EA045240B1 - CANCER TREATMENT METHODS - Google Patents

Description

Field of technology

The present invention describes methods of treating cancer, including myeloproliferative neoplasm (MPN), using an inhibitor of murine dual microprotein 2 homolog (MDM2).

BACKGROUND OF THE INVENTION p53 is a tumor suppressor and transcription factor that responds to cellular stress by activating the transcription of multiple genes involved in cell cycle arrest, apoptosis, aging, and DNA repair. Unlike normal cells, which infrequently induce p53 activation, tumor cells are subject to constant cellular stress from various insults, including hypoxia and proapoptotic oncogene activation. Thus, there is a strong selective advantage for inactivation of the p53 pathway in tumors, and it has been proposed that ablation of p53 function may be a prerequisite for tumor survival. In support of this notion, three groups of researchers used mouse models to demonstrate that the absence of p53 function is a persistent requirement for the maintenance of established tumors. When the researchers restored p53 function in tumors with inactivated p53, the tumors regressed.

p53 is inactivated by mutation and/or loss in 50% of solid tumors and 10% of liquid tumors. Other key players in the p53 pathway are also altered genetically or epigenetically in cancer. MDM2, an oncoprotein, suppresses p53 function and is activated by gene amplification at a frequency reported to be as high as 10%. MDM2 is in turn inhibited by another tumor suppressor, p14ARF. It has been suggested that changes in the chain of events downstream of p53 may be responsible for at least partial inactivation of the p53 pathway in p53 WT (p53 wild-type) tumors. In support of this concept, some p53WT tumors appear to exhibit a reduced capacity for apoptosis, although their ability to undergo cell cycle arrest remains unaffected. One cancer treatment strategy involves the use of small molecules that bind MDM2 and neutralize its interaction with p53. MDM2 inhibits p53 activity through three mechanisms: 1) acting as an E3 ubiquitin ligase, promoting p53 degradation; 2) binding to and blocking the transcription activation domain of p53; and 3) exporting p53 from the nucleus to the cytoplasm. All three mechanisms can be blocked by neutralizing the MDM2-p53 interaction. In particular, this therapeutic strategy can be applied to tumors that are p53 WT, and studies with small molecule MDM2 inhibitors have shown promise in reducing tumor growth both in vitro and in vivo. Additionally, in patients with p53-inactivated tumors, stabilization of wild-type p53 in normal tissues by MDM2 inhibition may provide selective protection of normal tissues from mitotic poisons. As used in the present invention, MDM2 refers to human MDM2 protein, and p53 refers to human p53 protein. It should be noted that human MDM2 may also be referred to as HDM2 or hMDM2. Several MDM2 inhibitors are in human clinical trials for the treatment of various types of cancer.

Myeloproliferative neoplasms (MPN), including but not limited to polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF), are clonal hematopoietic stem cell (HSC) disorders characterized by clonal proliferation of terminally differentiated myeloid cells. Approximately 1, 4, and 20% of patients with ET, PV, and PMF, respectively, enter the blast phase (BP), called MPN-BP, within a 10-year period from diagnosis. Cervantes F, et al., Acta Haematol, 1991; 85(3): 124-127. MPN-BP and de novo acute myeloid leukemia (AML) each have distinct mutational patterns and clinical courses. Rampal R, et al., Proc Natl Acad Sci USA. 2014; 111(50):E5401-10. Patients with MPN-BP have a particularly dismal prognosis, with a median survival of less than 6 months with currently available treatments.

The present invention relates to methods of treating a myeloproliferative neoplasm in humans with an MDM2 inhibitor, alone or in combination with one or more additional pharmaceutically active agents.

The present invention relates to the use of an MDM2 inhibitor, alone or in combination with one or more additional pharmaceutically active agents, for the treatment of myeloproliferative neoplasm (MPN).

Summary of the present invention

The present invention relates to a method of treating a myeloproliferative neoplasm (MPN), comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II):

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or a pharmaceutically acceptable salt thereof. In one embodiment, the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia, idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome (MDS), and systemic mast cell disease (SMCD). In one embodiment, the MPN is selected from the group consisting of polycythemia vera (PV), primary myelofibrosis, and essential thrombocythemia. In one embodiment, the MPN is polycythemia vera (PV). In one embodiment, the MPN is essential thrombocythemia. In one embodiment, the MPN is myelofibrosis. In one embodiment, the myelofibrosis is selected from primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF). In one embodiment, the person has not benefited from ruxolitinib therapy against any of the MPNs above. In one embodiment, a person has failed treatment with ruxolitinib for myelofibrosis. In one embodiment, a person has failed treatment with ruxolitinib for primary myelofibrosis (PMF). In one embodiment, a person has failed treatment with ruxolitinib for myelofibrosis following polycythemia vera (post-PV MF). In one embodiment, a person has failed treatment with ruxolitinib for post-essential thrombocythemia myelofibrosis (post-ET MF).

In one embodiment, the present invention provides a method of treating polycythemia vera (PV), comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a method of treating essential thrombocythemia, comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a method of treating primary myelofibrosis, comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a method of treating idiopathic myelofibrosis, comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a method of treating chronic myelogenous leukemia (CML), comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a method of treating acute myelogenous leukemia (AML), comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a method of treating Merkel cell carcinoma (MCC), comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof .

In one embodiment, the present invention provides a method of treating systemic mastocytosis (SM), comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to a method of treating chronic neutrophilic leukemia (CNL), comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or pharmaceutically acceptable salt.

In one embodiment, the present invention provides a method of treating myelodysplastic syndrome (MDS), comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a method of treating systemic mast cell disease (SMCD), comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof .

In one embodiment, the thrombocythemia is essential thrombocythemia (ET).

In one embodiment, the compound of formula (I) or formula (II) is in crystalline form. In one embodiment, the crystalline form is characterized by an X-ray powder diffraction pattern containing at least three peaks at diffraction angles of 2 theta degrees, selected from the group consisting of peaks at approximately 11.6, 12.4, 18.6, 19 .0, 21.6 and 23.6±0.1.

In one embodiment, the compound of formula (I) or formula (II) is in amorphous form.

In one embodiment, the compound of formula (I) or formula (II) is in free form

In one embodiment, the MDM2 inhibitor is a pharmaceutically acceptable salt of a compound of formula (I) or formula (II).

In one embodiment, a compound of formula (I) or formula (II) is administered once daily at a dose selected from the group consisting of 15, 25, 3θ, 50, 6θ, 75, 90, 100, 120, 150, 175 , 180, 200, 225, 240, 250, 275, 300, 325, 350, 360, 375 and 480 mg.

In one embodiment, a compound of formula (I) or formula (II) is administered twice daily at a dose selected from the group consisting of 15, 25, 30, 50, 60, 75, 90, 100, 120, 150, 175, 180, 200, 225, 240, 250, 275, 300, 325, 350, 360, 375 and 480 mg.

In one embodiment, a human is treated with a compound of formula (I) for a period selected from the group consisting of about 14 days, about 21 days, about 28 days, about 35 days, about 42 days, about 49 days, and about 56 days.

In one embodiment, a compound of formula (I) or formula (II) is administered orally.

An MDM2 inhibitor for use in the treatment of a myeloproliferative neoplasm (MPN), comprising the step of administering to a person in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

Brief description of drawings

The above summary as well as the following detailed description of the present invention will be better understood when read in conjunction with the accompanying drawings.

Fig. 1 shows an X-ray powder diffraction (XRPD) pattern of the compound of formula (I) in crystalline anhydrous form.

Fig. 2 shows that the compound of formula (I) inhibits the proliferation of p53 wild type (WT) tumor cells.

Fig. 3 shows that the compound of formula (I) inhibits in vivo growth of SJSA-1 (MDM2 amplified) tumors in a dose-dependent manner. ED ₅₀ =9.0 mg/kg.

Fig. 4 shows that the compound of formula (I) inhibits in vivo tumor growth of HCT116 (KRAS) in a dose-dependent manner. ED ₅₀ =31.6 mg/kg.

Fig. 5 shows that the compound of formula (I) inhibits in vivo A375sq2 (BRAF) tumor growth in a dose-dependent manner. ED ₅₀ =18 mg/kg.

Fig. 6 shows the p21 fold induction of the compound of formula (I) at various doses.

Fig. 7 shows that the compound of formula (I) inhibits AML tumor growth in a dose-dependent manner.

Fig. 8 shows that the compound of formula (I) inhibits AML tumor growth in a mouse model in a dose-dependent manner.

Fig. 9 shows that the compound of formula (I) inhibits the cell cycle.

Fig. 10 shows that the compound of formula (I) induces apoptosis in wild-type p53 AML cell lines.

Detailed Description of the Present Invention

Although preferred embodiments of the present invention are shown and described herein, these embodiments are presented by way of example only and are not intended to otherwise limit the scope of the present invention. Various alternatives to the described embodiments of the present invention can be used in the practice of the present invention.

Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one skilled in the art to which the present invention relates.

The terms administered in combination with and co-administered, as used herein, encompass the administration of two or more active pharmaceutical ingredients to a subject in such a manner that both agents and/or their metabolites are present in the subject simultaneously. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in compositions in which two or more agents are present.

The term effective amount or therapeutically effective amount refers to an amount of an active pharmaceutical ingredient or combination of active pharmaceutical ingredients as described herein that is sufficient to accomplish the intended use, including, but not limited to, the treatment of a disease. The therapeutically effective amount may vary depending on the intended use (in vitro or in vivo) or the subject and disease condition being treated (eg, weight, age, and sex of the subject), severity of the disease condition, route of administration, and other factors that can be readily determined. specialist in this field of technology. The term also refers to a dose that will produce a specific response in target cells (eg, decreased platelet adhesion and/or cell migration). The specific dosage will vary depending on the specific compounds selected, the dosage regimen to be followed, whether the compound is administered in combination with other compounds, the time of administration, the tissue into which it is administered, and the physical delivery system in which the compound is transported.

The terms enantiomerically enriched, enantiomerically pure and non-racemic, as used herein, refer to compositions in which the percentage by weight of one enantiomer is greater than the amount of that one enantiomer in the reference mixture of the racemic composition (eg, greater than 1:1 by weight). For example, an enantiomerically enriched preparation of the (S)enantiomer means a preparation of a compound containing more than 50% by weight of the (S)-enantiomer relative to the (R)-enantiomer, such as at least 75% by weight, such as at least at least 80% by weight. In some embodiments, the fortification may be significantly greater than 80% by weight, providing a substantially enantiomerically enriched, substantially enantiomerically pure, or substantially non-racemic drug, which refers to drug compositions that contain at least 85% by weight of one enantiomer relative to the other enantiomer such as at least 90% by weight, and such as at least 95% by weight. The terms diastereomerically enriched and diastereomerically pure, as used herein, refer to compositions in which the percentage by weight of one diastereomer is greater than the amount of that one diastereomer in the control mixture of diastereomers. In some embodiments, the enrichment may be significantly greater than 80% by weight, providing a substantially diastereomerically enriched or substantially diastereomerically pure drug, which refers to formulations of compositions that contain at least 85% by weight of one diastereomer relative to the other diastereomers, such as at least 90% by weight, and such as at least 95% by weight

In preferred embodiments, the enantiomerically enriched composition has a higher potency in terms of therapeutic benefit per unit weight than the racemic mixture of the composition. Enantiomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the preparation and crystallization of chiral salts; or the preferred enantiomers can be prepared by asymmetric synthesis. See, for example, Jacques, et al., Enantiomers, Racemates and Resolutions, Wiley Interscience, New York (1981); E. L. Eliel and S. H. Wilen, Stereochemistry of Organic Compounds, Wiley-Interscience, New York (1994).

Enantiomeric purity, as used herein, refers to the relative amounts, expressed as a percentage, of the presence of a particular enantiomer relative to another enantiomer. For example, if a compound that potentially has the (R) or (S) isomeric configuration is present as a racemic mixture, the enantiomeric purity is about 50% relative to either the (R) or (S) isomer. If in a given compound one isomeric form predominates over the other, for example, 80% (S) - and 20% (R) -, the enantiomeric purity of the compound relative to the (S) isomeric form is 80%. The enantiomeric purity of a compound can be determined by a number of methods known in the art, including, but not limited to, chromatography using a chiral support, polarimetric rotation measurement of polarized light, nuclear magnetic resonance spectroscopy using chiral shift reagents, which include, but are not limited to, These include chiral complexes containing lanthanides or Pirkle's alcohol, or derivatization of compounds using a chiral compound such as a Mosher acid followed by chromatography or nuclear magnetic resonance spectroscopy.

The term IC ₅₀ refers to the half-maximal inhibitory concentration, i.e. inhibition of 50% of the required activity. The term EC ₅₀ refers to the drug concentration at which the half-maximal response is achieved.

Isomers are different compounds that have the same molecular formula. Stereoisomers are isomers that differ only in the way the atoms are arranged in space, i.e. have different stereochemical configurations. Enantiomers are a pair of stereoisomers that are incompatible mirror images of each other. A 1:1 mixture of a pair of enantiomers is a racemic mixture. The term (±) is used to denote a racemic mixture where appropriate. Diastereoisomers are stereoisomers that contain at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is determined according to the Cahn-Ingold-Prelog R-S system. When a compound is a pure enantiomer, the stereochemistry of each chiral carbon atom may be indicated by R or S. Separated compounds whose absolute configuration is unknown may be designated (+) or (-) depending on the direction (dextro- or levorotatory), in which they rotate plane-polarized light at the wavelength of the sodium D line. Some of the compounds described in the present invention contain one or more asymmetric centers and, thus, can give enantiomers, diastereomers and other stereoisomeric forms, which can be defined in terms of absolute stereochemistry as (R) - or (S) -. The present chemical compounds, pharmaceutical compositions and methods are intended to include all given possible isomers, including racemic mixtures, optically pure forms and intermediate mixtures. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents or can be separated using conventional methods. When the compounds described in the present invention contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise indicated, the compounds are intended to include both E and Z geometric isomers.

MPN-BP refers to the blast phase (BP) of myeloproliferative neoplasms (MPNs) described herein.

A pharmaceutically acceptable carrier or pharmaceutically acceptable excipient is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, and absorption retarding agents. The use of these media and agents for active pharmaceutical ingredients is well known in the art. Unless any conventional vehicle or agent is incompatible with the active pharmaceutical ingredient, it is contemplated for use in the therapeutic compositions of the present invention. Additional active ingredients may also be included in the compositions described.

The term pharmaceutically acceptable salt refers to salts derived from a variety of organic and inorganic counterions known in the art. Pharmaceutically acceptable acid addition salts can be prepared with inorganic acids and organic acids. Inorganic acids from which salts can be prepared include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid. Organic acids from which salts can be prepared include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid and salicylic acid. Pharmaceutically acceptable base addition salts can be prepared with inorganic and organic bases. Inorganic bases from which salts can be prepared include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese and aluminum. Organic bases from which salts can be prepared include, for example, primary, secondary and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines and basic ion exchange resins. Specific examples include isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine and ethanolamine. In selected embodiments, the pharmaceutically acceptable base addition salt is selected from ammonium, potassium, sodium, calcium, and magnesium salts. The term cocrystal refers to a molecular complex derived from a number of cocrystal formers known in the art. Unlike a salt, a cocrystal typically does not involve proton transfer between the cocrystal and the drug, and instead involves intermolecular interactions such as hydrogen bonding, aromatic ring stacking, or dispersion forces between the cocrystal former and the drug in the crystal structure.

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The terms QD, qd or q.d. stand for quaque die, or once a day. The terms BID, bid or b.i.d. means bis in die, or twice a day. The terms TID, tid or t.i.d. stands for ter in die or three times a day. The terms QID, qid or q.i.d. denote quater in die or four times a day.

Solvate refers to a compound in the form of a physical associate with one or more molecules of a pharmaceutically acceptable solvent.

Therapeutic effect, as this term is used in the present invention, includes a therapeutic effect and/or a prophylactic effect, as described above. A prophylactic effect includes delaying or eliminating the onset of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, stopping or reversing the progression of a disease or condition, or any combination thereof.

When ranges are used in the present invention to describe, for example, physical or chemical properties such as molecular weight or chemical formulas, all combinations and subcombinations of ranges and specific embodiments are intended to be included therein. Applying the term approximately when referring to a number or numerical range means that the number or numerical range stated is approximate within experimental variability (or within statistical experimental error), and thus the number or numerical range can vary, for example, from 1 to 15% of the specified number or number range. The term comprising (and related terms such as contain or contains or having or including) includes embodiments such as, for example, an embodiment of any composition of a substance, method or process that consists of or consists essentially of the features described.

The compounds of the present invention also include crystalline and amorphous forms of the compound of formula (I) or formula (II), including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs and amorphous forms of the compounds, as well as the mixtures. Crystalline form and polymorph are intended to include all crystalline and amorphous forms of a compound, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs and amorphous forms, as well as mixtures thereof, unless a specific crystalline or amorphous form.

Methods of treating cancer.

The present invention relates to a method of treating cancer, comprising the step of administering to a human in need an inhibitor of murine dual microprotein homolog 2 (MDM2) or a pharmaceutically acceptable salt thereof, wherein the cancer is selected from the group consisting of carcinomas such as bladder, breast, colon cancer , rectum, kidney, liver, lung (small cell lung cancer and non-small cell lung cancer), esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate, head and neck, and skin (including basal and squamous cell cancer, melanoma, Merkel cell carcinoma, Kaposi's sarcoma, skin lymphoma); hematopoietic tumors of the lymphoid lineage (including leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma); hematopoietic tumors of myeloid origin (including acute and chronic myelogenous leukemia, myelodysplastic syndrome and promyelocytic leukemia); tumors of mesenchymal origin (including fibrosarcoma and rhabdomyosarcoma and other sarcomas, such as soft tissue and bone); tumors of the central and peripheral nervous system (including astrocytoma, neuroblastoma, glioma, glioblastoma and schwannomas); and other tumors (including melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderoma pigmentosa, keratoctantoma and follicular thyroid cancer). In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

The present invention also relates to the use of a murine dual microprotein homolog 2 (MDM2) inhibitor or a pharmaceutically acceptable salt thereof for the treatment of cancer in a human in need, wherein the cancer is selected from the group consisting of carcinomas such as bladder, breast, colon, rectum, kidney, liver, lung (small cell lung cancer and non-small cell lung cancer), esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate, head and neck, and skin (including basal and squamous cell carcinoma , melanoma, Merkel cell carcinoma, Kaposi's sarcoma, skin lymphoma); hematopoietic tumors of the lymphoid lineage (including leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkey's lymphoma tta); hematopoietic tumors of myeloid origin (including acute and chronic myelogenous leukemia, myelodysplastic syndrome and promyelocytic leukemia); tumors of mesenchymal origin (including fibrosarcoma and rhabdomyosarcoma and other sarcomas, such as soft tissue and bone); tumors of the central and peripheral nervous system (including astrocytoma, neuroblastoma, glioma, glioblastoma and schwannomas); and other tumors (including melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderoma pigmentosa, keratoctantoma and follicular thyroid cancer). In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

Methods for Treating Myeloproliferative Neoplasm (MPN) The present invention also provides a method for treating MPN, comprising the step of administering to a human in need an inhibitor of murine dual microprotein homolog 2 (MDM2) or a pharmaceutically acceptable salt thereof. In one embodiment, the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome ( MDS), systemic mast cell disease (SMCD). In one embodiment, the myelofibrosis is selected from the group consisting of primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF). In one embodiment, primary myelofibrosis (PMF) is selected from the group consisting of pre-fibrotic PFM/early stage PFM and advanced fibrotic stage PFM. In one embodiment, the MPN is selected from the group consisting of chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia, chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia (JMML), hypereosinophilic syndromes (HES) , and myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T). In one embodiment, polycythemia vera is phlebotomy-dependent polycythemia vera. In one embodiment, the individual is determined to be hydroxyurea (HU) intolerant (unacceptable side effects). In one embodiment, the human subject is determined to be unresponsive to hydroxyurea (HU) (inadequate response). In one embodiment, the human subject suffers from splenomegaly. In one embodiment, the human subject suffers from splenomegaly and is phlebotomy dependent. In one embodiment, the human subject is phlebotomy dependent without splenomegaly. In one embodiment, the person is refractory to ruxolitinib therapy. Failure of ruxolitinib therapy includes, but is not limited to, (i) failure to reduce the severity or progression of any MPN in a person receiving ruxolitinib, or (ii) relapse of any MPN in a person after ruxolitinib therapy. In one embodiment, ruxolitinib therapy failure is the failure to reduce the severity or progression of any MPN in a person receiving ruxolitinib. In one embodiment, failure of ruxolitinib therapy is the recurrence of any MPN in a person after therapy with ruxolitinib. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating MPN, comprising the step of administering to a human in need an inhibitor of murine dual microprotein 2 (MDM2) homolog, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, in wherein MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis and essential thrombocythemia (ET). In one embodiment, polycythemia vera is phlebotomy-dependent polycythemia vera. In one embodiment, the human subject is determined to be suffering from an intolerance

- 7 045240 hydroxyurea (HM) (unacceptable side effects). In one embodiment, the human subject is determined to be unresponsive to hydroxyurea (HU) (inadequate response). In one embodiment, the human subject suffers from splenomegaly. In one embodiment, the individual suffers from splenomegaly and is phlebotomy dependent. In one embodiment, the human subject is phlebotomy dependent without splenomegaly. In one embodiment, the human subject has failed prior therapy with ruxolitinib.

