Combination of ruxolitinib and incb057643 for treating myeloproliferative neoplasm

Abstract

To provide a new treatment method for a patient in need of treatment of myeloproliferative neoplasm.SOLUTION: The present application relates to treatment of myeloproliferative neoplasm in which Ruxolitinib being JAK1 / JAK2 inhibitor is used in combination with 2,2,4- trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridine-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazine-3(4H)-one being BET protein inhibitor, wherein the combination is unexpectedly synergistic.SELECTED DRAWING: None

Description

Technical Field

[0001] Priority Claim This application claims the benefit of U.S. Provisional Patent Application No. 63 / 034,214, filed on June 3, 2020, the entire disclosure of which is incorporated herein by reference.

[0002] This application relates to the treatment of myeloproliferative tumors using ruxolitinib, a JAK1 / JAK2 inhibitor, in combination with 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, a BET protein inhibitor, and this combination is unexpectedly synergistic.

Background Art

[0003] Compound 1 is a small molecule inhibitor of the acyl-binding bromodomains found within the bromodomain and extra-terminal domain (BET) proteins. Four family members (BRD2, 3, 4, T) bind to acetylated histones and promote transcription by recruiting transcriptional initiation and elongation complexes, thereby acting as a link between chromatin and transcriptional activation (see Shi, J., and Vakoc, C.R., “The Mechanisms Behind the Therapeutic Activity of BET Bromodomain Inhibition” Molecular Cell 2014;54,728-736, which is incorporated herein by reference in its entirety).

[0004] The BRD4 (and in some cases BRD3) gene is a component of chromosomal translocations that give rise to a highly malignant subtype of squamous cell carcinoma called "NUT midline" cancer. These midline cancers are defined by the t(15;19) translocation that generates the BRD4-NUT oncogene. The resulting fusion protein consists of the N-terminal BRD4 and the C-terminal NUT (nuclear protein of the testis). Removal of this protein from chromatin by either RNA silencing or small molecule inhibitors results in differentiation and growth arrest in these cancer types (see P., et al., Selective Inhibition of BET Bromodomains. Nature 2010;468,1067-1073, which is incorporated herein by reference in its entirety).

[0005] BET proteins are thought to exert a growth effect, in particular by promoting the transcription of oncogenes such as c-myc. BRD4 inhibitory compounds have been shown to prevent BRD4 from associating with chromatin containing the c-myc promoter in a multiple myeloma model. This dissociation results in a decrease in c-myc expression levels and a decrease in cell viability (see Delmore, J.E., et al., ‘‘BET Bromodomain Inhibition as a Therapeutic Strategy to Target c-MYC’’ Cell 2011;146,904-917, which is incorporated herein by reference in its entirety). BRD4 has recently been discovered to bind to "super-enhancers", which are extensive transcriptional regulatory elements within genes that are required for cell fate and survival. An example has been found in multiple myeloma, where the c-myc gene has been found to have a super-enhancer region that is bound by BRD4 and gives rise to high levels of c-myc transcription (see Loven, J., et al., ‘‘Selective Inhibition of Tumor Oncogenes by Disruption of Super-Enhancers’’ Cell 2013;153,320-334, which is incorporated herein by reference in its entirety).

[0006] The MPLW515L mutation is found in a small subset of patients with Philadelphia chromosome-negative myeloproliferative neoplasms (MPNs). This mutation causes constitutive activation of JAK2, leading to enhanced signaling through its downstream targets and subsequent increased levels of cell proliferation. Since there is substantial overlap between the JAK / STAT signaling pathway and the inflammatory pathways transcriptionally regulated by BET proteins, the combination of BET inhibitors and JAK inhibitors in MPNs may form the basis of a new treatment protocol.

[0007] New treatments are needed to improve patient outcomes. This application addresses this need and the like.

Summary of the Invention

[0008] This application provides, inter alia, a method of treating a myeloproliferative neoplasm in a patient in need thereof, comprising administering to the patient 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, and ruxolitinib or a pharmaceutically acceptable salt thereof.

[0009] This application also provides a method of treating a myeloproliferative neoplasm in a patient in need thereof, comprising administering to the patient 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, and ruxolitinib or a pharmaceutically acceptable salt thereof at a dose of from about 5 mg / day to about 60 mg / day on a free base basis.

[0010] This application further provides the use of a compound in the dosage described herein for the manufacture of a medicament for use in the treatment of myeloproliferative neoplasms.

[0011] This application also provides a dosage of the compounds described herein for use in the treatment of myeloproliferative neoplasms.

[0012] Details of one or more embodiments are set forth in the following description. Other features, objects, and advantages will be apparent from the description and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0013]

Figure 1A

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Mode for Carrying Out the Invention

[0014] For the terms “for example” and “etc.” and their grammatical equivalents, unless otherwise specified, they are understood to be followed by the phrase “but not limited to”.

[0015] As used herein, the singular forms “a”, “an” and “the” include plural referents unless the context clearly dictates otherwise.

[0016] As used herein, the term “about” means “approximately” (e.g., plus or minus about 10% of the indicated value).

[0017] As used herein, the term "solid form" refers to a compound provided herein in either an amorphous state or a crystalline state ("crystalline form" or "crystalline solid" or "crystalline solid form"), and a compound provided herein in a crystalline state may contain a solvent or water within the crystal lattice, for example, to form a solvated or hydrated crystal form. As used herein, the term "hydrate" means referring to a crystal form containing water molecules in the crystal lattice. Examples of "hydrate" crystal forms include hemihydrate, monohydrate, dihydrate, etc. Other hydrate forms such as channel hydrates are also included within the meaning of this term.

[0018] The present invention relates to the use of ruxolitinib, a JAK1 / JAK2 inhibitor, in combination with a BET protein inhibitor for treating myeloproliferative neoplasms. Ruxolitinib, (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile, is an inhibitor of JAK1 and JAK2. The IC 50 of ruxolitinib was measured with 1 mM ATP by the following Assay A and found to be less than 10 nM for JAK1 and JAK2. Ruxolitinib can be prepared by the procedure described in US7,598,257 (Example 67), filed on December 12, 2006, which is hereby incorporated by reference in its entirety. Ruxolitinib phosphate can be prepared as described in US2008 / 0312259, which is hereby incorporated by reference in its entirety.

[0019] It has been demonstrated that BET inhibition inhibits inflammatory signaling in myeloproliferative neoplasms (Kleppe M, Koche R, Zou L, et al., ‘‘Dual targeting of oncogenic activation and inflammatory signaling increases therapeutic efficacy in myeloproliferative neoplasms’’. Cancer Cell 2018;33:29-43.e7, which is incorporated herein by reference in its entirety). In the referenced study, BET inhibitors in combination with JAK inhibition reduced pathologic cytokine production and overall disease burden. Specifically, in an in vivo model of myeloproliferative neoplasms, combination therapy with ruxolitinib and JQ1, JAK1 / 2 and BET inhibitors, respectively, resulted in a decrease in inflammation and a decrease in disease burden. The treatment also eliminated fibrosis in mice with myelofibrosis, a response not previously observed with monotherapy JAK inhibitor therapy. Mechanistically, the synergistic effect of JAK and BET inhibitors is proposed to be mediated by blocking two master regulators of pathologic inflammatory signaling, JAK / STAT and NF-kB, respectively. Collectively, these data justify the evaluation of BET inhibitors in malignancies characterized by underlying inflammation such as myeloproliferative neoplasms including primary myelofibrosis.

[0020] The present application provides a method of treating a myeloproliferative neoplasm in a patient in need thereof, the method comprising administering to the patient 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one (Compound 1 below), an inhibitor of BET proteins such as BRD2, BRD3, BRD4, and BRD-t. Compound 1 can be prepared as described in U.S. Patent No. 9,540,368 or U.S. Patent No. 10,189,832, each of which is incorporated herein by reference in its entirety.

