COMPOSITIONS COMPRISING A DHODH INHIBITOR FOR THE TREATMENT OF ACUTE MYELOID LEUKEMIA

Abstract

The present invention relates to a method of treatment of acute myeloid leukemia (AML). In one embodiment, the present invention relates to a method of treating acute myeloid leukemia (AML) comprising administering to a subject in need thereof a dihydroorotate dehydrogenase (DHODH) inhibitor, alone in combination with at least one/ms-like tyrosine kinase 3 (FLT-3) inhibitor and/or a DNA polymerase inhibitor.

Claims

1. A method of treating acute myeloid leukemia (AML) comprising administering to a subject in need thereof a dihydroorotate dehydrogenase (DHODH) inhibitor alone or in combination with at least one FLT-3 inhibitor and/or at least one DNA polymerase inhibitor.

2. The method of claim 1, wherein the DHODH inhibitor is selected from leflunomide, teriflunomide, brequinar, dichloroallyl lawsone, maritimus (FK 778), redoxal, DSM265, ASLAN003, PTC299, BRD9185, ML390 and 2-(3′-butoxy-3-chloro-5-fluorobiphenyl-4-ylcarbamoyl)benzoic acid and pharmaceutically acceptable salts thereof and hydrates and solvates thereof.

3. The method of claim 1, wherein the DHODH inhibitor is 2-(3′-butoxy-3-chloro-5-fluorobiphenyl-4-ylcarbamoyl)benzoic acid or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof.

4. The method of claim 1, wherein the FLT-3 inhibitor is selected from midostaurin, gilteritinib, quizartinib, crenolanib, AKN-028, FF10101, SKLB1028, SKI-G-801, KW-2449, AMG-553, Clifutinib, CHMFL-FLT3-335,N-(4-(6-acetamidopyrimidin-4-yloxy)phenyl)-2-(2-(trifluoromethyl)phenyl)acetamide, SU5614, CG-806, symadex, and pharmaceutically acceptable salts thereof and hydrates and solvates thereof.

5. The method of claim 1 any one of claim 1, wherein the FLT-3 inhibitor is gilteritinib or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof.

6. The method of claim 1, wherein the DNA polymerase inhibitor is cytarabine or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof.

7. The method of claim 1, wherein the DHODH inhibitor is administered as a front-line therapy for the acute myeloid leukemia.

8. The method of claim 1, wherein the subject suffers from relapsed-refractory acute myeloid leukemia.

9. The method of claim 1, wherein the subject is a human.

10. The method of claim 1, wherein (i) the DHODH inhibitor is administered to the subject by the oral, intravenous, intramuscular, or intraperitoneal route; (ii) the FLT-3 inhibitor is administered to the subject by the oral, intravenous, intramuscular, or intraperitoneal route; and (iii) the DNA polymerase inhibitor is administered to the subject by the oral, intravenous, intramuscular, or intraperitoneal route.

11. The method of claim 10, wherein (i) the DHODH inhibitor is administered by the oral route; (ii) the FLT-3 inhibitor is administered by the oral route; and (iii) the DNA polymerase inhibitor is administered by the oral route.

12-15. (canceled)

16. The method of claim 1, wherein the DHODH inhibitor is administered at a dose of about 25 to about 1000 mg.

17-18. (canceled)

19. The method of claim 1, wherein the DHODH inhibitor is administered as a single dose or in divided doses.

20. The method of claim 1, further comprising administering one or more anti-cancer treatments, one or more cytostatic, cytotoxic or anticancer agents, targeted therapy, or any combination of any of the foregoing.

21. The method of claim 20, wherein the the DHODH inhibitor is administered together or sequentially with the one or more anti-cancer treatments, one or more cytostatic, cytotoxic or anticancer agents, targeted therapy, or any combination of any of the foregoing.

22. The method of claim 20, wherein the one or more anticancer agents are selected from DNA interactive agents; topoisomerase II inhibitors; topoisomerase I inhibitors; tubulin interacting agents; hormonal agents; thymidilate synthase inhibitors; and anti-metabolites; other tyrosine kinase inhibitors; angiogenesis inhibitors; EGF inhibitors; VEGF inhibitors; CDK inhibitors; SRC inhibitors; c-Kit inhibitors; Her1/2 inhibitors; monoclonal antibodies directed against growth factor receptors; other protein kinase modulators; CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone); R-CHOP (rituximab-CHOP); hyperCV AD (hyperfractionated cyclophosphamide, vincristine, doxorubicin, dexamethasone, methotrexate, cytarabine); R-hyperCV AD (rituximab-hyperCV AD); FCM (fludarabine, cyclophosphamide, mitoxantrone); R-FCM (rituximab, fludarabine, cyclophosphamide, mitoxantrone); bortezomib and rituximab; temsirolimus and rituximab; temsirolimus and bortezomib; Iodine-131 tositumomab and CHOP; CVP (cyclophosphamide, vincristine, prednisone); R-CVP (rituximab-CVP); ICE (iphosphamide, carboplatin, etoposide); R-ICE (rituximab-ICE); FCR (fludarabine, cyclophosphamide, rituximab); FR (fludarabine, rituximab); and D.T. PACE (Dexamethasone, Thalidomide, Cisplatin, Adriamycin, Cyclophosphamide, Etoposide) and any combination of any of the foregoing.