The present invention also provides a method of treating MPN (MPN-BP) in the blast phase, comprising the step of administering to a human in need an inhibitor of murine dual microprotein 2 homolog (MDM2) or a pharmaceutically acceptable salt thereof. In one embodiment, the MPN-BP is selected from the group consisting of blast-phase polycythemia vera (BP-PV), blast-phase myelofibrosis, blast-phase primary myelofibrosis, blast-phase thrombocythemia, blast-phase essential thrombocythemia (BP-ET), idiopathic blast-phase myelofibrosis phase, systemic blast phase mastocytosis (BP-SM), blast phase chronic neutrophilic leukemia (BP-CNL), blast phase myelodysplastic syndrome (BP-MDS) and blast phase systemic mast cell disease (BP-SMCD). In one embodiment, the blast-phase myelofibrosis is selected from the group consisting of blast-phase primary myelofibrosis (BP-PMF), blast-phase polycythemia vera myelofibrosis (BP-post-PV MF), and blast-phase essential thrombocythemia myelofibrosis (BP-post) -ET MF). In one embodiment, blast phase primary myelofibrosis (BP-PMF) is selected from the group consisting of pre-fibrotic PMF/early blast phase PMF and advanced fibrotic blast phase PMF. In one embodiment, the MPN-BP is selected from the group consisting of blast phase chronic neutrophilic leukemia (BP-CNL), blast phase chronic eosinophilic leukemia, blast phase chronic myelomonocytic leukemia (BP-CMML), blast phase atypical chronic myeloid leukemia (BP -aCML), blast phase juvenile myelomonocytic leukemia (BP-JMML), blast phase hypereosinophilic syndromes (BP-HES), and blast phase myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (BP-MDS/MPN-RS-T). In one embodiment, blast phase polycythemia vera is phlebotomy-dependent polycythemia vera. In one embodiment, the individual is determined to be hydroxyurea (HU) intolerant (unacceptable side effects). In one embodiment, the human subject is determined to be unresponsive to hydroxyurea (HU) (inadequate response). In one embodiment, the human subject suffers from splenomegaly. In one embodiment, the human subject suffers from splenomegaly and is phlebotomy dependent. In one embodiment, the human subject is phlebotomy dependent without splenomegaly. In one embodiment, the person is refractory to ruxolitinib therapy. Failure of ruxolitinib therapy includes, but is not limited to, (i) failure to reduce the severity or progression of any MPN in a person receiving ruxolitinib, or (ii) relapse of any MPN in a person after ruxolitinib therapy. In one embodiment, ruxolitinib therapy failure is the failure to reduce the severity or progression of any MPN-BP in a person receiving ruxolitinib. In one embodiment, ruxolitinib therapy failure is relapse of any MPN-BP in a person following ruxolitinib therapy. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is administered at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily day, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily , 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily once daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is administered to a human subject according to Dosage and Dosage Regimen.

In one embodiment, the myelofibrosis is selected from primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF).

In one embodiment, the myelofibrosis is selected from primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF), and the person has not benefited from ruxolitinib therapy for the treatment of PMF, post-PV MF or post ET MF.

In one embodiment, MPN is characterized by a CALR mutation (calreticulin located on chromosome 19p13.2), as described in Massie, et al., N Engl J. Med. 2013, 25: 2379-2390 and is incorporated herein by reference in its entirety.

In one embodiment, MPN is characterized by an MPL mutation (myeloproliferative leukemia virus oncogene; located on chromosome 1p34), as described in Pikman, et al., Plos Med. 2006; 3(7):e270 and is incorporated herein by reference in its entirety.

In one embodiment, MPN is characterized by the JAK2V617F mutation. JAK2V617F is a functional mutation promoting cytokine-independent growth of myeloid cells and constitutes the majority of myeloproliferative neoplasms (MPNs) as described by Nakatake et al (Oncogene, 2012, 31, 1323-1333) and is incorporated herein by reference in its entirety completeness.

In one embodiment, MPN is characterized by one or more mutations selected from the group consisting of JAK2V617F, MPL, CALR, and mixtures thereof.

In one embodiment, the present invention provides a method of treating polycythemia vera (PV) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II ).

In one embodiment, the present invention provides a method of treating polycythemia vera (PV) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC5998 4 , CHEMBL2386350, MK-8242, DS-3032, DS3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating polycythemia vera (PV) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating essential thrombocythemia (ET) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II ).

In one embodiment, the present invention provides a method of treating essential thrombocythemia (ET) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC5998 4 , CHEMBL2386350, MK-8242, DS3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating essential thrombocythemia (ET) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating primary myelofibrosis in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II).

In one embodiment, the present invention provides a method of treating primary myelofibrosis in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II ), RG7388, triptolide,

- 9 045240

HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8 242,DS- 3032, DS3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating primary myelofibrosis in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II ), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating post-polycythemia vera myelofibrosis (post-PV MF) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I ) or formula (II).

In one embodiment, the present invention provides a method of treating post-polycythemia vera myelofibrosis (post-PV MF) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of: formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin- 3b, sardomethane, NSC59984,

CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating post-polycythemia vera myelofibrosis (post-PV MF) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of: formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating post-essential thrombocythemia myelofibrosis (post-ET MF) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I ) or formula (II).

In one embodiment, the present invention relates to a method of treating post-essential thrombocythemia myelofibrosis (post-ET MF) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of: formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin- 3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention relates to a method of treating post-essential thrombocythemia myelofibrosis (post-ET MF) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of: formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating chronic neutrophilic leukemia (CNL) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I) or formula ( II).

In one embodiment, the present invention provides a method of treating chronic neutrophilic leukemia (CNL) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I) , formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating chronic neutrophilic leukemia (CNL) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I) , formulas (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601

- 10 045240 and their pharmaceutically acceptable salts.

In one embodiment, the present invention provides a method of treating chronic eosinophilic leukemia in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II).

In one embodiment, the present invention provides a method of treating chronic eosinophilic leukemia in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula ( II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEM BL2386350, MK -8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating chronic eosinophilic leukemia in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula ( II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating chronic myelomonocytic leukemia (CMML) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I) or formula ( II).

In one embodiment, the present invention provides a method of treating chronic myelomonocytic leukemia (CMML) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I) , formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating chronic myelomonocytic leukemia (CMML) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I) , formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating atypical chronic myeloid leukemia (aCML) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II).

In one embodiment, the present invention provides a method of treating atypical chronic myeloid leukemia (aCML) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I ), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane , NSC59984, CHEMBL2386350, MK8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention relates to a method of treating atypical chronic myeloid leukemia (aCML) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I ), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating juvenile myelomonocytic leukemia (JMML) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I) or formula ( II).

In one embodiment, the present invention provides a method of treating juvenile myelomonocytic leukemia (JMML) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I) , formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-11 045240

8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating juvenile myelomonocytic leukemia (JMML) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I) , formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating hypereosinophilic syndromes (HES) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II ).

In one embodiment, the present invention provides a method of treating hypereosinophilic syndromes (HES) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984 CHEMBL2386350 , MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating hypereosinophilic syndromes (HES) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention relates to a method of treating myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T) in a human, which includes the step of administering to the human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is a compound of formula (I) or formula (II).

In one embodiment, the present invention relates to a method of treating myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T) in a human, which includes the step of administering to the human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin- 3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention relates to a method of treating myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T) in a human, which includes the step of administering to the human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof .

In one embodiment, the present invention provides a method of treating MPN in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily day, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily , 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome ( MDS) and systemic mast cell disease (SMCD). In one embodiment, myelofibrosis is selected from the group consisting of primary

- 12 045240 myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF) and myelofibrosis after essential thrombocythemia (post-ET MF). In one embodiment, primary myelofibrosis (PMF) is selected from the group consisting of pre-fibrotic/early stage PMF and advanced fibrotic stage PMF. In one embodiment, the MPN is selected from the group consisting of chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia, chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia (JMML), hypereosinophilic syndromes (HES) , and myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T).

In one embodiment, the present invention provides a method of treating MPN in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily day, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily , 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily, wherein MPN is selected from the group consisting of polycythemia vera (PV), primary myelofibrosis, and essential thrombocythemia (ET).

In one embodiment, the present invention provides a method of treating MPN in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily day, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily , 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily, wherein MPN is selected from the group consisting of chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia, chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia leukemia (JMML), hypereosinophilic syndromes (HES), and myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T).

In one embodiment, the present invention provides a method of treating polycythemia vera (PV) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once per day. day, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily , 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily once daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily per day, 375 mg twice daily, and 480 mg twice daily.

In one embodiment, the present invention provides a method of treating primary myelofibrosis in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200

- 13 045240 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily , 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily.

In one embodiment, the present invention provides a method of treating post-polycythemia vera myelofibrosis (post-PV MF) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor, or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily day, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily , 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily.

In one embodiment, the present invention relates to a method of treating post-essential thrombocythemia myelofibrosis (post-ET MF) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily day, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily , 360 mg twice daily, 375 mg twice daily, and 480 mg BID.

In one embodiment, the present invention provides a method of treating essential thrombocythemia (ET) in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once per day. day, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily , 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily once daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily per day, 375 mg twice daily, and 480 mg twice daily.

In one embodiment, the present invention provides a method of treating MPN in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily , 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily once daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily

- 14 045240 day, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome ( MDS), systemic mast cell disease (SMCD). In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts. In one embodiment, the myelofibrosis is selected from the group consisting of primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF). In one embodiment, primary myelofibrosis (PMF) is selected from the group consisting of pre-fibrotic PFM/early stage PFM and advanced fibrotic stage PFM. In one embodiment, the MPN is selected from the group consisting of chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia, chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia (JMML), hypereosinophilic syndromes (HES) , and myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T).

In one embodiment, the present invention provides a method of treating MPN in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily , 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily once daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily day, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, and essential thrombocythemia (ET). In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts. In one embodiment, the myelofibrosis is selected from the group consisting of primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF). In one embodiment, primary myelofibrosis (PMF) is selected from the group consisting of pre-fibrotic/early stage PFM and advanced fibrotic stage PFM. In one embodiment, the MPN is selected from the group consisting of chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia, chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia (JMML), hypereosinophilic syndromes (HES) , and myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T).

In one embodiment, the present invention provides a method of treating polycythemia vera (PV) in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily per day, 250

- 15 045240 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily , 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239EE, RO8994 , nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, the present invention provides a method of treating primary myelofibrosis in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once per day. day, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily , 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily once daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily per day, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention relates to a method of treating post-polycythemia vera myelofibrosis (post-PV MF) in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group , consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily day, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily , 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily once daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, the present invention relates to a method of treating post-essential thrombocythemia myelofibrosis (post-ET MF) in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an inhibitor

- 16 045240

MDM2 or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily , 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily once daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily per day, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, Mx69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, the present invention provides a method of treating essential thrombocythemia (ET) in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily day, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily , 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239EE, RO8994 , nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, the present invention relates to a method of treating chronic neutrophilic leukemia (CNL) in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily day, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily , 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-17 045240

1601 and their pharmaceutically acceptable salts.

In one embodiment, the present invention provides a method of treating chronic eosinophilic leukemia in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily day, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily , 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily once daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI1601, and pharmaceutically acceptable salts.

In one embodiment, the present invention relates to a method of treating chronic myelomonocytic leukemia (CMML) in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily day, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily , 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention relates to a method of treating atypical chronic myeloid leukemia (aCML) in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily day, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily , 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838,

- 18 045240

MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984,

CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242,

DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention relates to a method of treating juvenile myelomonocytic leukemia (JMML) in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily day, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily , 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating hypereosinophilic syndromes (HES) in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily day, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily , 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI1601, and pharmaceutically acceptable salts.

In one embodiment, the present invention relates to a method of treating myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T) in a human, which includes the step of administering to said human a therapeutically effective amount of a composition comprising an MDM2 inhibitor or a pharmaceutically acceptable compound thereof. salt, at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily day, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily , 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily

- 19 045240 once daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239EE, RO8994 , nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of a myeloproliferative neoplasm (MPN), comprising the step of administering to a human one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof. In one embodiment, the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome ( MDS), systemic mast cell disease (SMCD). In one embodiment, the myelofibrosis is selected from the group consisting of primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF). In one embodiment, primary myelofibrosis (PMF) is selected from the group consisting of pre-fibrotic PFM/early stage PFM and advanced fibrotic stage PFM. In one embodiment, the MPN is selected from the group consisting of chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia, chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia (JMML), hypereosinophilic syndromes (HES) , and myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T).

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of a myeloproliferative neoplasm (MPN), comprising the step of administering one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MPN is selected from the group, consisting of polycythemia vera (PV), primary myelofibrosis and essential thrombocythemia (ET).

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of polycythemia vera (PV), wherein the treatment includes the step of administering to a human one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of essential thrombocythemia (ET), wherein the treatment includes the step of administering to a human one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of primary myelofibrosis, wherein the treatment includes the step of administering to a human one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of post-polycythemia vera myelofibrosis (post-PV MF), wherein the treatment includes the step of administering to a human one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof. salt.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of post-essential thrombocythemia myelofibrosis (post-ET MF), wherein the treatment includes the step of administering to a subject one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof. salt.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of chronic neutrophilic leukemia (CNL), wherein the treatment includes the step of administering to a human one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of chronic eosinophilic leukemia, wherein the treatment includes the step of administering to a human one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of

- 20 045240 chronic myelomonocytic leukemia (CMML), wherein the treatment includes the step of administering to a person one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of atypical chronic myeloid leukemia (aCML), wherein the treatment includes the step of administering to a human one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of juvenile myelomonocytic leukemia (JMML), wherein the treatment includes the step of administering to a human one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of hypereosinophilic syndromes (HES), wherein the treatment includes the step of administering to a human one or more doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to the use of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T), wherein the treatment includes the step of administering to a human one or more than doses of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof.

In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention relates to the use of a composition containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of a myeloproliferative neoplasm (MPN), comprising the step of administering to a human one or more doses of a composition containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof. salt. In one embodiment, the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome ( MDS), systemic mast cell disease (SMCD). In one embodiment, the myelofibrosis is selected from the group consisting of primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF). In one embodiment, primary myelofibrosis (PMF) is selected from the group consisting of pre-fibrotic PFM/early stage PFM and advanced fibrotic stage PFM. In one embodiment, the MPN is selected from the group consisting of chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia, chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia (JMML), hypereosinophilic syndromes (HES) , and myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T).

In one embodiment, the present invention relates to the use of a composition containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of a myeloproliferative neoplasm (MPN), comprising the step of administering to a human one or more doses of a composition containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof. salt, where MPN is selected from the group consisting of polycythemia vera (PV), primary myelofibrosis and essential thrombocythemia (ET). In one embodiment, the present invention relates to the use of a composition containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of polycythemia vera (PV), wherein the treatment includes the step of administering to a person one or more doses of a composition containing an MDM2 inhibitor or its pharmaceutically acceptable salt. In one embodiment, the present invention relates to the use of a composition containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of essential thrombocythemia (ET), wherein the treatment includes the step of administering to a person one or more doses of a composition containing an MDM2 inhibitor or its pharmaceutically acceptable salt. In one embodiment, the present invention relates to the use of a composition containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of primary myelofibrosis, wherein the treatment includes the step of administering to a human one or more doses of the composition containing

- 21 045240 MDM2 inhibitor or a pharmaceutically acceptable salt thereof. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239EE, RO8994 , nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for use in the treatment of myeloproliferative neoplasm (MPN). In one embodiment, the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome ( MDS), systemic mast cell disease (SMCD). In one embodiment, the myelofibrosis is selected from the group consisting of primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF). In one embodiment, primary myelofibrosis (PMF) is selected from the group consisting of pre-fibrotic PFM/early stage PFM and advanced fibrotic stage PFM. In one embodiment, the MPN is selected from the group consisting of chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia, chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia (JMML), hypereosinophilic syndromes (HES) , and myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T).

In one embodiment, the present invention provides an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for use in the treatment of polycythemia vera (PV). In one embodiment, the present invention provides an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for use in the treatment of primary myelofibrosis. In one embodiment, the present invention provides an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for use in the treatment of essential thrombocythemia (ET). In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI1601, and pharmaceutically acceptable salts.

The methods described above can be used as first-line cancer therapy, or after treatment with conventional chemotherapeutic active pharmaceutical ingredients, including cyclophosphamide, fludarabine (FC chemotherapy) and chlorambucil.

In one embodiment, the present invention provides a method of treating myelofibrosis selected from the group consisting of primary myelofibrosis (known as chronic idiopathic myelofibrosis) and primary myelofibrosis secondary to polycythemia vera or essential thrombocythemia, comprising the step of administering a therapeutically effective amount of an MDM2 inhibitor or pharmaceutically acceptable salt.

In one embodiment, the present invention relates to a method of treating myelofibrosis selected from the group consisting of primary myelofibrosis (known as chronic idiopathic myelofibrosis) and primary myelofibrosis secondary to polycythemia vera or essential thrombocythemia, comprising the step of administering a therapeutically effective amount of a composition containing an inhibitor MDM2 or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a method of treating MPN in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MPN is selected from the group consisting of polycythemia vera (PV), essential thrombocythemia (ET ) and primary myelofibrosis, wherein the therapeutically effective amount is selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily, 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, and 480 mg once daily. In one

- 22 045240 embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating MPN in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MPN is selected from the group consisting of polycythemia vera (PV), essential thrombocythemia (ET ) and primary myelofibrosis, where the therapeutically effective amount is 120 mg once daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, the present invention provides a method of treating MPN in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MPN is selected from the group consisting of polycythemia vera (PV), essential thrombocythemia (ET ) and primary myelofibrosis, wherein the therapeutically effective amount is selected from the group consisting of 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239EE, RO8994 , nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention provides a method of treating MPN in a human, which includes the step of administering to said human a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, wherein the MPN is selected from the group consisting of polycythemia vera (PV), essential thrombocythemia (ET ) and primary myelofibrosis, where the therapeutically effective amount is 120 mg twice daily. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI773, MX69, YH239-EE, RO8994 , nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

An MDM2 inhibitor or a pharmaceutically acceptable salt thereof may also be used in combination with radiation therapy, hormonal therapy, surgery and immunotherapy, where these treatments are well known to those skilled in the art, for the treatment of MPN selected from the group consisting of polycythemia vera (PV) , myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome (MDS), systemic mast cell disease (SMCD). In one embodiment, the myelofibrosis is selected from the group consisting of primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF). In one embodiment, primary myelofibrosis (PMF) is selected from the group consisting of pre-fibrotic

- 23 045240

PFM/PFM of the early stage and PFM of the advanced fibrous stage. In one embodiment, the MPN is selected from the group consisting of chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia, chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia (JMML), hypereosinophilic syndromes (HES) , and myelodysplastic/myeloproliferative neoplasms with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T).

