WO2024229440A2 - Methods of treating cancer using mdm2 inhibitors and compositions related thereto - Google Patents

Abstract

This disclosure relates to methods of treating cancer using MDM2 inhibitors disclosed herein, and pharmaceutical compositions related thereto. In certain embodiments, this disclosure relates to methods of treating cancer, such as hematological cancers comprising administering an effective amount of a MDM2 inhibitor to a subject in need thereof. In certain embodiments, the MDM2 inhibitor is sabizabulin and the hematological cancer is acute myeloid leukemia.

Description

METHODS OF TREATING CANCER USING MDM2 INHIBITORS AND COMPOSITIONS RELATED THERETO

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 63/463,695 filed May 3, 2023. The entirety of this application is hereby incorporated by reference for all purposes.

BACKGROUND

Acute myeloid leukemia (AML) is a hematological cancer in which the bone marrow produces abnormal myeloid blood cells. Myeloid blood cells typically develop into white blood cells that protect the body from pathogens. In AML, many of the myeloid blood cells do not develop normally, but instead become abnormal white, red blood cells, or platelets which build up in the bone marrow and blood stream. This often leads to complications such as difficulties in fighting infections, anemia, and spontaneous bleeding. AML is often treated using chemotherapy sometimes in combination with radiation therapy or a stem cell transplant. However, these therapies are not universally effective, and recurrence after ceasing chemotherapy is not uncommon. Thus, there is a need to identify improved therapies.

Chen et al. report the discovery of 2-aryl-4-benzoyl-imidazole analogues targeting tubulin polymerization as antiproliferative agents. J Med Chem, 2012, 55, 7285-7289. See also US Patent Nos. 10,525,037 and 10,865,196.

Deng et al. report VERU-111, suppresses triple-negative breast cancer tumor growth and metastasis and bypasses taxane resistance. Mol Cancer Ther, 2020, 19(2): 348-363.

Steiner et al. report a method of treating breast cancer with estrogen receptor modulators. U.S. Patent Application Publication No. US 2022/0257552.

Wang et al. report small-molecule MDM2 inhibitors in clinical trials for cancer therapy. European Journal of Medicinal Chemistry, 236 (2022) 114334.

References cited herein are not an admission for prior art.

SUMMARY

This disclosure relates to methods of treating cancer using MDM2 inhibitors disclosed herein, and pharmaceutical compositions related thereto. In certain embodiments, this disclosure relates to methods of treating cancer, such as hematological cancers comprising administering an effective amount of a MDM2 inhibitor to a subject in need thereof. In certain embodiments, the MDM2 inhibitor is sabizabulin and the hematological cancer is acute myeloid leukemia. In certain embodiments, the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5- tri ethoxyphenylj ethanone (sabizabulin), derivative, prodrug, metabolite, or salt thereof.

In certain embodiments, the MDM2 inhibitor is administered in combination with another anticancer agent. In certain embodiments, the MDM2 inhibitor is administered before or after receiving bone marrow transplant or blood stem cell transplant and optionally radiation therapy.

In certain embodiments, this disclosure relates to MDM2 inhibitors for use in the production of a medicament for treating cancer.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

Figure 1 A shows data on Spearman’s correlation between IC50 values of VERU-111 and expression levels of MDM2 in the 14 MDM2+ AML cell lines. Data indicates that the cytotoxicity of VERU-111 is MDM2-dep endent and p53 -independent in AML cell lines. Expression of MDM2 protein in 16 AML cell lines and NBMM was detected by Western blot.

Figure IB shows data form WST assays for comparison of VERU-111 cytotoxicity to MDM2 KO (sgMDM2) EU-1 and parental EU-1. Cells were treated with different concentrations of VERU-111 for 48 h.

Figure 1C shows from WST assays for dose-response of six of the 16 AML cell lines as indicated to VERU-111.

