Methods of treating cancer

Abstract

This invention relates to methods for treating cancer, providing methods for treating solid tumors, B-cell lymphomas, or cancers with abnormal H3-K27 methylation, the method comprising orally administering a dosage form of EPZ-6438 in a therapeutically effective amount to a subject in need. This invention also relates to pharmaceutical compositions comprising one or more inhibitors of the human histone methyltransferase EZH2, and methods for treating cancer using such one or more EZH2 inhibitors.

Description

[0001] This invention application is a divisional application of the invention patent application with application number 201580063476.2, application date November 17, 2015, and invention title "Solid Drug Formulation and Preparation Method Thereof".

[0002] Related applications

[0003] This application claims the benefit and priority of the following U.S. patent application numbers: 62 / 080,985, filed November 17, 2014; 62 / 166,572, filed May 26, 2015; and 62 / 251,903, filed November 6, 2015, the contents of each of which are hereby incorporated by reference in their entirety. Background Technology

[0004] EZH2 (a histone methyltransferase) has been linked to various cancers. Specifically, mutations and / or overactivity of EZH2 have been found in a range of cancers, such as lymphoma, leukemia, and breast cancer. There remains a need for new agents as EZH2 inhibitors for use in anticancer therapy. Summary of the Invention

[0005] This invention provides a method for treating cancer (e.g., solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation). The method comprises orally administering to a subject in need a dosage form having a chemical formula (I) or a pharmaceutically acceptable salt thereof in a therapeutically effective amount.

[0006] The therapeutically effective dose described herein is a single dose that provides a bioequivalence of a mean AUC(0-12) from about 337 ng*hr / ml to about 18882 ng*hr / ml (e.g., from about 1720 ng*hr / ml to about 18882 ng*hr / ml, or from about 7798 ng*hr / ml to about 18882 ng*hr / ml) after administration to the subject. As used herein, the expressions “compound having chemical formula (I)”, “compound 1”, and “EPZ-6438” all refer to the same compound and may be used interchangeably.

[0007] In another aspect, the present invention provides a method for treating cancer (e.g., advanced solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation), the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said therapeutically effective amount is provided to be bioequivalent to about 12 ng*hr / ml per 1 mg of EPZ-6438 (e.g., about 7 ng*hr / ml per 1 mg of EPZ-6438). The average AUC(0-12) of a single dose of EPZ-6438 is approximately 8 ng*hr / ml to approximately 12 ng*hr / ml, approximately 9 ng*hr / ml to approximately 12 ng*hr / ml, or approximately 9.7 ng*hr / ml to approximately 11.8 ng*hr / ml.

[0008] In yet another aspect, the present invention provides a method for treating cancer (e.g., advanced solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation), the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said therapeutically effective amount provides a mean Cmax that is bioequivalent to a mean Cmax from about 102 ng / ml to about 4125 ng / ml (e.g., from about 476 ng / ml to about 4125 ng / ml or from about 1730 ng / ml to about 4125 ng / ml).

[0009] In yet another aspect, the present invention provides a method for treating cancer (e.g., advanced solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation), the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said therapeutically effective amount is a single dose providing a mean Cmax that is bioequivalent to a mean Cmax of about 1.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438 (e.g., from about 2.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438).

[0010] In another aspect, the present invention provides a method for treating cancer (e.g., solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation), the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein the therapeutically effective amount provides a median Tmax from about 1 hour to about 2 hours.

[0011] The present invention also provides a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein the therapeutically effective amount provides a mean AUC (0-12) of at least about 1170 ng*hr / ml (e.g., at least about 4421 ng*hr / ml) after administration to the subject.

[0012] In addition, the present invention provides a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need of it a dosage form having a therapeutically effective amount, wherein the therapeutically effective amount is a single dose that provides a bioequivalence of a mean AUC(0-12) from about 337 ng*hr / ml to about 18882 ng*hr / ml (e.g., from about 1720 ng*hr / ml to about 18882 ng*hr / ml, or from about 7798 ng*hr / ml to about 18882 ng*hr / ml) after administration to the subject.