23. The method of claim 20, wherein the one or more anticancer treatments is selected from chemotherapy, radiation therapy, biological therapy, bone marrow transplantation, stem cell transplant, or any combination of any of the foregoing.

24-42. (canceled)

43. The method of claim 1, wherein the method comprises administering to the subject the DHODH inhibitor (S)-N-(2-chloro-6-fluorophenyl)-4-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl-5-fluoro-2-((1,1,1-trifluoropropan-2-yl)oxy)benzamide or a pharmaceutically acceptable salt thereof in combination with at least one FLT-3 inhibitor and/or a DNA polymerase inhibitor.

44-47. (canceled)

48. A pharmaceutical composition for use in the treatment of acute myeloid leukemia (AML) comprising a dihydroorotate dehydrogenase (DHODH) inhibitor alone or in combination with at least one FLT-3 inhibitor and/or a DNA polymerase inhibitor and a pharmaceutically acceptable carrier.

49. The pharmaceutical composition of claim 48, wherein DHODH inhibitor is leflunomide, teriflunomide, brequinar, dichloroallyl lawsone, maritimus (FK 778), Redoxal, DSM265, ASLAN003, PTC299, BRD9185, ML390 and 2-(3′-butoxy-3-chloro-5 -fluorobiphenyl-4-ylcarbamoyl)benzoic acid or a pharmaceutically acceptable salt thereof.

50. The pharmaceutical composition of claim 48, wherein the DHODH inhibitor is 2-(3′-butoxy-3-chloro-5-fluorobiphenyl-4-ylcarbamoyl)benzoic acid or a pharmaceutically acceptable salt thereof.

51. A pharmaceutical composition for use in the treatment of acute myeloid leukemia (AML) comprising (S)-N-(2-chloro-6-fluorophenyl)-4-(4-ethyl-3(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluoro-2-((1,1,1-trifluoropropan-2-yl)oxy)benzamide or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof in combination with at least one FLT-3 inhibitor and/or a DNA polymerase inhibitor and a pharmaceutically acceptable carrier.

52. The pharmaceutical composition of claim 48, wherein the composition further comprises one or more cytostatic, cytotoxic or anticancer agents.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0076] FIG. 1 is a bar graph depicting the anti-proliferative effect of compound A in combination with gilteritinib in AML cell line THP-1 according to the procedure described in Example 1B.

[0077] FIG. 2 is a bar graph depicting the anti-proliferative effect of compound A in combination with gilteritinib in AML cell line U937 according to the procedure described in Example 1C.

[0078] FIG. 3 is a bar graph depicting the effect of compound A on CD11b mRNA expression in THP-1 cell lines according to the procedure described in Example 2.

[0079] FIG. 4 is a bar graph depicting the effect of compound A on CD11b expression in THP-1 cell lines according to the procedure described in Example 2A.

[0080] FIG. 5 is a bar graph depicting the effect of compound A on CD11b expression in MV411 cell lines according to the procedure described in Example 2A.

[0081] FIG. 6 is a bar graph depicting the effect of compound A in combination with gilteritinib on p-Akt expression in THP-1 cell lines according to the procedure described in Example 3.

[0082] FIG. 7 is a bar graph depicting the effect of compound A in combination with gilteritinib on p-Erk 1/2 expression in THP-1 cell lines according to the procedure described in Example 3.

[0083] FIG. 8 is a bar graph depicting the effect of compound A in combination with cytarabine on tumor weight in MV411 xenograft model according to the procedure described in Example 4.

[0084] FIG. 9 is a line graph depicting the effect of compound A in combination with cytarabine on tumor volume in MV411 xenograft model according to the procedure described in Example 4.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

[0085] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood in the field to which the subject matter belongs. In the event that there is a plurality of definitions for terms herein, those in this section prevail. Where reference is made to a URL or other such identifier or address, it is understood that such identifiers generally change and particular information on the internet comes and goes, but equivalent information is found by searching the internet. Reference thereto evidences the availability and public dissemination of such information.

[0086] It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter.

[0087] In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, as used in the specification, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. In this application, the use of “or” means “and/or” unless stated otherwise. Furthermore, use of the term “including” as well as other forms, such as “include”, “includes,” and “included,” is not limiting.

[0088] Definition of standard chemistry and molecular biology terms may be found in reference works including, but not limited to, Carey and Sundberg “ADVANCED ORGANIC CHEMISTRY 4th edition” Vols. A (2000) and B (2001), Plenum Press, New York and “MOLECULAR BIOLOGY OF THE CELL 5th edition” (2007), Garland Science, New York. Unless otherwise indicated, conventional methods of mass spectroscopy, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology are contemplated within the scope of the embodiments disclosed herein.

[0089] Unless specific definitions are provided, the nomenclature employed in connection with, and the laboratory procedures and techniques of, analytical chemistry, and medicinal and pharmaceutical chemistry described herein are those generally used. In some embodiments, standard techniques are used for chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients. In other embodiments, standard techniques are used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipofection). In certain embodiments, reactions and purification techniques are performed e.g., using kits of manufacturer's specifications or as described herein. The foregoing techniques and procedures are generally performed of conventional methods and as described in various general and more specific references that are cited and discussed throughout the present specification.