Figure 2A shows data on the weight of SCID mice xenografted with human MOLM-14 AML cells and treated with different doses of VERU-111, as compared with the untreated control.

Figure 2B shows data from bioluminescence imaging showing intensity of total body leukemia developing in SCID mice treated without (control) or with different doses of VERU-111, i.e., EFS curves among each dose of VERU-111 -treated groups and the control group of xenografted SCID mice.

Figure 3 shows data indicating VERU-111 has anti -tumor activity against melanoma, colorectal cancer, and Ewing’s sarcoma cell lines. Expansion of A2058 (melanoma), TC106 (Ewing’s sarcoma), and HCT-116 (colorectal cancer) cell lines was inhibited in cultures treated with the indicated concentrations of VERU-111 for 72 hours. DETAILED DISCUSSION

Before the present disclosure is described in greater detail, it is to be understood that this disclosure is not limited to particular embodiments described, and as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described.

All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure. Further, the dates of publication provided could be different from the actual publication dates that may need to be independently confirmed.

As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present disclosure. Any recited method can be carried out in the order of events recited or in any other order that is logically possible.

An "embodiment" is an example, and not necessarily limited to such example. Embodiments of the present disclosure will employ, unless otherwise indicated, techniques of medicine, organic chemistry, biochemistry, molecular biology, pharmacology, and the like, which are within the skill of the art. Such techniques are explained fully in the literature. It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. In this specification and in the claims that follow, reference will be made to a number of terms that shall be defined to have the following meanings unless a contrary intention is apparent.

"Subject" refers to any animal, preferably a human patient, livestock, rodent, monkey, or domestic pet.

"Cancer" refers any of various cellular diseases with malignant neoplasms characterized by the proliferation of cells. It is not intended that the diseased cells must actually invade surrounding tissue and metastasize to new body sites. Cancer can involve any tissue of the body and have many different forms in each body area. Within the context of certain embodiments, whether "cancer is reduced" may be identified by a variety of diagnostic manners known to one skill in the art including, but not limited to, observation the reduction in size or number of tumor masses or if an increase of apoptosis of cancer cells observed, e.g., if more than a 5 % increase in apoptosis of cancer cells is observed for a sample compound compared to a control without the compound. It may also be identified by a change in relevant biomarker or gene expression profile, such as PSA for prostate cancer, HER2 for breast cancer, or others.

As used herein, the terms "prevent" and "preventing" include the prevention of the recurrence, spread or onset. It is not intended that the present disclosure be limited to complete prevention. In some embodiments, the onset is delayed, or the severity of the disease is reduced.

As used herein, the terms "treat" and "treating" are not limited to the case where the subject (e g., patient) is cured and the disease is eradicated. Rather, embodiments, of the present disclosure also contemplate treatment that merely reduces symptoms, and/or delays disease progression.

As used herein, the term "combination with" when used to describe administration with an additional treatment means that the agent may be administered prior to, together with, or after the additional treatment, or a combination thereof.

As used herein, the term “derivative” refers to a structurally similar compound that retains sufficient functional attributes of the identified analogue. The derivative may be structurally similar because it is lacking one or more atoms, substituted, a salt, in different hydration/oxidation states, or because one or more atoms within the molecule are switched, such as, but not limited to, replacing a oxygen atom with a sulfur atom, replacing an amino group with a hydroxyl group, replacing a nitrogen with a protonated carbon (CH) in an aromatic ring, replacing a bridging amino group (-NH-) with an oxy group (-O-), or vice versa. The derivative may be a prodrug. Derivatives may be prepared by any variety of synthetic methods or appropriate adaptations presented in synthetic or organic chemistry textbooks, such as those provide in March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Wiley, 6th Edition (2007) Michael B. Smith or Domino Reactions in Organic Synthesis, Wiley (2006) Lutz F. Tietze hereby incorporated by reference.