Chemical formula

[0021] The present application provides a method for treating a myeloproliferative neoplasm in a patient in need thereof, the method comprising administering to the patient 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, and ruxolitinib or a pharmaceutically acceptable salt thereof. Administration of the combination of Compound 1 and ruxolitinib can provide enhanced efficacy. As demonstrated by the examples provided herein, the combination of Compound 1 and ruxolitinib showed enhanced efficacy in an in vivo model and was significantly (p < 0.05) more effective than either ruxolitinib alone or Compound 1 alone. That is, the combination of ruxolitinib and Compound 1 provides better efficacy than the sum of the parts.

[0022] The present application provides a method for treating a myeloproliferative neoplasm in a patient in need thereof, the method comprising administering to the patient 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, and ruxolitinib or a pharmaceutically acceptable salt thereof.

[0023] The present application further provides a method for treating a myeloproliferative neoplasm in a patient in need thereof, the method comprising administering to the patient 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of ruxolitinib or a pharmaceutically acceptable salt thereof.

[0024] The present application further provides a method for treating a myeloproliferative neoplasm in a patient in need of treatment for a myeloproliferative neoplasm, the method comprising administering to the patient a 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, at a dose of about 2 mg / day to about 20 mg / day on a free base basis, and a therapeutically effective amount of ruxolitinib or a pharmaceutically acceptable salt thereof.

[0025] In some embodiments, the dose of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, is about 2 mg / day to about 18 mg / day on a free base basis.

[0026] In some embodiments, the dose of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, is about 2 mg / day to about 12 mg / day on a free base basis.

[0027] In some embodiments, the dose of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, is about 4 mg / day to about 8 mg / day on a free base basis.

[0028] For example, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 2, about 4, about 6, about 10, about 12, about 14, about 16, about 18, or about 20 mg / day on a free base basis. In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 2, about 4, about 6, about 8, about 10, about 12, about 14, about 16, about 18, or about 20 mg / day on a free base basis. In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 2, about 4, about 6, about 8, about 10, or 12 mg / day on a free base basis. In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 4, about 6, or about 8 mg / day on a free base basis. In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 8, about 10, about 12, about 14, or about 16 mg / day on a free base basis.In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 10, about 12, about 14, about 16 mg, or about 18 mg / day on a free base basis. In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof is about 2 mg / day on a free base basis. In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof is about 4 mg / day on a free base basis. In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof is about 6 mg / day on a free base basis. In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof is about 8 mg / day on a free base basis. In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof is about 10 mg / day on a free base basis.In some embodiments, the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, is about 12 mg / day on a free base basis.

[0029] In some embodiments, 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof and nilotinib or a pharmaceutically acceptable salt thereof are administered once daily (QD). In some embodiments, 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof and nilotinib or a pharmaceutically acceptable salt thereof are administered twice daily (BID).

[0030] In some embodiments, the dosage of nilotinib or a pharmaceutically acceptable salt thereof is from about 5 mg / day to about 60 mg / day on a free base basis. For example, the dosage of nilotinib or a pharmaceutically acceptable salt thereof is about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, or 60 mg / day on a free base basis.

[0031] In some embodiments, the dosage of nilotinib or a pharmaceutically acceptable salt thereof is from about 2.5 mg BID to about 30 mg BID on a free base basis. For example, the dosage of nilotinib or a pharmaceutically acceptable salt thereof is about 2.5, about 5, about 7.5, about 10, about 12.5, about 15, about 17.5, about 20, about 25, or about 30 mg BID on a free base basis.

[0032] In some embodiments, the pharmaceutically acceptable salt of ruxolitinib is ruxolitinib phosphate.

[0033] In some embodiments, 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, is selected from phosphate, dihydrochloride, hydrochloride, maleate, adipate, hydrobromide, (R)-(-)-mandelate, salicylate, benzoate, benzenesulfonate, L-pyroglutamate, methanesulfonate, (1S)-(+)-10-camphorsulfonate, fumarate, sulfate, L-tartrate, and the D-tartrate of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one. In some embodiments, 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one is the free base.

[0034] In some embodiments, 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one comprises one or more crystalline solid forms of an inhibitor of BET protein. A more detailed discussion of the crystalline forms of Compound 1 can be found in U.S. Patent No. 10,626,114, which is hereby incorporated by reference in its entirety and is briefly described herein. Typically, different crystalline forms of the same substance have different bulk properties, for example, with respect to hygroscopicity, solubility, stability, etc.

[0035] The crystalline solid forms of Compound 1 may contain a solvent such as water (e.g., hydrated forms) or may be substantially free of water and solvents (e.g., anhydrous formation). In some embodiments, the crystalline solid form is an anhydrate. In further embodiments, the crystalline solid form is hydrated. Compound 1 can be obtained in solid crystalline forms called Form I, Form II, Form III, Form IV, Form V, Form Va, Form VI, Form VII, Form VIII, Form IX, Form X, Form XI, Form XII, Form XIII, Form XIV, and Form XV, which are described in U.S. Patent No. 10,626,114, which is hereby incorporated by reference in its entirety.

[0036] Experimental data indicate that Form I is an anhydrate. Form I is characterized by its XRPD pattern and other solid-state properties. In some embodiments, Form I has one or more characteristic XRPD peaks selected from about 8.7, about 9.8, about 12.7, about 21.4, and about 23.3 degrees with respect to 2-theta.

[0037] Experimental data indicate that Form II is an anhydrate. Form II is characterized by its XRPD pattern and other solid-state properties. In some embodiments, Form II has one or more characteristic XRPD peaks selected from about 6.7, about 9.5, about 10.5, about 14.8, about 16.2, about 17.0, about 18.8, and about 19.3 degrees with respect to 2-theta.

[0038] In some embodiments, Form III has one or more characteristic XRPD peaks selected from about 7.8, about 12.4, about 13.1, about 15.2, about 15.5, about 16.9, about 17.5, and about 20.3 degrees with respect to 2-theta.

[0039] In some embodiments, Form IV has one or more characteristic XRPD peaks selected from about 11.2, about 16.3, about 18.7, and about 22.1 degrees with respect to 2-theta.

[0040] In some embodiments, Form V has one or more characteristic XRPD peaks selected from about 8.2, about 8.5, about 14.1, about 16.3, about 17.1, about 18.9, about 19.8, about 21.8, and about 22.7 degrees with respect to 2 theta.

[0041] In some embodiments, Form Va has one or more characteristic XRPD peaks selected from about 8.7, about 16.5, about 17.3, about 19.9, and about 21.6 degrees with respect to 2 theta.

[0042] In some embodiments, Form VI has one or more characteristic XRPD peaks selected from about 8.5, about 9.6, about 11.4, about 12.1, about 13.5, about 14.5, about 15.2, about 17.1, about 17.7, about 18.1, about 19.2 and about 20.7 degrees with respect to 2 theta.

[0043] In some embodiments, Form VII has one or more characteristic XRPD peaks selected from about 9.9, about 12.2, about 14.8, about 15.7, about 17.0, about 17.5, and about 18.8 degrees with respect to 2 theta.

[0044] In some embodiments, Form VIII has one or more characteristic XRPD peaks selected from about 8.1, about 8.5, about 16.2, about 16.6, about 17.0, about 17.5, about 18.0, about 18.9, about 19.6, and about 20.1 degrees with respect to 2 theta.

[0045] In some embodiments, Form IX has one or more characteristic XRPD peaks selected from about 8.6, about 9.1, about 11.4, about 13.4, about 15.2, about 18.2, about 22.1, about 22.8, and about 23.9 degrees with respect to 2 theta.

[0046] In some embodiments, Form X has one or more characteristic XRPD peaks selected from about 14.9, about 15.3, about 15.8, about 17.0, about 17.7, about 18.3, and about 19.7 degrees with respect to 2 theta.

[0047] In some embodiments, Form XI has one or more characteristic XRPD peaks selected from about 8.9, about 12.8, about 18.0, about 21.5, about 22.6, and about 23.3 degrees with respect to 2-theta.

[0048] In some embodiments, Form XII has one or more characteristic XRPD peaks selected from about 5.6, about 11.7, about 13.8, about 14.5, about 16.9, about 17.7, about 18.7, about 23.5, about 24.6, about 34.3, about 44.2, and 44.6 degrees with respect to 2-theta.