[0090] Pharmaceutically acceptable salts forming part of this invention include salts derived from inorganic bases such as Li, Na, K, Ca, Mg, Fe, Cu, Zn, and Mn; salts of organic bases such as N,N′-diacetylethylenediamine, glucamine, triethylamine, choline, hydroxide, dicyclohexylamine, metformin, benzylamine, trialkylamine, and thiamine; salts of chiral bases such as alkylphenylamine, glycinol, and phenyl glycinol; salts of natural amino acids such as glycine, alanine, valine, leucine, isoleucine, norleucine, tyrosine, cystine, cysteine, methionine, proline, hydroxy proline, histidine, omithine, lysine, arginine, and serine; quaternary ammonium salts of the compounds of invention with alkyl halides, alkyl sulphates such as Mel and (Me)2SO4; salts of non-natural amino acids such as D-isomers or substituted amino acids; salts of guanidine; and salts of substituted guanidine wherein the substituents are selected from nitro, amino, alkyl, alkenyl, alkynyl, ammonium or substituted ammonium salts and aluminum salts. Salts may include acid addition salts where appropriate which are sulphates, nitrates, phosphates, perchlorates, borates, hydrohalides, acetates, tartrates, maleates, citrates, fumarates, succinates, palmoates, methanesulphonates, benzoates, salicylates, benzenesulfonates, ascorbates, glycerophosphates, and ketoglutarates.

[0091] When ranges are used herein for physical properties, such as molecular weight, or chemical properties, all combinations and sub-combinations of ranges and specific embodiments therein are intended to be included. The term “about” when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range may vary from, for example, between 1% and 15% of the stated number or numerical range. The term “comprising” (and related terms such as “comprise” or “comprises” or “having” or “including”) includes those embodiments, for example, an embodiment of any composition of matter, composition, method, or process, or the like, that “consist of” or “consist essentially of” the described features.

[0092] Abbreviations used herein have their conventional meaning within the chemical and biological arts, unless otherwise indicated.

[0093] The term “effective amount” or “therapeutically effective amount” refers to that amount of a compound described herein that is sufficient to effect the intended application including, but not limited to, disease treatment, as defined below. The therapeutically effective amount may vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated, e.g., the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art. The term also applies to a dose that will induce a particular response in target cells, e.g., reduction of platelet adhesion and/or cell migration. The specific dose will vary depending on the particular compounds chosen, the dosing regimen to be followed, whether it is administered in combination with other compounds, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried.

[0094] As used herein, the terms “treatment” and “treating” refer to an approach for obtaining beneficial or desired results including, but not limited to, therapeutic benefit and/or a prophylactic benefit. By therapeutic benefit is meant eradication or amelioration of the underlying disorder being treated. Also, a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the patient, notwithstanding that the patient may still be afflicted with the underlying disorder. For prophylactic benefit, the compositions may be administered to a patient at risk of developing a particular disease, or to a patient reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease may not have been made.

[0095] The term “front-line treatment” refers to the first treatment given for a disease. It is often part of a standard set of treatments, such as surgery followed by chemotherapy and radiation. When used by itself, front-line therapy is generally the one accepted as the best treatment. If it does not cure the disease or it causes severe side effects, other treatment may be added or used instead. It is also called induction therapy, primary therapy, and primary treatment.

[0096] The term “relapsed” refers to disease that reappears or grows again after a period of remission.

[0097] The term “refractory” is used to describe when the cancer does not respond to treatment (meaning that the cancer cells continue to grow) or when the response to treatment does not last very long.

[0098] The term “subject” or “patient” refers to an animal, such as a mammal, for example a human The methods described herein can be useful in both human therapeutics and veterinary applications. In some embodiments, the patient is a mammal, and in some embodiments, the patient is human For veterinary purposes, the terms “subject” and “patient” include, but are not limited to, farm animals including cows, sheep, pigs, horses, and goats; companion animals such as dogs and cats; exotic and/or zoo animals; laboratory animals including mice, rats, rabbits, guinea pigs, and hamsters; and poultry such as chickens, turkeys, ducks, and geese.

[0099] The term “carrier,” as used herein, refers to relatively nontoxic chemical compounds or agents that facilitate the incorporation of an active compound into cells or tissues.

[0100] The terms “pharmaceutically acceptable carrier” and “pharmaceutically acceptable excipient” include, but are not limited to, any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, one or more suitable diluents, fillers, salts, disintegrants, binders, lubricants, glidants, wetting agents, controlled release matrices, colorants, flavorings, carriers, excipients, buffers, stabilizers, solubilizers, and any combination of any of the foregoing. Except insofar as any conventional media or agent is incompatible with the active ingredient(s), its use in the therapeutic compositions of the invention is contemplated. Supplementary active ingredients can also be incorporated into the compositions.

[0101] The term “diluent” refers to chemical compounds that are used to dilute the compound of interest prior to delivery. In some embodiments, diluents are used to stabilize compounds because they provide a more stable environment. Salts dissolved in buffered solutions (which also provide pH control or maintenance) are utilized as diluents, including, but not limited to a phosphate buffered saline solution.