[0013] In yet another aspect, the present invention also provides a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said therapeutically effective amount is a single dose providing a mean AUC(0-12) that is bioequivalent to a mean AUC(0-12) of ...

[0014] In yet another aspect, the invention also relates to a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said single dose provides a mean Cmax that is bioequivalent to a mean Cmax from about 102 ng / ml to about 4125 ng / ml (e.g., from about 476 ng / ml to about 4125 ng / ml or from about 1730 ng / ml to about 4125 ng / ml).

[0015] In another aspect, the present invention relates to a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said therapeutically effective amount is a single dose providing a mean Cmax that is bioequivalent to a mean Cmax of about 1.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438 (e.g., from about 2.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438).

[0016] The present invention also relates to a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein the therapeutically effective amount provides a median Tmax from about 1 hour to about 2 hours.

[0017] The present invention also relates to a method for treating advanced solid tumors or B-cell lymphomas, the method comprising orally administering a dosage form of EPZ-6438 having a therapeutically effective amount to a subject in need, wherein the therapeutically effective amount provides a mean AUC (0-12) of at least about 1170 ng*hr / ml (e.g., at least about 4421 ng*hr / ml) after administration to the subject.

[0018] The present invention also provides an oral dosage form for treating cancer (e.g., solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation), the oral dosage form comprising a therapeutically effective amount of EPZ-6438 and at least one pharmaceutically acceptable carrier or excipient, wherein the therapeutically effective amount is a single dose that provides a bioequivalence of a mean AUC(0-12) from about 337 ng*hr / ml to about 18882 ng*hr / ml (e.g., from about 1720 ng*hr / ml to about 18882 ng*hr / ml, or from about 7798 ng*hr / ml to about 18882 ng*hr / ml) after administration to a human subject.

[0019] In another aspect, this article provides an oral dosage form for the treatment of advanced solid tumors or B-cell lymphoma, comprising a therapeutically effective amount of EPZ-6438 and at least one pharmaceutically acceptable carrier or excipient, wherein the therapeutically effective amount is a single dose providing a mean AUC(0-12) that is bioequivalent to a mean AUC(0-12) of about 4 ng*hr / ml to about 12 ng*hr / ml per 1 mg of EPZ-6438 (e.g., about 7 ng*hr / ml to about 12 ng*hr / ml per 1 mg of EPZ-6438, about 8 ng*hr / ml to about 12 ng*hr / ml per 1 mg of EPZ-6438, about 9 ng*hr / ml to about 12 ng*hr / ml per 1 mg of EPZ-6438, or about 9.7 ng*hr / ml to about 11.8 ng*hr / ml per 1 mg of EPZ-6438).

[0020] In another aspect, this article provides an oral dosage form for the treatment of advanced solid tumors or B-cell lymphomas, comprising a therapeutically effective amount of EPZ-6438 and at least one pharmaceutically acceptable carrier or excipient, wherein the therapeutically effective amount provides a mean Cmax that is bioequivalent to a mean Cmax from about 102 ng / ml to about 4125 ng / ml (e.g., from about 476 ng / ml to about 4125 ng / ml or from about 1730 ng / ml to about 4125 ng / ml).

[0021] In yet another aspect, this document provides a dosage form for the treatment of advanced solid tumors or B-cell lymphomas, comprising a therapeutically effective amount of EPZ-6438 and at least one pharmaceutically acceptable carrier or excipient, wherein the therapeutically effective amount is a single dose providing a mean Cmax that is bioequivalent to a mean Cmax of about 1.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438 (e.g., about 2.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438).

[0022] This article also provides an oral dosage form for the treatment of advanced solid tumors or B-cell lymphomas, comprising a therapeutically effective amount of EPZ-6438 and at least one pharmaceutically acceptable carrier or excipient, wherein the therapeutically effective amount provides a mean AUC (0-12) of at least about 1170 ng*hr / ml (e.g., at least about 4421 ng*hr / ml) after administration to a human subject.