The term "substituted" refers to a molecule wherein at least one hydrogen atom is replaced with a substituent. When substituted, one or more of the groups are "substituents." The molecule may be multiply substituted. In the case of an oxo substituent ("=O"), two hydrogen atoms are replaced. Example substituents within this context may include halogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl, carbocycloalkyl, heterocarb ocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, -NRaRb, -NR_aC(=O)Rb, -NR_aC(=O)NR_aNRb, -NRaC(=O)ORb, - NRaSCLRb, -C(=O)Ra, -C(=O)ORa, -C(=O)NRaRb, -OC(=O)NRaRb, -ORa, -SR_a, -SORa, - S(=O)2Ra, -OS(=O)2Raand -S(=O)2ORa. Ra and Rb in this context may be the same or different and independently hydrogen, halogen hydroxyl, alkyl, alkoxy, alkyl, amino, alkylamino, dialkylamino, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl.

The term "prodrug" refers to an agent that is converted into a biologically active form in vivo. Prodrugs are often useful because, in some situations, they may be easier to administer than the parent compound. They may, for instance, be bioavailable by oral administration whereas the parent compound is not. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. A prodrug may be converted into the parent drug by various mechanisms, including enzymatic processes and metabolic hydrolysis. Typical prodrugs are pharmaceutically acceptable esters. Prodrugs include compounds wherein a hydroxy, amino or mercapto group is bonded to any group that, when the prodrug of the active compound is administered to a subject, cleaves to form a free hydroxy, free amino or free mercapto group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of an alcohol and the like, e.g., succinate esters, ethyl succinate esters, palmitate ester, stearate esters, phosphate esters; or acetamide, formamide and benzamide derivatives of an amine functional group in the active compound. Other contemplated prodrugs are esters of carboxylic acid groups such as alkyl carboxylate esters, i.e., methyl ester, ethyl ester, propyl ester, polyethylene glycol ester, or derivatives thereof.

Methods of treating cancer

In certain embodiments, this disclosure relates to methods of treating cancer comprising administering an effective amount of a MDM2 inhibitor to a subject in need thereof. In certain embodiments, cancer is a hematological cancer, e.g., the hematological cancer is acute myeloid leukemia. In certain embodiments, the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4- yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) and the hematological cancer is acute myeloid leukemia.

In certain embodiments, the MDM2 inhibitor is (2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5- bis(4-chlorophenyl)-4,5-dimethyl-4,5-dihydro-lH-imidazol-l-yl)(4-(3-(methylsulfonyl)propyl) piperazin- l-yl)methanone (RG7112), derivative, prodrug, metabolite, or salt thereof.

In certain embodiments, the MDM2 inhibitor is 4-(3-(3-chloro-2-fluorophenyl)-4-(4- chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoic acid (Idasanutlin or RG7388), derivative, prodrug, metabolite, ester, or salt thereof. In certain embodiments, the prodrug is a derivative conjugated to polyethylene glycol, e.g., polyethylene glycol ester (RO6839921).

In certain embodiments, the MDM2 inhibitor is 6-chloro-4'-(3-chloro-2-fluorophenyl)-N- (4-hydroxycyclohexyl)-2'-neopentyl-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (SAR405838), derivative, prodrug, ester, metabolite, or salt thereof.

In certain embodiments, the MDM2 inhibitor is N-(6-carbamoyl tetrahydro-2H-pyran-3- yl)-6"-chloro-4'-(2-chloro-3-fluoropyridin-4-yl)-4,4-dimethyl-2"-oxodispiro[cyclohexane-l,2^l- pyrrolidine-3',3"-indoline]-5'-carboxamide (DS-3032b), derivative, prodrug, ester, metabolite, or salt thereof e.g., 4-methylbenzene sulfonate hydrate.

In certain embodiments, the MDM2 inhibitor is 5-(5-chloro-l-methyl-2-oxo-l,2- dihydropyridin-3-yl)-6-(4-chlorophenyl)-2-(2,4-dimethoxypyrimidin-5-yl)-l-isopropyl-5,6- dihydropyrrolo[3,4-d]imidazol-4(lH)-one (HDM201), derivative, prodrug, ester, metabolite, or salt thereof.