[0102] The term “glidant” is a substance that use to increase the flowability of a powder. That means it promotes the flow of the tablet granules (or the powder). It does so by reducing the friction between these granules. Suitable glidants include, but are not limited to, fumed silicon dioxide, sodium aluminosilicate, calcium silicate, powdered cellulose, colloidal silicon dioxide, microcrystalline cellulose, corn starch, sodium benzoate, calcium carbonate, magnesium carbonate, talc, metallic stearates, calcium stearate, magnesium stearate, zinc stearate, magnesium lauryl sulfate, and magnesium oxide, or a mixture thereof.

[0103] The term “filler” refers to a substance that adds bulk to products making very small active ingredient components easy for consumer to take. Suitable fillers include, but are not limited to, calcium carbonate, dibasic calcium phosphate, lactose, magnesium carbonate, magnesium oxide, lactose anhydrous, microcrystalline cellulose, insomalt, mannitol and any mixtures thereof, more preferably isomalt and/or microcrystalline cellulose.

[0104] The term “lubricant” refers to a substance that is used to prevent the clumping of active ingredients and prevent the sticking of materials to machines in the manufacturing plant. Suitable lubricants include, but are not limited to, stearic acid, a salt of stearic acid, talc, sodium stearyl fumarate, calcium stearate, glyceryl behenate, magnesium silicate, magnesium trisilicate, hydrogenated castor oil or mixtures thereof.

[0105] The terms “disintegrant” and “disintegrator” refer to a substance that allows for breakdown of a capsule or tablet when wet. This ensures rapid breakdown to facilitate rapid absorption of a product. Suitable disintegrants include, but are not limited to, sodium starch glycolate, starch, croscarmellose sodium, crospovidone, carboxymethyl cellulose calcium, carboxymethylcellulose sodium, magnesium aluminium silicate or mixtures thereof.

[0106] The term “binder” refers to a substance that is used to hold ingredients together. They also give weight and allow small active ingredients to be combined into an easy to take capsule or tablet. Binders are typically sugar derivatives. Suitable binders include, but are not limited to, hydroxypropyl cellulose, polyvinylpyrrolidone k-30, hydroxypropyl cellulose (low-substituted), starch or mixtures thereof, more preferably hydroxypropyl cellulose (low-substituted).

[0107] The term “co-administration,” “administered in combination with,” and their grammatical equivalents, as used herein, encompasses administration of two or more agents to a subject so that both agents and/or their metabolites are present in the animal at the same time. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in a composition in which both agents are present.

METHODS OF TREATMENT AND USES

[0108] In any of the methods of treatment and uses described herein, one or more additional active agents can be administered with the compound of formula (A) or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof. For example, the compound of formula (A) or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof may be administered in combination (administered together or sequentially) with one or more known anti-cancer treatments such as chemotherapy, radiation therapy, biological therapy, bone marrow transplantation, stem cell transplant or any other anticancer therapy or with one or more cytostatic, cytotoxic or anticancer agents or targeted therapy either alone or in combination, such as but not limited to, for example, DNA interactive agents, such as fludarabine, cisplatin, chlorambucil, bendamustine or doxorubicin; alkylating agents, such as cyclophosphamide; topoisomerase II inhibitors, such as etoposide; topoisomerase I inhibitors such as CPT-11 or topotecan; tubulin interacting agents, such as paclitaxel, docetaxel or the epothilones (for example ixabepilone), either naturally occurring or synthetic; hormonal agents, such as tamoxifen; thymidilate synthase inhibitors, such as 5-fluorouracil; and anti-metabolites, such as methotrexate; other tyrosine kinase inhibitors such as gefitinib (Iressa®) and OSI-774; angiogenesis inhibitors; EGF inhibitors; VEGF inhibitors; CDK inhibitors; SRC inhibitors; c-Kit inhibitors; Her1/2 inhibitors, checkpoint kinase inhibitors and monoclonal antibodies directed against growth factor receptors such as erbitux (EGF) and herceptin (Her2); CD20 monoclonal antibodies such as rituximab, ublixtumab (TGR-1101), ofatumumab (HuMax; Intracel), ocrelizumab, veltuzumab, GA101(obinutuzumab), AME-133v (LY2469298, Applied Molecular Evolution), ocaratuzumab (Mentrik Biotech), PRO131921, tositumomab, hA20 (Immunomedics, Inc.), ibritumomab-tiuxetan, BLX-301 (Biolex Therapeutics), rituximab (Reditux®) (Dr. Reddy's Laboratories), and PRO70769 (described in WO2004/056312); other B-cell targeting monoclonal antibodies such as belimumab, atacicept or fusion proteins such as blisibimod and BR3-Fc, other monoclonal antibodies such as alemtuzumab and other protein kinase modulators.

[0109] The methods of treatment and uses described herein also include use of one or more additional active agents (or a regimen of one or more additional active agents) to be administered with the compound of formula (A) or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof. For example CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone); R-CHOP (rituximab-CHOP); hyperCV AD (hyperfractionated cyclophosphamide, vincristine, doxorubicin, dexamethasone, methotrexate, cytarabine); R-hyperCV AD (rituximab-hyperCV AD); FCM (fludarabine, cyclophosphamide, mitoxantrone); R-FCM (rituximab, fludarabine, cyclophosphamide, mitoxantrone); bortezomib and rituximab; temsirolimus and rituximab; temsirolimus and Velcade®; Iodine-131 tositumomab (Bexxar®) and CHOP; CVP (cyclophosphamide, vincristine, prednisone); R-CVP (rituximab-CVP); ICE (iphosphamide, carboplatin, etoposide); R-ICE (rituximab-ICE); FCR (fludarabine, cyclophosphamide, rituximab); FR (fludarabine, rituximab); and D.T. PACE (Dexamethasone, Thalidomide, Cisplatin, Adriamycin, Cyclophosphamide, Etoposide).