[0049] In some embodiments, Form XIII has one or more characteristic XRPD peaks selected from about 5.7, about 8.6, about 9.8, about 11.8, about 12.6, about 13.4, about 14.1, about 14.8, about 16.6, and about 19.1 degrees with respect to 2-theta.

[0050] In some embodiments, Form XIV has one or more characteristic XRPD peaks selected from about 4.0, about 11.2, about 11.9, about 14.1, about 14.8, and about 15.9 degrees with respect to 2-theta.

[0051] In some embodiments, Form XV has one or more characteristic XRPD peaks selected from about 7.4, about 9.6, about 12.4, about 13.4, about 15.5, about 16.9, about 17.7, about 19.0, about 19.5, about 20.6, and about 22.5 degrees with respect to 2-theta.

[0052] The present application provides a method for treating a myeloproliferative neoplasm in a patient in need of treatment thereof, the method comprising administering to the patient 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of ruxolitinib or a pharmaceutically acceptable salt thereof. In some embodiments, the myeloproliferative neoplasm is selected from polycythemia vera (PV), essential thrombocythemia (ET), primary myelofibrosis, chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), hypereosinophilic syndrome (HES), systemic mastocytosis with cutaneous involvement (SMCD), chronic neutrophilic leukemia (CNL), and chronic eosinophilic leukemia.

[0053] In some embodiments, the myeloproliferative neoplasm is polycythemia vera (PV).

[0054] In some embodiments, the myeloproliferative neoplasm is essential thrombocythemia (ET).

[0055] In some embodiments, the myeloproliferative neoplasm is primary myelofibrosis.

[0056] In some embodiments, the myeloproliferative neoplasm is myelofibrosis.

[0057] In some embodiments, oral administration is performed with ruxolitinib, or a pharmaceutically acceptable salt thereof, and 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof. In some embodiments, ruxolitinib or a pharmaceutically acceptable salt thereof is administered orally. In some embodiments, 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is administered orally.

[0058] In some embodiments, ruxolitinib or a pharmaceutically acceptable salt thereof and 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof are administered simultaneously or sequentially.

[0059] The foregoing embodiments are intended to be combined in any suitable combination as if the embodiments were multiple dependent claims (e.g., embodiments regarding individual dosages of ruxolitinib, embodiments regarding individual dosages of BET protein inhibitor (Compound 1), embodiments regarding salt forms, embodiments regarding individual types of myeloproliferative tumors, and embodiments regarding oral administration can be combined in any combination). The combinations are not separately listed herein merely for the sake of brevity.

[0060] All compounds and their pharmaceutically acceptable salts can be found together with, or isolated from, other substances such as water and solvents (e.g., hydrates and solvates). In the solid state, the compounds and salts described herein can exist in various forms, for example, they can take the form of solvates including hydrates. Since the compounds can be in any solid form such as polymorphs or solvates, unless otherwise specified, references to the compounds and their salts herein should be understood to encompass any solid form of the compounds.

[0061] As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0062] The present invention also includes pharmaceutically acceptable salts of the compounds described herein. The term "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds in which the parent compound is modified by converting an existing acidic or basic moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. Pharmaceutically acceptable salts of the present invention include, for example, non-toxic salts of the parent compounds formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or free base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or in a mixture of the two (generally, non-aqueous media such as ether, ethyl acetate, alcohols (e.g., methanol, ethanol, isopropanol, or butanol) or acetonitrile (MeCN) are preferred). A list of suitable salts can be found in Remington’s Pharmaceutical Sciences, 17th It can be found in Ed., (Mack Publishing Company, Easton, 1985), p. 1418, Berge et al., J. Pharm. Sci., 1977, 66(1), 1 - 19, and Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (Wiley, 2002). In some embodiments, the compounds described herein include N - oxide forms.

[0063] The dosages described herein are on a free base basis. "Free base basis" means that even when the actual active ingredient is a salt of a compound having a molecular weight different from that of the free base, the amount of the compound (e.g., ruxolitinib or its salt) in the dosage form is measured based only on the molecular weight of the free base of the compound. For example, the conversion factor of ruxolitinib phosphate to the free base is 0.7575.

[0064] The terms "subject" or "patient" used interchangeably refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, or primates, most preferably humans.

[0065] The phrase "therapeutically effective amount" refers to the amount of an active compound or pharmaceutical agent that elicits a biological or medical response in a tissue, system, animal, subject, or human that is sought by a researcher, veterinarian, physician, or other clinician.

[0066] The terms "treating" or "treatment" refer to one or more of: (1) inhibiting a disease, e.g., inhibiting a disease, condition or disorder in an individual who is experiencing or exhibiting the pathology or general symptoms of the disease, condition or disorder (i.e., preventing further progression of the pathology and / or general symptoms), (2) ameliorating a disease, e.g., ameliorating a disease, condition or disorder in an individual who is experiencing or exhibiting the pathology or general symptoms of the disease, condition or disorder, e.g., reducing the severity of the disease (i.e., reversing the pathology and / or symptoms). In one embodiment, treating or treatment includes preventing or reducing the risk of developing a disease, e.g., preventing or reducing the risk of developing a disease, condition or disorder in an individual who may have a predisposition to the disease, condition or disorder but who has not yet experienced or presented the pathology or general symptoms of the disease.

[0067] The term "BID" means twice a day.

[0068] The term "QD" means once a day.

[0069] Additional combinations Compound 1 can be used in further combination therapies in which Compound 1 is administered in combination with other treatments such as the administration of one or more additional therapeutic agents. The additional therapeutic agents are typically those commonly used to treat the particular condition being treated. Additional therapeutic agents can include, for example, chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressive agents, and Bcr-Abl, Flt-3, RAF, FAK, and JAK kinase inhibitors for treating BET protein-related diseases, disorders or conditions. One or more additional pharmaceuticals can be administered to the patient simultaneously or sequentially.

[0070] In some embodiments, Compound 1 can be used in combination with a therapeutic agent that targets an epigenetic regulator. Examples of epigenetic regulators include histone lysine methyltransferase, histone arginine methyltransferase, histone demethylase, histone deacetylase, histone acetylase, and DNA methyltransferase. Histone deacetylase inhibitors include, for example, vorinostat.

[0071] Compound 1 can be used in combination with chemotherapeutic agents or other anti-proliferative agents for treating cancer and other proliferative diseases. Compound 1 can also be used in combination with medical therapies such as surgery or radiation therapy, e.g., gamma rays, neutron beam radiation therapy, electron beam radiation therapy, proton beam therapy, brachytherapy, and systemic radioisotopes. Examples of suitable chemotherapeutic agents include abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, bleomycin, bortezomib, bortezomib, busulfan intravenous, busulfan oral, calusterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, deruxtecan, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, drostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib mesylate, interferon alpha 2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, mechlorethamine, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone phenylpropionate, nelarabine, nolatrexed, oxaliplatin, paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, luxolitinib, sorafenib,Any of streptozocin, sunitinib, sunitinib maleate, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, and zoledronic acid is included.

[0072] Compound 1 can be used in combination with loxoribine for treating cancer and other proliferative diseases.

[0073] Compound 1 can be used in combination with one or more immune checkpoint inhibitors. Exemplary immune checkpoint inhibitors include inhibitors against immune checkpoint molecules such as CD27, CD28, CD40, CD122, CD96, CD73, CD47, OX40, GITR, CSF1R, JAK, PI3KΔ, PI3Kγ, TAM, arginase, CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, VISTA, PD-1, PD-L1, and PD-L2. In some embodiments, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In some embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, and VISTA. In some embodiments, the compounds provided herein can be used in combination with one or more agents selected from KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors, and TGFRβ inhibitors.

[0074] In some embodiments, the inhibitor of the immune checkpoint molecule is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

[0075] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of PD-1, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), pidilizumab, SHR-1210, PDR001, or AMP-224. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD1 antibody is pembrolizumab.

