MM) or chronic myeloid leukaemia (CML) cell lines were seeded at 20,000 cells in 100 L of medium in 96-well format and treated with increasing dose of ASLAN003 for 48 hours. DMSO was used as control. The CellTiter-Glo Luminescent Cell Viability Assay, which is a homogeneous method of determining the number of viable cells in culture based on quantitation of the ATP present, i.e. an indicator of metabolically active cells was then performed on the cell lines.

[0301] FIGS. 16 to 23 show the results of the cell viability assay performed on the MM cell lines. FIGS. 24 & 25 show the results of the cell viability assay performed on the K562 and KCL-22 CML cell lines, respectively.

[0302] The results indicate that ASLAN003 has the ability to reduce the viability of MM and CML cells, suggesting that ASLAN003 may be useful for treating both of these haematological cancers.