[0110] The DHODH compounds described herein are also useful in combination (administered together or sequentially) with one or more steroidal anti-inflammatory drugs, non-steroidal anti-inflammatory drugs (NSAIDs) or immune selective anti-inflammatory Derivatives (ImSAIDs).

[0111] According to one embodiment, the compound of formula (A) or a hydrate, a pharmaceutically acceptable salt or a solvate thereof can also be administered in combination with one or more other active principles useful in one of the pathologies mentioned above, for example an anti-emetic, analgesic, anti-inflammatory or anti-cachexia agent.

[0112] It is also possible to combine any of the methods, uses and/or compounds described herein with a radiation treatment.

[0113] It is also possible to combine any of the methods, uses and/or compounds described herein with surgery including either pre, post, or during period of surgery.

[0114] These treatments can be administered simultaneously, separately, sequentially and/or spaced in time.

PHARMACEUTICAL COMPOSITIONS

[0115] Any of the pharmaceutical compositions described herein may comprise a DHODH inhibitor (such as Compound (A) or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof) and optionally one or more pharmaceutically acceptable carriers or excipients.

[0116] In one embodiment, the pharmaceutical compositions described herein may comprise a DHODH inhibitor or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof and at least one FLT-3 inhibitor and optionally one or more pharmaceutically acceptable carriers or excipients.

[0117] In another embodiment, the pharmaceutical compositions described herein may comprise a DHODH inhibitor or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof and at least one DNA polymerase inhibitor and optionally one or more pharmaceutically acceptable carriers or excipients.

[0118] In one embodiment, the pharmaceutical composition comprises a therapeutically effective amount of a DHODH inhibitor, such as Compound (A) or a hydrate, pharmaceutically acceptable salt or solvate thereof. The pharmaceutical composition may include one or more additional active ingredients, as described according to any embodiment herein.

[0119] In another embodiment, the pharmaceutical composition comprises a therapeutically effective amount of a DHODH inhibitor, such as compound (A) or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof, and an FLT-3 inhibitor such as gilteritinib. The pharmaceutical composition may include one or more additional active ingredients, as described herein.

[0120] In another embodiment, the pharmaceutical composition includes a therapeutically effective amount of a DHODH inhibitor, such as compound (A) or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof, and a DNA polymerase inhibitor such as cytarabine or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof. The pharmaceutical composition may include one or more additional active ingredients, as described herein.

[0121] Suitable pharmaceutical carriers and excipients may be selected from diluents, fillers, salts, disintegrants, binders, lubricants, glidants, wetting agents, controlled release matrices, colorants, flavorings, buffers, stabilizers, solubilizers, and any combination of any of the foregoing.

[0122] The pharmaceutical compositions described herein can be administered alone or in combination with one or more other active agents. Where desired, the DHODH inhibitor and other agent(s) may be mixed into a preparation or both components may be formulated into separate preparations to use them in combination separately or at the same time.

[0123] The pharmaceutical compositions described herein can be administered alone or in combination with one or more other active agents. Where desired, the DHODH inhibitor and FLT-3 inhibitor and optionally other agent(s) may be mixed into a preparation or both components may be formulated into separate preparations to use them in combination separately or at the same time.

[0124] The pharmaceutical compositions described herein can be administered alone or in combination with one or more other active agents. Where desired, the DHODH inhibitor and DNA polymerase inhibitor and optionally other agent(s) may be mixed into a preparation or both components may be formulated into separate preparations to use them in combination separately or at the same time.

[0125] The pharmaceutical compositions described herein can be administered together or in a sequential manner with one or more other active agents. Where desired, the DHODH inhibitor and other agent(s) may be co-administered or both components may be administered in a sequence to use them as a combination.

[0126] The pharmaceutical compositions described herein can be administered together or in a sequential manner with one or more other active agents. Where desired, the DHODH inhibitor and FLT-3 inhibitor and other agent(s) may be co-administered, or all the components may be administered in a sequence to use them as a combination.

[0127] The pharmaceutical compositions described herein can be administered together or in a sequential manner with one or more other active agents. Where desired, the DHODH inhibitor and DNA polymerase inhibitor and optionally other agent(s) may be co-administered or all the components may be administered in a sequence to use them as a combination.

[0128] The DHODH inhibitor alone or in combination with FLT-3 inhibitor and/or DNA polymerase inhibitor and its pharmaceutical compositions described herein can be administered by any route that enables delivery of the DHODH inhibitor to the site of action, such as orally, intranasally, topically (e.g., transdermally), intraduodenally, parenterally (including intravenously, intraarterially, intramuscularally, intravascularally, intraperitoneally or by injection or infusion), intradermally, by intramammary, intrathecally, intraocularly, retrobulbarly, intrapulmonary (e.g., aerosolized drugs) or subcutaneously (including depot administration for long term release e.g., embedded-under the-splenic capsule, brain, or in the cornea), sublingually, anally, rectally, vaginally, or by surgical implantation (e.g., embedded under the splenic capsule, brain, or in the cornea).