[0076] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of PD-L1, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is BMS-935559, MEDI4736, MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments, the anti-PD-L1 monoclonal antibody is MPDL3280A or MEDI4736.

[0077] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CTLA-4, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab.

[0078] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of LAG3, such as an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016 or LAG525.

[0079] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of GITR, such as an anti-GITR antibody. In some embodiments, the anti-GITR antibody is TRX518 or MK-4166.

[0080] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of OX40, such as an anti-OX40 antibody or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is MEDI0562. In some embodiments, the OX40L fusion protein is MEDI6383.

[0081] Compound 1 can be used in combination with one or more agents for treating diseases such as cancer. In some embodiments, the agent is an alkylating agent, a proteasome inhibitor, a corticosteroid, or an immunomodulatory agent. Examples of alkylating agents include cyclophosphamide (CY), melphalan (MEL), bendamustine, and the like. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulatory agent is lenalidomide (LEN) or pomalidomide (POM).

[0082] To treat autoimmune or inflammatory conditions, Compound 1 can be administered in combination with a corticosteroid such as triamcinolone, dexamethasone, fluocinolone, cortisone, prednisone, or flumetholone.

[0083] To treat autoimmune or inflammatory conditions, Compound 1 can be administered in combination with an immunosuppressant such as fluocinolone acetonide (Retisert®), rimxolone (AL-2178, Vexol, Alcon) or cyclosporine (Restasis®).

[0084] To treat autoimmune or inflammatory conditions, Compound 1 is combined with Dehydrex (trademark) (Holles Labs), Cibamide (Opko), Sodium Hyaluronate (Vismed, Lantibio / TRB Chemedia), Cyclosporine (ST-603, Sirion Therapeutics), ARG101(T) (testosterone, Argentis), AGR1012(P) (Argentis), Ecabet Sodium (Senju-Ista), Gefarnate (Santen), 15-(s)-Hydroxyicosatetraenoic acid (15(S)-HETE), Sevirermin, Doxycycline (ALTY-0501, Alacrity), Minocycline, iDestrin (trademark) (NP50301, Nascent Pharmaceuticals), Cyclosporine A (Nova 22007, Novagali), Oxytetracycline (Duramycin, MOLI1901, Lantibio), CF101 (2S,3S,4R,5R)-3,4-Dihydroxy-5-[6-[(3-iodophenyl)methylamino]purin-9-yl]-N-methyl-oxolane-2-carbamyl, Can-Fite Biopharma), Vercosporin (LX212 or LX214, Lux Biosciences), ARG103 (Agentis), RX-10045 (synthetic resolvin analog, Resolvyx), DYN15 (Dyanmis Therapeutics), Riboglitazone (DE011, Daiichi Sanko), TB4 (RegeneRx), OPH-01 (Ophtalmis Monaco), PCS101 (Pericor Science), REV1-31 (Evolutec), Lacritin (Senju), Levamiside (Otsuka-Novartis), OT-551 (Othera), PAI-2 (University of Pennsylvania and Temple University), Pilocarpine, Tacrolimus, Pimecrolimus (AMS981, Novartis), Loteprednol Etabonate, Rituximab, Diclofasol Tetrasodium (INS365,It can be administered in combination with one or more additional agents selected from Inspire), KLS-0611 (Kissei Pharmaceuticals), dehydroepiandrosterone, anakinra, anakinra, efalizumab, mycophenolate sodium, etanercept (Embrel®), hydroxychloroquine, NGX267 (Torrey Pines Therapeutics), or thalidomide.,

[0085] In some embodiments, Compound 1 can be administered in combination with one or more agents selected from antibiotics, antiviral agents, antifungal agents, anesthetics, anti-inflammatory agents including steroid and non-steroid anti-inflammatory drugs, and anti-allergy agents. Examples of suitable pharmaceuticals include aminoglycosides such as amikacin, gentamicin, tobramycin, streptomycin, netilmicin, and kanamycin; fluoroquinolones such as ciprofloxacin, norfloxacin, ofloxacin, trovafloxacin, lomefloxacin, levofloxacin, and enoxacin; naphthyridine; sulfonamide; polymyxin; chloramphenicol; neomycin; paromomycin; colistimethate; bacitracin; vancomycin; tetracycline; rifampin and its derivatives ("rifamycins"); cycloserine; beta-lactam; cephalosporin; amphotericin; fluconazole; flucytosine; natamycin; miconazole; ketoconazole; corticosteroid; diclofenac; flurbiprofen; ketorolac; suprofen; cromolyn; lodoxamide; levocabastine; naphazoline; antazoline; pheniramine; or azalide antibiotics.

[0086] Other examples of agents that may be combined with the provided compounds include the treatment of Alzheimer's disease such as donepezil and rivastigmine; the treatment of Parkinson's disease such as L-DOPA / carbidopa, entacapone, ropinirole, pramipexole, bromocriptine, pergolide, trihexyphenidyl and amantadine; therapeutic agents for multiple sclerosis (MS) such as beta-interferon (e.g., Avonex® and Rebif®), glatiramer acetate, and mitoxantrone; the treatment of asthma such as albuterol and montelukast; antipsychotic agents such as ziprasidone, risperidone, seroquel, and haloperidol; anti-inflammatory agents such as dexamethasone, or corticosteroids such as prednisone, TNF blockers, IL-1RA, azathioprine, cyclophosphamide, sulfasalazine; immunomodulators including immunosuppressants such as cyclosporine, tacrolimus, rapamycin, mycophenolate mofetil, interferon, corticosteroids, cyclophosphamide, azathioprine and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferon, anticonvulsants, ion channel blockers, riluzole, or antiparkinson agents; agents for treating cardiovascular diseases such as beta blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers, or statins; therapeutic agents for liver diseases such as corticosteroids, cholestyramine, interferon, and antiviral agents; agents for treating blood disorders such as corticosteroids, antileukemic agents, or growth factors; or agents for treating immunodeficiency disorders such as gamma globulin.

[0087] In some embodiments, Compound 1 is administered in combination with a JAK kinase inhibitor (e.g., ruxolitinib, tofacitinib, baricitinib, CYT387, GLPG0634, lestaurtinib, pacritinib, TG101348, or a JAK1-selective inhibitor), a Pim kinase inhibitor (including an inhibitor of one or more of PIM1, PIM2, and PIM3), a PI3 kinase inhibitor including a PI3K delta-selective and broad-spectrum PI3K inhibitor, a MEK inhibitor, a cyclin-dependent kinase inhibitor, a b-RAF inhibitor, an mTOR inhibitor, a proteasome inhibitor (e.g., bortezomib, carfilzomib), an HDAC inhibitor (e.g., panobinostat, vorinostat), a DNA methyltransferase inhibitor, dexamethasone, melphalan, or an immunomodulatory agent (e.g., lenalidomide, pomalidomide).

[0088] When two or more pharmaceuticals are administered to a patient, they can be administered simultaneously, sequentially, or in combination (e.g., for three or more agents).

[0089] Composition The compounds can be administered in the form of pharmaceutical compositions. These compositions can be prepared by methods well known in the pharmaceutical art and can be administered by various routes depending on whether local or systemic treatment is indicated and on the region to be treated. Administration can be local (including transdermal, epidermal, ocular, and mucosal (including intranasal, vaginal, and rectal delivery)), pulmonary (e.g., inhalation or insufflation of powder or aerosol, including by nebulizer; intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular injection or infusion, or intracranial, e.g., intrathecal or intraventricular administration. Parenteral administration can be in the form of a single bolus injection or, e.g., by continuous perfusion pump. Pharmaceutical compositions and formulations for local administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder, or oily bases, thickeners, etc. may be necessary or desirable.