In certain embodiments, the MDM2 inhibitor is 4-amino-l-b-D-arabinofuranosyl-2(lH)- pyrimidinone (MK-8242), derivative, prodrug, ester, metabolite, or salt thereof. In certain embodiments, the MDM2 inhibitor is 2-(5-(3-chlorophenyl)-6-(4-chlorophenyl)- l-(l-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid

(AMG232), derivative, prodrug, ester, metabolite, or salt thereof.

In certain embodiments, the MDM2 inhibitor is l-(4-chlorophenyl)-7-isopropoxy-6- m ethoxy -2-(4-(methyl((4-(4-methyl-3 -oxopiperazin- l-yl)cy cl ohexyl)methyl)amino)phenyl)- 1,4- dihydroisoquinolin-3(2H)-one (NVP-CGM097) derivative, prodrug, ester, metabolite, or salt thereof.

In certain embodiments, the MDM2 inhibitor is 4-(6"-chloro-4'-(3-chloro-2-fluorophenyl)- r-ethyl-2"-oxodispiro[cyclohexane-l,2'-pyrrolidine-3',3"-indoline]-5'-carboxamido)bicyclo [2.2.2]octane-l -carboxylic acid (APG-115), derivative, prodrug, ester, metabolite, or salt thereof.

In certain embodiments, the MDM2 inhibitor is (2S,5S,8S,HS,14S,17S,20S,32R,E)-8- ((lH-indol-3-yl)methyl)-32-((R)-2-((2S,3R)-2-((R)-2-acetamido-4-methylpentanamido)-3- hydroxybutanamido)-3-phenylpropanamido)-N-((2R,5S,8S,l lS,14S,17S)-l-amino-2,5,8,l 1,14- pentamethyl-l,4,7,10,13,16-hexaoxo-3,6,9,12, 15-pentaazaoctadecan-17-yl)-14-(3-amino-3- oxopropyl)-5-(4-hydroxybenzyl)-17-isobutyl-2,l l,20,32-tetramethyl-3,6,9,12,15,18,33- heptaoxo- 1,4, 7, 10, 13,16, 19-heptaazacyclotritriacont-24-ene-20-carboxamide (ALRN-6924), derivative, prodrug, ester, metabolite, or salt thereof.