[0129] The pharmaceutical compositions described herein can be administered in solid, semi-solid, liquid or gaseous form, or may be in dried powder, such as lyophilized form. The pharmaceutical composition can be packaged in forms convenient for delivery, including, for example, solid dosage forms such as capsules, sachets, cachets, gelatins, papers, tablets, suppositories, pellets, pills, troches, and lozenges. The type of packaging will generally depend on the desired route of administration. Implantable sustained release formulations are also contemplated, as are transdermal formulations.

[0130] The pharmaceutical composition may, for example, be in a form suitable for oral administration as a tablet, capsule, pill, powder, sustained release formulations, solution, suspension, for parenteral injection as a sterile solution, suspension or emulsion, for topical administration as an ointment or cream or for rectal administration as a suppository. The pharmaceutical composition may be in unit dosage forms suitable for single administration of precise dosages.

[0131] Oral solid dosage forms are described in, e.g., Remington's Pharmaceutical Sciences, 20th Ed., Lippincott Williams & Wilkins., 2000, Chapter 89, “Solid dosage forms include tablets, capsules, pills, troches or lozenges, and cachets or pellets”. Also, liposomal or proteinoid encapsulation may be used to formulate the compositions (as, for example, proteinoid microspheres reported in U.S. Pat. No. 4,925,673). Liposomal encapsulation may include liposomes that are derivatized with various polymers (e.g., U.S. Pat. No. 5,013,556).

[0132] The pharmaceutical compositions described herein may include a DHODH inhibitor and inert ingredients which protect against degradation in the stomach and which permit release of the biologically active material in the intestine.

[0133] The amount of the DHODH inhibitor, such as Compound (A) or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof to be administered is dependent on the mammal being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound and the discretion of the prescribing physician. In certain embodiments, an effective dosage is in the range of about 0.001 to about 100 mg per kg body weight per day, preferably about 1 to about 35 mg/kg/day, in single or divided doses. For a 70 kg human, this would amount to about 0.05 to about 7 g/day, preferably about 0.05 to about 2.5 g/day. An effective amount of a DHODH inhibitor, such as compound (A) or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof of the invention may be administered in either single or multiple doses (e.g., two or three times a day).

[0134] The amount of the FLT-3 inhibitor, such as gilteritinib or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof, or the amount of the DNA polymerase inhibitor, such as cytarabine or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof to be administered is dependent on the mammal being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound and the discretion of the prescribing physician. However, an effective dosage for each inhibitor may be in the range of about 0.001 to about 100 mg per kg body weight per day, preferably about 1 to about 35 mg/kg/day, in single or divided doses. For a 70 kg human, this would amount to about 0.05 to about 7 g/day, preferably about 0.05 to about 2.5 g/day. An effective amount of the FLT-3 inhibitor and/or the DNA polymerase inhibitor may be administered in either single or multiple doses (e.g., two or three times a day).

[0135] More preferably, in any of the methods and uses described herein, the DHODH inhibitor is a compound of Formula (A) or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof.

[0136] A further embodiment of the present invention relates to a method of treating AML comprising administering a therapeutically effective amount of a pharmaceutical composition according to any of the embodiments described herein to a subject in need thereof (preferably, a human subject) in need thereof.

[0137] A further embodiment of the present invention relates to the use of a pharmaceutical composition according to any of the embodiments described herein in the preparation of a medicament for treating haematological and solid cancers, e.g., AML.

[0138] The following general methodology described herein provides the manner and process of using the DHODH inhibitor alone or in combination with FLT-3 inhibitor and/or DNA polymerase inhibitor and are illustrative rather than limiting. Further modification of provided methodology and additionally new methods may also be devised in order to achieve and serve the purpose of the invention. Accordingly, it should be understood that there may be other embodiments which fall within the spirit and scope of the invention as defined by the specification hereto

Preparation of Compound A

Intermediate 1: 3′-butoxy-3-chloro-5-fluorobiphenyl-4-amine

[0139] The title compound (3′-butoxy-3-chloro-5-fluorobiphenyl-4-amine) (0.190 g) was prepared from 4-bromo-2-chloro-6-fluoroaniline (0.2 g, 0.89 mmol) and 3-butoxyphenylboronic acid (0.224 g, 1.16 mmol) by using a Suzuki coupling reaction in the presence of tetrakis(triphenylphosphine)palladium(0) (0.08 eq.) and potassium carbonate (3.3 eq.). The mixture was degassed with N.sub.2 for 30 min. and refluxed until both the starting materials disappeared as monitored by TLC. Work-up (H.sub.2O/AcOEt) and purification gave the desired product as a yellow solid (0.19 g). .sup.1H-NMR (δ ppm, DMSO-d.sub.6, 400 MHz): 7.44-7.41 (m, 2H), 7.27 (t, J 7.9, 1H), 7.17-7.10 (m, 2H), 6.81-6.84 (m, 1H), 5.50 (s, 2H), 4.01 (t, J 5.3, 2H), 1.72-1.65 (m, 2H), 1.50-1.41 (m, 2H), 0.93 (t, J 7.4, 3H).