Treatments for chronic myelogenous leukemia (CML) and acute myelogenous leukemia (AML).

The present invention relates to a method of treating chronic myelogenous leukemia (CML), comprising the step of administering to a human in need an inhibitor of murine dual microprotein homologue 2 (MDM2), or a pharmaceutically acceptable salt thereof. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI1601, and pharmaceutically acceptable salts. In one embodiment, the MDM2 inhibitor is administered at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily day, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily , 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily once daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is administered to a human subject according to Dosage and Dosage Regimen.

The present invention relates to the use of a murine dual microprotein homolog 2 (MDM2) inhibitor, or a pharmaceutically acceptable salt thereof, for the treatment of chronic myelogenous leukemia (CML) in a human in need. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is administered at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily day, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily , 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily once daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is administered to a human subject according to Dosage and Dosage Regimen.

The present invention relates to a method of treating acute myelogenous leukemia (AML), comprising the step of administering to a human in need an inhibitor of murine dual microprotein homologue 2 (MDM2) or a pharmaceutically acceptable salt thereof. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239EE, RO8994 , nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one version

- 24 045240 implementation, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is administered at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily day, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily , 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily once daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is administered to a human subject according to Dosage and Dosage Regimen.

The present invention relates to the use of a murine dual microprotein homolog 2 (MDM2) inhibitor or a pharmaceutically acceptable salt thereof for the treatment of acute myelogenous leukemia (AML) in a human in need. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is administered at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily day, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily , 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily once daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is administered to a human subject according to Dosage and Dosage Regimen.

Treatment options for chronic Merkel cell carcinoma.

The present invention relates to a method of treating Merkel cell carcinoma (MCC), comprising the step of administering to a human in need an inhibitor of murine dual microprotein homolog 2 (MDM2) or a pharmaceutically acceptable salt thereof. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239EE, RO8994 , nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is administered at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily day, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily , 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily once daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is administered to a human subject according to Dosage and Dosage Regimen.

- 25 045240

The present invention relates to the use of a murine dual microprotein homolog 2 (MDM2) inhibitor or a pharmaceutically acceptable salt thereof for the treatment of Merkel cell carcinoma (MCC) in a human in need. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts. In one embodiment, the MDM2 inhibitor is administered at a dosage selected from the group consisting of 15 mg once daily, 25 mg once daily, 30 mg once daily, 50 mg once daily, 60 mg once daily day, 75 mg once daily, 90 mg once daily, 100 mg once daily, 120 mg once daily, 150 mg once daily, 175 mg once daily, 180 mg once daily , 200 mg once daily, 225 mg once daily, 240 mg once daily, 250 mg once daily, 275 mg once daily, 300 mg once daily, 325 mg once daily, 350 mg once daily, 360 mg once daily, 375 mg once daily, 480 mg once daily, 15 mg twice daily, 25 mg twice daily, 30 mg twice daily, 50 mg twice daily, 60 mg twice daily, 75 mg twice daily, 90 mg twice daily, 100 mg twice daily, 120 mg twice daily, 150 mg twice daily, 175 mg twice daily, 180 mg twice daily, 200 mg twice daily, 225 mg twice daily, 240 mg twice daily, 250 mg twice daily, 275 mg twice daily, 300 mg twice daily once daily, 325 mg twice daily, 350 mg twice daily, 360 mg twice daily, 375 mg twice daily, and 480 mg twice daily. In one embodiment, the MDM2 inhibitor is administered to a human subject according to Dosage and Dosage Regimen.

MDM2 inhibitor.

The compound of formula (I) has the structure and name shown below.

2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2yl)-3-methyl -2-oxopiperidin-3-yl)acetic acid:

N ₃ с^у ^сн з

(I)

The synthesis of the compound of formula (I) is given in international applications: WO 2011/153509 and WO 2014/200937; US patents No. 8569341; 9593129; 9296736; 9623018; 9757367; 9801867; 9376386; and 9855259, the description of which is incorporated herein by reference in its entirety.

In one embodiment, the compound of formula (I) or formula (II) is in amorphous form. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) in crystalline form.

In one embodiment, the MDM2 inhibitor is a compound of formula (I) in crystalline anhydrous form. In one embodiment, the MDM2 inhibitor is a compound of formula (I) in crystalline anhydrous form, characterized by a powder x-ray diffraction pattern containing peaks at 2 theta diffraction angles in degrees of approximately 11.6, 12.4, 18.6, 19.0, 21.6 and 23.6. In one embodiment, the MDM2 inhibitor is a compound of formula (I) in crystalline anhydrous form having an x-ray diffraction pattern substantially as shown in FIG. 1. The method for obtaining this crystalline form is described in international application WO 2014200937, the description of which is incorporated by reference in its entirety.

In one embodiment, the MDM2 inhibitor is a compound of formula (II) having the structure and name shown below.

- 26 045240

4-(2-((3R,5R,6S)-1-((S)-2-(tert-butylsulfonyl)-1-cyclopropylethyl)-6-(4-chloro-3-fluorophenyl)-5-(3chlorophenyl )-3-methyl-2-oxopiperidin-3-yl)acetamido)-2-methoxybenzoic acid.

The preparation of the compound of formula (II) is described in US Pat. No. 8,952,036, the disclosure of which is incorporated herein by reference in its entirety.

RG7388 (idazanutlin).

In one embodiment, the MDM2 inhibitor is RG7388. RG7388 has a chemical structure and name shown as:

4-[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2dimethylpropyl)pyrrolidine -2-carbonyl]amino]-3-methoxybenzoic acid

Triptolide (PG490)

In one embodiment, the MDM2 inhibitor is triptolide. Triptolide has a chemical structure and name shown as: (5bS,6aS,7aS,8R,8aR,9aS,9bS,10aS,10bS)-8hydroxy-8a-isopropyl-10b-methyl-2,5,5b,6,6a. 8,8a,9a,9b,10b-decahydrotris(oxyreno) [2',3':4b,5;2,3:6,7;2',3':8a,9] phenanthro[1,2-c ]furan-3(1H)-one

Nutlin-3a.

In one embodiment, the MDM2 inhibitor is nutlin-3a. Nutlin-3a has a chemical structure and name shown as: 4-[(4S,5R)-4,5-bis(4-chlorophenyl)-2-(4-methoxy2-propan-2-yloxyphenyl)-4,5- Dihydroimidazol-1-carbonyl]piperazin-2-one

HDM201.

In one embodiment, the MDM2 inhibitor is HDM201. HDM201 has a chemical structure and name shown as: (4S)-5-(5-chloro-1-methyl-2-oxopyridin-3-yl)-4-(4chlorophenyl)-2-(2,4-dimethoxypyrimidin-5 -yl)-3-propan-2-yl-4H-pyrrolo[3,4-d]imidazol-6-one

RG7112.

In one embodiment, the inhibitor

MDM2 is an RG7112. RG7112 has

- 27 045240 chemical structure and name shown as: [(4S,5R)-2-(4-tert-butyl-2-ethoxyphenyl)-4,5-bis(4chlorophenyl)-4,5-dimethylimidazol-1-yl ]-[4-(3-methylsulfonylpropyl)piperazin-1-yl]methanone

CGM097A.

In one embodiment, the MDM2 inhibitor is CGM097A. CGM097A has a chemical structure and name shown as: (1S)-1-(4-chlorophenyl)-6-methoxy-2-[4-[methyl[[4-(4-methyl-3-oxopiperazin-1-yl) cyclohexyl]methyl]amino]phenyl]-7-propan-2-yloxy-1,4dihydroisoquinolin-3-one

Nutlin-3.

In one embodiment, the MDM2 inhibitor is nutlin-3. Nutlin-3 has a chemical structure and name shown as: 4-[4,5-bis(4-chlorophenyl)-2-(4-methoxy-2-propan-2yloxyphenyl)-4,5-dihydroimidazol-1-carbonyl] piperazin-2-one

SJ-172550.

In one embodiment, the MDM2 inhibitor is SJ-172550. SJ-172550 has a chemical structure and name shown as: methyl 2-[2-chloro-6-ethoxy-4-[(3-methyl-5-oxo-1phenylpyrazol-4-ylidene)methyl]phenoxy] acetate

SAR405838 (MI-77301).

In one embodiment, the MDM2 inhibitor is SAR405838. SAR405838 has a chemical structure and name shown as: (2'R,3R,3'S,5'S)-6-chloro-3'-(3-chloro-2fluorophenyl)-5'-(2,2-dimethylpropyl)-N- (4-hydroxycyclohexyl)-2-oxospiro[1H-indole-3,4'pyrrolidine]-2'-carboxamide

- 28 045240

MI-773.

In one embodiment, the MDM2 inhibitor is MI-773. MI-773 has a chemical structure and name shown as: (2'R,3S,3'S,5'R)-6-chloro-3'-(3-chloro-2-fluorophenyl)-5'-(2,2dimethylpropyl )-N-(4-hydroxycyclohexyl)-2-oxospiro[1H-indole-3,4'-pyrrolidine]-2'-carboxamide

MX69.

In one embodiment, the MDM2 inhibitor is MX69. MX69 has a chemical structure and name shown as: 4-[8-[(3,4-dimethylphenyl)sulfamoyl]-3a,4,5,9btetrahydro-3H-cyclopenta[c]quinolin-4-yl]benzoic acid

YH239-EE.

In one embodiment, the MDM2 inhibitor is YH239-EE. YH239-EE has a chemical structure and name shown as: ethyl 3-[2-(tert-butylamino)-1-[(4chlorophenyl)methylformylamino]-2-oxoethyl]-6-chloro-1H-indole-2-carboxylate

RO8994.

In one embodiment, the MDM2 inhibitor is RO8994. RO8994 has a chemical structure and name shown as: (2'R,3R,3'S,5'S)-N-(4-carbamoyl-2-methoxyphenyl)-6chloro-3'-(3-chloro-2-fluorophenyl)-5 '-(2,2-dimethylpronyl)-2-oxospiro[1H-indole-3,4'-nirrolidine]-2'carboxamide

Nutlin-Z.

In one embodiment, the MDM2 inhibitor is nutlin-3b. Nutlin-3b has a chemical structure and name shown as: 4-[(4R,5S)-4,5-bis(4-chlorophenyl)-2-(4-methoxy2-propan-2-yloxyphenyl)-4,5- Dihydroimidazol-1-carbonyl]piperazin-2-one

- 29 045240

Serdemegan (JNJ-26854165).

In one embodiment, the MDM2 inhibitor is Serdemethane. Serdemethane has a chemical structure and name shown as: 1-N-[2-(1H-indol-3-yl)ethyl]-4-N-pyridin-4ylbenzene-1,4-diamine

NSC59984.

In one embodiment, the MDM2 inhibitor is NSC59984. NSC59984 has a chemical structure and name shown as: (E)-1-(4-methylpiperazin-1-yl)-3-(5nitrofuran-2-yl)prop-2-en-1-one o

CHEMBL2386350.

In one embodiment, the MDM2 inhibitor is CHEMBL2386350. CHEMBL2386350 has a chemical structure and name shown as: 2-[4-[(4S,5R)-2-(4-tert-butyl-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)-4,5-dimethylimidazole -1-carbonyl]piperazin-1-yl]-1morpholin-4-ylethanone

C.I.

CGM0970B.

In one embodiment, the MDM2 inhibitor is CGM0970B. CGM0970B has a chemical structure and name shown as: (1R)-1-(4-chlorophenyl)-6-methoxy-2-[4-[methyl[[4-(4-methyl-3-oxopiperazin-1-yl) cyclohexyl]methyl]amino]phenyl]-7-propan-2-yloxy-1,4dihydroisoquinolin-3-one

MK-8242.

In one embodiment, the MDM2 inhibitor is MK-8242. MK-8242 has the chemical structure and name shown (hydroxymethyl)oxolan-2-yl]pyrimidin-2-one

4-amino-1-[(2R,3S,4S,5R)-3,4-dihydroxy-5as:

DS-3032.

In one embodiment, the MDM2 inhibitor is DS-3032. DS-3032 has a chemical structure and name shown as: (3'R,4'S,5'R)-N-((3R,6S)-6-carbamoyltetrahydro-2Hpyran-3-yl)-6-chloro-4' -(2-chloro-3-fluoropyridin-4-yl)-4,4-dimethyl-2-oxodispiro[cyclohexane-1,2'pyrrolidin-3',3-indoline]-5'-carboxamide

- 30 045240

DS-3032B.

In one embodiment, the MDM2 inhibitor is DS-3O32B. DS-3032B has a chemical structure and name shown as: (3'R,4'S,5'R)-N-((3R,6S)-6-carbamoyltetrahydro2H-pyran-3-yl)-6-chloro-4' -(2-chloro-3-fluoropyridin-4-yl)-4,4-dimethyl-2-oxodispiro[cyclohexane-1,2'pyrrolidine-3',3-indoline]-5'-carboxamide 4-methylbenzenesulfonate

HDM201.

In one embodiment, the MDM2 inhibitor is HDM201. HDM201 has a chemical structure and name shown as: (4S)-5-(5-chloro-1-methyl-2-oxopyridin-3-yl)-4-(4chlorophenyl)-2-(2,4-dimethoxypyrimidin-5 -yl)-3-propan-2-yl-4H-pyrrolo[3,4-d]imidazol-6-one

APG-115.

In one embodiment, the MDM2 inhibitor is APG-115. APG-115 has a chemical structure and name shown as: 4-((3'R,4'S,5'R)-6-Chloro-4'-(3-chloro-2-fluorophenyl)-1'ethyl-2- oxodispiro[cyclohexane-1,2'-pyrrolidine-3',3-indoline]-5'-carboxamido)bicyclo[2,2,2]octane1-carboxylic acid

MI-1061.

In one embodiment, the MDM2 inhibitor is APG-115. APG-115 has a chemical structure and name shown as: 4-((3'R,4'S,5'R)-6-chloro-4'-(3-chloro-2-fluorophenyl)-2oxodispiro[cyclohexane-1. 2'-pyrrolidine-3',3-indoline]-5'-carboxamido)benzoic acid

In some embodiments, the present invention relates to pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, for the treatment of cancer, wherein the cancer is selected from the group consisting of carcinomas such as bladder, breast, colon, rectal, kidney cancer , liver, lungs (small cell lung cancer, non-small cell lung cancer

- 31 045240 whom), esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate, head and neck, and skin (including basal and squamous cell carcinoma, cutaneous melanoma, Merkel cell carcinoma, Kaposi's sarcoma, skin lymphoma); hematopoietic tumors of lymphoid origin (including leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma) hematopoietic tumors of myeloid origin origin (including acute and chronic myelogenous leukemia, myelodysplastic syndrome and promyelocytic leukemia); tumors of mesenchymal origin (including fibrosarcoma and rhabdomyosarcoma, and other sarcomas, such as soft tissue and bone); tumors of the central and peripheral nervous system (including astrocytoma, neuroblastoma, glioma, glioblastoma and schwannomas); and other tumors (including melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderoma pigmentosa, keratoctantoma and follicular thyroid cancer). In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of a myeloproliferative neoplasm (MPN), wherein the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome (MDS), systemic mast cell disease (SMCD), wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC5998 4 , CHEMBL2386350, MK8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of chronic myelogenous leukemia (CML), wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), rG7388 , triptolide, HDM201, RG7112, cGm097A, CGM0970B, SJ172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL23863 50, MK-8242, DS -3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of acute myelogenous leukemia (AML), wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388 , triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL23863 50, MK-8242 , DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of Merkel cell carcinoma (MCC), wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, Mx69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL 2386350, MK- 8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of a myeloproliferative neoplasm (MPN), wherein the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome (MDS), systemic mast cell disease (SMCD), wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In some embodiments, the present invention relates to pharmaceuticals

- 32 045240 tic compositions containing a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, for the treatment of myeloproliferative neoplasm (MPN), where MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome (MDS), systemic mast cell disease (SMCD).

In some embodiments, the present invention provides pharmaceutical compositions containing a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof, for the treatment of myeloproliferative neoplasm (MPN), wherein the MPN is selected from the group consisting of polycythemia vera (PV) , essential thrombocythemia (ET), and primary myelofibrosis.

In some embodiments, the present invention provides pharmaceutical compositions containing a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof, for the treatment of polycythemia vera (PV).

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor, or a pharmaceutically acceptable salt thereof, for the treatment of polycythemia vera (PV), wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350 , MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of polycythemia vera (PV), wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In some embodiments, the present invention provides pharmaceutical compositions containing a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof, for the treatment of essential thrombocythemia (ET).

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of essential thrombocythemia (ET), wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350 , MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of essential thrombocythemia (ET), wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In some embodiments, the present invention provides pharmaceutical compositions containing a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof for the treatment of primary myelofibrosis.

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of primary myelofibrosis, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201 , RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032 , DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In some embodiments, the present invention provides pharmaceutical compositions containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of primary myelofibrosis, wherein the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112 , CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, the myelofibrosis is selected from primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF), and myelofibrosis after essential thrombocythemia (post-ET MF).

Pharmaceutical compositions are typically formulated to provide a therapeutically effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof. If necessary, the pharmaceutical compositions contain a pharmaceutically acceptable salt and/or coordinate thereof

- 33 045240 tion complex, and one or more pharmaceutically acceptable excipients, carriers, including inert solid diluents and excipients, diluents, including sterile aqueous solution and various organic solvents, penetration enhancers, solubilizers and adjuvants. If desired, other ingredients in addition to the MDM2 inhibitor or a pharmaceutically acceptable salt thereof may be mixed in the formulation, or both components may be formulated as separate formulations for use in combination separately or at the same time.

In selected embodiments, the concentration of MDM2 inhibitor or pharmaceutically acceptable salt thereof provided in the pharmaceutical compositions of the present invention is less than, for example, 100, 90, 80, 70, 60, 50, 40, 30, 20, 19, 18, 17 , 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.4, 0.3, 0.2, 0 ,1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004 , 0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007, 0.0006, 0.0005, 0.0004, 0.0003, 0.0002 or 0.0001% w/w, wt ./about. or v/v

In selected embodiments, the concentration of MDM2 inhibitor or pharmaceutically acceptable salt thereof provided in the pharmaceutical compositions of the present invention is independently greater than 90, 80, 70, 60, 50, 40, 30, 20, 19.75, 19.50, 19 ,25, 19, 18.75, 18.50, 18.25, 18, 17.75, 17.50, 17.25, 17, 16.75, 16.50, 16.25, 16, 15.75 , 15.50, 15.25, 15, 14.75, 14.50, 14.25, 14, 13.75, 13.50, 13.25, 13, 12.75, 12.50, 12.25 , 12, 11.75, 11.50, 11.25, 11, 10.75, 10.50, 10.25, 10, 9.75, 9.50, 9.25, 9, 8.75, 8 .50, 8.25, 8, 7.75, 7.50, 7.25, 7, 6.75, 6.50, 6.25, 6, 5.75, 5.50, 5.25, 5 , 4.75, 4.50, 4.25, 4, 3.75, 3.50, 3.25, 3, 2.75, 2.50, 2.25, 2, 1.75, 1.50 , 125, 1, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0 , 03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007, 0.0006, 0.0005, 0 .0004, 0.0003, 0.0002 or 0.0001% w/w, w/v or v/v

In selected embodiments, the concentration of the MDM2 inhibitor or pharmaceutically acceptable salt thereof independently ranges from about 0.0001 to about 50%, from about 0.001 to about 40%, from about 0.01 to about 30%, from about 0.02 to about 29%, from about 0.03 to about 28%, from about 0.04 to about 27%, from about 0.05 to about 26%, from about 0.06 to about 25%, from about 0.07 to about 24%, from about 0.08 to about 23%, from about 0.09 to about 22%, from about 0.1 to about 21%, from about 0.2 to about 20%, from about 0.3 to about 19%, from about 0.4 to about 18%, from about 0.5 to about 17%, from about 0.6 to about 16%, from about 0.7 to about 15%, from about 0.8 to about 14%, from about 0.9 to about 12%, or from about 1 to about 10% w/w, w/v. or v/v

In selected embodiments, the concentration of the MDM2 inhibitor or pharmaceutically acceptable salt thereof is independently in the range of from about 0.001 to about 10%, from about 0.01 to about 5%, from about 0.02 to about 4.5%, from about 0 .03 to about 4%, about 0.04 to about 3.5%, about 0.05 to about 3%, about 0.06 to about 2.5%, about 0.07 to about 2% , from about 0.08 to about 1.5%, from about 0.09 to about 1%, from about 0.1 to about 0.9% w/w, w/v. or v/v

In selected embodiments, the amount of MDM2 inhibitor or pharmaceutically acceptable salt thereof is independently equal to or less than 10, 9.5, 9.0, 8.5, 8.0, 7.5, 7.0, 6.5, 6 ,0, 5.5, 5.0, 4.5, 4.0, 3.5, 3.0, 2.5, 2.0, 1.5, 1.0, 0.95, 0.9 , 0.85, 0.8, 0.75, 0.7, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0 .25, 0.2, 0.15, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01 , 0.009, 0.008, 0.007, 0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007, 0.0006, 0.0005, 0.0004, 0.0003, 0.0002 or 0.0001 g.