In certain embodiments, the MDM2 inhibitor is administered in combination with another anticancer agent. A “chemotherapy agent,” “chemotherapeutic,” “anti-cancer agent,” or the like, refer to molecules that are recognized to aid in the treatment of a cancer. Contemplated examples include the following molecules or derivatives such as abemaciclib, abiraterone acetate, methotrexate, paclitaxel, adriamycin, acalabrutinib, brentuximab vedotin, ado-trastuzumab emtansine, aflibercept, afatinib, netupitant, palonosetron, imiquimod, aldesleukin, alectinib, alemtuzumab, pemetrexed disodium, copanlisib, melphalan, brigatinib, chlorambucil, amifostine, aminolevulinic acid, anastrozole, apalutamide, aprepitant, pamidronate disodium, exemestane, nelarabine, arsenic trioxide, ofatumumab, atezolizumab, bevacizumab, avelumab, axicabtagene ciloleucel, axitinib, azacitidine, carmustine, belinostat, bendamustine, inotuzumab ozogamicin, bevacizumab, bexarotene, bicalutamide, bleomycin, blinatumomab, bortezomib, bosutinib, brentuximab vedotin, brigatinib, busulfan, irinotecan, capecitabine, fluorouracil, carboplatin, carfilzomib, ceritinib, daunorubicin, cetuximab, cisplatin, cladribine, cyclophosphamide, clofarabine, cobimetinib, cabozantinib-S-malate, dactinomycin, crizotinib, ifosfamide, ramucirumab, cytarabine, dabrafenib, dacarbazine, decitabine, daratumumab, dasatinib, defibrotide, degarelix, denileukin diftitox, denosumab, dexamethasone, dexrazoxane, dinutuximab, docetaxel, doxorubicin, durvalumab, rasburicase, epirubicin, elotuzumab, oxaliplatin, eltrombopag olamine, enasidenib, enzalutamide, eribulin, vismodegib, erlotinib, etoposide, everolimus, raloxifene, toremifene, panobinostat, fulvestrant, letrozole, filgrastim, fludarabine, flutamide, pralatrexate, obinutuzumab, gefitinib, gemcitabine, gemtuzumab ozogamicin, glucarpidase, goserelin, propranolol, trastuzumab, topotecan, palbociclib, ibritumomab tiuxetan, ibrutinib, ponatinib, idarubicin, idelalisib, imatinib, talimogene laherparepvec, ipilimumab, romidepsin, ixabepilone, ixazomib, ruxolitinib, cabazitaxel, palifermin, pembrolizumab, ribociclib, tisagenlecleucel, lanreotide, lapatinib, olaratumab, lenalidomide, lenvatinib, leucovorin, leuprolide, lomustine, trifluridine, olaparib, vincristine, procarbazine, mechlorethamine, megestrol, trametinib, temozolomide, methylnaltrexone bromide, midostaurin, mitomycin C, mitoxantrone, plerixafor, vinorelbine, necitumumab, neratinib, sorafenib, nilutamide, nilotinib, niraparib, nivolumab, tamoxifen, romiplostim, sonidegib, omacetaxine, pegaspargase, ondansetron, osimertinib, panitumumab, pazopanib, interferon alfa- 2b, pertuzumab, pomalidomide, mercaptopurine, regorafenib, rituximab, rolapitant, rucaparib, siltuximab, sunitinib, thioguanine, temsirolimus, thalidomide, thiotepa, trabectedin, valrubicin, vandetanib, vinblastine, vemurafenib, vorinostat, zoledronic acid, or combinations thereof such as cyclophosphamide, methotrexate, 5 -fluorouracil (CMF); doxorubicin, cyclophosphamide (AC); mustine, vincristine, procarbazine, prednisolone (MOPP); adriamycin, bleomycin, vinblastine, dacarbazine (ABVD); cyclophosphamide, doxorubicin, vincristine, prednisolone (CHOP); bleomycin, etoposide, cisplatin (BEP); epirubicin, cisplatin, 5 -fluorouracil (ECF); epirubicin, cisplatin, capecitabine (ECX); methotrexate, vincristine, doxorubicin, cisplatin (MVAC).

In certain embodiments, the method of administration is in a subject with a lymphodepleted environment due to prior or concurrent administration of lymphodepl eting agents. In certain embodiments, lymphodepleting agents (e.g., cyclophosphamide and fludarabine).

In certain embodiments, the anticancer agent is cytarabine (ara-C) and an anthracycline drug. In certain embodiments, the anthracycline drug is daunorubicin or idarubicin. In certain embodiments, the anticancer agent is cladribine, fludarabine, or etoposide.

In certain embodiments, the subject is diagnosed with leukemia cells having an FLT3 gene mutation and the MDM2 inhibitor is administered in combination with administering midostaurin. In certain embodiments, the subject is diagnosed with leukemia cells having a high expression of CD33 and the MDM2 inhibitor is administered in combination with administering gemtuzumab ozogamicin.

In certain embodiments, the chemotherapy agent is an anti-PD-1, anti-PD-Ll anti-CTLA4 antibody or combinations thereof, such as an anti-CTLA4 (e.g., ipilimumab, tremelimumab) and anti-PDl (e.g., nivolumab, pembrolizumab, cemiplimab) and anti-PD-Ll (e.g., atezolizumab, avelumab, durvalumab).