[0090] The pharmaceutical composition can contain a compound or a pharmaceutically acceptable salt thereof as an active ingredient, in combination with one or more pharmaceutically acceptable carriers (excipients). In some embodiments, the composition is suitable for topical administration. In the manufacture of the composition, the active ingredient is typically mixed with the excipient, diluted by the excipient, or encapsulated in such a carrier in the form of, for example, capsules, sachets, paper or other containers. When the excipient acts as a diluent, the excipient can be a solid, semi-solid or liquid material that acts as a vehicle, carrier or medium for the active ingredient. Thus, the composition can be in the form of tablets, pills, powders, troches, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or liquid medium), for example, ointments containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

[0091] When preparing the formulation, the active compound can be milled to obtain an appropriate particle size before being combined with other ingredients. If the active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh. If the active compound is substantially water-soluble, the particle size can be adjusted by milling to provide a substantially uniform distribution in the formulation, for example, about 40 mesh.

[0092] The compound can be milled using known milling procedures such as wet milling to obtain a particle size suitable for tablet formation and other dosage forms. The finely divided (nanoparticle) preparations of the compounds of the present invention can be prepared by processes known in the art, see, for example, WO2002 / 000196.

[0093] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulations can include lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl benzoate and propyl hydroxybenzoate; sweetening and flavoring agents. The compositions of the present invention can be formulated to provide rapid, sustained, or delayed release of the active ingredient after administration to a patient by using procedures known in the art.

[0094] In some embodiments, the pharmaceutical composition comprises silicified microcrystalline cellulose (SMCC) and at least one compound described herein, or a pharmaceutically acceptable salt thereof. In some embodiments, the silicified microcrystalline cellulose comprises about 98% microcrystalline cellulose and about 2% silicon dioxide w / w.

[0095] In some embodiments, the composition is a sustained release composition comprising at least one compound described herein or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier. In some embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof and at least one component selected from microcrystalline cellulose, lactose monohydrate, hydroxypropyl methylcellulose, and polyethylene oxide. In some embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof, microcrystalline cellulose, lactose monohydrate, and hydroxypropyl methylcellulose. In some embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof, microcrystalline cellulose, lactose monohydrate, and polyethylene oxide. In some embodiments, the composition further comprises magnesium stearate or silicon dioxide. In some embodiments, the microcrystalline cellulose is Avicel PH102™. In some embodiments, the lactose monohydrate is Fast-flo 316™. In some embodiments, the hydroxypropyl methylcellulose is hydroxypropyl methylcellulose 2208 K4M (e.g., Methocel K4 M Premier™) and / or hydroxypropyl methylcellulose 2208 K100LV (e.g., Methocel K00LV™). In some embodiments, the polyethylene oxide is polyethylene oxide WSR 1105 (e.g., Polyox WSR 1105™).

[0096] The components used to formulate the pharmaceutical composition are of high purity and substantially free of potentially harmful contaminants (e.g., at least of national food grade, generally at least of analytical grade, and more typically at least of pharmaceutical grade). Particularly for human consumption, the composition is preferably manufactured or formulated under the standards of Good Manufacturing Practice as defined by the applicable regulations of the U.S. Food and Drug Administration. For example, suitable formulations can be sterile and / or substantially isotonic and / or can fully comply with all Good Manufacturing Practice regulations of the U.S. Food and Drug Administration.

[0097] The active compounds can be effective over a wide dosage range and are generally administered in a therapeutically effective amount. However, it will be understood that the actual amount of the compound administered will usually be determined by the physician according to relevant circumstances including the condition being treated, the selected route of administration, the actual compound being administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, etc.

[0098] The therapeutic dosage of the compounds of the present invention can vary, for example, according to the particular use for which the treatment is carried out, the mode of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compounds of the present invention in the pharmaceutical composition can vary depending on many factors including the dosage, chemical properties (e.g., hydrophobicity), and route of administration.

[0099] To prepare solid compositions such as tablets, the main active ingredient is mixed with pharmaceutical excipients to form a solid preliminary preparation composition containing a homogeneous mixture of the compounds of the present invention. When these preliminary preparation compositions are referred to as homogeneous, the active ingredient is typically uniformly dispersed throughout the composition so that the composition can be easily subdivided into equally effective unit dosage forms, such as tablets, pills and capsules. This solid preliminary preparation is then subdivided into unit dosage forms of the above type containing, for example, from about 0.1 to about 1000 mg of the active ingredient of the present invention.

[0100] The tablets or pills of the present invention can be formulated by coating or other methods to provide a dosage form with the advantage of long-acting. For example, the tablets or pills can contain an inner dosage amount and an outer dosage amount component, and the latter is in the form of an envelope over the former. The two components can be separated by an enteric layer that acts to resist disintegration in the stomach and allows the inner component to enter the duodenum intact or allows the release to be delayed. Various materials can be used for such an enteric layer or coating, and such materials include a number of polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0101] Liquid forms in which the compounds and compositions of the present invention can be incorporated for oral or injectable administration include aqueous solutions, properly flavored syrups, aqueous or oil suspensions, and flavored emulsions containing edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0102] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, as well as powders. The liquid or solid compositions can contain the appropriate pharmaceutically acceptable excipients described above. In some embodiments, the composition is administered via the oral or nasal respiratory route for local or systemic effects. The composition can be nebulized by the use of an inert gas. The nebulized solution can be breathed directly from the nebulizing device, or the nebulizing device can be attached to a face mask, tent, or intermittent positive pressure breathing apparatus. The solution, suspension, or powder composition can be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0103] The topical preparation can include one or more conventional carriers. In some embodiments, the ointment can contain water and one or more hydrophobic carriers selected, for example, from liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white Vaseline®, etc. The carrier composition of the cream can be based on water in combination with glycerol and one or more other components, such as glycerol monostearate, PEG-glycerol monostearate, and cetylstearyl alcohol. The gel can be formulated using isopropyl alcohol and water in suitable combination with other components, such as glycerol, hydroxyethyl cellulose, etc.

[0104] The amount of the compound or composition administered to the patient will vary depending on what is administered, the purpose of administration such as prophylaxis or treatment, the condition of the patient, the mode of administration, etc. For therapeutic use, the composition can be administered to a patient already suffering from the disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose depends on the judgment of the attending physician according to the disease state being treated, as well as factors such as the severity of the disease, the age, weight, and general condition of the patient.

[0105] The composition administered to the patient can be in the form of the above pharmaceutical compositions. These compositions can be sterilized by conventional sterilization techniques or can be sterile filtered. The aqueous solution can be packaged for use as is or can be lyophilized, and the lyophilized preparation is combined with a sterile aqueous carrier prior to administration. The pH of the compound preparation is typically from 3 to 11, more preferably from 5 to 9, and most preferably from 7 to 8. It will be understood that the use of certain of the foregoing excipients, carriers, or stabilizers can result in the formation of pharmaceutical salts.

[0106] Kit This application also includes a useful pharmaceutical kit comprising one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound or any of its embodiments. Such kits can further include one or more of various conventional pharmaceutical kit components, such as, for example, containers containing one or more pharmaceutically acceptable carriers, additional containers, etc. Instructions can also be included in the kit in the form of an insert or label indicating the amount of the component to be administered, instructions for administration, and / or instructions for mixing the components.

[0107] The present invention will be described in more detail using specific examples. The following examples are provided for illustrative purposes only and are in no way intended to limit the present invention. Those skilled in the art will readily recognize various non-essential parameters that can be changed or modified to produce essentially the same results. The compounds of the examples were found to be BET protein kinase inhibitors by at least one assay described herein.

Example

[0108] Example 1: Characterization of Compound 1 in a xenograft model of JAK2V617F-MPN / AML Compounds and formulations: Compound 1 was used in these tests and was manufactured by Incyte Corporation. Ruxolitinib was used in these tests and was manufactured by Wilmington PharmaTech.

[0109] Methods: Approximately 5- to 6-week-old female SCID mice were subcutaneously inoculated with 107 SET-2 cells in Matrigel. Administration was initiated when the tumor size reached approximately 200 mm3. Dosing for these tests was performed by forced oral administration.

[0110] For efficacy studies, tumor measurements were taken every 2 - 3 days using a digital caliper. Tumor volume was calculated using the formula: volume = (length × width2) / 2, where width was the smaller dimension. Body weight was also monitored.

[0111] Animals were sacrificed at (1) the end of the study, (2) when tumor size reached 10% of total body weight, or (3) when body weight decreased by 20%.