Compound A: 2-(3′-butoxy-3-chloro-5-fluorobiphenyl-4-ylcarbamoyl)benzoic acid

[0140] Intermediate 1 (90 mg, 0.31 mmol) was dissolved in ˜2 ml of acetic acid Phthalic anhydride (90 mg, 0.6 mmol) was added to the mixture and stirred at room temperature overnight. The solid that separated out was filtered and washed with petroleum ether and dried under vacuum to obtain the title compound (39 mg) as a white solid. M.P.: 128-130° C. .sup.1H-NMR (δ ppm, DMSO-d.sub.6, 400 MHz): 13.02 (s, 1H), 10.23 (s, 1H), 7.82 (d, J 7.9, 1H), 7.73 (s, 1H), 7.60-7.57 (m, 4H), 7.37 (t, J 7.9, 1H), 7.32-7.25 (m, 2H), 6.99-6.96 (m, 1H), 4.06 (t, J 6.4, 2H), 1.73-1.68 (m, 2H), 1.45 (h, J 7.5, 2H), 0.94 (t, J 7.4, 3H). MS (m/z): 440.19 ([M-H].sup.−).

[0141] The present invention is now further illustrated by means of the following biological examples.

BIOLOGICAL EXAMPLES

[0142] Provided below are illustrative examples of the use of a DHODH inhibitor alone or in combination with a FLT-3 inhibitor or a DNA polymerase inhibitor which provides and establishes a synergic effect for the combination when compared to the effect of the individual DHODH inhibitor or the FLT-3 inhibitor or DNA polymerase inhibitor alone.

Example 1

Anti-Proliferative Effect of Compound A in AML Cell Lines (MTT Assay)

[0143] Compound A was tested across a panel of AML cell lines (U937, HL-60, THP-1, KG-1 and MV411). Cells were plated in 96-well plates and incubated with desired concentrations of Compound A for 72 hours (h). At the end of the incubation period, MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)) was added. The plates were placed on a shaker for 5 min to mix the formazan and the optical density at 560 nM was measured on a spectrophotometer. Data were plotted using Graphpad prism for calculation of the GI.sub.50 concentrations.

[0144] Results: All the AML cell lines tested were sensitive to Compound A with GI.sub.50 ranging between 2.4 to 7.6 μM. (See Table-1).

TABLE-US-00001 TABLE 1 Compound A GI50 (μM) in AML Cell Lines Cell Line U937 HL60 THP-1 KG-1 MV411 GI50 (μM) 2.4 3.5 2.5 7.6 2.5

Example 1A

Anti-Proliferative Effect of Compound A in the Presence of Uridine Rescue in AML Cell Lines (MTT Assay)

[0145] Compound A was tested in the absence of Uridine (U937, HL-60, THP-1 and MV411 cell lines) or in the presence of Uridine (100 μM for U937, HL-60 and MV411 and 300 μM for THP-1). Cells were plated in 96-well plates and incubated with desired concentrations of Compound A for 72 h. At the end of the incubation period, MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)) was added. The plates were placed on a shaker for 5 min to mix the formazan and the optical density at 560 nM was measured on a spectrophotometer. Data were plotted using Graphpad prism for calculation of the GI.sub.50 concentrations.

[0146] Results: Addition of 100 μM or 300 μM of Uridine caused a rightward shift in the activity of Compound A with GI.sub.50>10 μM. (See Table-1A).

TABLE-US-00002 TABLE 1A Compound A GI50 (μM) shift with Uridine Rescue in AML Cell Lines Cell Line −Uridine +Uridine U937 3.2 >10 THP-1 2.0 >10 HL-60 3.6 >10 MV411 2.6 >10

[0147] Conclusion. Compound A inhibited growth of AML cell lines with GI.sub.50 between 2-3.2 μM and addition of uridine caused rightward shift with GI50>10 μM

Example 1B

Anti-Proliferative Effect of Compound A in Combination with Gilteritinib in AML Cell Line THP-1 (MTT Assay)

[0148] Compound A (at 3 μM) and Gilteritinib (0.25 μM) was tested in AML cell line THP-1. Cells were plated in 96-well plates and incubated with desired concentrations of Compound A for 72 h. At the end of the incubation period, MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)) was added. The plates were placed on a shaker for 5 min to mix the formazan and the optical density at 560 nM was measured on a spectrophotometer. Data were plotted using Graphpad prism for calculation of the percentage inhibition to determine the effect of Compound A as a single agent or in combination with gilteritinib.

[0149] Results: Compound A potentiated (p<0.05) the activity of gilteritinib by inhibiting the cell growth in THP-1 cell lines (See FIG. 1).

TABLE-US-00003 TABLE 1B Compound % inhibition Compound A 35.83 Gilteritinib 40.10 Compound A + Gilteritinib 50.20

Example 1C

Anti-Proliferative Effect of Compound A in Combination with Gilteritinib in AML Cell Line U937 (MTT Assay)

[0150] Compound A (at 3 μM) and gilteritinib (1.5 μM) was tested in AML cell line U937. Cells were plated in 96-well plates and incubated with desired concentrations of Compound A for 72 h. At the end of the incubation period, MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)) was added. The plates were placed on a shaker for 5 min to mix the formazan and the optical density at 560 nM was measured on a spectrophotometer. Data were plotted using Graphpad prism for calculation of the percentage inhibition to determine the effect of Compound A as a single agent or in combination with gilteritinib.