In certain embodiments, the MDM2 inhibitor is administered before or after receiving bone marrow transplant or blood stem cell transplant and optionally radiation therapy.

In certain embodiments, the subject is a human patient.

In certain embodiments, the subject is diagnosed with cancer or hematological malignancy. In certain embodiments, the hematological malignancy is multiple myeloma, leukemia, or lymphoma. In certain embodiments, the hematological malignancy is acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myelogenous leukemia, acute monocytic leukemia (AMOL), Hodgkin's lymphomas, and non-Hodgkin's lymphomas such as Burkitt lymphoma, B- cell lymphoma.

In certain embodiments, the cancer is selected from bladder cancer, lung cancer, breast cancer, colon cancer, rectal cancer, endometrial cancer, pancreatic cancer, kidney cancer, prostate cancer, thyroid cancer, brain cancer, multiple myeloma, lymphoma, or leukemia.

In certain embodiments, the disclosure also provides the use of MDM2 inhibitors disclosed herein for the preparation of a medicament useful for the treatment of a cancer or neoplasm in a human patient or other mammal. In certain embodiment, this disclosure relates to methods for the treatment a subject at risk of, exhibiting symptoms of, suspected of, or diagnosed with a cancer or neoplasm selected from skin cancer, melanoma, Barret's adenocarcinoma; biliary tract carcinomas; breast cancer; cervical cancer; cholangiocarcinoma; central nervous system tumors including primary CNS tumors such as glioblastomas, astrocytomas (including glioblastoma multiforme) and ependymomas, and secondary CNS tumors (i.e., metastases to the central nervous system of tumors originating outside of the central nervous system), colorectal cancer, including large intestinal colon carcinoma; gastric cancer; carcinoma of the head and neck including squamous cell carcinoma of the head and neck; hematologic cancers including leukemias and lymphomas such as acute lymphoblastic leukemia, acute myelogenous leukemia (AML), myelodysplastic syndromes, chronic myelogenous leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, megakaryoblastic leukemia, multiple myeloma and erythroleukemia; hepatocellular carcinoma; lung cancer including small cell lung cancer and non-small cell lung cancer; ovarian cancer; endometrial cancer; pancreatic cancer; pituitary adenoma; prostate cancer; renal cancer; sarcoma; and thyroid cancers.

The MDM2 inhibitors disclosed herein can be used alone in the treatment of each of the foregoing conditions or can be used to provide additive or potentially synergistic effects with certain existing chemotherapies, radiation, biological or immunotherapeutics (including monoclonal antibodies) and vaccines. The MDM2 inhibitors disclosed herein may be useful for restoring effectiveness of certain existing chemotherapies and radiation and or increasing sensitivity to certain existing chemotherapies and/or radiation.

In certain embodiments, the subject is a human subject is 2, 12, or 16 years old or older. In certain embodiments, the subject is a human subject is 2, 12, or 15 years old or less than 2, 12, or 16 years old. In certain embodiments, the subject is a human subject is 55 or 65 years old or older.

In certain embodiments, the subject is a human subject is an infant, e.g., from one month to two years of age. In certain embodiments, the subject is a human subject is a child, e.g., from one two to twelve years of age. In certain embodiments, the subject is a human subject is an adolescent, e.g., from twelve to sixteen years of age. In certain embodiments, the subject is a human subject sixteen years of age or older.

In certain embodiments, the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4- yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily in an amount of about 10 mg to 500 mg.

In certain embodiments, the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4- yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily for less than 14 days in an amount of about 10 mg to 500 mg.

In certain embodiments, the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4- yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily in an amount of about 500 mg to 1000 mg. In certain embodiments, the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4- yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily for less than 7 days in an amount of about 10 mg to 500 mg.

In certain embodiments, the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4- yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily for less than 7 days in an amount of about 500 mg to 1000 mg.

In certain embodiments, the subject is 55 years or 65 years or older, and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily in an amount of about 10 mg to 500 mg.

In certain embodiments, the subject is 55 years or 65 years or older, and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily for less than 14 days in an amount of about 10 mg to 500 mg.

In certain embodiments, the subject is 55 years or 65 years or older, and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily in an amount of about 500 mg to 1000 mg.

In certain embodiments, the subject is 55 years or 65 years or older, and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily for 7 days or more in an amount of about 10 mg to 500 mg.

In certain embodiments, the subject is 55 years or older, and the MDM2 inhibitor is (2- (lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily for 7 days or more in an amount of about 500 mg to 1000 mg.

In certain embodiments, the subject is an infant, child, or adolescent and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily in an amount of about 10 mg to 500 mg.

In certain embodiments, the subject is an infant, child, or adolescent and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone

(sabizabulin) administered daily for 14 days or more in an amount of about 10 mg to 500 mg.

In certain embodiments, the subject is an infant, child, or adolescent and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone

(sabizabulin) administered daily in an amount of about 500 mg to 1000 mg. In certain embodiments, the subject is an infant, child, or adolescent and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily for 7 days or more in an amount of about 10 mg to 500 mg.

In certain embodiments, the subject is an infant, child, or adolescent and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily for 7 days or more in an amount of about 500 mg to 1000 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a subject in need thereof and administered daily in an amount of about 10 mg to 500 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a subject in need thereof and administered daily for 14 days or 25 days or more in an amount of about 10 mg to 500 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a subject in need thereof and administered daily in an amount of about 500 mg to 1000 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a subject in need thereof and administered daily for 7 days or more in an amount of about 10 mg to 500 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a subject in need thereof and administered daily for 7 days or more in an amount of about 500 mg to 1000 mg. In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a subject in need thereof and the subject is 55 years or 65 years or older, and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily in an amount of about 10 mg to 500 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a subject in need thereof and the subject is 55 years or 65 years or older, and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily for 14 days or 25 days or more in an amount of about 10 mg to 500 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a subject in need thereof and the subject is 55 years or 65 years or older, and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily in an amount of about 500 mg to 1000 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a subject in need thereof and the subject is 55 years or 65 years or older, and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily for less than 7 days in an amount of about 10 mg to 500 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone

(sabizabulin) to a subject in need thereof and the subject is 55 years or older, and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone

(sabizabulin) administered daily for 7 days or more in an amount of about 500 mg to 1000 mg. In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a subject in need thereof and the subject is an infant, child, or adolescent and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily in an amount of about 10 mg to 500 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a human subject in need thereof and the human subject is an infant, child, or adolescent and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5- trimethoxyphenyl)methanone (sabizabulin) administered daily for 14 days or 25 days or more in an amount of about 10 mg to 500 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a human subject in need thereof and the human subject is an infant, child, or adolescent and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5- trimethoxyphenyl)methanone (sabizabulin) administered daily in an amount of about 500 mg to 1000 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a human subject in need thereof and the human subject is an infant, child, or adolescent and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5- trimethoxyphenyl)methanone (sabizabulin) administered daily for more than 14 day or more than 25 days in an amount of about 10 mg to 500 mg.

In certain embodiments, this disclosure relates to a method of treating leukemia or acute myelogenous leukemia (AML) comprising administering an effective amount of a MDM2 inhibitor, e.g., (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) to a human subject in need thereof and the human subject is an infant, child, or adolescent and the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol-4-yl)(3,4,5- trimethoxyphenyl)methanone (sabizabulin) administered daily for more than 14 days or more than 25 days in an amount of about 500 mg to 1000 mg.