[0112] Statistical analysis was performed using Prism Graphpad software. All comparisons were made using two - way ANOVA with p < 0.05 considered significant.

[0113] Efficacy of Compound 1 in the JAK2 V617F - expressing HEL xenograft model of erythroleukemia The purpose of this study was to investigate the ability of the BET inhibitor Compound 1 to inhibit tumor growth in an MPN - like AML model expressing the JAK2 V617F mutation. Female SCID mice were subcutaneously inoculated with 10 7 HEL cells in Matrigel. When tumors reached approximately 160 mm 3 Compound 1 was orally administered to the mice (n = 10 per group) for 14 days as follows. 1. Either 1 or 3 mg / kg, or vehicle control BID 2. 3, 10, or 30 mg / kg QD

[0114] As shown in Figure 1A, administration of Compound 1 inhibited tumor growth in a dose - related manner, and the mean tumor volume from each Compound 1 - treated group was statistically significantly decreased from the vehicle group (p < 0.02 for each group, two - way ANOVA). The 30 mg / kg QD dose was the most effective, resulting in a significant tumor regression. The data indicate that since QD administration is as effective as BID administration in this model, administration of Compound 1 in a BID regimen can be considered but is not necessary. As shown in Figure 1B, all doses were tolerated as determined by the absence of weight loss.

[0115] Efficacy from the combination of Compound 1 and ibrutinib in a xenograft model of SET-2 Ten SET-2 cells in Matrigel were subcutaneously inoculated into female SCID mice. 7 When the tumors reached approximately 175 mm 3 in size, mice (n = 8 per group) were orally administered for 14 days as follows. 1. Vehicle control or ibrutinib at 30 mg / kg BID 2. Compound 1 at 10 mg / kg QD 3. Combination of ibrutinib and Compound 1

[0116] For optimal readout of possible combination effects, Compound 1 and ibrutinib were dosed sub-optimally. As seen in Figure 2A, both monotherapies significantly inhibited tumor growth compared to vehicle control (p < 0.02 each, two-way ANOVA). Furthermore, the combination of Compound 1 and ibrutinib gave enhanced significant efficacy compared to the results predicted based on the efficacy of the single agents (p < 0.007 for the combination vs. both single-agent arms, two-way ANOVA). There was a ~15% body weight loss in the combination group (Figure 2B), but this nadir was within the limits of tolerability.

[0117] Efficacy from the combination of Compound 1 and ibrutinib in a xenograft model of SET-2 Ten SET-2 cells in Matrigel were subcutaneously inoculated into female SCID mice. 7 When the tumors reached approximately 150 mm 3 in size, mice (n = 8 per group) were orally administered for 11 days as follows. 1. Vehicle control or ibrutinib at 60 mg / kg BID 2. Compound 1 at 10 mg / kg QD 3. Combination of ibrutinib and Compound 1

[0118] In this study, ruxolitinib was administered at doses higher than 1.2, resulting in significant tumor growth inhibition as a single agent (Figure 3A). Administration of compound 1 with ruxolitinib did not increase the tumor growth inhibition seen with either single-agent ruxolitinib or single-agent compound 1. The combination of compound 1 and ruxolitinib resulted in an acceptable weight loss (about 12% in Figure 3B), but all doses were tolerated.

[0119] The BET inhibitor compound 1 is effective at acceptable doses in two models of JAK2V617F AML. The combination of compound 1 and the sub-optimal dose of the JAK1 / 2 inhibitor ruxolitinib (30 mg / kg BID) results in a significant increase in tumor growth inhibition compared to single-agent therapy. These data demonstrate the potential of BET inhibitors in combination with JAK inhibitors in JAK2V617F-driven myeloid malignancies.

[0120] Example 2: Efficacy of compound 1 in combination with ruxolitinib in a mouse model of MPLW515L-driven MPN Compounds and formulations: Compound 1 and ruxolitinib were used in these studies and were manufactured by Wilmington PharmaTech.

[0121] Methods: One week before bone marrow transplantation, 8-week-old female Balb / c mice (Charles River Laboratories) were injected intraperitoneally once with 150 mg / kg of 5-fluorouracil. Five days later, these mice were sacrificed and bone marrow was collected by aspiration of the femurs. Red blood cells were lysed using BD Pharma Lyse buffer (BD Biosciences) and then washed with PBS. The remaining bone marrow was seeded in 10% FBS / RPMI overnight in standard cell culture incubation. The next day, 1 ml of virus was added to 106 cells, and then an ecotropic retrovirus (MSCV puromycin backbone) expressing the MPLW515L gene was allowed to infect the bone marrow by centrifugation at 1800 rpm, 37 °C for 90 minutes in a 6-well plate. After centrifugation, the infected cells were maintained in an incubator until injection into the recipient the next day.

[0122] The bone marrow of recipient mice was ablated using 50 mg / kg of 5-fluorouracil 5 days before transplantation with Balb / c bone marrow expressing MPLW515L via retroviral infection (approximately 2×10 5 cells per recipient mouse) in 8-week-old Balb / c mice. Blood counts were performed 7 days after transplantation, and four cohorts with equal mean platelet counts (n = 10 / cohort) were created for treatment. Administration was started on day 8 after transplantation and continued for 13 days. On day 14 after transplantation, the mice were sacrificed, blood was collected by cardiac puncture, and the spleen was weighed as a surrogate for disease burden.

[0123] Blood was collected once a week by retro-orbital sinus, and complete blood count values (CBC) were determined using a blood analyzer (Abaxis, model HM5).

[0124] Statistical analysis was performed using Prism Graphpad software. All comparisons were made using an unpaired t-test, and p < 0.05 was considered significant.

[0125] Efficacy of Compound 1 in combination with ruxolitinib in the MPLW515L model of mouse MPN To evaluate the ability of BET inhibitor Compound 1 to improve the activity of ruxolitinib alone in a preclinical MPN model, female Balb / c mice were transplanted with MPLW515L-expressing bone marrow. On day 7 after transplantation, the mice were randomized into cohorts for dosing based on platelet levels determined by CBC. The next day, Compound 1 at 10 mg / kg, QD, ruxolitinib at 60 mg / kg, BID, the combination of Compound 1 and ruxolitinib, or vehicle control were administered to the mice to initiate oral dosing. Dosing was continued for 14 days, after which the mice were bled for CBC, the spleens were harvested and weighed as a surrogate for disease burden. Two vehicle-treated mice died before the end of the study on day 14 after dosing, one on day 12 and the other on day 13 after dosing.

[0126] While each single agent was able to reduce spleen size, the combination gave the highest efficacy. The spleens of mice treated with the Compound 1 / ruxolitinib combination were significantly smaller than those of either single agent alone (Figure 4A). Ruxolitinib had a greater effect on reducing white blood cell counts than Compound 1, and there was little difference in WBC counts between ruxolitinib-treated mice and those receiving combination therapy (Figure 4B).

[0127] Statistics were determined by unpaired t-tests performed using Prism Graphpad software ( ** p<0.01, **** p<0.0001).

[0128] At the doses selected for this study, both Compound 1 and ruxolitinib were able to delay the expansion of white blood cells and suppress splenomegaly induced by MPLW515L-expressing bone marrow. The combination of Compound 1 and ruxolitinib resulted in a significant reduction in disease burden compared to either monotherapy as measured by spleen weight. These data demonstrate the potential of BET inhibitors in combination with JAK inhibitors in myeloproliferative neoplasms.