[0151] Results: Compound A potentiated (p<0.05) the activity of gilteritinib by inhibiting the cell growth in U937 cell lines (See FIG. 2).

TABLE-US-00004 TABLE 1C Compound % inhibition Compound A 41.36 Gilteritinib 41.66 Compound A + Gilteritinib 60.13

Example-2

Effect of Compound A on CD11b mRNA Expression in THP-1 Cell Lines

[0152] THP-1 cells were plated in complete media at pre-determined density in 6-well plates and the cells were treated with Compound A for 72 hours. mRNA was isolated using TRI reagent according to the manufacturer's protocol (TRI Reagent from Sigma). cDNA was synthesized using cDNA Synthesis kit according to manufacturer's protocol (First Strand cDNA Synthesis kit) and Real-Time PCR was performed. Data was calculated using delta delta Ct method. Fold change in mRNA expression was plotted using GraphPad Prism (Version 5.02)

[0153] Results: Compound A caused differentiation in THP-1 by inducing the CD11b gene expression by 80-fold at 3 μM (See FIG. 3).

Example-2A

Effect of Compound A on CD11b Expression in THP-1 and MV411 Cell Lines

[0154] Cells were plated in complete media at pre-determined density in 6-well plates and the cells were treated with Compound A 96 hours for THP-1 cell lines and 72 hours for MV411 cell lines) in the presence or absence of uridine. The cells were stained with CD11b Antibody PE according to manufacturer's protocol (CD11b Monoclonal Antibody (ICRF44), PE, eBioscience) and the cells were acquired and analysed by flow cytometry (Guava Easycyte).

[0155] Results: Compound A caused differentiation in THP-1 by inducing the CD11b cell surface expression in 40% of the cell population at 5 μM and addition of uridine reduced the CD11b expression to 15% (See FIG. 4).

[0156] Compound A caused differentiation in MV411 by inducing the CD11b cell surface expression in 35% of the cell population at 3 μM (See FIG. 5).

Example-3

Effect of Compound A in Combination with Gilteritinib on p-Akt and p-Erk 1/2 Expressions of AML Cell Line

[0157] THP-1 cells were plated in 1% FBS media at pre-determined density in 6-well plates and cells were incubated with Compound A and in combination with gilteritinib for 3 hours. Cells were pelleted, washed with PBS and lysed with lysis buffer (1M Tris-HCl pH 7.5, 1 M NaCl, 0.5 M EDTA pH 8.0, 0.1 M EGTA pH 8.0, protease inhibitor (10×), sodium fluoride, sodium orthovanadate, 200 mM PMSF). Protein estimation was performed using Bradford reagent (ThermoScientific). Samples were denatured, 20 μg of protein was loaded in 7.5% resolving gel for p-Akt and p-Erk 1/2 and SDS Page was performed. Resolved protein was transferred on to the PVDF membrane and probed with anti-rabbit p-Akt and p-Erk 1/2 (1:1000 dilution) primary antibody for overnight at 4° C. Membrane was probed with Ani-rabbit HRP linked IgG secondary antibody at room temperature for 1 hour and ECL substrate was added to the membrane. Membrane was exposed and images were taken in iBright western blot imaging systems. Intensity of the bands were determined using ImageJ 1.42q (NIH, USA) and normalized to β-Actin (loading control). Fold change or percent inhibitions were plotted using GraphPad Prism (Version 5.02).

[0158] Results: The combination of Compound A (3 μM) and gilteritinib (0.1 μM) reduced AKT phosphorylation by 54% and p-Erk 1/2 phosphorylation by 58% when compared to gilteritinib alone in the THP-1 cell line (See FIGS. 6 and 7).

Example-4

Effect of Compound A in Combination with Cytarabine on MV411 Mouse Xenograft Model

[0159] The effect of Compound A was determined in a MV411 mouse xenograft model. Briefly, 5×10.sup.6 cells were injected into the right flank region by Subcutaneous administration under sterile condition. The oral administration of Compound (A) at 30 mg/kg/BID for 21 days. The tumors were measured using a calliper in two dimensions, length (a) and width (b). Tumor volumes were estimated from measurements of the two diameters of the individual tumors as follows: Tumor Volume (mm3)=(a×b2)/2. At the end of the study period, animals were sacrificed and the tumors harvested.

[0160] Results: At the dose tested, Compound A demonstrated significant (P<0.001) anti-tumor activity both as a single agent and in combination with cytarabine at 20 mg/Kg with tumor growth inhibitions of 37 and 73% respectively. No adverse events or body weight changes were observed throughout the study period.

[0161] Conclusion: Compound A demonstrated potential in animal models of AML as a single agent or in combination with cytarabine as shown in FIGS. 8 and 9 and the data indicates a therapeutic potential of the Compound A in treatment of AML.

[0162] Although the invention herein has been described with reference to particular embodiments, it is to be understood that these embodiments are merely illustrative of the principles and applications of the present invention. It is therefore to be understood that numerous modifications may be made to the illustrative embodiments and that other arrangements may be devised without departing from the spirit and scope of the present invention as described above. It is intended that the appended claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

[0163] All publications, patents and patent applications cited in this application are herein incorporated by reference to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference.