Combination of Ruxolitinib and INCB057643 for the Treatment of Myeloproliferative Neoplasms

Abstract

This application relates to the treatment of myeloproliferative neoplasms using the JAK1 / JAK2 inhibitor ruxolitinib in combination with the BET protein inhibitor 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 combination that is unexpectedly synergistic.

Description

[Technical field]

[0001] Priority claim This application claims priority to U.S. Provisional Patent Application No. 63 / 034,214, filed June 3, 2020, which is incorporated by reference in its entirety.

[0002] The present application relates to the treatment of myeloproliferative neoplasms using the JAK1 / JAK2 inhibitor ruxolitinib in combination with the BET protein inhibitor 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, which combination is unexpectedly synergistic. [Background technology]

[0003] Compound 1 is a small molecule inhibitor of the acyl-binding bromodomain found in bromodomain and extracellular domain (BET) proteins. Four family members (BRD2, 3, 4, T) promote transcription by binding to acetylated histones and recruiting transcription initiation and elongation complexes, thereby acting as a link between chromatin and transcriptional activation (see Shi, J., and Vakoc, CR, "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 occasionally BRD3) genes are components of chromosomal translocations that result in an aggressive subtype of squamous cell carcinoma called "NUT midline" carcinomas. These midline carcinomas are defined by the t(15;19) translocation that gives rise to the BRD4-NUT oncogene. The resulting fusion protein consists of BRD4 at the N-terminus and NUT (nuclear protein of the testis) at the C-terminus. 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, incorporated herein by reference in its entirety).

[0005] BET proteins are believed to exert their proliferative effects by promoting the transcription of oncogenes, such as c-myc, among others. BRD4 inhibitor compounds have been shown to prevent BRD4 from associating with chromatin, including on the c-myc promoter, in a multiple myeloma model. This dissociation leads to reduced c-myc expression levels and reduced cell viability (see Delmore, JE, 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 found to bind to ``super enhancers,'' which are numerous transcriptional regulatory elements within genes required for cell fate and survival. One example is found in multiple myeloma, where the c-myc gene was found to have a super-enhancer region that is bound by BRD4 and results in 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, incorporated herein by reference in its entirety).

[0006] The MPLW515L mutation is found in a small proportion 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. Because there is much overlap between the JAK / STAT signaling pathway and inflammatory pathways that are transcriptionally regulated by BET proteins, the combination of BET and JAK inhibitors in MPNs may form the basis of a new therapeutic treatment protocol.

[0007] New therapies are needed to improve patient outcomes. This application addresses this need and others. Summary of the Invention

[0008] The present application provides, inter alia, a method of treating myeloproliferative neoplasms 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] The application also provides a method of treating a myeloproliferative neoplasm in a patient in need of such treatment, 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 about 5 mg / day to about 60 mg / day on a free base basis.

[0010] The application further provides the use of a compound, in the dosage amounts described herein, for use in the manufacture of a medicament for use in the treatment of a myeloproliferative neoplasm.

[0011] The application also provides compounds, in the doses described herein, for use in treating myeloproliferative neoplasms.

[0012] The details of one or more embodiments are set forth in the description below. Other features, objects, and advantages will become apparent from the description and the claims. [Brief description of the drawings]

[0013] [Figure 1A] FIG. 1 is a graph showing efficacy of Compound 1 in a HEL xenograft model, tumor volume (mm3) versus days post-inoculation. [Figure 1B] FIG. 1 is a graph showing the tolerability of Compound 1 in a HEL xenograft model, % tumor-corrected body weight change versus days post-inoculation. [Figure 2A] FIG. 1 is a graph showing efficacy of the combination of Compound 1 and ruxolitinib in the SET-2 model, tumor volume (mm3) versus days post-inoculation. [Figure 2B] FIG. 1 is a graph showing the tolerability of the combination of Compound 1 and ruxolitinib in the SET-2 model, tumor-corrected body weight change (%) versus days post-inoculation. [Figure 3A] FIG. 1 is a graph showing efficacy of the combination of Compound 1 and ruxolitinib in the SET-2 model, tumor volume (mm3) versus days post-inoculation. [Figure 3B] FIG. 1 is a graph showing the tolerability of the combination of Compound 1 and ruxolitinib in the SET-2 model, tumor-corrected body weight change (%) versus days post-inoculation. [Figure 4A] Graph showing spleen weight and white blood cell count as a measure of efficacy of Compound 1 and ruxolitinib in the MPLW515L mouse model of MPN. [Figure 4B] Graph showing spleen weight and white blood cell count as a measure of efficacy of Compound 1 and ruxolitinib in the MPLW515L mouse model of MPN. [Diagram 5]1 is a table (Table 3) showing a summary of Compound 1 pharmacokinetic parameters for Compound 1 as monotherapy (Parts 1 and 2) at steady state (Cycle 1, Day 8). [Figure 6] 1 is a graph showing inhibition of cMyc protein expression in a pharmacodynamic spiked cell assay versus plasma levels of Compound 1. [Figure 7] 1 is a graph showing steady-state plasma concentrations (mean±standard deviation) of Compound 1 in participants following once-daily administration of Compound 1 as monotherapy. [Figure 8] 1 is a graph showing probability of response (hyperglycemia) versus steady-state AUC of Compound 1 following once daily administration of Compound 1 as monotherapy. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The terms "for example" and "etc.", as well as their grammatical equivalents, are understood to be followed by the phrase "including but not limited to," unless otherwise noted.

[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" (eg, plus or minus about 10% of the indicated value).

[0017] As used herein, the phrase "solid form" refers to a compound provided herein in either an amorphous or 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 in the crystal lattice, for example to form a solvated or hydrated crystal form. The term "hydrated" as used herein is meant to refer to a crystal form that contains water molecules in the crystal lattice. Examples of "hydrated" crystal forms include hemihydrate, monohydrate, dihydrate, etc. Other hydrated forms, such as channel hydrates, are also included within the meaning of this term.

[0018] The present invention relates to the use of the JAK1 / JAK2 inhibitor, ruxolitinib, in combination with a BET protein inhibitor to treat 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 of ruxolitinib 50 was measured at 1 mM ATP by Assay A below and found to be less than 10 nM for JAK1 and JAK2. Ruxolitinib can be made by the procedures described in US 7,598,257, filed December 12, 2006 (Example 67), which is incorporated herein by reference in its entirety. Ruxolitinib phosphate can be prepared as described in US 2008 / 0312259, which is incorporated herein 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, incorporated herein by reference in its entirety). In the referenced study, BET inhibitors in combination with JAK inhibition reduced pathological cytokine production and overall disease burden. Specifically, in an in vivo model of myeloproliferative neoplasms, combination treatment with ruxolitinib and JQ1, a JAK1 / 2 and BET inhibitor, respectively, resulted in reduced inflammation, reduced disease burden. Treatment also eliminated fibrosis in myelofibrotic mice, a response not previously observed with single-agent JAK inhibitor therapy. Mechanistically, we propose that the synergistic effects of JAK and BET inhibitors are mediated by blocking two master regulators of pathological 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 myeloproliferative neoplasms 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 (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. Pat. No. 9,540,368 or U.S. Pat. No. 10,189,832, each of which is incorporated herein by reference in its entirety. [ka]

[0021] The present application provides a method of treating myeloproliferative neoplasms in a patient in need of such treatment, 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 pharma- ceutically acceptable salt thereof, and ruxolitinib, or a pharma- ceutically acceptable salt thereof. Administering a 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 or Compound 1 alone. Thus, the combination of ruxolitinib and Compound 1 provides efficacy that is better than the sum of its parts.

[0022] The present application provides a method of treating myeloproliferative neoplasms 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.

[0023] The application further provides a method of treating myeloproliferative neoplasms 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 pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of ruxolitinib, or a pharma- ceutically acceptable salt thereof.

[0024] The application further provides a method of treating myeloproliferative neoplasms 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 at a dose of about 2 mg / day to about 20 mg / day on a free base basis, or a pharma- ceutical acceptable salt thereof, and a therapeutically effective amount of ruxolitinib or a pharma-ceutical acceptable salt thereof.

[0025] 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 pharma- ceutically acceptable salt thereof, is from about 2 mg / day to about 18 mg / day on a free base basis.

[0026] 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 pharma- ceutically acceptable salt thereof, is from 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 pharma- ceutically 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 pharma- ceutically 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 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 pharma- ceutically 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 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 pharma- ceutically acceptable salt thereof is about 2, about 4, about 6, about 8, about 10, or about 12 mg / day on a free base basis. 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 pharma- ceutically acceptable salt thereof is about 4, about 6, or about 8 mg / day on a free base basis. 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 pharma- ceutically 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 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 pharma- ceutically acceptable salt thereof is about 10, about 12, about 14, about 16, or about 18 mg / day on a free base basis. 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 pharma- ceutically 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 pharma- ceutically 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 pharma-ceutically 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 pharma- ceutically 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 pharma-ceutically 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 pharma- ceutically 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 ruxolitinib or a pharmaceutically acceptable salt thereof are administered once a day (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 ruxolitinib or a pharmaceutically acceptable salt thereof are administered twice a day (BID).

[0030] In some embodiments, the dose of ruxolitinib or a pharma- ceutically acceptable salt thereof is about 5 mg / day to about 60 mg / day on a free base basis. For example, the dose of ruxolitinib or a pharma- ceutically 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 dose of ruxolitinib or a pharma- ceutically acceptable salt thereof is about 2.5 mg BID to about 30 mg BID on a free base basis. For example, the dose of ruxolitinib or a pharma- ceutically 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 pharma- ceutically 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 pharma- ceutically acceptable salt thereof, is selected from the group consisting of phosphate, dihydrochloride, hydrochloride, maleate, adipate, hydrobromide, (R)-(−)-mandelate, salicylate, benzoate, and the like. The salt is selected from the group consisting of an anthracene salt, a benzenesulfonate salt, an L-pyroglutamate salt, a methanesulfonate salt, a (1S)-(+)-10-camphorsulfonate salt, a fumarate salt, a sulfate salt, an L-tartrate salt, and a D-tartrate salt 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, the 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 a 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 a BET protein. A more detailed discussion of the crystalline forms of Compound 1 can be found in U.S. Pat. No. 10,626,114, which is incorporated herein by reference in its entirety and is briefly described herein. Typically, different crystalline forms of the same substance have different bulk properties, e.g., with respect to hygroscopicity, solubility, stability, etc.

[0035] The crystalline solid form of Compound 1 may contain solvents such as water (e.g., hydrated forms) or may be substantially free of water and solvents (e.g., anhydrous forms). In some embodiments, the crystalline solid form is anhydrous. In further embodiments, the crystalline solid form is hydrated. Compound 1 can be obtained in solid crystalline forms designated 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 X, as described in U.S. Pat. No. 10,626,114, the entirety of which is incorporated herein by reference.

[0036] Experimental data indicates that Form I is anhydrous. 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 2-theta.

[0037] Experimental data indicates that Form II is anhydrous. 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 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 two-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 two-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 two-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 two-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 two-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 two-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 two-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 two-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 two-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 two-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 two-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 two-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 two-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 two-theta.

[0052] The present application provides a method of treating myeloproliferative neoplasms 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 pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of ruxolitinib, or a pharma- ceutically 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 (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, ruxolitinib, or a pharma- ceutically 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 pharma- ceutically acceptable salt thereof, are orally administered. In some embodiments, ruxolitinib, or a pharma- ceutically acceptable salt thereof, is orally administered. 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 pharma- ceutically acceptable salt thereof, is orally administered.

[0058] In some embodiments, ruxolitinib, or a pharma- ceutically 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 pharma-ceutically 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 directed to individual doses of ruxolitinib, embodiments directed to individual doses of a BET protein inhibitor (Compound 1), embodiments directed to salt forms, embodiments directed to individual types of myeloproliferative neoplasms, and embodiments directed to oral administration may be combined in any combination). The combinations are not recited separately herein merely for the sake of brevity.

[0060] All compounds and their pharma- ceutically acceptable salts can be found together with or isolated from other substances such as water and solvents (e.g., hydrates and solvates). When in the solid state, the compounds and their salts described herein can exist in various forms, for example, in the form of solvates, including hydrates. The compounds may be in any solid form, such as polymorphs or solvates, so unless otherwise specified, references to compounds and their salts herein should be understood to include any solid form of the compounds.

[0061] The phrase "pharmacologically acceptable" is used herein to refer to compounds, materials, compositions and / or dosage forms that are suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, within the scope of sound medical judgment, commensurate with a reasonable benefit / risk ratio.

[0062] The present invention also includes pharma- ceutically acceptable salts of the compounds described herein. The term "pharma- ceutically acceptable salts" refers to derivatives of the disclosed compounds in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharma- ceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, and the like. The pharma- ceutically acceptable salts of the present invention include non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharma- ceutically acceptable salts of the present invention can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. In general, 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 with non-aqueous media such as ether, ethyl acetate, alcohol (e.g., methanol, ethanol, isopropanol, or butanol) or acetonitrile (MeCN) being preferred). A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th 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 dosage described herein is based on free base." Based on free base " refers to the amount of compound (such as ruxolitinib or its salt) in dosage form, even if the actual active ingredient is a salt of a compound with different molecular weight from free base, the amount is measured based only on the molecular weight of the free base of the compound.For example, the conversion factor of ruxolitinib phosphate to free base is 0.7575.

[0064] The terms "individual" or "patient", used interchangeably, refer to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, or primate, and most preferably a human.

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

[0066] The term "treating" or "treatment" refers to one or more of: (1) inhibiting the disease, e.g., inhibiting the disease, condition or disorder in an individual experiencing or exhibiting the pathology or symptomology of the disease, condition or disorder (i.e., arresting further progression of the pathology and / or symptomology); (2) ameliorating the disease, e.g., ameliorating the disease, condition or disorder in an individual experiencing or exhibiting the pathology or symptomology 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 has not yet experienced or exhibited the pathology or symptomology of the disease.

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

[0068] The term "QD" means once daily.

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

[0070] In some embodiments, compound 1 can be used in combination with therapeutic agents that target epigenetic regulators.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 to treat 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 radiation, neutron beam radiation therapy, electron beam radiation therapy, proton therapy, brachytherapy, and total body 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, calsterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clotrimazole, cyclosporine ... arabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, deleukin, denileukin diphytoxin, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fentanyl citrate, Rudarabine, 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 phenpropionate, nelarabine, nofetumomab, oxaliplatin, paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, sorafenib,Streptozocin, sunitinib, sunitinib maleate, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, and zoledronate.

[0072] Compound 1 can be used in combination with ruxolitinib to treat 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 a KIR inhibitor, a TIGIT inhibitor, a LAIR1 inhibitor, a CD160 inhibitor, a 2B4 inhibitor, and a TGFRβ inhibitor.

[0074] In some embodiments, the inhibitor of an 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 an 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, prednisolone, or flumetholone.

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

[0084] Compound 1 was administered in combination with Dehydrex™ (Holles Labs), civamide (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)-hydroxyeicosatetraenoic acid (15(S)-HETE), seviremine, doxycycline (ALTY-0501, Alacrity), minocycline, iDestrin™ (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), voclosporin (LX212 or LX214, Lux Biosciences), ARG103 (Agentis), RX-10045 (synthetic resolvin analog, Resolvyx), DYN15 (Dyanmis Therapeutics), rivoglitazone (DE011, Daiichi Sanko), TB4 (RegeneRx), OPH-01 (Ophtalmis Monaco), PCS101 (Pericor Science), REV1-31 (Evolutec), lacritin (Senju), rebamipide (Otsuka-Novartis), OT-551 (Othera), PAI-2 (University of Pennsylvania and Temple University), pilocarpine, tacrolimus, pimecrolimus (AMS981, Novartis), loteprednol etabonate, rituximab, diquafosol tetrasodium (INS365,It may be administered in combination with one or more additional agents selected from the group consisting of anakinra, efalizumab, mycophenolate sodium, etanercept (Embrel®), hydroxychloroquine, NGX267 (TorreyPines Therapeutics), or thalidomide.

[0085] In some embodiments, Compound 1 may be administered in combination with one or more agents selected from antibiotics, antivirals, antifungals, anesthetics, anti-inflammatory agents, including steroidal and nonsteroidal anti-inflammatory agents, and anti-allergy agents. Examples of suitable pharmaceutical agents include aminoglycosides, such as amikacin, gentamicin, tobramycin, streptomycin, netilmicin, and kanamycin; fluoroquinolones, such as ciprofloxacin, norfloxacin, ofloxacin, trovafloxacin, lomefloxacin, levofloxacin, and enoxacin; naphthyridine; sulfonamides; polymyxins; chloramphenicol; neomycin; paramomycin; colistimethate; bacitracin; bacitracin; These include: cyclosporine; tetracyclines; rifampin and its derivatives ("rifampins"); cycloserine; beta-lactams; cephalosporins; amphotericin; fluconazole; flucytosine; natamycin; miconazole; ketoconazole; corticosteroids; diclofenac; flurbiprofen; ketorolac; suprofen; cromolyn; lodoxamide; levocabastine; naphazoline; anthrazoline; pheniramine; or azalide antibiotics.

[0086] Other examples of drugs with which the provided compounds may be combined include treatments for Alzheimer's disease, such as donepezil and rivastigmine; treatments for Parkinson's disease, such as L-DOPA / carbidopa, entacapone, ropinirole, pramipexole, bromocriptine, pergolide, trihexyphenidyl, and amantadine; treatments for multiple sclerosis (MS), such as beta-interferons (e.g., Avonex® and Rebif®), glatiramer acetate, and mitoxantrone; treatments for asthma, such as albuterol and montelukast; treatments for schizophrenia, such as Zyprexa, rispedal, seroquel, and haloperidol; anti-inflammatory agents, such as corticosteroids, such as dexamethasone, or prednisone, TNF blockers, IL-1RA, azathioprine, cyclophosphamide, and 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, interferons, anticonvulsants, ion channel blockers, riluzole, or anti-Parkinson's agents; agents for treating cardiovascular disease, such as beta blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers, or statins; agents for treating liver disease, such as corticosteroids, cholestyramine, interferons, and antivirals; agents for treating blood disorders, such as corticosteroids, anti-leukemia agents, or growth factors; or agents for treating immune deficiency 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 one or more inhibitors 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 immunomodulator (e.g., lenolidomide, pomalidomide).

[0088] When two or more pharmaceutical agents are administered to a patient, they may be administered simultaneously, sequentially, or in combination (eg, for three or more agents).

[0089] composition The compounds can be administered in the form of pharmaceutical compositions. These compositions can be prepared in a manner 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 area to be treated. Administration can be topical (including transdermal, epidermal, ocular and mucosal (including intranasal, vaginal and rectal delivery)), pulmonary (e.g., inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal or intranasal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal intramuscular or injection or infusion, or intracranial, e.g., intrathecal or intraventricular administration. Parenteral administration can be in the form of a single bolus dose or can be, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

[0090] The pharmaceutical composition may contain the compound or its pharma- ceutically acceptable salt as an active ingredient in combination with one or more pharma- ceutically acceptable carriers (excipients). In some embodiments, the composition is suitable for topical administration. In preparing the composition, the active ingredient is typically mixed with an excipient, diluted by an excipient, or enclosed within such a carrier, for example in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it may be a solid, semi-solid or liquid material that acts as a vehicle, carrier or medium for the active ingredient. Thus, the composition may be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid or liquid media), for example ointments containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injection solutions and sterile packaged powders.

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

[0092] The compounds can be milled using known milling procedures, such as wet milling, to obtain a particle size suitable for tablet formation and other formulation types. Finely divided (nanoparticulate) preparations of the compounds of the 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, gum acacia, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose.The formulation may include lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl benzoate and propyl hydroxybenzoate; sweeteners and flavoring agents.The composition of the present invention can be formulated to provide quick, sustained or delayed release of 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 pharma- ceutically 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 pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable carrier. In some embodiments, the composition comprises at least one compound described herein or a pharma- ceutically 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 pharma- ceutically acceptable salt thereof and microcrystalline cellulose, lactose monohydrate, and hydroxypropyl methylcellulose. In some embodiments, the composition comprises at least one compound described herein or a pharma- ceutically acceptable salt thereof and 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 ingredients used to formulate pharmaceutical compositions are of high purity and substantially free of potentially harmful contaminants (e.g., at least national food grade, generally at least analytical grade, and more typically at least pharmaceutical grade). Particularly for human consumption, the compositions are preferably manufactured or formulated under Good Manufacturing Practice standards as defined in applicable regulations of the US Food and Drug Administration. For example, suitable formulations may be sterile and / or substantially isotonic, and / or in full compliance with all Good Manufacturing Practice regulations of the US Food and Drug Administration.

[0097] The active compounds may be effective over a wide dosage range and are generally administered in a therapeutically effective amount. However, it will be understood that the amount of compound actually administered will usually be determined by the physician depending on the 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, and the like.

[0098] The therapeutic dosage of a compound of the invention can vary according to, for example, the particular application for which the treatment is made, the manner of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of a compound of the invention in a pharmaceutical composition can vary depending on a number of factors, including the dosage, chemical characteristics (e.g., hydrophobicity), and the route of administration.

[0099] To prepare solid compositions such as tablets, the principal active ingredient is mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogenous mixture of the compound of the present invention. When these preformulation compositions are referred to as homogenous, the active ingredient is typically dispersed evenly throughout the composition so that the composition can be readily subdivided into equally effective unit dosage forms, such as tablets, pills, and capsules. This solid preformulation is then subdivided into unit dosage forms of the type described above, containing, for example, 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 coated or otherwise compounded to provide a dosage form that provides the advantage of long-acting action. For example, the tablets or pills can include an inner dosage and an outer dosage component, the latter 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 allow the inner component to enter the duodenum intact or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, including a number of polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0101] Liquid forms into which the compounds and compositions of the present invention can be incorporated for administration orally or by injection include aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, and flavored emulsions including 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 pharma- ceutically acceptable aqueous or organic solvents, or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharma- ceutically acceptable excipients as described above. In some embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect. The compositions can be nebulized by the use of inert gases. Nebulized solutions 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 machine. Solution, suspension, or powder compositions can be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0103] Topical formulations can include one or more conventional carriers. In some embodiments, ointments can contain water and one or more hydrophobic carriers, such as selected from liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white Vaseline® (petrolatum), etc. The carrier composition of creams can be based on water combined with glycerol and one or more other ingredients, such as glycerol monostearate, PEG-glycerol monostearate, and cetylstearyl alcohol. Gels can be formulated using isopropyl alcohol and water, appropriately combined with other ingredients, such as glycerol, hydroxyethylcellulose, etc.

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

[0105] The compositions administered to a patient may be in the form of pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or may be sterile filtered. Aqueous solutions may be packaged for use as is or lyophilized, the lyophilized preparations being combined with a sterile aqueous carrier prior to administration. The pH of the compound preparations is typically 3-11, more preferably 5-9, most preferably 7-8. It will be appreciated that the use of certain of the aforementioned excipients, carriers, or stabilizers may result in the formation of pharmaceutical salts.

[0106] kit The present application also includes useful pharmaceutical kits that include 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 a variety of conventional pharmaceutical kit components, such as, for example, a container containing one or more pharma- ceutically acceptable carriers, additional containers, etc., as would be readily apparent to one of skill in the art. Instructions, either as an insert or label, indicating the amounts of components to be administered, directions for administration, and / or directions for mixing the components, can also be included in the kit.

[0107] The present invention will be described in more detail with specific examples.The following examples are provided for illustrative purposes and are not intended to limit the present invention in any way.Those skilled in the art will easily recognize various non-critical parameters that can be changed or modified to produce essentially the same results.The compounds of the examples have been found to be BET protein kinase inhibitors by at least one of the assays described herein. EXAMPLES

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

[0109] method: Female SCID mice, approximately 5-6 weeks of age, were inoculated subcutaneously with 107 SET-2 cells in Matrigel. Dosing began when tumor size reached approximately 200 mm3. Dosing for these studies was by oral gavage.

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

[0111] Animals were sacrificed when: (1) the end of the study; (2) tumor size reached 10% of total body weight; or (3) they lost 20% of their body weight.

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

[0113] Efficacy of Compound 1 in a JAK2 V617F-expressing HEL xenograft model of erythroleukemia The objective 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 injected with 10 7 HEL cells were inoculated subcutaneously. The tumors were approximately 160 mm 3 Once this was reached, mice (n=10 per group) were orally dosed with Compound 1 for 14 days as follows: Either 1.1 or 3 mg / kg BID or vehicle control 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, with the mean tumor volume from each Compound 1 treatment group being statistically significantly reduced 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 one statistical tumor regression. The data indicate that QD administration is as effective as BID administration in this model, so administration of Compound 1 in a BID regimen may be considered but is unnecessary. As shown in Figure 1B, all doses were tolerated as determined by the lack of weight loss.

[0115] Efficacy from the combination of Compound 1 and ruxolitinib in the SET-2 xenograft model Female SCID mice were cultured in Matrigel for 10 min. 7 SET-2 cells were inoculated subcutaneously. The tumors were approximately 175 mm 3 Once this was reached, mice (n=8 per group) were orally dosed for 14 days as follows: 1. Vehicle control or 30 mg / kg ruxolitinib BID 2.10 mg / kg of Compound 1 QDs 3. Combination of ruxolitinib and compound 1

[0116] Compound 1 and ruxolitinib were suboptimally dosed for optimal readout of potential combination efficacy. As seen in Figure 2A, both monotherapies significantly inhibited tumor growth relative to vehicle controls (p<0.02, respectively, two-way ANOVA). Furthermore, the combination of Compound 1 and ruxolitinib provided enhanced and significant efficacy compared to the results expected based on the efficacy of the single agents (p<0.007 for combination vs. both single agent arms, two-way ANOVA). Approximately 15% weight loss occurred in the combination group (Figure 2B), but this nadir was within the limits of tolerability.

[0117] Efficacy from the combination of Compound 1 and ruxolitinib in the SET-2 xenograft model Female SCID mice were cultured in Matrigel for 10 min. 7 SET-2 cells were inoculated subcutaneously. The tumors were approximately 150 mm 3 Once this was reached, mice (n=8 per group) were orally dosed for 11 days as follows: 1. Vehicle control or 60 mg / kg ruxolitinib BID 2.10 mg / kg of Compound 1 QDs 3. Combination of ruxolitinib and compound 1

[0118] In this study, ruxolitinib was administered at doses higher than 1.2 and resulted in significant tumor growth inhibition as monotherapy (Figure 3A). Administration of compound 1 with ruxolitinib did not increase the tumor growth inhibition seen with single-agent ruxolitinib or single-agent compound 1. All doses were tolerated, although the combination of compound 1 and ruxolitinib resulted in tolerable weight loss (approximately 12% in Figure 3B).

[0119] The BET inhibitor compound 1 is effective at tolerated doses in two models of JAK2V617F AML. Combination of compound 1 with a suboptimal dose of the JAK1 / 2 inhibitor ruxolitinib (30 mg / kg BID) results in a significant increase in tumor growth inhibition compared to monotherapy. These data demonstrate the potential of BET inhibitors in combination with JAK inhibitors in JAK2V617F-driven myeloid neoplasms.

[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] method: One week before bone marrow transplantation, 8-week-old female Balb / c mice (Charles River Laboratories) were injected once intraperitoneally with 150 mg / kg 5-fluorouracil. Five days later, the mice were sacrificed and bone marrow was harvested by femoral aspiration. 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, bone marrow was infected with an ecotropic retrovirus (MSCV puromycin backbone) expressing the MPLW515L gene by adding 1 ml of virus to 106 cells and then centrifuging in a 6-well plate at 1800 rpm, 37°C, 90 minutes. After centrifugation, the infected cells were kept in an incubator until injection into the recipient the next day.

[0122] The bone marrow of the recipient mice was retrovirally infected (approximately 2 × 10 per recipient mouse) in 8-week-old Balb / c mice. 5 Mice were ablated with 50 mg / kg 5-fluorouracil 5 days prior to transplantation with Balb / c bone marrow expressing MPLW515L via IL-16 (IL-16) (IL-16) (IL-16). Blood counts were performed 7 days after transplantation, and four cohorts (n=10 / cohort) with equal mean platelet counts were generated for treatment. Dosing began 8 days after transplantation and continued for 13 days. On day 14 after transplantation, mice were sacrificed, blood was collected by cardiac puncture, and spleens were weighed as a surrogate for disease burden.

[0123] Blood was collected weekly via the orbital sinus and complete blood counts (CBC) were determined by hematology counter (Abaxis, model HM5).

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

[0125] Efficacy of Compound 1 in combination with ruxolitinib in the MPLW515L model of murine MPN To evaluate the ability of the 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. Seven days after transplantation, mice were randomized into cohorts for dosing based on platelet levels determined by CBC. The next day, mice were administered oral dosing with Compound 1 at 10 mg / kg QD, ruxolitinib at 60 mg / kg BID, a combination of Compound 1 and ruxolitinib, or vehicle control to begin treatment. Dosing continued for 14 days, after which mice were bled for CBC and spleens were harvested and weighed as a surrogate for disease burden. Two vehicle-treated mice died before the end of the study at 14 days post-dosing, one died at 12 days post-dosing, and the other died at 13 days post-dosing.

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

[0127] Statistics were performed using Prism Graphpad software ( ** p<0.01, **** p<0.0001) was determined by an unpaired t-test performed using the

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

[0129] Example 3: Statistical Demonstration of Synergy Between Compound 1 and Ruxolitinib Synergy between Compound 1 and ruxolitinib was assessed using the Chou-Talalay formula (see Chou, Drug Combination Studies and Their Synergy Quantification Using the Chou-Talalay Method, Cancer Res; 70(2)January 15, 2010, incorporated herein by reference). Synergy indicates a greater effect of the combination than would be expected based on how the individual compounds are acting. Table 1 provides data from two experiments described herein. The first dasa set listed is the MPLW515L model, using spleen weight as a disease progression marker (i.e., larger spleen = worse disease). MPL SPL indicates the size of the spleen in grams from the MPLW515L mouse MPN model. Fa indicates the fraction affected, which is the percent change compared to vehicle. The expected value row indicates the expected effect of the combination based on the Chou-Talalay synergy index. Since the number is smaller than the effect in the affected fraction, this demonstrates a greater effect than expected from combining compound 1 with ruxolitinib, demonstrating that a synergistic effect occurs between the two compounds, resulting in an unexpected effect. The second experiment is SET-2, a JAK2V617F expressing cell line used in xenograft tumor models. 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, indicating an unexpected synergistic effect. In both experiments, the Chou-Talalay formula gives values ​​smaller than the effect seen with the combination of ruxolitinib + compound 1, which is interpreted as a combination that is synergistic in each experiment.

[0130] [Table 1]

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

[0132] The study will be divided into 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. Study drug will be self-administered once daily for 28 days, which is one cycle. Participants will continue to take study drug unless benefit is achieved and discontinuation criteria are met.

[0133] If the starting dose of 4 mg QD is deemed to be tolerated in Part 1, then the starting dose in Part 2 will be 4 mg QD, otherwise the starting dose in Part 2 will be 2 mg QD. Part 2 will also administer Compound 1 as monotherapy.

[0134] AEs will be continuously monitored throughout the study, in both parts 1 and 2. If the cumulative incidence of grade 3 or 4 drug-related AEs occurs in >40% of participants, the study will be stopped. Similarly, if there is >1 fatal drug-related event, the study will be stopped. 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 advanced 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 cutoff date (September 23, 2019), 25, 101, and 8 participants who received 8 mg, 12 mg, and 16 mg across Part 1 and Part 2 (treated as monotherapy), respectively, have been evaluated for PK. The PK parameters of 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 for Compound 1 as monotherapy (Parts 1 and 2) at Cycle 1, Day 1. Table 3 shows a summary of the pharmacokinetic parameters of Compound 1 for Compound 1 as monotherapy (Parts 1 and 2) at steady state (Cycle 1, Day 8).

[0136] [Table 2]

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

[0138] Following multiple dose administration in the fasted state, plasma concentrations of Compound 1 typically increased 1–2 hours after administration (T max The median value of max ) and then exhibits a monophasic decay with a steady-state geometric mean t 1 / 2 The steady state AUC was approximately 10 hours and was not dose-dependent. Based on the effective half-life of 10 hours, steady state was predicted to be achieved after 2 days of once-daily dosing. 0-24 First dose AUC 0-24Accumulation of Compound 1 exposure is minimal (<10%) by comparison with the steady-state C max and AUC 0-24 The increase in C was dose-proportional, i.e., Compound 1 exhibited nearly linear pharmacokinetics over the dose range tested. Dose-normalized PK parameters (C) using either full studies or pairwise comparisons (see Table 2) were max , ss and AUC ss、0~24 ) ANOVA with dose-normalized C max , ss or AUC ss、0~24 Compound 1 demonstrated a geometric mean CL of 9.92 to 10.7 L / h that was not statistically significantly different across or between doses (P>0.05). ss Low steady-state oral clearance with IVF and a geometric mean V of 138 to 197 L z At a dose of 12 mg QD, t 1 / 2 , steady state C max , and AUC 0-24 The geometric means (CV%) of were 9.17h (49.7%), 272nM (40.9%), and 2740h*nM (43.2%), respectively.

[0139] Estimated steady-state AUC of compound 1 0-24 is the geometric mean AUC of 12 mg QD ss,0-24 Based on linear PK extrapolation from , the PK data for Compound 1 at 4 mg QDs is 457 h*nM and 913 h*nM for 2 mg QDs and 4 mg QDs, respectively. The simulated PK data for Compound 1 at 4 mg QDs is shown in FIG.

[0140] Pharmacodynamics for the treatment of compound 1 as monotherapy Pharmacodynamic analysis was performed using an ex vivo assay measuring protein levels of cMyc, a BRD4 target gene, in KMS12BM cells, which were spiked into participant plasma samples taken pre- and at various time points post-dose. In preliminary PD analysis following oral dosing of 8 mg, 12 mg, and 16 mg, compound 1 demonstrated inhibition of total cMyc protein expression, with maximum inhibition occurring between 1 and 4 hours. Mean cMyc inhibition at steady state (day 8, 8 hours from pre-dose) was 30%, 44%, and 65% for the 8 mg (n=11), 12 mg (n=51), and 16 mg (n=7) doses, respectively. Maximum 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, 8 days pre-dose) 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 nonlinear regression curve fitting (see Figure 6). Ex vivo IC50 coverage for compound 1 at steady state concentrations is shown in Figure 7.

[0141] Relationship between Compound 1 steady-state AUC and treatment-emergent adverse events Selected clinical safety endpoints deemed appropriate (e.g., frequently occurring TEAEs (treatment-emergent adverse events), incidence of all AEs >20%, and clinically significant adverse events) were used to assess exposure-safety relationships. Safety data from a total of 106 participants (including both solid tumors and hematological malignancies) receiving monotherapy treatment with compound 1 in parts 1 and 2 (n=10 (8 mg), n=89 (12 mg), and n=8 (16 mg)) were used for the analysis. Frequent TEAEs (all grades and causality) in participants who received at least one dose of 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%), INR increased (8.5%), and gastrointestinal bleeding (1.9%).

[0142] No statistically significant correlations were identified between Compound 1 AUC at steady state and any 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 of Compound 1 steady state AUC and probability of hyperglycemia. The predicted probability of hyperglycemia occurrence at the 2 mg and 4 mg doses is 8.0% and 9.2%, respectively, based on the relationship between AUC and hyperglycemia (Note: open squares indicate Compound 1 AUC ss,0-24 The first (893-2035h*nM), second (2074-2605nM), third (2622-3564nM) and fourth (3606-9788nM). AUC of compound 1 ss,0-24 The lack of correlation between and TEAEs may be due to the narrow dose range investigated in this study (8, 12, or 16 mg) and the small sample size in the 8 mg and 16 mg arms of the study (the majority of participants received 12 mg).

[0143] Table 4 lists the AUC and estimated parameters for grade 3 increases in INR (international normalized ratio) and grade 4 gastrointestinal bleeding of Compound 1. Two grade 3 events of INR occurred in participants taking warfarin concomitantly. The steady-state AUC of these two participants was 6990 h after administration of 12 mg QD and 16 mg QD of Compound 1, respectively. * The AUCs were 9770 h*nM and 9770 h*nM, which 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 receiving the 12 mg QD dose; the steady-state AUC for these two participants was 3470 h*nM and 3610 h*nM, respectively, an AUC slightly higher than the geometric mean steady-state AUC for 12 mg QD.

[0145] [Table 3]

[0146] Example 5: Efficacy of Compound 1 in Combination with Ruxolitinib in Participants with Myelofibrosis Preliminary efficacy (reduction in spleen length and / or volume) was observed in two of three myelofibrosis participants treated in the study 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 three participants had previously received ruxolitinib for six months. The first and second participants had relapsed myelofibrosis 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 Compound 1 monotherapy 12 mg QD and had a 92.5% reduction in spleen length by spleen palpation (nadir compared to baseline). The second participant with recurrent myelofibrosis received Compound 1 monotherapy 8 mg QD. The second participant on the lower dose of Compound 1 only had disease progression, and the second participant remained on the study for 24 days. The third participant received Compound 1 8 mg QD 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). To put the positive results of the first and third participants in context, when ruxolitinib monotherapy was used in clinical trials, 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 a BET inhibitor (CPI-0610) in myelofibrosis participants was also reported in the MANIFEST trial (NCT02158858, incorporated herein by reference in its entirety; Hoffman et al. 2019, Mascarenhas et al. 2019). Enrolled myelofibrosis participants were either refractory, intolerant, or ineligible for ruxolitinib (in the CPI-0610 monotherapy cohort) or had received ruxolitinib but experienced suboptimal response or progression of myelofibrosis (in the CPI-0610 and ruxolitinib combination cohort). Participants experienced best spleen volume reductions ranging from 6%-44% in 10 evaluable participants (monotherapy and combination).

[0149] In addition to those described herein, 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 cited in this application, including but not limited to all patents, patent applications, and publications, is incorporated herein by reference in its entirety. The present application also includes the following aspects. [Aspect 1] 1. A method of treating myeloproliferative neoplasms in a patient in need of such treatment, 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. [Aspect 2] The method of embodiment 1, wherein the ruxolitinib or a pharma- ceutically acceptable salt thereof is ruxolitinib phosphate. [Aspect 3] The method of embodiment 1 or 2, wherein the 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. [Aspect 4] The method of embodiment 3, wherein the solid form is anhydrous. [Aspect 5] The method of embodiment 3, wherein the solid form is Form I. [Aspect 6] The method of embodiment 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 two-theta. [Aspect 7] The method of embodiment 3, wherein the solid form has Form II. [Aspect 8] The method of embodiment 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 two-theta. [Aspect 9] The method according to any one of aspects 1 to 8, wherein the dose of ruxolitinib or a pharma- ceutically acceptable salt thereof is from about 5 mg / day to about 60 mg / day. [Aspect 10] The method of any one of aspects 1-8, wherein the dose of ruxolitinib or a pharma- ceutically acceptable salt thereof is from about 2.5 mg BID to about 30 mg BID. [Aspect 11] The method of any one of aspects 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof, is from about 2 mg / day to about 20 mg / day. [Aspect 12] The method of any one of aspects 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof, is from about 2 mg / day to about 18 mg / day. [Aspect 13] The method of any one of aspects 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof, is from about 2 mg / day to about 12 mg / day. [Aspect 14] The method of any one of aspects 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof, is about 4 mg / day to about 8 mg / day. [Aspect 15] 11. The method of any one of aspects 1-10, wherein 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 pharma- ceutically 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. [Aspect 16] The method of any one of aspects 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof, is about 2 mg / day on a free base basis. [Aspect 17] The method of any one of aspects 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof, is about 4 mg / day on a free base basis. [Aspect 18] The method of any one of aspects 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof, is about 6 mg / day on a free base basis. [Aspect 19] The method of any one of aspects 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof, is about 8 mg / day on a free base basis. [Aspect 20] The method of any one of aspects 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof, is about 10 mg / day on a free base basis. [Aspect 21] The method of any one of aspects 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof, is about 12 mg / day on a free base basis. [Aspect 22] 22. The method according to any one of aspects 1 to 21, wherein 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 (SMCD), chronic neutrophilic leukemia (CNL), and chronic eosinophilic leukemia. [Aspect 23] 23. The method of embodiment 22, wherein said myeloproliferative neoplasm is polycythemia vera (PV). [Aspect 24] 23. The method of embodiment 22, wherein said myeloproliferative neoplasm is essential thrombocythemia (ET). [Aspect 25] 23. The method of embodiment 22, wherein the myeloproliferative neoplasm is primary myelofibrosis. [Aspect 26] The method of any one of aspects 1 to 25, wherein ruxolitinib, or a pharma- ceutically 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 pharma- ceutically acceptable salt thereof, are orally administered. [Aspect 27] The method according to any one of aspects 1 to 26, wherein ruxolitinib, or a pharma- ceutically 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 pharma- ceutically acceptable salt thereof, are orally administered simultaneously or sequentially.

Claims

1. A pharmaceutical composition for treating myeloproliferative neoplasms in a patient in need thereof, comprising 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 pharmaceutical acceptable salt thereof, and ruxolitinib or a pharmaceutical acceptable salt thereof.

2. The pharmaceutical composition of claim 1 , wherein the ruxolitinib or a pharma- ceutically acceptable salt thereof is ruxolitinib phosphate.

3. 2. The pharmaceutical composition of claim 1, wherein the 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.

4. The pharmaceutical composition of claim 3 , wherein the solid form is anhydrous.

5. 4. The pharmaceutical composition of claim 3, wherein the solid form is Form I.

6. 6. The pharmaceutical composition of 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 two-theta.

7. 4. The pharmaceutical composition of claim 3, wherein the solid form has Form II.

8. 8. The pharmaceutical composition of 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 two-theta.

9. The pharmaceutical composition of any one of claims 1 to 8, wherein the dose of ruxolitinib or a pharma- ceutically acceptable salt thereof is from about 5 mg / day to about 60 mg / day.

10. The pharmaceutical composition of any one of claims 1 to 8, wherein the dose of ruxolitinib or a pharma- ceutically acceptable salt thereof is from about 2.5 mg BID to about 30 mg BID.

11. The pharmaceutical composition of any one of claims 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof is from about 2 mg / day to about 20 mg / day.

12. 11. The pharmaceutical composition of any one of claims 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof is from about 2 mg / day to about 18 mg / day.

13. 11. The pharmaceutical composition of any one of claims 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof is from about 2 mg / day to about 12 mg / day.

14. 11. The pharmaceutical composition of any one of claims 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof is from about 4 mg / day to about 8 mg / day.

15. 11. The pharmaceutical composition of any one of claims 1 to 10, wherein 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 pharma- ceutically acceptable salt thereof is about 2 mg / day, about 4 mg / day, about 6 mg / day, about 8 mg / day, about 10 mg / day, about 12 mg / day, about 14 mg / day, about 16 mg / day, about 18 mg / day, or about 20 mg / day on a free base basis.

16. 11. The pharmaceutical composition of 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 pharma- ceutically acceptable salt thereof is about 2 mg / day on a free base basis.

17. 11. The pharmaceutical composition of 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 pharma- ceutically acceptable salt thereof is about 4 mg / day on a free base basis.

18. 11. The pharmaceutical composition of 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 pharma- ceutically acceptable salt thereof is about 6 mg / day on a free base basis.

19. 11. The pharmaceutical composition of 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 pharma- ceutically acceptable salt thereof is about 8 mg / day on a free base basis.

20. 11. The pharmaceutical composition of 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 pharma- ceutically acceptable salt thereof is about 10 mg / day on a free base basis.

21. 11. The pharmaceutical composition of 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 pharma- ceutically acceptable salt thereof is about 12 mg / day on a free base basis.

22. The pharmaceutical composition according to any one of claims 1 to 21, wherein 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 (SMCD), chronic neutrophilic leukemia (CNL), and chronic eosinophilic leukemia.

23. 23. The pharmaceutical composition of claim 22, wherein the myeloproliferative neoplasm is polycythemia vera (PV).

24. 23. The pharmaceutical composition of claim 22, wherein the myeloproliferative neoplasm is essential thrombocythemia (ET).

25. 23. The pharmaceutical composition of claim 22, wherein the myeloproliferative neoplasm is primary myelofibrosis.

26. 26. The pharmaceutical composition of any one of claims 1 to 25, comprising ruxolitinib, or a pharma- ceutically 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 pharma- ceutically acceptable salt thereof, formulated for oral administration.

27. 27. The pharmaceutical composition of any one of claims 1 to 26, wherein ruxolitinib, or a pharma- ceutically 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 pharma- ceutically acceptable salt thereof, are formulated for simultaneous or sequential administration.

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