[0129] Example 3: Statistical Demonstration of the Synergistic Effect between Compound 1 and Ruxolitinib The synergistic effect between Compound 1 and ruxolitinib was evaluated using the Chou-Talalay equation (see Chou, ‘‘Drug Combination Studies and Their Synergy Quantification Using the Chou-Talalay Method’’ Cancer Res;70(2) January 15, 2010, which is incorporated herein by reference). A synergistic effect shows a greater effect by the combination than would be expected based on how the individual compounds act. Table 1 has data from two experiments described herein. The first dasa set listed is the MPLW515L model, which uses spleen weight as a disease progression marker (i.e., a larger spleen = worse disease). MPL SPL indicates the size of the spleen in grams from the MPLW515L mouse MPN model. Fa indicates the percentage change compared to the vehicle, showing the fraction affected. The row of expected values shows the expected effect of the combination based on the Chou-Talalay synergy index. Since the numbers are smaller than the effect in the fraction affected, this demonstrates a greater effect than would be expected from combining Compound 1 and ruxolitinib, demonstrating that a synergistic effect occurs between the two compounds and results in an unexpected effect. The second experiment is SET-2, a JAK2V617F-expressing cell line used in the xenograft tumor model. The Fa for SET-2 is the tumor growth inhibition level for each dose and combination used. Again, the expected value is smaller than the actual effect, showing an unexpected synergistic effect. In both experiments, the Chou-Talalay equation gives a value smaller than the effect seen with the combination of ruxolitinib + Compound 1, which is interpreted as a synergistic combination in each experiment.

[0130] [Table 1]

[0131] Example 4: Clinical Trial Protocol of Compound 1 as Monotherapy in Participants with Myelofibrosis Study Design This is a Phase 1 open-label, two-part study of Compound 1 as monotherapy in participants with relapsed or refractory myelofibrosis (for further details, see ‘‘Safety and Tolerability Study of INCB057643 in Participants With Myelofibrosis’’, ClinicalTrials.gov Identifier: NCT04279847, which is incorporated herein by reference in its entirety). INCB057643 is also referred to as Compound 1 in the present disclosure. Participants have received at least one prior treatment, including ruxolitinib, and have no further available treatment known to provide clinical benefit and have an intermediate-2 or high-risk category according to DIPSS. Participants will receive 4 mg of Compound 1 QD continuously.

[0132] The study will be conducted in two parts. Part 1 will evaluate the initial safety and tolerability of 4 mg QD of Compound 1 in participants with relapsed or refractory myelofibrosis. The study drug will be self-administered once daily for 28 days, which is one cycle. Participants will continue taking the study drug as long as they are deriving benefit and do not meet the discontinuation criteria.

[0133] If the starting dose of 4 mg QD is considered acceptable in Part 1, the starting dose for Part 2 will be 4 mg QD; otherwise, the starting dose for Part 2 will be 2 mg QD. Part 2 will also administer Compound 1 as monotherapy.

[0134] Throughout the study, AEs will be continuously monitored in both Part 1 and Part 2. If the cumulative incidence of grade 3 or 4 drug-related AEs occurs in more than 40% of the participants, the study will be discontinued. Similarly, if there is more than one fatal drug-related event, the study will be discontinued. The study may only be resumed after discussion with the FDA.

[0135] Pharmacokinetics of Compound 1 as Monotherapy The clinical PK of Compound 1 in participants with progressive malignancies (solid and hematological malignancies) was evaluated in a Phase 1 / 2 open-label, dose-escalation / dose-expansion, safety and tolerability study. Participants received continuous QD doses of Compound 1. As of the data cut-off date (September 23, 2019), 25, 101, and 8 participants who received 8 mg, 12 mg, and 16 mg of Compound 1 (treated as monotherapy) across Parts 1 and 2, respectively, had been evaluated for PK. The PK parameters of the parent Compound 1 are summarized in Tables 2 and 3 (see Figure 5). Table 2 shows a summary of the pharmacokinetic parameters of Compound 1 as monotherapy (Parts 1 and 2) on Day 1 of Cycle 1. Table 3 shows a summary of the pharmacokinetic parameters of Compound 1 as monotherapy (Parts 1 and 2) at steady state (Day 8 of Cycle 1).

[0136]

Table 2

[0137] Blood samples for determination of plasma concentrations of Compound 1 in Parts 1 and 2 were drawn at the lead-in dose on Day 1 of Cycle 1, Day 2 of Cycle 1, and Day 8 of Cycle 1, and at 0.5, 1, 2, 4, 6, and 8 hours on Day 1 of Cycle 1 and Day 8 of Cycle 1. Plasma samples of Compound 1 were assayed by a validated LC-MS / MS method.

[0138] Following multiple-dose administration in the fasting state, plasma concentrations of Compound 1 typically peak at 1 - 2 hours (T max median) after dosing (C max ), then show monoexponential decay, and the geometric mean t 1 / 2 at steady state is approximately 10 hours and is not dose-dependent. Steady state was predicted to be achieved after 2 days with once-daily dosing based on a 10-hour effective half-life. Steady state AUC 0-24 divided by the AUC of the first dose 0-24By comparison, the accumulation of compound 1 exposure was minimal (<10%). Within the dose range of 8 to 16 mg QD, the steady-state C max and AUC 0-24 increased proportionally to the dose, that is, compound 1 showed approximately linear pharmacokinetics over the tested dose range. ANOVA of the dose-normalized PK parameters (C max , ss and AUC ss、0~24 ) using all tests or pairwise comparisons (see Table 2) demonstrated that the dose-normalized C max , ss or AUC ss、0~24 did not differ statistically significantly over or between doses (P>0.05). Compound 1 had a low steady-state oral clearance with a geometric mean CL ss / F of 9.92 - 10.7 L / h and a moderate volume of distribution with a geometric mean V z / F of 138 - 197 L. At a dose of 12 mg QD, the geometric mean (CV%) of t 1 / 2 , steady-state C max , and AUC 0-24 was 9.17 h (49.7%), 272 nM (40.9%), and 2740 h*nM (43.2%), respectively.

[0139] The estimated steady-state AUC 0-24 of compound 1 was 457 h*nM and 913 h*nM for 2 mg QD and 4 mg QD, respectively, based on linear PK extrapolation from the 12 mg QD geometric mean AUC ss,0-24 . The simulated PK data of compound 1 at 4 mg QD are shown in Figure 7.

[0140] Pharmacodynamics for the treatment of compound 1 as monotherapy Pharmacodynamic analysis was performed using an ex vivo assay that measures the protein level of cMyc, a BRD4 target gene in KMS12BM cells, spiked into participant plasma samples collected at various time points before and after dosing. In preliminary PD analysis after oral dosing of 8 mg, 12 mg, and 16 mg, Compound 1 showed inhibition of total cMyc protein expression, with maximal inhibition occurring between 1 and 4 hours. Mean cMyc inhibition at steady state (Day 8, 8 hours post-dose) was 30%, 44%, and 65% at doses of 8 mg (n = 11), 12 mg (n = 51), and 16 mg (n = 7), respectively. The maximal individual peak inhibition of total cMyc protein expression ranged from 16% to 77% at 8 mg, 20% to 92% at 12 mg, and 61% to 97% at the 16 mg dose level. Inhibition of cMyc decreased to less than 10% at trough (Cycle 1, pre-dose 8 days) in the 8 mg and 12 mg cohorts and to approximately 30% in the 16 mg cohort. Composite PK PD curves were plotted for 70 participants, and an IC50 value of 202.4 nM was determined by non-linear regression curve fitting (see Figure 6). The ex vivo IC50 coverage of Compound 1 at steady state concentration is shown in Figure 7.

[0141] Relationship between steady state AUC of Compound 1 and treatment-emergent adverse events The exposure-safety relationship was evaluated using selected clinically relevant safety endpoints considered to be appropriate (e.g., frequently occurring TEAEs (treatment-emergent adverse events), incidence of all AEs (adverse events) > 20%, and clinically significant adverse events). Safety data from a total of 106 participants (including both solid tumors and hematological malignancies) receiving monotherapy with Compound 1 in Parts 1 and 2 (n = 10 (8 mg), n = 89 (12 mg), and n = 8 (16 mg)) were used in the analysis. Frequently occurring TEAEs (all grades and causality) in participants who received at least one dose of the study drug included nausea (47.9%), fatigue (45.5%), anorexia (35.5%), thrombocytopenia (28.9%), vomiting (28.1%), anemia (27.3%), diarrhea (27.3%), constipation (20.7%), and dysgeusia (19.8%), as well as clinically significant AEs including hyperglycemia (17.4%), epistaxis (9.9%), increased INR (8.5%), and gastrointestinal hemorrhage (1.9%).

[0142] No statistically significant correlations were identified between the steady-state AUC of Compound 1 and any of the TEAEs or clinically significant AEs evaluated in this analysis, except for hyperglycemia (p value < 0.05). Figure 8 shows the model-predicted versus observed relationship between the steady-state AUC of Compound 1 and the probability of hyperglycemia. The predicted probabilities of hyperglycemia at doses of 2 mg and 4 mg are 8.0% and 9.2%, respectively, based on the relationship between AUC and hyperglycemia (Note: open squares are for Compound 1 AUC ss,0-24 of the 1st (893 - 2035 h*nM), 2nd (2074 - 2605 nM), 3rd (2622 - 3564 nM), and 4th (3606 - 9788 nM). The lack of correlation between the AUC of Compound 1 and TEAEs ss,0-24 may be due to the narrow dose range (8, 12, or 16 mg) investigated in this study and the small sample sizes in the 8 mg and 16 mg groups of the study (the majority of participants received 12 mg).

[0143] Table 4 lists the estimated parameters for the increase in AUC of Compound 1 and grade 3 INR (International Normalized Ratio) and grade 4 gastrointestinal bleeding. Two grade 3 events of INR occurred in participants taking warfarin concomitantly. The steady-state AUCs of these two participants were 6990 h * nM and 9770 h*nM for 12 mg QD and 16 mg QD of Compound 1, respectively, and the AUCs were much higher than the geometric mean steady-state AUCs at 12 mg QD (2740 h*nM) and 16 mg QD (3610 h*nM), respectively.

[0144] Two grade 4 events of gastrointestinal bleeding were observed in two participants who received the 12 mg QD dose, and the steady-state AUCs of these two participants were 3470 h*nM and 3610 h*nM, respectively, and the AUCs were slightly higher than the geometric mean steady-state AUC at 12 mg QD.

[0145] [Table 3]

[0146] Example 5: Efficacy of Compound 1 in combination with ruxolitinib in participants with myelofibrosis Preliminary efficacy (decrease in spleen length and / or volume) was observed in 2 out of 3 myelofibrosis participants treated in the trial referred to herein (for further details, see ‘‘Safety and Tolerability Study of INCB057643 in Participants With Myelofibrosis’’, ClinicalTrials.gov Identifier: NCT04279847, which is incorporated herein by reference in its entirety). All 3 participants had received ruxolitinib 6 months prior. The first and second participants had relapsed myelofibrosis that was treated with compound 1 monotherapy. The third participant had been treated with single-agent ruxolitinib but had a suboptimal response. In response to a suboptimal response, the third participant was subsequently treated with compound 1 in combination with ruxolitinib.

[0147] The first participant received 12 mg QD of compound 1 monotherapy and had a 92.5% reduction in spleen length by spleen palpation (nadir compared to baseline). A second participant with myelofibrosis received 8 mg QD of compound 1 monotherapy. The second participant on the low dose of compound 1 only experienced disease progression and remained in the study for 24 days. The third participant was administered 8 mg QD of compound 1 in combination with ruxolitinib and experienced a 77% reduction in spleen length by spleen palpation and a 44% reduction in spleen volume by imaging (nadir compared to baseline). In the context of the positive results of the first and third participants, when ruxolitinib monotherapy was used in a clinical trial, a reduction in spleen size of approximately 35% was observed (for further details, see Verstovsek S., Morgan G. ‘‘Results of COMFORT-I, a randomized double-blind phase III trial of JAK 1 / 2 inhibitor INCB18424(424) vs placebo(PB) for patients with myelofibrosis(MF)’’, Abstract #6500. 2011 American Society of Clinical Oncology Annual Meeting, which is incorporated herein by reference in its entirety).

[0148] Further preliminary efficacy of the BET inhibitor (CPI-0610) in myelofibrosis participants was also reported in the MANIFEST trial (NCT02158858; Hoffman et al 2019, Mascarenhas et al 2019, which are incorporated herein by reference in their entirety). Enrolled myelofibrosis participants were refractory, intolerant, or ineligible for ruxolitinib or had received ruxolitinib but experienced suboptimal response or progression of myelofibrosis (in the CPI-0610 and ruxolitinib combination cohort). Participants experienced a best spleen volume reduction in the range of 6% - 44% in 10 evaluable participants (monotherapy and combination).

[0149] In addition to what is described in this specification, various modifications of the present invention will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to be included within the scope of the appended claims. Each reference, including but not limited to all patents, patent applications, and publications cited in this application, is hereby incorporated by reference in its entirety into this specification.

Claims

**Claim 1** A method for treating a myeloproliferative neoplasm in a patient in need thereof, comprising administering to said patient 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, and ruxolitinib or a pharmaceutically acceptable salt thereof. A method for treating a myeloproliferative neoplasm in a patient in need thereof. **Claim 2** The method according to claim 1, wherein the ruxolitinib or a pharmaceutically acceptable salt thereof is ruxolitinib phosphate. **Claim 3** The method according to claim 1 or 2, wherein 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one is in solid form. **Claim 4** The method according to claim 3, wherein the solid form is an anhydrate. **Claim 5** The method according to claim 3, wherein the solid form is Form I. **Claim 6** The method according to claim 5, wherein Form I has one or more characteristic XRPD peaks selected from about 8.7, about 9.8, about 11.6, about 12.7, about 14.7, about 15.7, about 20.0, about 21.4, about 23.3 and about 27.1 degrees with respect to 2 theta. **Claim 7** The method according to claim 3, wherein the solid form has Form II. **Claim 8** The method according to claim 7, wherein Form II has one or more characteristic XRPD peaks selected from about 6.7, about 9.5, about 10.5, about 14.8, about 16.2, about 17.0, about 18.8, and about 19.3 degrees with respect to 2 theta. **Claim 9** The method according to any one of claims 1 to 8, wherein the dosage of ruxolitinib or a pharmaceutically acceptable salt thereof is about 5 mg / day to about 60 mg / day. **Claim 10** The method according to any one of claims 1 to 8, wherein the dosage of ruxolitinib or a pharmaceutically acceptable salt thereof is about 2.5 mg BID to about 30 mg BID. **Claim 11** The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, is from about 2 mg / day to about 20 mg / day.

12. The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, is from about 2 mg / day to about 18 mg / day.

13. The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, is from about 2 mg / day to about 12 mg / day.

14. The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, is from about 4 mg / day to about 8 mg / day.

15. The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 2, about 4, about 6, about 8, about 10, about 12, about 14, about 16, about 18, or about 20 mg / day on a free base basis.

16. The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 2 mg / day on a free base basis.

17. The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 4 mg / day on a free base basis.

18. The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 6 mg / day on a free base basis.

19. The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 8 mg / day on a free base basis.

20. The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 10 mg / day on a free base basis.

21. The method according to any one of claims 1 to 10, wherein the dosage of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one or a pharmaceutically acceptable salt thereof is about 12 mg / day on a free base basis.

22. The method according to any one of claims 1 to 21, wherein the myeloproliferative tumor is selected from polycythemia vera (PV), essential thrombocythemia (ET), primary myelofibrosis, chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), hypereosinophilic syndrome (HES), systemic mastocytosis with cutaneous involvement (SMCD), chronic neutrophilic leukemia (CNL), and chronic eosinophilic leukemia.

23. The method according to claim 22, wherein the myeloproliferative tumor is polycythemia vera (PV).

24. The method according to claim 22, wherein the myeloproliferative tumor is essential thrombocythemia (ET).

25. The method according to claim 22, wherein the myeloproliferative tumor is primary myelofibrosis.

26. The method according to any one of claims 1 to 25, wherein ruxolitinib, or a pharmaceutically acceptable salt thereof, and 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, are orally administered.

27. The method according to any one of claims 1 to 26, wherein ruxolitinib, or a pharmaceutically acceptable salt thereof, and 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptable salt thereof, are orally administered simultaneously or sequentially.

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