In selected embodiments, the amount of MDM2 inhibitor or pharmaceutically acceptable salt thereof is independently greater than 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0 ,0009, 0.001, 0.0015, 0.002, 0.0025, 0.003, 0.0035, 0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0, 0085 , 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0 .08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55 , 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.5, 2, 2.5, 3, 3.5 , 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 g.

The MDM2 inhibitor or a pharmaceutically acceptable salt thereof is effective over a wide range of dosages. For example, in the treatment of adults, a dosage independently ranging from 0.01-1000 mg, 0.5-100 mg, 1-50 mg per day, and 5-40 mg per day are examples of dosages that can be used. The exact dosage will depend on the route of administration, the form in which the compound is administered, the sex and age of the patient being treated, the body weight of the patient being treated, and the preference and experience of the attending physician.

Described below are non-limiting exemplary pharmaceutical compositions and methods for their preparation.

Pharmaceutical compositions for oral administration.

In selected embodiments, the present invention provides a pharmaceutical composition for oral administration comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, and a pharmaceutical excipient suitable for oral administration.

In selected embodiments, the present invention provides such pharmaceutical compositions for oral administration comprising: (i) an effective amount of an MDM2 inhibitor, or a pharmaceutically acceptable salt thereof, in combination and (ii) a pharmaceutical excipient suitable for oral administration. In selected embodiments, the composition further comprises (iii) an effective amount of at least one additional active ingredient.

In selected embodiments, the pharmaceutical composition may be a liquid pharmaceutical composition suitable for oral consumption. The pharmaceutical compositions of the present invention suitable for oral administration may be presented in the form of discrete dosage forms such as capsules, sachets or tablets, or liquids or aerosol sprays, each containing a predetermined amount of the active ingredient in the form of a powder or granules, in the form of a solution or a suspension in an aqueous or non-aqueous liquid, an oil-in-water emulsion, or a liquid water-in-oil emulsion. These dosage forms can be prepared by any of the methods, but all methods include the step of combining the active ingredient(s) with a carrier that represents one or more of the required ingredients. Typically, the compositions are prepared by uniformly and thoroughly mixing the active ingredient(s) with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired form. For example, a tablet may be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing in a suitable device the active ingredient in bulk form, such as powder or granules, optionally mixed with an excipient such as, but not limited to, a binder, a lubricant, an inert diluent and/or a surfactant or dispersant. Molded tablets can be prepared by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

The present invention further includes anhydrous pharmaceutical compositions and dosage forms, since water may promote the degradation of some compounds. For example, water can be added (eg, 5%) in the pharmaceutical field as a means of simulating long-term storage to determine characteristics such as shelf life or stability of formulations over time. The anhydrous pharmaceutical compositions and dosage forms of the present invention can be prepared using anhydrous or low-moisture ingredients and low-moisture or low-humidity conditions. The pharmaceutical compositions and dosage forms of the present invention that contain lactose can be made anhydrous if significant exposure to moisture and/or humidity is expected during manufacturing, packaging and/or storage. The anhydrous pharmaceutical composition can be prepared and stored so as to maintain its anhydrous properties. Accordingly, anhydrous compositions can be packaged using materials known to prevent exposure to water so that they can be included in suitable formulation kits. Examples of suitable packaging include, but are not limited to, sealed foil, plastic and the like, unit dose containers, blister packs and strip packs.

The MDM2 inhibitor or a pharmaceutically acceptable salt thereof can be formulated with a pharmaceutical carrier according to conventional methods for preparing pharmaceutical formulations. The carrier may take a variety of forms depending on the form of preparation required for administration. When preparing compositions for oral dosage form, any of the usual pharmaceutical vehicles may be used as carriers, such as, for example, water, glycols, oils, alcohols, flavorings, preservatives, colorings and the like in the case of oral liquid preparations (such as suspensions, solutions and elixirs) or aerosols; or carriers, such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders and disintegrants, can be used in the case of oral solid preparations, in some embodiments, without the use of lactose. For example, suitable carriers include powders, capsules and solid oral tablets. If desired, the tablets can be coated by standard aqueous or non-aqueous methods.

Binding agents suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic gums such as gum arabic, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (for example, ethylcellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropylmethylcellulose, microcrystalline cellulose and mixtures thereof.

Examples of suitable excipients for use in the pharmaceutical compositions and dosage forms described herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dexrates, kaolin, mannitol , silicic acid, sorbitol, starch, pre-gelatinized starch and mixtures thereof.

Disintegrants can be used in the compositions of the present invention to produce tablets that disintegrate when exposed to an aqueous environment. Too much baking powder can result in tablets that disintegrate in the bottle. Too small an amount may not be sufficient for disintegration to occur, altering the rate and extent of release of the active ingredients from the dosage form. Thus, a sufficient amount of disintegrant that is neither too little nor too much to detrimentally alter the release of the active ingredient(s) can be used to prepare dosage forms of the compounds described in the present invention. The amount of disintegrant used may vary depending on the type of formulation and method of administration and can be readily recognized by those skilled in the art. From about 0.5 to about 15 weight percent disintegrant or from about 1 to about 5 weight percent disintegrant can be used in the pharmaceutical composition. Disintegrants that can be used to prepare the pharmaceutical compositions and dosage forms of the present invention include, but are not limited to, agaragar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pregelatinized starch, other starches, clays, other algins, other celluloses, gums or mixtures thereof

Lubricants that can be used to form the pharmaceutical compositions and dosage forms of the present invention include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, lauryl sulfate sodium, talc, hydrogenated vegetable oil (such as peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil and soybean oil), zinc stearate, ethyl oleate, ethyl ureate, agar, or mixtures thereof. Additional lubricants include, for example, silica gel, coagulated synthetic silica aerosol, or mixtures thereof. Optionally, a lubricant can be added in an amount of less than about 1% by weight of the pharmaceutical composition.

When aqueous suspensions and/or elixirs are desired for oral administration, the main active ingredient therein may be mixed with various sweeteners or flavoring agents, coloring agents or dyes and, if necessary, emulsifying and/or suspending agents, together with diluents such as water, ethanol , propylene glycol, glycerin and various combinations thereof.

Tablets may be uncoated or coated by known methods to slow down disintegration and absorption in the gastrointestinal tract and thus provide sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be used. Oral formulations may also be presented as hard gelatin capsules, in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules, in which the active ingredient is mixed with water or an oily vehicle. , such as peanut oil, liquid paraffin or olive oil.

Surfactants that can be used to prepare the pharmaceutical compositions and dosage forms of the present invention include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. That is, a mixture of hydrophilic surfactants can be used, a mixture of lipophilic surfactants can be used, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant can be used.

A suitable hydrophilic surfactant may typically have an HLB value of at least 10, while suitable lipophilic surfactants may typically have an HLB value equal to or less than 10. An empirical parameter used to characterize the relative hydrophilicity and hydrophobicity of nonionic amphiphilic compounds represent the hydrophilic-lipophilic balance (HLB value). Surfactants with lower HLB values are more lipophilic or hydrophobic and have greater solubility in oils, while surfactants with higher HLB values are more hydrophilic and have greater solubility in aqueous solutions. Hydrophilic surfactants are generally considered to be those compounds that have an HLB value greater than about 10, as well as anionic, cationic, or zwitterionic compounds for which the HLB scale is not generally applicable. Likewise, lipophilic (i.e., hydrophobic) surfactants are compounds having an HLB value equal to or less than about 10. However, a surfactant's HLB value is only an approximate guideline typically used for formulation of industrial, pharmaceutical, and

- 36 045240 cosmetic emulsions.

Hydrophilic surfactants can be ionic or non-ionic. Suitable ionic surfactants include, but are not limited to, alkyl ammonium salts; fusidic acid salts; derivatives of fatty acids and amino acids, oligopeptides and polypeptides; glyceride derivatives of amino acids, oligopeptides and polypeptides; lecithins and hydrogenated lecithins; lysolecithins and hydrogenated lysolecithins; phospholipids and their derivatives; lysophospholipids and their derivatives; salts of carnitine esters of fatty acids; alkyl sulfate salts; fatty acid salts; docusate sodium; acylactylates; mono- and diacetylated esters of mono- and diglycerides and tartaric acid; succinylated mono- and diglycerides; esters of mono-diglycerides and citric acid; and mixtures thereof.

In the above group, ionic surfactants include, for example, lecithins, lysolecithin, phospholipids, lysophospholipids and derivatives thereof; salts of carnitine esters of fatty acids; alkyl sulfate salts; fatty acid salts; docusate sodium; acylactylates; mono- and diacetylated esters of mono- and diglycerides and tartaric acid; succinylated mono- and diglycerides; esters of mono- and diglycerides and citric acid; and mixtures thereof.

Ionic surfactants may be ionized forms of lecithin, lysolecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerone, lysophosphatidic acid, lysophosphatidylserine, PEG-phosphatidylethanolamine, PVPphosphate idylethanolamine, lactic acid fatty acid esters, stearoyl -2-lactylate, stearoyl lactylate, succinylated monoglycerides, mono/diacetylated tartaric acid esters of mono/diglycerides, citric acid esters of mono/diglycerides, cholylsarcosine, caproate, caprylate, caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate , linolenate, stearate, lauryl sulfate, teracecyl sulfate, docusate, lauroylcarnitines, palmitoylcarnitines, myristoylcarnitines and their salts and mixtures.

Hydrophilic nonionic surfactants may include, but are not limited to, alkyl glucosides; alkyl maltosides; alkylthioglucosides; lauryl macrogol glycerides; polyoxyalkylene alkyl ethers such as polyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols such as polyethylene glycol alkylphenols; polyoxyalkylene alkylphenol fatty acid esters such as polyethylene glycol fatty acid monoesters and polyethylene glycol fatty acid diesters; esters of polyethylene glycol glycerol and fatty acids; esters of polyglycerol and fatty acids; polyoxyalkylene sorbitan fatty acid esters, such as polyethylene glycol sorbitan fatty acid esters; hydrophilic transesterification products of a polyol with at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; polyoxyethylene sterols, their derivatives and analogues; polyoxyethylated vitamins and their derivatives; block copolymers of polyoxyethylene and polyoxypropylene; and mixtures thereof; polyethylene glycol sorbitan fatty acid esters and hydrophilic transesterification products of a polyol with at least one member of the group consisting of triglycerides, vegetable oils and hydrogenated vegetable oils. The polyol may be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol or a saccharide.

Other hydrophilic nonionic surfactants include, but are not limited to, PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-oleate 20, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-glyceryl trioleate 25, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30 glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate, PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryl laurate, PEG-40 palm kernel oil , PEG- 50 hydrogenated castor oil, PEG-40 castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenated castor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6 caprate/caprylate glycerides, PEG-8 caprate/caprylate glycerides, polyglyceryl laurate-10, PEG-30 cholesterol, PEG-25 phytosterol, PEG30 soy sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitan laurate, polysorbate 20, polysorbate 80, POE- 9 lauryl ether, POE-23 lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearyl ether, PEG-100 tocopheryl succinate, PEG-24 cholesterol, polyglyceryl-10 oleate, Tween 40, Tween 60, monostearate sucrose, sucrose monolaurate, sucrose monopalmitate, PEG 10-100 nonylphenol series, PEG 15-100 octylphenol series and poloxamers.

Suitable lipophilic surfactants include, by way of example only: fatty alcohols; esters of glycerol and fatty acids; esters of acetylated glycerol and fatty acids; esters of lower alcohols and fatty acids; propylene glycol esters of fatty acids; sorbitan fatty acid esters; polyethylene glycol sorbitan fatty acid esters; sterols and sterol derivatives; polyoxyethylated sterols and sterol derivatives; polyethylene glycol alkyl ethers; sugar esters; sugar esters; derivatives of mono- and diglycerides and lactic acid;

- 37 045240 products of hydrophobic transesterification of a polyol with at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; fat-soluble vitamins/vitamin derivatives; and mixtures thereof. In this group, preferred lipophilic surfactants include glycerol fatty acid esters, propylene glycol fatty acid esters and mixtures thereof, or are hydrophobic transesterification products of a polyol with at least one member of the group consisting of vegetable oils, hydrogenated vegetable oils and triglycerides.

In one embodiment, the composition may contain a solubilizer to ensure good solubilization and/or dissolution of the compound of the present invention and to minimize precipitation of the compound of the present invention. This may be particularly important for non-oral compositions, such as injectable compositions. A solubilizer may also be added to increase the solubility of the hydrophilic drug and/or other components, such as surfactants, or to maintain the composition as a stable or homogeneous solution or dispersion.

Examples of suitable solubilizers include, but are not limited to, the following: alcohols and polyols such as ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediols and its isomers, glycerin, pentaerythritol, sorbitol, mannitol, transcutol, dimethyl isosorbide, polyethylene glycol, polypropylene glycol , polyvinyl alcohol, hydroxypropyl methcellulose and other cellulose derivatives, cyclodextrins and cyclodextrin derivatives; polyethylene glycol ethers having an average molecular weight of from about 200 to about 6000, such as tetrahydrofurfuryl alcohol ether and PEG (glycofurol) or methoxy PEG; amides and other nitrogen-containing compounds such as 2-pyrrolidone, 2-piperidone, ε-caprolactam, N-alkylpyrrolidone, Nhydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrolidone; esters such as ethyl propionate, tributyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, triethyl citrate, ethyl oleate, ethyl caprylate, ethyl butyrate, triacetin, propylene glycol monoacetate, propylene glycol diacetate, epsilon-caprolactone and its isomers, δ-valerolactone and its isomers, β-butyrolac tone and its isomers; and other solubilizers known in the art, such as dimethylacetamide, dimethylisosorbide, N-methylpyrrolidones, monooctanoin, diethylene glycol monoethyl ether and water.

Mixtures of solubilizers can also be used. Examples include, but are not limited to, triacetin, triethyl citrate, ethyl oleate, ethyl caprylate, dimethyl acetamide, N-methyl pyrrolidone, N-hydroxyethyl pyrrolidone, polyvinyl pyrrolidone, hydroxypropyl methylcellulose, hydroxypropyl cyclodextrins, ethanol, polyethylene glycol 200-100, glycofurol, transcutol, propylene glycol and dimethyl isosorbide. Particularly preferred solubilizers include sorbitol, glycerin, triacetin, ethyl alcohol, PEG-400, glycofurol and propylene glycol.

The amount of solubilizer that may be included is not particularly limited. The amount of said solubilizer may be limited to a biologically acceptable amount, which can be readily determined by one skilled in the art. In some cases, it may be useful to include amounts of solubilizers well in excess of bioacceptable amounts, for example, to maximize drug concentration, with excess solubilizer being removed before providing the composition to the patient using conventional techniques such as distillation or evaporation. Thus, if present, the solubilizer may be present in a weight ratio of 10, 25, 50, 100%, or up to about 200% by weight, based on the total weight of the drug and other excipients. If necessary, very small amounts of solubilizer can also be used, for example 5, 2, 1% or even less. Typically, the solubilizer may be present in an amount of from about 1% to about 100%, more typically from about 5 to about 25% by weight.

The composition may further contain one or more pharmaceutically acceptable additives and excipients. These additives and excipients include, but are not limited to, detackifying agents, antifoaming agents, buffering agents, polymers, antioxidants, preservatives, chelating agents, viscosity modulators, tonicity modifiers, flavoring agents, colorants, flavoring agents, opacifying agents, suspending agents, binders, fillers, plasticizers, lubricants and mixtures thereof.

In addition, an acid or base may be included in the composition to facilitate processing, to improve stability, or for other reasons. Examples of pharmaceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium bicarbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminosilicate, synthetic aluminum silicate, synthetic hydrocalcite, magnesium-aluminum hydroxide, diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine, triethylamine, triisopropanolamine, trimethylamine, tris(hydroxymethyl)aminomethane (TRIS) and the like. Also suitable are bases that are salts of a pharmaceutically acceptable acid such as acetic acid.

- 38 045240 slots, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinosulfonic acid acid, isoascorbic acid, lactic acid, maleic acid, oxalic acid, p-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid and the like. Salts of polyprotic acids such as sodium phosphate, disodium hydrogenphosphate and sodium dihydrogenphosphate can also be used. When the base is a salt, the cation may be any conventional and pharmaceutically acceptable cation such as ammonium, alkali metals and alkaline earth metals. Examples may include, but are not limited to, sodium, potassium, lithium, magnesium, calcium and ammonium.

Suitable acids are pharmaceutically acceptable organic or inorganic acids. Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, boric acid, phosphoric acid and the like. Examples of suitable organic acids include acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acids, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid acid, hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid , toluenesulfonic acid and uric acid.

Pharmaceutical compositions for injection.

In selected embodiments, the present invention provides an injectable pharmaceutical composition comprising an MDM2 inhibitor or a pharmaceutically acceptable salt thereof and an injectable pharmaceutical excipient. The components and amounts of agents in the compositions are as described in the present invention.

Forms into which the compositions of the present invention may be included for injection include aqueous or oleaginous suspensions or emulsions with sesame oil, corn oil, cottonseed oil or peanut oil, as well as elixirs, mannitol, dextrose or sterile aqueous solution, and similar pharmaceutical vehicles.

Aqueous solutions in saline are also commonly used for injection. Ethanol, glycerin, propylene glycol and liquid polyethylene glycol (and suitable mixtures thereof), cyclodextrin derivatives and vegetable oils may also be used. Suitable fluidity can be maintained, for example, by applying a coating such as lecithin to maintain the desired particle size in the event of dispersion and the use of surfactants. The action of microorganisms can be prevented by using various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid and thimerosal.

Sterile injection solutions are prepared by incorporating the MDM2 inhibitor or a pharmaceutically acceptable salt thereof in the required amounts into a suitable diluent with various other ingredients listed above, if necessary, followed by sterilization by filtration. Typically, dispersions are prepared by incorporating various sterilized active ingredients into a sterile vehicle that contains the basic dispersion medium and other necessary ingredients listed above. In the case of sterile powders for the preparation of sterile injectable solutions, some desirable methods of preparation are vacuum drying and freeze drying methods, which yield a powder of the active ingredient plus any additional required ingredient from a pre-sterilized solution thereof.

Administration of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of these compounds can be accomplished by any method that provides delivery of the compounds to the site of action. These routes include oral routes, intraduodenal routes, parenteral injection (including intravenous, intraarterial, subcutaneous, intramuscular, intravascular, intraperitoneal or infusion), topical (eg, transdermal application), rectal administration, via local delivery by catheter or stent, or by inhalation . The combination of compounds can also be administered intraadiposally or intrathecally.

Examples of forms for parenteral administration include solutions or suspensions of the active compound in sterile aqueous solutions, for example, aqueous solutions of propylene glycol or dextrose. If necessary, these dosage forms can be suitably buffered.

The present invention also relates to kits. The kits contain an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, either alone or in combination with suitable packaging, and written materials, which may include instructions for use, clinical discussion

- 39 045240 studies and list of side effects. These kits may also include information, such as scientific literature references, package inserts, clinical trial results and/or summaries thereof, and the like, that indicate or establish the activities and/or benefits of the composition, and/or that describe dosing , administration, side effects, drug interactions, or other information useful to the healthcare provider. This information may be based on the results of various studies, for example, studies using experimental animals, including in vivo models and studies based on clinical trials in humans. The kit may additionally contain another active pharmaceutical ingredient. Suitable packaging and additional items for use (eg, liquid measuring cup, foil wrapper to minimize air exposure, and the like) are known in the art and may be included in the kit. The kits described in the present invention can be provided, sold and/or promoted to healthcare providers, including doctors, nurses, pharmacists, formulary officials and the like. The kits may also, in selected embodiments, be sold directly to the consumer. In one embodiment, the present invention provides a kit containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for use in the treatment of MPN. In one embodiment, the MPN is selected from the group consisting of polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis. In one embodiment, the present invention provides a kit containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for use in the treatment of chronic myelogenous leukemia (CML). In one embodiment, the present invention provides a kit containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for use in the treatment of acute myelogenous leukemia (AML). In one embodiment, the present invention provides a kit containing an MDM2 inhibitor or a pharmaceutically acceptable salt thereof for use in the treatment of Merkel cell carcinoma (MCC).

Dosages and dosage regimens.

The amount of MDM2 inhibitor or pharmaceutically acceptable salt thereof administered will depend on the individual being treated, the severity of the disorder or condition, the rate of administration, clearance of the compounds, and the discretion of the treating physician. However, an effective dosage ranges from about 0.001 to about 100 mg per kg of body weight per day, such as from about 1 to about 35 mg/kg/day, in a single or divided dose. For a 70 kg person this would be from about 0.05 to 7 g/day, for example from about 0.05 to about 2.5 g/day. In some cases, dose levels below the lower end of the above range may be more than adequate, while in other cases even higher doses can be administered without causing any harmful side effects - for example, by dividing given large doses into several smaller doses for administration throughout the day.

In some embodiments, the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered as a single dose. Typically this administration will be by injection, for example an intravenous injection, to rapidly administer the agents. However, if necessary, other ways can be used. A single dose of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof may also be used to treat an acute condition.

In some embodiments, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered in multiple doses to treat MPN. In one embodiment, the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered in multiple doses. In one embodiment, the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered in multiple doses by injection - for example, by intravenous injection. In one embodiment, dosing may be one, two, three, four, five, six, or more than six times per day. In one embodiment, the dosage may be selected from the group consisting of once daily, twice daily, three times daily, four times daily, five times daily, six times daily, once every other day, once every week, twice a week, three times a week, four times a week, twice a week and monthly. In other embodiments, the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered from about once per day to about six times per day. In some embodiments, the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered once daily, while in other embodiments, the MDM2 inhibitor or pharmaceutically acceptable salt thereof is administered twice daily, and in other embodiments, the MDM2 inhibitor or pharmaceutically acceptable salt thereof is administered three times. in a day. In some embodiments, the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered three times per week, including every Monday, Wednesday, and Friday.

Administration of the MDM2 inhibitor or a pharmaceutically acceptable salt thereof may be continued for as long as necessary. In some embodiments, the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered for more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 , 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 or more days. In some embodiments, the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, an MDM2 inhibitor or pharmaceutical thereof

- 40 045240 a highly acceptable salt is administered for about 14 days, about 21 days, about 28 days, about 35 days, about 42 days, about 49 days, or about 56 days. In some embodiments, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered chronically, for example, to treat chronic effects. In another embodiment, administration of the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is continued for less than about 7 days. In yet another embodiment, administration is continued for more than about 6, 10, 14, 28 days, two months, three months, four months, five months, six months, seven months, eight months, nine months, ten months, eleven months or one year. In some embodiments, administration is continued for more than about one year, two years, three years, four years, or five years. In some embodiments, continuous dosing is achieved and maintained as long as needed.

In some embodiments, the effective dosage of the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is in the range of from about 1 to about 500 mg, from about 10 to about 300 mg, from about 20 to about 250 mg, from about 25 to about 200 mg, from about 10 to about 200 mg, about 20 to about 150 mg, about 30 to about 120 mg, about 10 to about 90 mg, about 20 to about 80 mg, about 30 to about 70 mg, about 40 to about 60 mg, from about 45 to about 55 mg, from about 48 to about 52 mg, from about 50 to about 150 mg, from about 60 mg to about 140 mg, from about 70 mg to about 130 mg, from about 80 to about 120 mg, from about 90 to about 110 mg, from about 95 to about 105 mg, from about 150 to about 250 mg, from about 160 to about 240 mg, from about 170 to about 230 mg, from about 180 to about 220 mg, about 190 to about 210 mg, about 195 to about 205 mg, or about 198 to about 202 mg. In some embodiments, the effective dosage of the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is about 15 mg, about 25 mg, about 30 mg, about 50 mg, about 50 mg, about 75 mg, about 90 mg, about 100 mg, about 120 mg , approximately 125 mg, approximately 150 mg, approximately 175 mg, approximately 180 mg, approximately 200 mg, approximately 225 mg, approximately 240 mg, approximately 250 mg, approximately 275 mg, approximately 300 mg, approximately 325 mg, approximately 350 mg, approximately 360 mg, approximately 375 mg, approximately 400 mg, approximately 425 mg, approximately 450 mg, approximately 475 mg, approximately 480 mg or approximately 500 mg. In some embodiments, the effective dosage of the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is 15, 25, 30, 50, 60, 75, 90, 100, 120, 150, 175, 180, 200, 225, 240, 250, 275, 300 , 325, 350, 360, 375 and 480 mg.

In some embodiments, the effective dosage of the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is in the range of from about 0.01 to about 4.3 mg/kg, from about 0.15 to about 3.6 mg/kg, from about 0.3 to about 3.2 mg/kg, from about 0.35 to about 2.85 mg/kg, from about 0.15 to about 2.85 mg/kg, from about 0.3 to about 2.15 mg/kg , from about 0.45 to about 1.7 mg/kg, from about 0.15 to about 1.3 mg/kg, from about 0.3 to about 1.15 mg/kg, from about 0.45 to about 1 mg/kg, from about 0.55 to about 0.85 mg/kg, from about 0.65 to about 0.8 mg/kg, from about 0.7 to about 0.75 mg/kg, from about 0 .7 to about 2.15 mg/kg, about 0.85 to about 2 mg/kg, about 1 to about 1.85 mg/kg, about 1.15 to about 1.7 mg/kg, from about 1.3 to about 1.6 mg/kg, about 1.35 to about 1.5 mg/kg, about 2.15 to about 3.6 mg/kg, about 2.3 to about 3, 4 mg/kg, about 2.4 to about 3.3 mg/kg, about 2.6 to about 3.15 mg/kg, about 2.7 to about 3 mg/kg, about 2.8 to about 3 mg/kg or from about 2.85 to about 2.95 mg/kg. In some embodiments, the effective dosage of the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is about 0.35 mg/kg, about 0.7 mg/kg, about 1 mg/kg, about 1.4 mg/kg, about 1.8 mg /kg, approximately 2.1 mg/kg, approximately 2.5 mg/kg, approximately 2.85 mg/kg, approximately 3.2 mg/kg, or approximately 3.6 mg/kg.

In some embodiments, the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered at a dosage of 10-500 mg twice daily, including dosages of 15, 25, 30, 50, 60, 75, 90, 100, 120, 150, 175, 180, 200, 225, 240, 250, 275, 300, 325, 350, 360, 375 and 480 mg twice daily.

In some embodiments, the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered at a dosage of 10-500 mg once daily, including dosages of 15, 25, 30, 50, 60, 75, 90, 100, 120, 150, 175, 180, 200, 225, 240, 250, 275, 300, 325, 350, 360, 375, and 480 mg once daily.

- 41 045240

An effective amount of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof can be administered as a single or multiple dose by any of the conventional routes of administration of agents having similar uses, including buccal, sublingual and transdermal routes, intra-arterial injection, intravenous, parenteral, intramuscular, subcutaneous or oral.

In some embodiments, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a subject periodically, which is known as intermittent administration. By intermittent administration is meant a period of administration of a therapeutically effective dose of an MDM2 inhibitor or a pharmaceutically acceptable salt thereof, followed by a period of dosage cessation, followed by another administration period, and so on. During each dosing period, the dosing frequency can be independently selected from: three times a day, twice a day, daily, once a week, twice a week, three times a week, four times a week, five times a week, six times a week week or monthly. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, Mx69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

By discontinuation period or rest period is meant a period of time during which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is discontinued. The discontinuation time period may be longer or shorter than the administration period, or the same as the administration period. During the discontinuation period, therapeutic agents other than the MDM2 inhibitor or a pharmaceutically acceptable salt thereof may be administered.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of a myeloproliferative neoplasm (MPN) during a first administration period, followed by a withdrawal period, followed by a second administration period, etc., in which the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome (MDS), systemic mast cell disease (SMCD). The first administration period, the second administration period and the rest period are independently selected from the group consisting of more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 , 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, one month, five weeks, six weeks, seven weeks, two months, nine weeks, ten weeks, eleven weeks , three months, thirteen weeks, fourteen weeks, fifteen weeks, four months or more days, in which an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject three times a day, twice a day, daily, once a week, twice a week , three times a week, four times a week, five times a week, six times a week or monthly. In one embodiment, the first administration period has the same duration as the second administration period. In one embodiment, the first administration period is shorter than the second administration period. In one embodiment, the first administration period is longer than the second administration period. In one embodiment, the first administration period and the second administration period are approximately one week, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112,

- 42 045240

CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of polycythemia vera (PV) during a first administration period, which is then followed by a rest period, which is then followed by a second administration period, etc. First the administration period, the second administration period and the rest period are independently selected from the group consisting of more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, one month, five weeks, six weeks, seven weeks, two months, nine weeks, ten weeks, eleven weeks, three months, thirteen weeks, fourteen weeks, fifteen weeks, four months or more days in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject three times a day, twice a day, daily, weekly, twice a week, three times a week , four times a week, five times a week, six times a week or monthly. In one embodiment, the first administration period has the same duration as the second administration period. In one embodiment, the first administration period is shorter than the second administration period. In one embodiment, the first administration period is longer than the second administration period. In one embodiment, the first administration period and the second administration period are approximately one week, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of essential thrombocythemia (ET) during a first administration period, which is then followed by a rest period, which is then followed by a second administration period, etc. First the administration period, the second administration period and the rest period are independently selected from the group consisting of more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, one month, five weeks, six weeks, seven weeks, two months, nine weeks, ten weeks, eleven weeks, three months, thirteen weeks, fourteen weeks, fifteen weeks, four months or more days in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject three times a day, twice a day, daily, weekly, twice a week, three times a week , four times a week, five times a week, six times a week or monthly. In one embodiment, the first administration period has the same duration as the second administration period. In one embodiment, the first administration period is shorter than the second administration period. In one embodiment, the first administration period is longer than the second administration period. In one embodiment, the first administration period and the second administration period are approximately one week, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered

- 43 045240 to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of primary myelofibrosis during a first administration period, which is then followed by a rest period, which is then followed by a second administration period, etc. The first administration period, the second administration period and the rest period are independently selected from the group consisting of more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, one month, five weeks, six weeks, seven weeks, two months, nine weeks, ten weeks, eleven weeks, three months, thirteen weeks, fourteen weeks, fifteen weeks, four months or more days on which an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject three times a day, twice a day, daily, weekly, twice a week, three times a week, four times per week, five times a week, six times a week or monthly. In one embodiment, the first administration period has the same duration as the second administration period. In one embodiment, the first administration period is shorter than the second administration period. In one embodiment, the first administration period is longer than the second administration period. In one embodiment, the first administration period and the second administration period are approximately one week, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of chronic myelogenous leukemia (CML) during a first administration period, which is then followed by a rest period, which is then followed by a second administration period, etc. The first administration period, the second administration period and the rest period are independently selected from the group consisting of more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 , 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, one month, five weeks, six weeks, seven weeks, two months, nine weeks, ten weeks, eleven weeks , three months, thirteen weeks, fourteen weeks, fifteen weeks, four months or more days in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject three times a day, twice a day, daily, weekly, twice a week, three times a day week, four times a week, five times a week, six times a week or monthly. In one embodiment, the first administration period has the same duration as the second administration period. In one embodiment, the first administration period is shorter than the second administration period. In one embodiment, the first administration period is longer than the second administration period. In one embodiment,

- 44 045240 the first administration period and the second administration period are approximately one week, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of acute myelogenous leukemia (AML) during a first administration period, which is then followed by a rest period, which is then followed by a second administration period, etc. The first administration period, the second administration period and the rest period are independently selected from the group consisting of more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 , 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, one month, five weeks, six weeks, seven weeks, two months, nine weeks, ten weeks, eleven weeks , three months, thirteen weeks, fourteen weeks, fifteen weeks, four months or more days in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject three times a day, twice a day, daily, weekly, twice a week, three times a day week, four times a week, five times a week, six times a week or monthly. In one embodiment, the first administration period has the same duration as the second administration period. In one embodiment, the first administration period is shorter than the second administration period. In one embodiment, the first administration period is longer than the second administration period. In one embodiment, the first administration period and the second administration period are approximately one week, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of Merkel cell carcinoma (MCC) during a first administration period, which is then followed by a rest period, which is then followed by a second administration period, etc. .The first administration period, the second administration period and the rest period are independent

- 45 045240 mo selected from a group consisting of more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, one month, five weeks, six weeks, seven weeks, two months, nine weeks, ten weeks, eleven weeks, three months, thirteen weeks, fourteen weeks, fifteen weeks, four months or more days in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject three times a day, twice a day, daily, weekly, twice a week, three times a week, four times a week, five times a week, six times a week or monthly. In one embodiment, the first administration period has the same duration as the second administration period. In one embodiment, the first administration period is shorter than the second administration period. In one embodiment, the first administration period is longer than the second administration period. In one embodiment, the first administration period and the second administration period are approximately one week, in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately three weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject daily; and the rest period is approximately two weeks. In one embodiment, the first administration period and the second administration period are approximately four weeks in which the MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject weekly; and the rest period is approximately two weeks. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a subject in need for the treatment of cancer on days 1-7 of a 21-day cycle (no MDM2 inhibitor is administered on days 8-21) for a period selected from 3 weeks, 6 weeks, 9 weeks, 12 weeks, 15 weeks, 18 weeks, 21 weeks, 24 weeks, 27 weeks, 30 weeks, 33 weeks, 36 weeks, 39 weeks, 42 weeks, 45 weeks, 48 weeks, 51 weeks, 54 weeks, 57 weeks , 60 weeks, 63 weeks, 66 weeks, 69 weeks, 72 weeks, 75 weeks, 78 weeks, 81 weeks, 84 weeks, 87 weeks, 90 weeks, 93 weeks, 96 weeks, 99 weeks, 102 weeks, 105 weeks, 108 weeks, 111 weeks, 114 weeks, 117 weeks, 120 weeks, 123 weeks, 126 weeks, 129 weeks, 132 weeks, 135 weeks, 138 weeks, 141 weeks, 144 weeks, 147 weeks, 150 weeks, 153 weeks and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin -3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof, wherein the cancer is selected from the group consisting of carcinomas , such as cancer of the bladder, breast, colon, rectum, kidney, liver, lung (small cell lung cancer and non-small cell lung cancer), esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate , head and neck, and skin (including basal and squamous cell carcinoma, melanoma, Merkel cell carcinoma, Kaposi's sarcoma, skin lymphomas); hematopoietic tumors of the lymphoid lineage (including leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma); hematopoietic tumors of myeloid origin (including acute and chronic myelogenous leukemia, myelodysplastic syndrome and promyelocytic leukemia); tumors of mesenchymal origin (including fibrosarcoma and rhabdomyosarcoma and other sarcomas, such as soft tissue and bone); tumors of the central and peripheral nervous system (including astrocytoma, neuroblastoma, glioma, glioblastoma and schwannomas); and other tumors (including melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderoma pigmentosa, keratoctantoma and follicular thyroid cancer). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI1601, and pharmaceutically acceptable salts.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof

- 46 045240 is administered to a subject in need for the treatment of MPN on days 1-7 of a 21-day cycle (no MDM2 inhibitor is administered on days 8-21) for a time period selected from 3 weeks, 6 weeks, 9 weeks, 12 weeks, 15 weeks , 18 weeks, 21 weeks, 24 weeks, 27 weeks, 30 weeks, 33 weeks, 36 weeks, 39 weeks, 42 weeks, 45 weeks, 48 weeks, 51 weeks, 54 weeks, 57 weeks, 60 weeks, 63 weeks, 66 weeks, 69 weeks, 72 weeks, 75 weeks, 78 weeks, 81 weeks, 84 weeks, 87 weeks, 90 weeks, 93 weeks, 96 weeks, 99 weeks, 102 weeks, 105 weeks, 108 weeks, 111 weeks, 114 weeks, 117 weeks, 120 weeks, 123 weeks, 126 weeks, 129 weeks, 132 weeks, 135 weeks, 138 weeks, 141 weeks, 144 weeks, 147 weeks, 150 weeks, 153 weeks, and 156 weeks, where an MDM2 inhibitor is selected from the group, consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin- 3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof, wherein MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome (MDS), systemic mast cell disease (SMCD). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a subject in need for the treatment of primary myelofibrosis on days 1-7 of a 21-day cycle (on days 821 no MDM2 inhibitor is administered) for a time period selected from 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 132, 135, 138, 141, 144, 147, 150, 153 and 156 weeks, where the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin- 3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a subject in need for the treatment of polycythemia vera on days 1-7 of a 21-day cycle (no MDM2 inhibitor is administered on days 8-21) for a time period selected from 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 132, 135, 138, 141, 144, 147, 150, 153 and 156 weeks wherein the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of phlebotomy-dependent polycythemia vera on days 1-7 of a 21-day cycle (no MDM2 inhibitor is administered on days 8-21) for a period of time selected from 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 132, 135, 138, 141, 144, 147, 150, 153 and 156 weeks, where the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239- EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984,

CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) and a pharmaceutically acceptable salt thereof; and MDM 2 inhibitor is administered orally at a dose of 120 mg or 240 mg once daily (QD). In one embodiment, the human subject suffers from hydroxyurea (HU) intolerance (unacceptable side effects). In one embodiment, the human subject is refractory to hydroxyurea (HU) (inappropriate response). In one embodiment, the human subject suffers from splenomegaly. In one embodiment, the human subject suffers from splenomegaly and is phlebotomy dependent. In one embodiment, the human subject is phlebotomy dependent without splenomegaly. In one version

- 47 045240 implementation, ruxolitinib therapy did not help the person.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a subject in need for the treatment of essential thrombocythemia on days 1-7 of a 21-day cycle (no MDM2 inhibitor is administered on days 8-21) for a time period selected from 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 132, 135, 138, 141, 144, 147, 150, 153, and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994 , nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032, DS-3032B, APG115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a subject in need for the treatment of chronic myelogenous leukemia (CML) on days 1-7 of a 21-day cycle (no MDM2 inhibitor is administered on days 8-21) for a period of time selected from 3. 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 132, 135, 138, 141, 144, 147, 150, 153 and 156 weeks, where the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a subject in need for the treatment of acute myelogenous leukemia (AML) on days 1-7 of a 21-day cycle (no MDM2 inhibitor is administered on days 8-21) for a time period selected from 3. 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 132, 135, 138, 141, 144, 147, 150, 153 and 156 weeks, where the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a subject in need for the treatment of Merkel cell carcinoma (MCC) on days 1-7 of a 21-day cycle (no MDM2 inhibitor is administered on days 8-21) for a time period selected from 3 , 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78 , 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 132, 135, 138, 141, 144, 147, 150, 153 and 156 weeks, where the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE , RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof.

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of cancer on days 1-7 of a 28-day cycle (no MDM2 inhibitor is administered on days 8-28) for a time period selected from 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152 and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS -3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof, wherein the cancer is selected from the group consisting of carcinomas such as bladder, breast, colon, rectal, kidney, liver, lung (small cell) lung and non-small cell lung cancer), esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate, head and neck, and skin (including basal and squamous cell carcinoma, melanoma, Merkel cell carcinoma, Kaposi's sarcoma , skin lymphoma); hematopoietic tumors

- 48 045240 lymphoid lineages (including leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma); hematopoietic tumors of myeloid origin (including acute and chronic myelogenous leukemia, myelodysplastic syndrome and promyelocytic leukemia); tumors of mesenchymal origin (including fibrosarcoma and rhabdomyosarcoma and other sarcomas, such as soft tissue and bone); tumors of the central and peripheral nervous system (including astrocytoma, neuroblastoma, glioma, glioblastoma and schwannomas); and other tumors (including melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderoma pigmentosa, keratoctantoma and follicular thyroid cancer). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) and a pharmaceutically acceptable salt thereof; and MDM 2 inhibitor is administered orally at a dose of 120 mg or 240 mg once daily (QD).

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need to treat MPN on days 1-7 of a 28-day cycle (no MDM2 inhibitor is administered on days 8-28) for a time period selected from 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152, and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201 , RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032 , DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof, wherein the MPN is selected from the group consisting of polycythemia vera (PV), myelofibrosis, primary myelofibrosis, thrombocythemia, essential thrombocythemia (ET), idiopathic myelofibrosis, systemic mastocytosis (SM), chronic neutrophilic leukemia (CNL), myelodysplastic syndrome (MDS), systemic mast cell disease (SMCD). In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) and a pharmaceutically acceptable salt thereof; and MDM 2 inhibitor is administered orally at a dose of 120 mg or 240 mg once daily (QD).

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of primary myelofibrosis on days 1-7 of a 28-day cycle (no MDM2 inhibitor is administered on days 8-28) for a time period selected from 4, 8, 12 , 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104, 108, 112 , 116, 120, 124, 128, 132, 136, 140, 144, 148, 152 and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112 , CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS-3032 , D.S. -3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) and a pharmaceutically acceptable salt thereof; and MDM 2 inhibitor is administered orally at a dose of 120 mg or 240 mg once daily (QD).

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a person in need for the treatment of polycythemia vera on days 1-7 of a 28-day cycle (no MDM2 inhibitor is administered on days 8-28) for a time period selected from 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152 and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS -3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) and a pharmaceutically acceptable salt thereof; and MDM 2 inhibitor is administered orally at a dose of 120 mg or 240 mg once daily (QD).

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of essential thrombocythemia on days 1-7 of a 28-day cycle (no MDM2 inhibitor is administered on days 8-28) for a period of time selected from 4, 8,

- 49 045240

12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152 and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242, DS -3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) and a pharmaceutically acceptable salt thereof; and MDM 2 inhibitor is administered orally at a dose of 120 mg or 240 mg once daily (QD).

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of chronic myelogenous leukemia (CML) on days 1-7 of a 28-day cycle (no MDM2 inhibitor is administered on days 8-28) for a period of time selected from 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100 , 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152 and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388 , triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin3a, nutlin-3b, serdemethane, NSC59984, CHEMBL238635 0, MK-8242, DS -3032, DS-3032B, APG-115, MI1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) and a pharmaceutically acceptable salt thereof; and MDM 2 inhibitor is administered orally at a dose of 120 mg or 240 mg once daily (QD).

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of acute myelogenous leukemia (AML) on days 1-7 of a 28-day cycle (no MDM2 inhibitor is administered on days 8-28) for a period of time selected from 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100 , 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152 and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388 , triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin3b, serdemethane, NSC59984, CHEMBL238635 0, MK-8242, DS -3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) and a pharmaceutically acceptable salt thereof; and MDM 2 inhibitor is administered orally at a dose of 120 mg or 240 mg once daily (QD).

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of Merkel cell carcinoma (MCC) on days 1-7 of a 28-day cycle (no MDM2 inhibitor is administered on days 8-28) for a period of time selected from 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152 and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin-3a, nutlin3b, serdemethane, NSC59984, CHEMBL2 386350, MK-8242, DS-3032, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112, CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and their pharmaceutically acceptable salts. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) and a pharmaceutically acceptable salt thereof; and MDM 2 inhibitor is administered orally at a dose of 120 mg or 240 mg once daily (QD).

In one embodiment, an MDM2 inhibitor or a pharmaceutically acceptable salt thereof is administered to a human subject in need for the treatment of phlebotomy-dependent polycythemia vera on days 1-7 of a 28-day cycle (no MDM2 inhibitor is administered on days 8-28) for a period of time selected from 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104, 1θ8, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152 and 156 weeks, wherein the MDM2 inhibitor is selected from the group consisting of a compound of formula (I), formula (II), RG7388, triptolide, HDM201, RG7112, CGM097A, CGM0970B, SJ-172550, SAR405838, MI-773, MX69, YH239-EE, RO8994, nutlin-3, nutlin3a, nutlin-3b, serdemethane, NSC59984, CHEMBL2386350, MK-8242,DS- 3032, DS-3032B, APG-115, MI1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is selected from the group consisting of compounds of formula (I), formula (II), RG7388, HDM201, RG7112,

CGM097A, CGM0970B, SAR405838, MK-8242, DS-3032B, APG-115, MI-1601 and pharmaceutically acceptable salts thereof. In one embodiment, the MDM2 inhibitor is a compound of formula (I) or formula (II) and a pharmaceutically acceptable salt thereof; and MDM 2 inhibitor is administered orally at a dose of 120 mg or 240 mg once daily (QD).

In one embodiment, a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof is administered to a human in need daily at a dose of 120 mg for the treatment of phlebotomy-dependent polycythemia vera on days 1-7 of a 28-day cycle (on days 8-28 no MDM2 inhibitor is administered) for a period of time selected from 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152 and 156 weeks. In one embodiment, the human subject suffers from hydroxyurea (HU) intolerance (unacceptable side effects). In one embodiment, the human subject is refractory to hydroxyurea (HU) (inappropriate response). In one embodiment, the human subject suffers from splenomegaly. In one embodiment, the human subject suffers from splenomegaly and is phlebotomy dependent. In one embodiment, the human subject is phlebotomy dependent without splenomegaly. In one embodiment, the person did not benefit from ruxolitinib therapy.

In one embodiment, a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof is administered to a human in need daily at a dose of 240 mg for the treatment of phlebotomy-dependent polycythemia vera on days 1-7 of a 28-day cycle (on days 8-28 no MDM2 inhibitor is administered) for a period of time selected from 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152 and 156 weeks. In one embodiment, the human subject is determined to suffer from hydroxyurea (HM) intolerance (unacceptable side effects). In one embodiment, the human subject is refractory to hydroxyurea (HU) (inappropriate response). In one embodiment, the human subject suffers from splenomegaly. In one embodiment, the human subject suffers from splenomegaly and is phlebotomy dependent. In one embodiment, the human subject is phlebotomy dependent without splenomegaly. In one embodiment, the person did not benefit from ruxolitinib therapy.

In one embodiment, a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof is administered to a human in need daily at a dose of 120 mg for the treatment of phlebotomy-dependent polycythemia vera on days 1-7 of a 21-day cycle (on days 8-21 an inhibitor MDM2 is not administered) for a period of time selected from 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60 , 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 132, 135 , 138, 141, 144, 147, 150, 153 and 156 weeks. In one embodiment, the human subject suffers from hydroxyurea (HU) intolerance (unacceptable side effects). In one embodiment, the human subject is refractory to hydroxyurea (HU) (inappropriate response). In one embodiment, the human subject suffers from splenomegaly. In one embodiment, the human subject suffers from splenomegaly and is phlebotomy dependent. In one embodiment, the human subject is phlebotomy dependent without splenomegaly. In one embodiment, the person did not benefit from ruxolitinib therapy.

In one embodiment, a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof is administered to a human in need daily at a dose of 240 mg for the treatment of phlebotomy-dependent polycythemia vera on days 1-7 of a 21-day cycle (on days 8-21 inhibitor MDM2 is not administered) for a period of time selected from 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60 , 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 132, 135 , 138, 141, 144, 147, 150, 153 and 156 weeks. In one embodiment, the human subject suffers from hydroxyurea (HU) intolerance (unacceptable side effects). In one embodiment, the human subject is refractory to hydroxyurea (HU) (inappropriate response). In one embodiment, the human subject suffers from splenomegaly. In one embodiment, the human subject suffers from splenomegaly and is phlebotomy dependent. In one embodiment, the human subject is phlebotomy dependent without splenomegaly. In one embodiment, the person did not benefit from ruxolitinib therapy.

Examples

The embodiments described in the present invention are further described with reference to the following examples. These examples are provided for purposes of illustration only, and the description given in the present invention should in no way be construed as being limited to these examples, but rather should be construed as including any and all variations that become apparent as a result of the teachings set forth in the present invention.

Example 1: effect of the compound of formula (I) on cells with polycythemia vera.

The method for testing the effect of compounds of formula (I) on polycythemia vera cells follows that described in Lu et al., Blood, 2012, 120(15); 3098-3105, the contents of which are included in this

- 51 045240 document by reference. Below is a brief description of the methodology

Obtaining cells.

Peripheral blood is collected from patients with polycythemia vera (PV). Appropriate approval was obtained from the institutional review board. Informed consent was obtained before the start of the study. All patients will meet the World Health Organization diagnostic criteria for polycythemia vera (PV). Peripheral blood samples were layered onto Ficoll-Hypaque (1.077 g/ml; GE Healthcare) and low-density mononuclear cells were separated by centrifugation. CD34+ cells were isolated using a human CD34+ cell selection kit (StemCell Technologies) according to the manufacturer's instructions. The purity of the CD34+ cell population was analyzed using a FACSCalibur flow cytometer (BD Biosciences); and at least 85% is required for all experiments. Fresh normal human bone marrow CD34+ cells were purchased from ALLCELLS as a control.

LDC tests.

The effects of a compound of formula (I) on patients with polycythemia vera (PV) can be assessed by the HPC tests described in Lu et al., Blood, 2012, 3098-3105, the contents of which are incorporated by reference. Briefly, CD34+ cells were cultured in serum-free medium (StemCell Technologies) containing 50 ng/ml stem cell factor (SCF), 50 ng/ml thrombopoietin (TPO), 50 ng/ml fins-like tyrosine kinase ligand 3 (Flt-3), and 50 ng /ml IL-3, and treated with various doses of the compound of formula (I) for 4 days. After 4 days of treatment, CD34+ cells are examined in semisolid media as described in Bruno et al., Blood, 2006, 3128-3134, the contents of which are incorporated by. Briefly, 5 x 10 ² CD34+ cells are plated in duplicate cultures containing 1 ml IMDM with 1.1% methylcellulose and 20% FBS, to which are added SCF, TPO, Flt-3 ligand, IL-3 and GM-CSF, each which are contained in an amount of 50 ng/ml, and 2 units/ml of erythropoietin (EPO). Colonies were counted after 14 days of incubation, and individual colonies were recovered and genotyped for JAK2V617F.

Two-round allele specific PCR for JAK2V617F-positive colonies

Genomic DNA was isolated from randomly collected colonies using an Extract-N-Amp blood PCR kit (Sigma-Aldrich). JAK2V617F is detected using a two-round allele specific PCR described in Bruno et al., Blood, 2006, 3128-3134, the contents of which are incorporated by reference. The final PCR products were analyzed on 2.0% agarose gels. The 279-bp product indicates positivity for allele-specific JAK2V617F, whereas the 229-bp product indicates negativity for JAK2V617F. Colonies are classified as homozygous for JAK2V617F if they contain only the 279-bp band, whereas heterozygous colonies are identified based on the presence of both the 279-bp and 229-bp bands.

Apoptosis test.

Treated cells were harvested and washed with PBS for Annexin-V staining (BD Biosciences); staining methods are performed in accordance with the protocols provided by the manufacturer. Data are acquired on a FACSCalibur flow cytometer (BD Biosciences), and at least 10,000 live cells are acquired for each assay (BD FACS Diva software; BD Biosciences).

Western blot analysis.

CD34+ cells are purified from the peripheral blood of patients with polycythemia vera (PV) and cultured in serum-free medium containing SCF, FL-3 ligand, IL-3, and TPO. The cells are treated with various doses of the compound of formula (I) for 4 hours. Cells were harvested, and whole-cell protein extracts were prepared with RIPA lysis buffer (Boston BioProducts) for Western blotting.

To prepare cytoplasmic and nuclear protein fractions from polycythemia vera (PV) patient cells, CD34+ cells are expanded in serum-free medium containing SCF, FL-3 ligand, and IL-3 for 10 days. CD34+ cells are then repurified and treated with various doses of the compound of formula (I) for 48 hours in the presence of SCF, FL-3 ligand, IL-3 and TPO. Protein extracts were prepared using NE-PER nuclear and cytoplasmic extraction reagent (Thermo Scientific) according to the manufacturer's instructions.

Before Western blotting, all samples were denatured with Laemmli SDS sample buffer (Boston BioProducts) by heating at 95°C for 5 min; each sample was separated on SDS-PAGE gels and transferred to polyvinyldifluoridine membranes (Bio-Rad). Phospho-p53, p53, MDM2, p21, p-STAT1, PUMA, and Bak were visualized using antibodies (Cell Signaling Technologies) and ECL Western blotting reagents (Denville Scientific).

Statistical analysis.

Results are presented as the mean ± standard deviation of individual data points obtained from different numbers of experiments. Statistical significance was determined using Student's t test or paired samples t test.

Example 2: Effects of a compound of formula (I) on cells with essential thrombocythemia.

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Experiments were carried out as in Example 1, except that essential thrombocythemia cells were used instead of polycythemia vera (PV) cells.

Example 3: Effects of the compound of formula (I) on primary myelofibrosis cells.

Experiments were performed as in Example 1, except that primary myelofibrosis cells were used instead of polycythemia vera (PV) cells.

Example 4: compound of formula (I) as monotherapy for patients with polycythemia vera.

The purpose of this study is to examine the safety and effectiveness of the compound of formula (I) in patients with polycythemia vera (PV). A similar clinical trial is underway with RG7388 (NCT02407080). The study included 30 patients with polycythemia vera (PV) who were prescribed a compound of formula (I) at a dose of 120 mg once daily (QD) on days 1-7 of a 21-day cycle (on days 8-21, without administration of a compound of formula (I)) for two years. The inclusion criteria are as follows.

JAK2V617F-positive polycythemia vera (PV).

Not previously treated with at least one other agent (hydroxyurea, interferon, anagrelide) for >18 years.

Acceptable pre-screening organ function, defined as: total bilirubin <1.5 times the upper limit of normal (ULN) unless due to Gilbert's disease or hemolysis, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) <2, 5x ULN, serum creatinine <1.5xULN.

Women of childbearing potential and men must consent to the use of adequate contraception (eg, hormonal or barrier birth control; abstinence) prior to enrollment and for the duration of study participation. If a female subject becomes pregnant or is suspected of becoming pregnant while participating in this study, she should be immediately withdrawn from the study.

During or at the end of the study, each patient with polycythemia vera (PV) is assessed for the following parameters to determine the safety and effectiveness of the compound of formula (I): 1) hematological response; 2) decreased load of the JAK2V617F allele; 3) changes in histopathological abnormalities of the bone marrow; 4) decrease in the initial level of reticulin/collagen fibrosis; 5) frequency of venous and arterial thrombosis; and 5) changes in MPN-related symptoms as assessed by the MPN-SAF.

Example 5: compound of formula (I) as monotherapy for patients with essential thrombocythemia.

The study was conducted in accordance with the methods described in Example 4, except that patients with essential thrombocythemia (ET) were included instead of patients with polycythemia vera (PV).

Example 6: compound of formula (I) as monotherapy for patients with primary myelofibrosis.

The study was conducted in accordance with the methods described in Example 4, except that patients with primary myelofibrosis were included instead of patients with polycythemia vera (PV).

Example 7: Efficacy of the compound of formula (I) against cancer.

A method for testing the effectiveness of a compound of formula (I) against cancer is described in Canon et al., Molecular Cancer Therapeutics, 2015; 649-658 and Rew, et al., J. Med. Chem. 2012; 55; 4936-54, the contents of which are incorporated by reference. The method is described briefly below.

Cells.

SJSA-1, HCT116, ACHN, NCI-H460, MOLM-13, RKO, MCF7, 22RV1, HT-29, PC-3, NCI-H82, NCISNU1 and MG-63 cells were purchased from ATCC and have since been confirmed short tandem repeat analysis (PowerPlex 18D Kit from Promega). NCI-H2452, SW982, C32, SK-HEP-1, A375, RT4, RPMI-2650, MDA-MB-134-VI, NCI-H2347 and A427 cells were purchased from ATCC and used for 6 months. IGR-1 and CML-T1 cells were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ) and used for 6 months. A375sq2 cells were generated in vivo by passage of A375 cells in mice. HCT116 ^p53'L cells were obtained from Bert Vogelstein. KS-1 and SNG-M were purchased from the Japanese Collection of Research Bioresources (HSRRB) and administered for 6 months. G-401, G-361, LS174T cells (purchased from ATCC), EOL-1 cells (purchased from DSMZ) and KP-4 cells (purchased from HSRRB) were used 6 months after purchase.

Binding assay using surface plasmon resonance (SPR) spectroscopy

Materials: Biacore T100 instrument (GE Healthcare), CM5 sensor chip (BR-1000-12), amine condensation kit (BR-1000-50) containing 1-ethyl-3-(3-dimethylaminepropyl)carbodiimide hydrochloride (EDC), N -Hydroxysuccinimide (NHS) and ethanolamine-HCl, GST capture kit (BR-1002-23), HBS-N buffer (BR-1006-70) were from GE Healthcare. All buffers were filtered through a 0.2 μM nylon membrane (VWR 87006-076). Inhibitor kinetics study on Biacore: the CM5 chip was pretreated with 10 mM NaOH, 10 mM HCl and 0.1% SDS, each time for 10

- 53 045240 second injection twice at a rate of 100 µl/min in HBS-N buffer. The immobilization and capture steps were performed as described: 1. An equal volume of 0.4 M EDC and 0.1 M NHS was mixed and injected onto the surface of the chip to activate it for 10 min at 10 μL/min; 2. Anti-GST antibody (30 μg/ml in 10 mM sodium acetate, pH 5) was immobilized on the chip by surface injection over 15 minutes at a rate of 8 μl/min; 3. To deactivate excess reactive groups and wash away unbound anti-GST antibodies, 1 M ethanolamine-HCl pH 8.5 was pumped over the surface for 10 min at a rate of 8 μl/min. The resulting level of antibody immobilization was approximately 18,000 RU; 4. 10 μg/ml GST in HBS-N buffer was injected into one flow cell for 15 minutes at a rate of 5 μl/min to create a 1500 RU control surface; 5. The sample surface was generated by injecting 20 μg/mL MDM2 into another flow cell for 15 minutes twice at a rate of 5 μL/min to achieve a capture level of about 2000 RU. Kinetic binding studies of inhibitors (molecular weight range 450-600) were carried out at 25°C with a constant flow of 90 μl/min. Sample assay buffer consisted of 25 mM Tris, pH 7.5, 150 mM NaCl, 0.005% (v/v) Tween 20, 0.2 mM TCEP plus 2.5% (v/v) DMSO. Inhibitors were diluted from 1 mM stock solution (in 100% DMSO) in sample assay buffer to 12 nM. The inhibitors were then serially diluted 2 times to obtain 8 spots. Diluted inhibitors were introduced onto the control surface and the surface with the sample for 1-2 minutes and dissociation was observed within 3-4 minutes. A buffer containing 2.3-3.5% DMSO was added to generate a DMSO correction curve. All sensorgrams were processed using Biacore Evaluation software (GE Healthcare) with a dual control procedure and DMSO calibration. KD values were determined using a 1:1 binding model including a mass transfer limitation condition. SJSA-1 cell proliferation test (Click-iT EdU HCS test) SJSA-1 cells were seeded at a density of 2.8 x 103 cells/well in 384-well cell culture plates (Perkin Elmer, #6007460) in 40 µl growth medium (RPMI 1640 supplemented with 10 mM HEPES, 1 mM sodium pyruvate, 1X penicillin-streptomycin, 2 mM glutamine and 10% fetal bovine serum). Cells were cultured for 24 hours at 37°C and 5% CO ₂ . The next day, cells were treated with MDM2mu inhibitor for 16 hours in the presence of 10% human serum. On day 3, the Click-iT EdU assay procedure was performed according to the manufacturer's instructions with assay volumes reduced to 25 μL to adjust for 384-well formats (Invitrogen, #C10357). Briefly, EdU (5ethynyl-2′-deoxyuridine) was added to cells to a final concentration of 10 μM and incubated for 1 h. After labeling, the cells were fixed with 4% formaldehyde and permeabilized with 0.1% Triton-X 100. After washing, the cells were incubated with Click-iT reaction buffer, and then the nuclei were stained. Cells were then washed and visualized using the Opera High Content Screening System (Perkin Elmer). The percentage of EdU incorporation was calculated and used to calculate the _IC50 . IC ₅₀ values were determined using Hill's four-parameter logistic (4PL) model.

BrdU Proliferation Test (HCT116 Specificity Test) The effectiveness of the MDM2ob inhibitor was also determined by analyzing the effect on cell proliferation by quantifying the extent of 5-bromo-2-deoxyuridine (BrdU) incorporation into compound-treated cells compared to DMSO-treated control cells. HCT116 p53WT or ^p53'L cells were seeded at a density of 6x103 cells/well in 96-well cell culture plates in 100 μl of growth medium (McCoy's 5A, 1X PSQ and 10% fetal bovine serum (all from Invitrogen)). Cells were initially cultured for 24 hours at 37°C and 5% CO ₂ before adding compound. MDM2 inhibitors were serially diluted in DMSO (Sigma #D2650), then re-diluted in assay media (McCoy's 5A, 1X PSQ, 10% human serum (Bioreclamation #HMSRM)) to a final DMSO concentration of 1%. Cells were incubated in the presence of inhibitor at 37°C and 5% CO ₂ for 16 hours, and then treated with diluted BrdU labeling reagent (final dilution 1:100, Invitrogen # 00-0103) for 1 hour at 37°C and 5 % CO ₂ . After BrdU labeling, the medium was removed and the cells were fixed and stained for BrdU incorporation. The extent of BrdU incorporation was determined using flow cytometry or a Cellomics Array Scan Vti plate reader with Target Activation bioequipment. The percentage of BrdU-positive cells in DMSO-treated control wells was used to normalize the signal and calculate the percent inhibition for each compound-treated well. Dose-response curves were generated using XLFit software to calculate IC50 values for each inhibitor tested.

HCT116 p21 TaqMan® test (HCT116 specificity test).

This assay was performed exactly as described above for the SJSA-1 p21 assay, with the following exceptions: growth medium for HCT116 p53WT or ^p53'L consisted of McCoy's 5A, 1X PSQ, and 10% fetal bovine serum (all from Invitrogen).

Biochemical (HTRF) test.

Materials: Human MDM2 (GST-thrombin-hMDM2 (1-188)) was prepared independently. Expressed in E. coli and purified using a Glutathione Sepharose 4B, Q-HP and Superdex 200 column. Human p53 (Avi-TrxA-6His-Thrombin-S-tag-EK-p53 (1-83)) was expressed in E. coli and passed through Ni-NTA, hydroxyapatite, Superdex 75 columns to achieve purity greater than 80%. Human serum

- 54 045240 obtained from Bioreclamation (HMSRM, unfiltered). Anti-GST monoclonal antibodies labeled with europium cryptate (Eu-anti-GST, 61GSTLB) and SA-Xlent (611SAXLB) were obtained from Cisbio. DTT, BSA, KH ₂ PO ₄ , Na ₂ HPO ₄ , DMSO, NaCl and KF were from Sigma. The serum-free reaction buffer consisted of 1.06 mM KH2PO4, 2.96 mM Na ₂ HPO ₄ , 0.155 M NaCl, 0.1% BSA, and 1 mM DTT. The serum HTRF test used a reaction buffer supplemented with 15% human serum. The HTRF test plate was White 384 Opti plates from Perkin Elmer (6007299). Envison (Perkin Elmer) was set to excitation at 320 nm. Emission was measured at 665 and 615 nm, and the Em665/Em615 ratio represented the MDM2-p53 interaction. Fluorescence was measured with a time resolution of 50 flashes for both detectors with a delay of 60 μs after each excitation. Read time 300 µs. Vprep was a product of Velocity11. The Wellmate microplate dispenser was from Thermo Scientific.

Serial Killer was produced in-house. Methods (determining inhibitor potency in HTRF assay): 20 µL of 1.5 mM inhibitor was serially diluted to 20 µL DMSO for 22 spots using Serial Killer. 1 μl of this diluted inhibitor was transferred to a reaction plate with 9 μl of Vprep reaction buffer. 10 μl of 1 nM MDM2 was added to the reaction plate and incubated with the inhibitor for 20 minutes before adding 20 μl of 1.25 nM p53. After 60 min, detection mixture (10 μL, 1 nM SA-Xlent, 3 nM Eu-anti-GST, and 0.5 M KF) was added to the reaction mixture, and the plate was read on the Envision after 18 h of incubation. The total reaction volume was 50 μl. MDM2, p53 and detection mixture were transferred to Wellmate assay plates. IC50 was determined from replicate data. For the HTRF assay in 15% serum: the addition sequence was the same as in the serum-free assay. 10 μl of 12.5 nM MDM2 diluted in reaction buffer containing 30% human serum was added to 10 μl of the buffer-compound mixture to obtain 15% serum. 20 nM p53 was diluted in 15% human serum buffer. The detection buffer was 15% serum with 10 nM SA-Xlent, 3 nM Eu-anti-GST, and 0.5 M KF.

Immunoblot analysis.

Tumor cells were treated with DMSO (0.1%) or a compound of formula (I) (0.1, 1 or 10 mmol/L). After 24 h, protein lysates were collected, electrophoresed, and transferred to polyvinylidene difluoride membranes (Life Technologies). Primary antibodies: p53 (DO-1; Calbiochem), MDM2 (BD Pharmingen), p21 (R&D Systems), PUMA (Abcam), or β-actin-HRP (Sigma).

Cell viability test (72 h).

Cell lines were seeded into 96- or 384-well plates at the optimal initial seeding density to ensure that cells did not reach confluence by the end of the assay. Cells were treated with DMSO control or the compound of formula (I) at various concentrations for 72 hours. CellTiter-Glo (Promega) or ATPlite 1step Luminescent (PerkinElmer) cell viability assay kits were used to determine the number of viable cells. Luminescence was measured using an EnVision Multilabel Reader (PerkinElmer) for each cell line at time zero (V0) before adding compounds, and after 72 hours of compound treatment. Growth inhibition (GI) was calculated on a 200-point scale according to the following equations, where V72 is the luminescence of the DMSO control after 72 hours, and T72 is the luminescence of the compound-treated sample: if T72>V0, then GI=100x(1-((T72 -V0)/(V72-V0))); if T72<V0, then GI=100x(1-((T72-V0)/V0)). GI values of 0, 100, and 200 represented uninhibited cell growth (i.e., DMSO control), cell stagnation, and complete cell killing, respectively. Dose-response curves were generated using XLFit software (IDBS) to calculate IC50 values for the compound of formula (I) in each cell line tested.

Animal studies.

All experimental methods on animals were carried out in accordance with the standards of the Association for the Assessment and Accreditation of Laboratory Animal Sciences. All studies used female athymic nude mice, 4 to 6 weeks of age (Harlan Laboratories, Hsd: Athymic Nude-Foxnlnu). Mice were housed five per cage with a filter in a sterile, environmentally controlled room (temperature 23 ± 2°C, relative humidity 50 ± 20%) with a 12-h light/dark cycle. Mice were fed a commercial rodent chow (Harlan Laboratories, #2920X) and provided with filtered tap water ad libitum. Mice were individually identified using microarrays (Bio Medic Data Systems) that were implanted subcutaneously at least 2 days before the study.

Pharmacodynamic analyses.

Tumor cells (SJSA-1: 5x106 cells, HCT 116: 2x106 cells) were injected subcutaneously into the flank of female athymic nude mice at a cell to Matrigel ratio of 2:1 (BD Bioscience). The compound of formula (I) was administered via gastric tube when the average tumor size reached approximately 300-450 mm ³ (n=4/group). Tumors were collected at 1, 2, 4, 8, and 24 hours post-dose (SJSA-1) or 6 hours post-dose (HCT116) and snap frozen in liquid nitrogen. Total RNA was purified using the Qiagen RNeasy 96 kit (Qiagen). Levels of p21 and the housekeeping gene GAPDH were determined from total RNA for each sample in technical duplicates using qRT-PCR. qRT-PCR reactions

- 55 045240 was analyzed on an Applied Biosystems Prism 7900HT instrument, and data were analyzed using Applied Biosystems SDS2.2 software. The SDS2.2 program calculated the copy numbers of p21 and GAPDH in each tumor sample. p21 copy number was normalized to GAPDH copy number, and the fold increase in normalized p21 levels relative to the media control was calculated for each sample. For MIC-1 determination, plasma was collected at the time of sacrifice, and MIC1 was determined using the R&D Quantikine Human MIC-1 Immunoassay (cat. no. DGD150) according to the manufacturer's instructions. The ELISA assay was performed using a Spectramax M5 microplate reader using Softmax pro v4 (Molecular Devices).

Xenograft studies.

SJSA-1 cells (5x10 ⁶ cells with Matrigel in a 2:1 ratio), NCI-H46O cells (5x10 ⁶ cells with Matrigel in a 2:1 ratio), A375sq2 (5x10 ⁶ cells with Matrigel in a 2:1 ratio) or HCT116 (2x10 ⁶ cells) were injected subcutaneously into the flank of female athymic nude mice (n=10/group). Treatment began when the tumor had developed and was approximately 200 mm ³ . The compound of formula (I) was administered once daily via gastric tube. Tumor sizes were assessed twice weekly using a Pro-Max electronic digital caliper (Sylvac) and tumor volume was calculated using the formula: length x width x height and expressed in ^mm3 . Data were expressed as mean ± standard error of the mean. Body weight was recorded twice weekly to assess tolerability (data not shown). Analysis of p21 mRNA at the end of xenograft studies was performed as described for pharmacodynamic analysis of p21.

Detection of BrdUrd and cleaved caspase-3 in xenografts.

Tumors were collected 6 hours after the last treatment, fixed in formalin and treated with paraffin. Two hours before collection, mice were intraperitoneally injected with BrdUrd (50 mg/kg). Tumor sections were immunostained for either BrdUrd or cleaved capsase-3 using commercial antibodies and counterstained with hematoxylin. Sections were scanned at ±20 with an Aperio digital scanner and positive nuclear density was determined using Visiomorph image analysis software.

Statistical analysis.

For in vivo efficacy studies of a compound of Formula (I), repeated measures ANOVA (RMANOVA) followed by Dunnett's post hoc test for multiple comparisons was used to assess the statistical significance of observed differences.

Results.

A broader panel of tumor cell lines, including wild-type (n=23) and mutant (n=7) p53 lines representing a range of tumor types and genetic backgrounds, were analyzed and the effect of treatment with a compound of formula (I) on cell growth over a 72-year period was determined. hour period. In 23 wild-type p53 cell lines evaluated, treatment with a compound of formula (I) inhibited cell growth with IC ₅₀ values ranging from 0.1 to 1 mmol/L (FIG. 2).

The antitumor activity of the compound of formula (I) was evaluated in xenograft models representing different genetic backgrounds and different tumor types. All tumor cell lines used in xenograft models contained wild-type p53. Daily oral administration of a compound of formula (I) resulted in significant tumor growth inhibition (TGI) in all models (FIGS. 3-5).

SJSA-1, an osteosarcoma model with MDM2 amplification, was most sensitive to treatment with the compound of formula (I) with an ED50 of 9.1 mg/kg (Fig. 3). In the highest dose group, 75 mg/kg, 10 of 10 tumors completely regressed and were undetectable after 10 days of treatment. Treatment with the compound of formula (I) was stopped in this group after 25 days, and the mice were observed for an additional 50 days. None of the mice showed regrowth of the SJSA-1 tumor. Additional xenograft models have demonstrated the range of in vivo antitumor activity of a compound of formula (I) or formula (II). In the HCT116 (KRAS mutant) colorectal cancer model, the highest dose of compound (I) resulted in 86% TGI compared to control, and the ED50 was 31 mg/kg (Fig. 4). Treatment with a compound of formula (I) in a BRAF A375sq2 mutant melanoma model resulted in a 97% TGI with an ED50 of 18 mg/kg (FIG. 5).

Tumors were collected at the end of each xenograft study to determine the effect of treatment with a compound of formula (I) on p53 pathway activity. Treatment with a compound of formula (I) resulted in a dose- and time-dependent induction of p21 mRNA compared to media-treated tumors (FIG. 6). The level of p21 induction in each tumor model was related to the grade of TGI, with p21 levels being highest in tumors whose growth was most suppressed. However, the maximum level of p21 induction varied depending on the tumor model.

Example 8: clinical trials.

A two-stage, randomized, open-label, multicenter phase 2a/2b study of the efficacy, safety, pharmacokinetics and pharmacodynamics of the compound of formula (I) compared with ruxolitinib in patients with phlebotomy-dependent polycythemia vera.

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Polycythemia vera (PV) is classified as a myeloproliferative neoplasm (MPN). Long-term survival is poor, with a survival rate of 18% after 20 years. There are limited therapeutic options for patients with PV, especially those intolerant/resistant to hydroxyurea (HU), and these patients are often treated with HU despite unacceptable side effects and poor response. The response rate for ruxolitinib is 21%, indicating a high unmet need for effective treatments in this subgroup of patients with PV. A compound of formula (I) or formula (II) is an orally bioavailable, small molecule, cytotoxic chemotherapeutic agent that binds to murine dual microprotein 2 (MDM2) and inhibits the MDM2/tumor protein 53 (p53) protein-protein interaction. The compound of formula (I) has been shown to inhibit the growth of p53 wild type (p53WT) tumor cells in vitro and tumor xenografts in vivo. p53 is a tumor suppressor and transcription factor that responds to cellular stress by activating the transcription of numerous genes involved in cell cycle arrest, apoptosis, aging, and deoxyribonucleic acid (DNA) repair. Preclinically, a key determinant of sensitivity to MDM2 inhibition is the p53 mutation status of the cells, with p53WT cells being sensitive to these inhibitors. MDM2 is a negative regulator of p53 and is overexpressed in CD34+ myeloproliferative neoplasm cells harboring p53WT. MDM2 destroys and inactivates p53, allowing clonal hematopoietic stem cells to proliferate.

Study design.

The study consisted of two parts. In Part A, the initial proof-of-concept phase, three treatment arms were evaluated for efficacy and safety in subjects with PV. If the overall response rate in Part A was at least 40% in subjects with splenomegaly, Part B was initiated, whereby PV subjects with splenomegaly were randomly assigned to treatment with either the recommended dose and schedule from Part A or ruxolitinib administered according to label instructions.

All Part A and Part B subjects continued treatment with a compound of formula (I) until disease progression or intolerance. Disease progression is defined as:

Indications for phlebotomy (after week 8): confirmed hematocrit (HCT) >45%, which is at least 3 percentage points higher than NBT obtained at baseline, or confirmed HBT >48%, with confirmation carried out 2 to 14 days after initial observation.

Splenic progression: volumetric assessment that there is an increase >25% over the volume determined at the time of the best documented splenic volume reading using centralized image review.

Bone marrow fibrosis/transformation: Increased bone marrow fibrosis from baseline and/or transformation to myelofibrosis (MF), myelodysplastic syndrome (MDS) or acute leukemia.

Part A: proof of concept/dose determination of compound of formula (I). Part A assessed the safety and effectiveness of two different doses and two different schedules of the compound of formula (I) in three treatment groups in subjects with PV. A total of 75 randomized subjects were evaluated in Part A. Subjects were randomly assigned to one of three treatment groups:

group 1, N=15 with splenomegaly, plus N=10 without splenomegaly: 120 mg once daily for 1-7 days, no treatment for 8-21 days (21-day cycle) group 2, N=15 with splenomegaly, plus N=10 without splenomegaly: 240 mg once daily for 1-7 days, no treatment for 8-21 days (21-day cycle) group 3, N=15 with splenomegaly, plus N=10 without splenomegaly: 120 mg once daily for 1-7 days, without treatment for 8-28 days (28-day cycle)

Part B: Compound of Formula (I) at the recommended dose and schedule from Part A versus ruxolitinib in subjects with PV: Part B evaluated 220 randomized subjects with splenomegaly. Subjects in Part B were randomly assigned to treatment with the recommended dose of compound of formula (I) and schedule determined by the DMC of A, or ruxolitinib according to the label directions:

group 1, N=110: subjects with splenomegaly were treated with a compound of formula (I) at the recommended dose and according to the schedule from part A group 2, N=110: subjects with splenomegaly were treated with ruxolitinib according to the label instructions.

Approximately 295 randomized subjects were planned: Part A: N=75 Part B: N=220 (110 subjects received the compound of formula (I), and 110 subjects received ruxolitinib)

Research objectives:

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Primary Objectives of Part A Final evaluation criteria/measurement result To determine effectiveness compounds of formula (I) in subjects with PV splenomegaly who are dependent on phlebotomy. Primary analysis effectiveness for Part A is based on a modified analysis according to intended treatment only for Part A subjects with splenomegaly who are p53WT. Proportion of participants with splenomegaly achieving a response at week 32, with response defined as achieving both of the following: No indication for phlebotomy starting at the week 8 visit and continuing through week 32, with no more than one indication for performing phlebotomy after randomization and before the week 8 visit. · Reduction in spleen volume as assessed by MRI (or CT) >35% from baseline at 32 weeks. To determine the dose of a compound of formula (I) in PV subjects with splenomegaly that will be evaluate compared to ruxolitinib in part B Dose selection of a compound of formula (I) for PV subjects with splenomegaly who have an overall response rate of >40% in part A Primary objectives of part B Final evaluation criteria Measurement result demonstrate the superiority of a compound of formula (I) over ruxolitinib in PV subjects with splenomegaly who are resistant/intolerant to hydroxyurea. Proportion of participants with splenomegaly achieving response at week 32, with response defined as achieving both of the following: No indication for phlebotomy starting at week 8 and continuing through week 32, with no more than one indication for phlebotomy after randomization and until the week 8 visit. • Reduction in spleen volume as assessed by MRI (or CT) >35% from baseline at 32 weeks.

Exclusion criteria.

1. adults > 18 years old.

2. Documents confirming that the patient meets the revised 2016 World Health Organization (WHO) criteria for the diagnosis of PV.

3. Subjects must be dependent on phlebotomy. Definition of phlebotomy dependency: hematocrit level of 40-45% with two phlebotomies or separated by at least 4 weeks within 24 weeks before screening or Hematocrit level greater than 45% with at least one phlebotomy within 16 weeks before screening .

4. Subjects with splenomegaly (defined as splenic volume >450 cc ^cm ) and without splenomegaly on MRI (or CT) are eligible for Part A. Part B only admits patients with splenomegaly on MRI or CT.

5. Previous treatment with hydroxyurea (HU) (parts A and B) or interferon (part A only). If previously treated with HU, the subject must be resistant/intolerant to HU according to

- 58 045240 with the following criteria:

HU resistance is defined as:

dose >2 g/day or maximum tolerated dose <2 g/day for at least 12 weeks, resulting in the need for phlebotomy to maintain hematocrit <45% or platelet count >400 x 10 ⁹ /L and white blood cell count (WBC) >10 x 10 ⁹ / l.

HU intolerance is defined as:

ANC <1.0 x 10 ⁹ /L or platelet count <100 x 10 ⁹ /L or hemoglobin <100 g/L (i.e. 10 g/dL) at the minimum dose of HU required to achieve a response (hematocrit <45% without phlebotomy and /or all 3 of platelet count <400x10 ⁹ /l, leukocyte count <10x10 ⁹ /l and non-palpable spleen) or

Presence of leg ulcers or other unacceptable non-hematologic toxic effects associated with HU (eg, mucocutaneous manifestations, gastrointestinal symptoms, pneumonitis, or fever at any dose of HU), defined as: Common Terminology Criteria for Adverse Events (CTCAE) version 5.0 grade 3-4 adverse events (AE) or > 1 week CTCAE version 5.0 grade 2 AE, or permanent discontinuation of HU, or interruption of HU until toxicity resolves, or hospitalization due to HU toxicity.

6. Eastern Cooperative Oncology Group (ECOG) performance status of 0, 1, or 2.

7. Women of childbearing age and men who have partners of childbearing age must consent to the use of an effective method of contraception during the study. In addition, men should continue to use contraception for 3 months after the last dose of study drug, and women should continue to use contraception for 1 week after taking the last dose of study drug. Effective birth control includes (a) combined hormonal contraceptives containing estrogen and progestogen (oral, intravaginal, transdermal); (b) progestogen-only hormonal contraceptives (oral, injectable, implantable); (c) intrauterine device; (d) intrauterine hormone releasing system; (f) bilateral occlusion of the fallopian tubes; (f) post-vasectomy partner; and (g) sexual abstinence.

Exclusion criteria.

1. Meets the criteria for post-PV myelofibrosis as defined by the International Working Group on Research and Treatment of Myeloproliferative Neoplasms (IWG-MRT).

2. >10% blast cells.

3. Clinically significant thrombosis within 3 months after the examination.

4. Inadequate liver or kidney function:

A. Renal failure (estimated creatinine clearance <45 ml/min according to Cockcroft-Gault):

(140-age) x weight (in kilograms) x [0.85 if female] iC'Cr -- ^v ' 72 x serum creatinine (in mg/dl)

b. Known history of hepatocellular disease (eg, hepatitis B or C, cirrhosis, or other hepatocellular disease)

With. Total bilirubin >2x upper limit of laboratory normal (ULN), excluding Gilbert's syndrome.

d. Alanine aminotransferase (ALT) >2.5χULN

5. Part B only: prior treatment with a JAK inhibitor.

6. Previous treatment with histone deacetylase (HDAC) inhibitors or BCL-2 inhibitors.

7. Patients previously treated with MDM2 antagonist therapy, p53-targeted therapy, or patients receiving interferon-alpha, anagrelide, or ruxolitinib for 28 days or approximately 5 half-lives, or hydroxyurea for 1 day, or patients receiving any other cytoreductive or investigational agents for 28 days or 5 half-lives of the initial dose. Aspirin is permitted according to PV treatment guidelines unless there are medical contraindications.

8. Absolute neutrophil count < ^1.5x109 /L before dosing on day 1 of cycle 1.

9. Platelet count <150x10 ⁹ /L before dosing on day 1 of cycle 1.

10. Irradiation of the spleen for 3 months before the first dose of the compound of formula (I).

11. Pregnant or breastfeeding women.

12. History of major organ transplantation.

13. Uncontrolled intercurrent illness, including, but not limited to, acute hepatitis A; known history of human immunodeficiency virus (HIV); clinically significant heart disease (New York Heart Association class III or IV); symptomatic congestive heart failure; unstable angina; ventricular arrhythmia; or mental illness/social situations that limit study eligibility.

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14. Subjects with clinically significant bacterial, fungal, parasitic or viral infection requiring treatment. Patients with acute bacterial infections requiring antibiotics should defer screening/enrollment until completion of antibiotic therapy.

15. Other malignancies within the last 3 years, except treated basal cell or squamous cell skin cancer, cervical carcinoma in situ, organ-confined or treated non-metastatic prostate cancer with normal prostate specific antigen, breast carcinoma in situ after completion of surgical resection or superficial transitional cell carcinoma of the bladder.

16. QT prolongation grade 2 or greater (>480 milliseconds, according to NCICCTCAE criteria, version 5.0).

Randomization method.

In Part A, subjects are randomly assigned to one of three treatment groups. In Part B, subjects are randomly assigned to a compound of formula (I) or ruxolitinib. Subjects in Part B only are stratified based on HU status: HU intolerance (unacceptable side effects) versus HU tolerance (inadequate response). Subjects are classified as HU-tolerant or HU-intolerant at the screening visit, and this categorization is used to allow subject stratification at randomization (study day 1).

Statistical analysis.

This study consists of two parts. In Part A, the initial proof of concept phase, test subjects are randomly assigned to 1 of 3 treatment groups: compound of formula (I) at a dose of 120 mg for a 21-day treatment cycle, compound of formula (I) at a dose of 240 mg for 21 -day treatment cycle or a compound of formula (I) at a dose of 120 mg over a 28-day treatment cycle. All three groups are examined in the same way and assessed in the same way. The primary efficacy analysis for Part A is based on the mITT analysis only in subjects with baseline splenomegaly who are p53WT. After all subjects with baseline splenomegaly in Part A have had the opportunity to complete week 32, the DMC will recommend a dose/schedule of compound of formula (I) for Part B. For Part B, subjects with baseline splenomegaly are randomized to receive treatment with either the recommended dose/regimen of a compound of formula (I) from Part A, or ruxolitinib administered according to the label instructions. Results of statistical analyses, descriptive summary statistics, and supporting lists are presented by study part (A or B), treatment group, and dose. All analyses, summaries and lists are obtained using SAS software (version 9.4 or higher). The detailed methodology for summary and statistical analysis of the data collected in this study is documented in a statistical analysis plan (SAP), which will be completed prior to database locking. SAP may change the data analysis plans specified in the protocol; and if this is the case, then it is clearly documented in SAP. Any significant changes to study design or study endpoints and/or analysis are also reflected in the protocol amendment.

Duration of the study.

The study is considered completed 2 years after the last subject is enrolled, at which time subjects who continue to receive study treatment are assessed for eligibility for the replication study.

Example 9: Open-label Phase 2a/2b study of a compound of formula (I) in subjects with primary myelofibrosis (PMF), MF after polycythemia vera (post-PV-MF) or MF after essential thrombocythemia (post-ET-MF) who failed with ruxolitinib.

There is a significant unmet need for improved treatments for patients with myelofibrosis (MF) who have primary resistance, suboptimal responses, or have relapsed after treatment with ruxolitinib. The compound of formula (I) is an orally bioavailable, small molecule cytotoxic chemotherapeutic agent that binds to murine dual microprotein 2 (MDM2) and inhibits the MDM2/tumor protein 53 (p53) protein-protein interaction. The compound of formula (I) has been shown to inhibit the growth of p53 wild type (p53WT) tumor cells in vitro and tumor xenografts in vivo. p53 is a tumor suppressor and transcription factor that responds to cellular stress by activating the transcription of numerous genes involved in cell cycle arrest, apoptosis, aging, and deoxyribonucleic acid (DNA) repair. Preclinically, a key determinant of sensitivity to MDM2 inhibition is the mutational status of p53 cells, with p53WT cells being sensitive to these inhibitors. MDM2 is a negative regulator of p53 and is overexpressed in CD34+ myeloproliferative neoplasm cells harboring p53WT. MDM2 destroys and inactivates p53, allowing clonal hematopoietic stem cells to proliferate.

Study design.

It is an open-label, two-part (Part A and Part B) phase study

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2a/2b compounds (Formula I) in subjects with PMF, post-PV-MF or post-ET-MF who have failed treatment with ruxolitinib. Approximately 190 subjects are included in the study (90 in Part A and 100 in Part B).

Part A (N=90): In Part A of the study, subjects were randomly assigned to 1-3 treatment groups:

group 1, N=30 subjects: compound of formula (I) at a dose of 120 mg once daily for 1-7 days, without treatment for -8-21 days (21-day cycle);

group 2, N=30 subjects: compound of formula (I) at a dose of 240 mg once daily for 1-7 days, without treatment for 8-21 days (21-day cycle);

group 3, N=30 subjects: compound of formula (I) at a dose of 240 mg once daily for 1-7 days, without treatment for 8-28 days (28-day cycle);

Part B (N=100): Approximately 100 subjects enrolled in Part B and treated according to the recommended dose and schedule from Part A. A Data Monitoring Committee (DMC) met every 3 months for Part A and Part B during the study for review safety data for a clinical trial. The DMC was also collected after all subjects in Part A had the opportunity to complete the 24-week assessment. The DMC determines the recommended dose and schedule of a compound of Formula (I) for Part B based on the efficacy and safety data from Part A. In Part A and Part B, subjects received a compound of Formula (I) by mouth (PO) QD on days 1-7 each 21-day or 28-day cycle. Dose reductions are allowed for hematological and non-hematological toxicity. All subjects should be treated until disease progression or intolerance. Determination of disease progression is based on imaging and modified ELN criteria.

Increase in splenic volume >25% from nadir during MRI (or CT) examination as determined by central image review.

Leukemic transformation, confirmed by the number of blasts in the bone marrow >20% or

Peripheral blood blast content >20%, associated with an absolute blast count >1x10 ⁹ /L, which lasts for at least 2 weeks.

Objectives of the study

Primary Goals Final criteria assessment/measurement result Determination of the spleen reaction Proportion of subjects achieving spleen volume reduction >35% from baseline to 24 weeks, as assessed by magnetic resonance imaging (MRI) or computed tomography (CT).

Exclusion criteria.

Subjects in both Part A and Part B must meet all of the following criteria to be eligible to participate in the study:

1. Age >18 years.

2. Palpable splenomegaly at least 5 cm below the left rib margin.

3. Confirmed diagnosis of PMF, post-PV-MF or post-ET-MF, as assessed by the treating physician according to World Health Organization (WHO) criteria

4. High risk, intermediate risk 2 or intermediate risk 1, as determined by the Dynamic International Prognostic System (DIPSS).

5. General condition indicator on the ECOG scale 0-2.

6. Adequate hematological, hepatic and renal function (as defined by the protocol and for 14 days prior to the first dose of the compound of formula (I))

Hematological: ANC >1.0x10 ⁹ /l in the absence of growth factors during the previous 7 days; platelet count >100x10 ⁹ /l; peripheral blood blast count <10%.

Hepatic: total bilirubin <2.0 times the upper limit of normal (ULN), excluding Gilbert's syndrome; aspartate transaminase/serum glutamic oxaloacetic transaminase (AST/SGOT) and alanine transaminase/serum glutamic pyruvic transaminase (ALT/SGPT) <2.5 ULN.

Renal: Cockcroft-Gault estimated creatinine clearance >45 mL/min:

I (140-age) x weight (in kilograms) x [0.85 if female] x serum creatinine (in mg/dl)

7. Women of childbearing age and men who have partners of childbearing age,

- 61 045240 must consent to the use of an effective method of contraception during the study. In addition, men should continue to use contraception for 3 months after the last dose of study drug, and women should continue to use contraception for 1 week after taking the last dose of study drug. Effective birth control includes (a) combined hormonal contraceptives containing estrogen and progestogen (oral, intravaginal, transdermal); (b) progestogen-only hormonal contraceptives (oral, injectable, implantable); (c) intrauterine device; (d) intrauterine hormone releasing system; (f) bilateral occlusion of the fallopian tubes; (f) post-vasectomy partner; and (g) sexual abstinence.

Subjects in Part A must meet the following ruxolitinib treatment failure criteria to be eligible for the study:

Part A failure of ruxolitinib must meet criteria (a) or (b) below.

a) either absence of splenic response, defined as completion of at least 12 weeks of treatment with ruxolitinib, or the presence of both of the following:

persistent splenomegaly on physical examination palpable >5 cm below the lower costal margin (LCM) and TSS >10 on the MPN-SAF TSS 2.0 or patients with a single symptom score >5 or two symptoms >3, including only left upper quadrant pain symptoms , bone pain, itching or night sweats;

b) or progressive disease at any time during treatment with ruxolitinib, as defined by any of the following:

increase in splenic volume >25% from nadir as assessed by MRI or CT;

the appearance of new splenomegaly that is palpable at least 5 cm below the LCM;

>100% increase in palpable distance below the LCM with initial splenomegaly from 5 to 10 cm increase in palpable distance below the LCM by >50% with initial splenomegaly >10 cm.

Subjects in Part B must meet the following ruxolitinib treatment failure criteria to be eligible for the study:

Treatment failure with ruxolitinib in Part B must meet criteria (a) or (b) below:

a) or absence of splenic response, defined as completion of at least 12 weeks of treatment with ruxolitinib, and the presence of at least one of the following:

for subjects undergoing MRI or CT to evaluate ruxolitinib treatment, absence of splenic volume reduction of at least >35%;

initial splenomegaly before ruxolitinib treatment, palpable 5-10 cm below the LCM, but remains palpable;

baseline splenomegaly before ruxolitinib treatment that is palpable >10 cm below the LCM but does not improve by at least 50%;

baseline splenomegaly before ruxolitinib treatment, palpable <5 cm below the LCM, cannot be considered a failure of ruxolitinib treatment;

b) or progressive disease at any time during treatment with ruxolitinib, as defined by any of the following:

increase in splenic volume >25% from nadir on MRI or CT.

The appearance of new splenomegaly that is palpable at least 5 cm below the LCM;

increase in palpable distance below the LCM by >100% for initial splenomegaly from 5 to 10 cm increase in palpable distance below the LCM by >50% for initial splenomegaly >10 cm.

Exclusion criteria.

Subjects in both Part A and Part B who meet any of the following criteria will not be allowed to participate in the study:

1. Participation in another interventional clinical trial within the last 4 weeks after taking the first dose of a compound of formula (I) (participation in observational studies is permitted).

2. Recent/concomitant treatment, such as major surgery, chemotherapy, immunomodulatory therapy, biological therapy, radiation therapy, or investigational therapy within 4 weeks or approximately 5 half-lives of the first dose of the compound of formula (I).

3. Previous splenectomy.

4. Irradiation of the spleen for 3 months before the first dose of the compound of formula (I).

5. Previous allogeneic stem cell transplantation or indication for allogeneic stem cell transplantation.

6. Previous treatment with histone deacetylase (HDAC) inhibitors or BCL-2 inhibitors.

7. Previous therapy with MDM2 inhibitors or p53-targeted therapy.

8. Pregnant or breastfeeding women.

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9. History of major organ transplantation.

10. Uncontrolled intercurrent illness, including, but not limited to, acute hepatitis A; known anemia of human immunodeficiency virus (HIV); clinically significant heart disease (New York Heart Association class III or IV); symptomatic congestive heart failure; unstable angina, ventricular arrhythmia; or mental illness/social situations that limit study eligibility.

11. Subjects with clinically significant bacterial, fungal, parasitic or viral infection requiring treatment. Subjects with acute bacterial infections requiring antibiotics should defer screening/enrollment until completion of antibiotic therapy.

12. Other malignancies within the last 3 years other than curable basal cell or squamous cell skin cancer, cervical carcinoma in situ, organ-confined or treated non-metastatic prostate cancer with normal prostate-specific antigen, breast carcinoma in situ after complete surgical resection, or superficial transitional cellular bladder cancer.

13. QT prolongation grade 2 or greater (>480 milliseconds, according to NCICCTCAE criteria, version 5.0).

14. Hematopoietic growth factors (eg, erythropoietin (Epo), granulocyte colony-stimulating factor (GCSF), romiplostim) for 28 days before receiving the first dose of the compound of formula (I).

15. Active or chronic bleeding within 4 weeks prior to the first dose of a compound of formula (I).

Randomization method.

Part A: Subjects are randomized in a 1:1:1 allocation design to one of three treatment groups. The contracted clinical provider developed the Part A randomization schedule, and the actual randomization assignment was made through a secure Interactive Randomization Technology (IRT) system. Part B: subjects are randomized to the compound of formula (I), dose and schedule recommended by the DMC.

Statistical analysis.

The DMC meets every 3 months for Part A and Part B during the study to review the safety data for the clinical trial. The DMC was collected after all subjects in Part A had the opportunity to complete the 24-week assessment. The DMCs determined the recommended dosage and schedule of a compound of formula (I) for Part B based on the efficacy and safety data from Part A. Results of statistical analyses, descriptive summary statistics, and supporting lists are presented by study part (A or B) and in Part A (by groups).

Duration of the study.

The study is considered completed 2 years after the last subject is enrolled, at which time subjects continuing to receive study treatment are assessed for eligibility for the replication study.

Example 10: Effect of the compound of formula (I) on MPN-BP stem cells.

MPN-BP cell proliferation.

CD3+ cell-depleted mononuclear cells (MNCs) from 1 MPN-BP patient who had the TP53 WT gene have now been shown to consistently engraft and induce leukemia in NSG mice. To collect a sufficient number of cells to evaluate the effect of the compound of formula (I) on MPN-BP stem cells, MPN-BP cells collected from the bone marrow (BM) or spleen of NSG mice were passaged into NSG mice through serial transplantation. Mutational patterns and karyotypic abnormalities present in cells after sequential transplantation were determined by next generation sequencing (NGS) and fluorescence in situ hybridization (FISH).

Effect of the compound of formula (I) on MPN-BP stem cells.

To study the effects of compounds of formula (I) on MPN-BP stem cells, 0.5-2x10 ⁶ cells/mouse collected from NSG mice treated with MPN-BP cells were transplanted into sublethally irradiated (220 cGy) 8-9 week old NSG mice . The mice were then monitored daily for their general condition and their body weight was measured weekly. Twenty-eight days after transplantation, peripheral blood was collected from recipient mice and analyzed by complete blood count (CBC) and flow cytometric analysis to determine whether human MPN-BP had developed in these mice. These mice were used in the following studies.

High dose study of the compound of formula (I).

Mice that developed MPN-BP and had a similar leukemic load in the peripheral blood were randomly assigned to 2 groups of 3-4 mice. These two groups of mice were administered medium or a compound of formula (I) (high dose, 100 or 150 mg/kg) by gavage once daily for 7 days. After treatment, the amount of blast was monitored weekly

Claims (9)

samples of peripheral blood using flow cytometric analysis. Treatment tolerability was assessed by daily body weight (BW) measurements. These analyzes determined the time course of MPN-BP return after each drug treatment, which was used to determine the treatment-free interval for the following survival and combination treatment studies. Survival studies for the compound of formula (I). Mice that developed MPN-BP and had a similar leukemic load in the peripheral blood were randomly assigned to 3 groups of 4-5 mice and treated with a compound of formula (I) by oral gavage at a high dose (100 or 150 mg/kg) and low dose (30-50 mg/kg) once a day for 7 days. These cycles were repeated for up to 3 cycles. After treatment, overall survival rate and disease progression rate were monitored by blood count and flow cytometry analysis of mouse blood every 14 days. Mice were sacrificed if any of the following criteria were met: manifestation of disease symptoms, such as weight loss of more than 20%, hunched back, decreased activity; palpable splenomegaly extending along the midline; severe anemia, thrombocytopenia or leukopenia. Mice were monitored until they met criteria for sacrifice. In addition, 2-3 mice receiving a high or low dose of the compound of formula (I) were sacrificed after completion of 3 treatment cycles, even if they did not show symptoms of the disease. After mice were sacrificed, cells were recovered from bone marrow, spleen, and peripheral blood, and the presence of human CD45+, CD34+, CD33+, CD14+, Gly A ⁺ , CD41a ⁺ , CD19+, and CD3 ⁺ cells in these organs was determined by mAb staining and flow cytometry. analysis. These assays determined whether treatment with a compound of formula (I) resulted in increased survival and decreased numbers of leukemic cells in mouse BM and decreased spread of leukemic cells into the spleen and peripheral blood of the mouse. The remaining BMCs and spleen cells after flow cytometry analysis were cryopreserved to quantify MPN-BP cell load and to evaluate the target effects of the compound of formula (I). hCD33 ⁺ cells from BM recipient mice were selected using a FACSAria cell sorter (BD) and analyzed using FISH for marker chromosomal abnormalities as well as variant allele frequencies of mutated myeloid malignancy genes. CLAIM 1. A method of treating a myeloproliferative neoplasm (MPN), comprising the step of administering to a human subject in need a therapeutically effective amount of an MDM2 inhibitor, wherein the MDM2 inhibitor is a compound of formula (I): (I) or a pharmaceutically acceptable salt thereof, wherein the MPN is polycythemia vera (PV), phlebotomy-dependent polycythemia vera, thrombocythemia, essential thrombocythemia (ET), or myelofibrosis. 2. The method of claim 1, wherein the MPN is polycythemia vera (PV). 3. The method of claim 2, wherein the MPN is phlebotomy-dependent polycythemia vera. 4. The method of claim 1, wherein the MPN is thrombocythemia. 5. The method of claim 4, wherein the thrombocythemia is essential thrombocythemia (ET). 6. The method of claim 1, wherein the MPN is myelofibrosis. 7. The method of claim 6, wherein the myelofibrosis is selected from primary myelofibrosis (PMF), myelofibrosis after polycythemia vera (post-PV MF) and myelofibrosis after essential thrombocythemia (post-ET MF). 8. The method of claim 7, wherein the human subject has not responded to prior therapy with ruxolitinib. 9. The method according to any one of claims 1 to 8, wherein the MPN is characterized by the JAK2V617F mutation. -