Sabizabulin (VERU-111) to treat acute myeloid leukemia (AML)

Acute myeloid leukemia (AML) is an aggressive form of leukemia that is initiated in immature forms of blood-forming cells commonly known as myeloid cells. Myeloid cells are found in bone marrow. AML is the most common type of aggressive leukemia found in older adults with a median age of diagnosis at 66 years. AML is reported to have the lowest survival rates of all types of leukemia. It is the second most common type of leukemia diagnosed in adults and children.

VERU-111/sabizabulin is a microtubule inhibitor. It has been discovered that VERU-111 also had inhibitory activity against MDM2, an oncogene that is frequently overexpressed in AML and has been associated with disease progression and poor treatment outcomes. VERU-111 also has therapeutic activity in AML cell culture and xenograft models.

Experiments indicate VERU-111 inhibits MDM2. Western blot shows pi tubulin inhibition, activation of caspase-3 and -9 as well as cleavage of death substrate PARP in AML cell lines THP1 and M0LM14 (Human leukemia cell line) following treatment with VERU-111 for 24 h. Western blot assays were used to evaluate MDM2 expression in AML cell lines (with different p53 status) that were treated for 24 h with 50 nM of either VERU-111 or PTX. The western blot assays indicate dose-response and time-course MDM2 inhibition by VERU-111 in M0LM14 cells.

Experiments indicate that VERU-111 inhibits MDM2 through inducing protein degradation. A M0LM14 cells were treated with or without VERU-111 (10 nM for 8h) and 10 pM MG 132 (cell-permeable proteasome inhibitor) for additional 4 h. Western blots was performed for expression of proteins. CHX chase assay was used for detection of MDM2 and p53 turnover in M0LM14 cells treated with VERU-111 for 4 h. IP and Western blot assay were performed using anti-MDM2 and anti -ubiquitin antibodies, to detect effects of VERU-111 (10 nM) on ubiquitination of endogenous MDM2 in M0LM14 cells.

Claims

1. A method of treating a hematological cancer comprising administering an effective amount of a MDM2 inhibitor to a subject in need thereof.

2. The method of claim 1, wherein the MDM2 inhibitor is sabizabulin.

3. The method of claim 2, wherein the hematological cancer is acute myeloid leukemia,

4. The method of claim 3, wherein the MDM2 inhibitor is administered in combination with another anti cancer agent.

5. The method of claim 4, wherein the anticancer agent is cytarabine (ara-C) and an anthracycline drug.

6. The method of claim 5, wherein the anthracycline drug is daunorubicin or idarubicin.

7. The method of claim 4 wherein the subject is diagnosed with leukemia cells having an FLT3 gene mutation and administering midostaurin.

8. The method of claim 4, wherein the anticancer agent is cladribine, fludarabine, or etoposide.

9. The method of claim 4 wherein the subject is diagnosed with leukemia cells having a high expression of CD33 and administering gemtuzumab ozogamicin.

10. The method of claim 3, wherein the MDM2 inhibitor is administered before or after receiving bone marrow transplant or blood stem cell transplant and optionally radiation therapy.

11. The method of claim 3, wherein the subject is a human subject is 12 years old or less.

12. The method of claim 3, wherein the subject is a human subject is 55 years old or more.

13. The method of claim 3, wherein the subject is a human subject is a child or adolescent.

14. The method of claim 3, wherein the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol- -yl)(3, 4, 5 -trimethoxy phenyl)methanone (sabizabulin) administered daily in an amount of about 10 mg to 500 mg.

15. The method of claim 3, wherein the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol- -yl)(3,4,5-trimethoxyphenyl)methanone (sabizabulin) administered daily in an amount of about 500 mg to 1000 mg.

15. The method of claim 3, wherein the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol- -yl)(3, 4, 5 -trimethoxy phenyl)methanone (sabizabulin) administered daily.

16. The method of claim 3, wherein the MDM2 inhibitor is (2-(lH-indol-3-yl)-lH-imidazol- -yl)(3, 4, 5 -trimethoxy phenyl)methanone (sabizabulin) administered daily for less than 14 days.