[0023] The present invention also relates to a solid pharmaceutical formulation comprising a therapeutic agent and one or more pharmaceutically acceptable excipients, wherein the therapeutic agent is compound 1, its salts, or combinations thereof, and the concentration of the therapeutic agent in the formulation is equal to about 35-65 wt.% of compound 1. In some embodiments, the concentration of the therapeutic agent in the formulation is equal to about 1-99 wt.%, 10-90%, 20-80%, 30-70%, or 35-65 wt.% of compound 1. In some embodiments, the concentration of the therapeutic agent in the formulation is equal to about 50 wt.%, 55 wt.%, or 60 wt.% of compound 1. In one embodiment, the pharmaceutical formulation is in a solid unit dosage form. In one embodiment, the pharmaceutical formulation is an oral unit dose formulation. In one embodiment, the pharmaceutical formulation is in tablet form.

[0024] The present invention also relates to a solid pharmaceutical formulation comprising a therapeutic agent (e.g., compound 1 or a salt thereof, or a combination thereof) and one or more pharmaceutically acceptable excipients selected from sodium starch glycolate, carboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium, or low-substituted hydroxypropyl cellulose, and combinations thereof. In one embodiment, the excipients are selected from sodium starch glycolate, carboxymethyl cellulose, calcium carboxymethyl cellulose, or croscarmellose sodium, and combinations thereof. In another embodiment, the excipients are selected from sodium starch glycolate, or carboxymethyl cellulose, and combinations thereof. In yet another embodiment, the solid pharmaceutical formulation further comprises lactose, hydroxypropyl cellulose, or magnesium stearate, or combinations thereof.

[0025] The present invention also relates to a solid pharmaceutical composition comprising a therapeutic agent and means for achieving immediate release of the therapeutic agent, wherein the therapeutic agent is selected from compound 1, its salts, and combinations thereof.

[0026] In another aspect, the present invention relates to a method for preparing the pharmaceutical formulations or compositions disclosed herein. The method comprises a) mixing a therapeutic agent, a diluent, a disintegrant, and optionally a lubricant to form a first mixture, wherein the therapeutic agent is selected from the group consisting of: compound 1, its salts, and combinations thereof. The method optionally includes one or more of the following steps:

[0027] b) Add an aqueous solution containing an adhesive, or an organic solvent-based solution (e.g., IPA, EtOH, etc.), or an organic / aqueous mixture (e.g., 1:1 EtOH:water) to the first mixture to form a second mixture;

[0028] c) Granulate the second mixture to form wet granules;

[0029] d) Dry these wet particles to form dried particles;

[0030] e) These dried particles are screened by size to obtain particles that meet the size requirements;

[0031] f) Mix these appropriately sized particles with a lubricant and a second disintegrant to form a third mixture;

[0032] g) Compress the third mixture to form tablets; and

[0033] h) Apply a coating suspension to these tablets to produce film-coated tablets.

[0034] In some embodiments, the present invention relates to a method for preparing the pharmaceutical formulation disclosed herein. The method includes a) mixing a therapeutic agent, a diluent, and a disintegrant to form a first mixture, wherein the therapeutic agent is selected from the group consisting of: compound 1, its salts, and combinations thereof.

[0035] The method optionally includes one or more of the following steps:

[0036] b) Granulate the first mixture into dry particles;

[0037] e) These dry particles are sorted by size;

[0038] f) Mix these appropriately sized particles with a lubricant and a second disintegrant to form a third mixture;

[0039] g) Compress the third mixture to form tablets; and

[0040] h) Apply a coating suspension to these tablets to produce film-coated tablets.

[0041] Although similar or equivalent methods and materials may be used in the practice or testing of this invention, suitable methods and materials are described below. All disclosures, patent applications, patents, and other references mentioned herein are incorporated herein by reference. No prior art to the claimed invention is acknowledged in connection with any references cited herein. In case of conflict, this specification, including its definitions, shall prevail. Furthermore, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0042] Any of the above aspects and embodiments can be combined with any other aspect or embodiment.

[0043] Other features and advantages of the invention will become apparent from the following detailed description and claims. Attached Figure Description

[0044] Figure 1A and 1BThe graph shows the average plasma concentration of EPZ-6438 over time after twice-daily administration of EPZ-6438: (A) on day 1 and (B) on day 15.

[0045] Figure 2 This is a graph showing the average molar ratio of metabolite exposure to the parent (ER-897387 / EPZ-6438) after a single dose (day 1) and multiple doses (day 15) administered twice daily.

[0046] Figure 3A It is a graph showing the maximum changes in tumor size versus dosage, and Figure 3B This is a graph showing the maximum changes in the tumor and its exposure at homeostasis (day 15).

[0047] Figure 4 This is a graph showing the average percentage of H3K27Me3-positive cells in skin biopsy samples (C1D1 = Cycle 1, Day 1, baseline before administration; C2D1 = Cycle 2, Day 1).

[0048] Figure 5 This is a graph showing the change in the percentage of H3K27Me3-positive cells from baseline in skin biopsies versus the EPZ-6438 dosage.

[0049] Figure 6 This is a graph showing the maximum percentage change in tumor size and the percentage change from baseline in H3K27Me3-positive cells in skin biopsies.

[0050] Figure 7 This is a graph showing the correlation between the percentage change from baseline in H3K27Me3-positive cells in skin biopsy samples and EPZ-6438 exposure. (The lines represent the fit to the inhibitory PK / PD model).

[0051] Figure 8 This is a flowchart of an embodiment of a method for manufacturing film-coated EPZ-6438 tablets.

[0052] Figure 9 This is a graph showing the dissolution profiles of EPZ-6438 50mg, 100mg, and 200mg tablets (n=6, average), using the paddle plate method (USP). <711> Instrument 2), pH 4.5 acetate buffer, 900 ml, 50 rpm.

[0053] Figure 10 This is a graph showing the dissolution profiles of EPZ-6438 50mg, 100mg, and 200mg tablets (n=6, average), using the paddle plate method (USP). <711> Instrument 2), 0.1N HCl, 900ml, 50rpm.

[0054] Figure 11This graph shows the dissolution curves of different formulations of EPZ-6438 (n=2, average value), using the paddle plate method (USP). <711> Instrument 2), 0.1N HCl, 900ml, 50rpm.

[0055] Figure 12 This graph shows the dissolution curves of different formulations of EPZ-6438 (n=2, average value), using the paddle plate method (USP). <711> Instrument 2), 0.1N HCl, 900ml, 50rpm.

[0056] Figure 13 This graph shows the dissolution curves of different formulations of EPZ-6438 (n=2, average value), using the paddle plate method (USP). <711> Instrument 2), pH 4.5 acetate buffer, 900 ml, 50 rpm.

[0057] Figure 14 This graph shows the dissolution curves of different formulations of EPZ-6438 (n=2, average value), using the paddle plate method (USP). <711> Instrument 2), pH 4.5 acetate buffer, 900 ml, 50 rpm.

[0058] Figure 15A This is a series of graphs showing the average plasma concentration of EPZ-6438 versus time distribution after twice-daily administration of EPZ-6438 on day 1 and day 15.

[0059] Figure 15B It is a series of images showing the pharmacokinetics of EPZ-6438 in the skin.

[0060] Figure 15C This is a graph showing the correlation between the inhibition of histone methylation and EPZ-6438 exposure.

[0061] Figure 16A-16F This is a series of graphs or images from a high-level image analysis of H3K27 trimethylation in the skin of subjects who were given the EZH2 inhibitor Tazemetostat. Detailed Implementation

[0062] Histone methyltransferases (HMTs) play a crucial role in the regulation of gene expression. Specifically, HMTs are involved in the regulation of cell division and differentiation. HMTs mediate the methylation of histones associated with specific genes. Depending on the methylated amino acid residues, methylation events can indicate either silencing or activation events of the relevant gene. Examples of silencing markers include trimethylation of H3K27; while trimethylation of H3K4 generates a gene activation signal. Many cell cycle checkpoint regulators and tumor suppressor genes exist in a “bivalent” state, which includes both activating histone modifications (e.g., H3K27me3) and repressive histone modifications (e.g., H3K4me3). Bivalent genes are prepared to undergo activation or repression depending on external factors. EZH2 regulates bivalent genes involved in B cell differentiation and maturation, including CDKN1, PRDM1, and IRF4.

[0063] EZH2 is a histone methyltransferase and the catalytic subunit of the PRC2 complex, which catalyzes mono- to trimethylation of lysine 27 (H3-K27) on histone H3. Histone H3-K27 trimethylation is a mechanism that inhibits the transcription of specific genes near histone modification sites. This trimethylation is known to be a cancer marker of altered expression in cancers such as prostate cancer (see, for example, U.S. Patent Application Publication No. 2003 / 0175736; incorporated herein by reference in its entirety). Other studies have provided evidence of a functional link between abnormal regulation of EZH2 expression, transcriptional repression, and tumorigenic transformation. Varambally et al. (2002) Nature 419(6907):624-9. Kleer et al. (2003) Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci USA) 100(20):11606-11.

[0064] EZH2 methylation activity plays a crucial role in the regulation and activation of B cells in germinal centers. EZH2 protein levels increase after B cell activation. Following activation, B cells reside in the germinal centers of lymphoid organs, where they undergo high-frequency somatic mutations—a process associated with the inhibition of anti-apoptotic genes and checkpoint regulators. EZH2 methylation events target genes involved in B cell proliferation, differentiation, and maturation, including CDKN1A (which plays a role in cell proliferation), PRDM1 (which plays a role in B cell differentiation), and IRF4 (which plays a role in B cell differentiation).

[0065] After B cells mature and leave the germinal center, the level of EZH2 within B cells decreases. However, the presence and activity of EZH2 after B cell maturation are associated with several lymphomas, including germinal center B-cell lymphoma. Aberrant activation of EZH2 has been found in three common subtypes of germinal cell lymphoma: follicular lymphoma (FL), germinal center B-cell-like diffuse large B-cell lymphoma (GCB DLBCL), and Burkitt lymphoma. Aberrant activation of EZH2 has also been found in primary mediastinal large B-cell lymphoma (PMBCL).

[0066] Genetic alterations within the EZH2 gene are associated with changes in histone methylation patterns. For example, certain point mutations in EZH2 are associated with altered H3K4 methylation in DLBCL; furthermore, chromosomal translocations and fusions SSX:SS18 are associated with altered H3K27 methylation in synovial sarcoma. EZH2 mutations that lead to the conversion of amino acid Y641 (equivalent to Y646, the catalytic domain) to F, N, H, S, or C induce H3K27 hypertrimethylation and drive lymphoma formation. Other genetic alterations affecting H3K27 methylation include EZH2 SET-domain mutations, EZH2 overexpression, overexpression of other PRC2 subunits, loss-of-function mutations in histone acetyltransferases (HAT), and loss of function of MLL2. H3K27 hypertrimethylation is induced in EZH2 Y646 mutant heterozygous cells relative to EZH2 homozygous wild-type (WT) cells, or relative to Y646 mutant homozygous cells.

[0067] EPZ-6438 (compound 1) is a small molecule inhibitor of EZH2, the catalytic subunit of the multicomb inhibitory complex 2 of methylated H3K27. Hypertrimethylation of H3K27 (H3K27Me3) has been shown to be tumorigenic in various malignancies, including a subclass of non-Hodgkin's lymphoma (NHL) with mutant EZH2. Inhibition of H3K27Me3 with EPZ-6438 resulted in the killing of EZH2 mutant lymphoma cells, and other EZH2 inhibitors showed activity in mutant and WT EZH2 NHL models. Additionally, tumors with loss of INI1 (a subunit of the SWI-SNF chromatin remodeling complex) appear to be EZH2-dependent. EPZ-6438 was shown to induce apoptosis and differentiation in an INI1-deficient malignant rhabdomyosarcoma (MRT) model in vitro and in mice carrying MRT xenografts.

[0068] The present invention is based, in at least part, on the discovery that Zeste enhancer homolog 2 (EZH2) inhibitors can effectively treat one or more cancers, such as one or more cancers characterized by aberrant H3-K27 methylation.

[0069] One aspect of the invention relates to a method for treating or alleviating cancer (e.g., solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation). The method comprises orally administering to a subject in need a dosage form of a compound having chemical formula (I) or a pharmaceutically acceptable salt thereof in a therapeutically effective amount.

[0070] The therapeutically effective dose is a single dose that provides a bioequivalence of a mean AUC(0-12) from about 337 ng*hr / ml to about 18882 ng*hr / ml (e.g., from about 1720 ng*hr / ml to about 18882 ng*hr / ml, or from about 7798 ng*hr / ml to about 18882 ng*hr / ml) after administration to the subject.

[0071] As used herein, the expressions “compound having chemical formula (I)”, “compound 1”, and “EPZ-6438” all refer to the same compound and can be used interchangeably.

[0072] In another aspect, the present invention provides a method for treating cancer (e.g., advanced solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation), the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said therapeutically effective amount is provided to be bioequivalent to about 12 ng*hr / ml per 1 mg of EPZ-6438 (e.g., about 7 ng*hr / ml per 1 mg of EPZ-6438). The average AUC(0-12) of a single dose of EPZ-6438 is approximately 8 ng*hr / ml to approximately 12 ng*hr / ml, approximately 9 ng*hr / ml to approximately 12 ng*hr / ml, or approximately 9.7 ng*hr / ml to approximately 11.8 ng*hr / ml.

[0073] In yet another aspect, the present invention provides a method for treating cancer (e.g., advanced solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation), the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said therapeutically effective amount provides a mean Cmax that is bioequivalent to a mean Cmax from about 102 ng / ml to about 4125 ng / ml (e.g., from about 476 ng / ml to about 4125 ng / ml or from about 1730 ng / ml to about 4125 ng / ml).

[0074] In yet another aspect, the present invention provides a method for treating cancer (e.g., advanced solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation), the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said therapeutically effective amount is a single dose providing a mean Cmax that is bioequivalent to a mean Cmax of about 1.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438 (e.g., from about 2.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438).

[0075] In another aspect, the present invention provides a method for treating cancer (e.g., solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation), the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein the therapeutically effective amount provides a median Tmax from about 1 hour to about 2 hours.

[0076] The present invention also provides a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein the therapeutically effective amount provides a mean AUC (0-12) of at least about 1170 ng*hr / ml (e.g., at least about 4421 ng*hr / ml) after administration to the subject.

[0077] In addition, the present invention provides a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need of it a dosage form having a therapeutically effective amount, wherein the therapeutically effective amount is a single dose that provides a bioequivalence of a mean AUC(0-12) from about 337 ng*hr / ml to about 18882 ng*hr / ml (e.g., from about 1720 ng*hr / ml to about 18882 ng*hr / ml, or from about 7798 ng*hr / ml to about 18882 ng*hr / ml) after administration to the subject.

[0078] In yet another aspect, the present invention also provides a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said therapeutically effective amount is a single dose providing a mean AUC(0-12) that is bioequivalent to a mean AUC(0-12) of ...

[0079] In yet another aspect, the invention also relates to a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said single dose provides a mean Cmax that is bioequivalent to a mean Cmax from about 102 ng / ml to about 4125 ng / ml (e.g., from about 476 ng / ml to about 4125 ng / ml or from about 1730 ng / ml to about 4125 ng / ml).

[0080] In another aspect, the present invention relates to a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein said therapeutically effective amount is a single dose providing a mean Cmax that is bioequivalent to a mean Cmax of about 1.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438 (e.g., from about 2.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438).

[0081] The present invention also relates to a method for inhibiting the histone methyltransferase activity of EZH2 or its mutants, the method comprising orally administering to a subject in need a dosage form of EPZ-6438 having a therapeutically effective amount, wherein the therapeutically effective amount provides a median Tmax from about 1 hour to about 2 hours.

[0082] The present invention also relates to a method for treating advanced solid tumors or B-cell lymphomas, the method comprising orally administering a dosage form of EPZ-6438 having a therapeutically effective amount to a subject in need, wherein the therapeutically effective amount provides a mean AUC (0-12) of at least about 1170 ng*hr / ml (e.g., at least about 4421 ng*hr / ml) after administration to the subject.

[0083] The present invention also provides an oral dosage form for treating cancer (e.g., solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation), the oral dosage form comprising a therapeutically effective amount of EPZ-6438 and at least one pharmaceutically acceptable carrier or excipient, wherein the therapeutically effective amount is a single dose that provides a bioequivalence of a mean AUC(0-12) from about 337 ng*hr / ml to about 18882 ng*hr / ml (e.g., from about 1720 ng*hr / ml to about 18882 ng*hr / ml, or from about 7798 ng*hr / ml to about 18882 ng*hr / ml) after administration to a human subject.

[0084] In another aspect, this article provides an oral dosage form for the treatment of advanced solid tumors or B-cell lymphoma, comprising a therapeutically effective amount of EPZ-6438 and at least one pharmaceutically acceptable carrier or excipient, wherein the therapeutically effective amount is a single dose providing a mean AUC(0-12) that is bioequivalent to a mean AUC(0-12) of about 4 ng*hr / ml to about 12 ng*hr / ml per 1 mg of EPZ-6438 (e.g., about 7 ng*hr / ml to about 12 ng*hr / ml per 1 mg of EPZ-6438, about 8 ng*hr / ml to about 12 ng*hr / ml per 1 mg of EPZ-6438, about 9 ng*hr / ml to about 12 ng*hr / ml per 1 mg of EPZ-6438, or about 9.7 ng*hr / ml to about 11.8 ng*hr / ml per 1 mg of EPZ-6438).

[0085] In another aspect, this article provides an oral dosage form for the treatment of advanced solid tumors or B-cell lymphomas, comprising a therapeutically effective amount of EPZ-6438 and at least one pharmaceutically acceptable carrier or excipient, wherein the therapeutically effective amount provides a mean Cmax that is bioequivalent to a mean Cmax from about 102 ng / ml to about 4125 ng / ml (e.g., from about 476 ng / ml to about 4125 ng / ml or from about 1730 ng / ml to about 4125 ng / ml).

[0086] In yet another aspect, this document provides a dosage form for the treatment of advanced solid tumors or B-cell lymphomas, comprising a therapeutically effective amount of EPZ-6438 and at least one pharmaceutically acceptable carrier or excipient, wherein the therapeutically effective amount is a single dose providing a mean Cmax that is bioequivalent to a mean Cmax of about 1.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438 (e.g., about 2.2 ng / ml to about 2.6 ng / ml per 1 mg of EPZ-6438).

[0087] This article also provides an oral dosage form for the treatment of advanced solid tumors or B-cell lymphomas, comprising a therapeutically effective amount of EPZ-6438 and at least one pharmaceutically acceptable carrier or excipient, wherein the therapeutically effective amount provides a mean AUC (0-12) of at least about 1170 ng*hr / ml (e.g., at least about 4421 ng*hr / ml) after administration to a human subject.

[0088] The terms "bioequivalent" or "bioequivalence" are field terms and are intended to be defined according to the 34th edition of Approved Drug Products with Therapeutic Equivalence Evaluations, published by the U.S. Department of Health and Human Services and commonly known as the "Orange Book." Bioequivalence of different formulations of the same drug involves equivalence in terms of the rate and extent of drug absorption. The extent and rate of absorption of the test formulation are compared to a reference formulation to determine whether the two formulations are bioequivalent. Standard bioequivalence studies are conducted in a crossover manner through extensive testing, and then measuring blood or plasma levels of the drug over time. Extensive testing involves administering a single dose of both the test drug and the reference drug to multiple volunteers (typically 12 to 24 healthy, normal adults). Detailed guidance for establishing bioequivalence between formulations and reference formulations has been published by the FDA Office of Generic Drugs, Division of Bioequivalence.

[0089] Two dosage forms whose absorption rates and extent differ by -20% / +25% or less are generally considered “bioequivalent.” Another method for average bioequivalence involves calculating a 90% confidence interval for the ratio of the average (total geometric mean) of the measurements of the test product and the reference product. To establish BE, the calculated confidence interval should generally fall within 80%–125% of the ratio of the product averages. In addition to this general method, other methods (including (1) logarithmic transformation of pharmacokinetic data, (2) methods for assessing sequence effects, and (3) methods for assessing outlier data) can be used to establish bioequivalence. For example, in (1) above, the confidence interval should generally fall within 80%–125% of the difference in the average of the logarithmically transformed PK parameters.

[0090] This invention relates to a pharmaceutical formulation comprising a therapeutic agent and one or more pharmaceutically acceptable excipients, wherein the therapeutic agent is compound 1:

[0091] (“Compound 1”) or its salt, or a combination thereof, and the concentration of the therapeutic agent in the preparation is equal to about 35-65 wt.% of Compound 1.

[0092] This invention relates to an immediate-release formulation comprising compound 1 or a salt thereof. In embodiments, the immediate-release formulation releases the therapeutic agent within a short time (e.g., after 60 minutes, at least 80% of the therapeutic agent contained in the formulation is released). In some embodiments, the immediate-release formulation releases at least 90%, at least 80%, at least 70%, or at least 60% of the therapeutic agent after 60 minutes in a medium with a pH range between 1 and 6.8 (e.g., pH = 1.2). In some embodiments, the immediate-release formulation releases at least 90%, at least 80%, at least 70%, or at least 60% of the therapeutic agent after 45 minutes in a medium with a pH range between 1 and 6.8 (e.g., pH = 1.2). In some embodiments, the immediate-release formulation is in tablet form.

[0093] When applicable, the formulations of the present invention may include one or more of the following features:

[0094] For example, the concentration of the therapeutic agent in the formulation is equal to about 40 wt.% to about 60 wt.% of compound 1.

[0095] For example, the concentration of the therapeutic agent in the formulation is equal to about 45 wt.% to about 55 wt.% of compound 1.

[0096] For example, the concentration of the therapeutic agent in the formulation is equal to about 47 wt.% to about 50 wt.% of compound 1.

[0097] For example, the therapeutic agent is a salt of compound 1, such as a hydrobromide (HBr) salt, like monohydrobromide.

[0098] For example, one or more pharmaceutically acceptable excipients include one or more diluents, one or more disintegrants, and one or more binders.

[0099] For example, the formulation contains about 10 wt.% to about 20 wt.% (e.g., about 10 wt.%, 11 wt.%, 12 wt.%, 13 wt.%, 14 wt.%, 15 wt.%, 16 wt.%, 17 wt.%, 18 wt.%, 19 wt.%, or 20 wt.%) of diluent.

[0100] For example, the total concentration of the therapeutic agent and diluent is about 60-80 wt.%, such as about 65 wt.%, 67.5 wt.%, 70 wt.%, 72.5 wt.%, 75 wt.%, or 80 wt.%.

[0101] For example, the diluent is lactose monohydrate.

[0102] For example, the formulation contains about 15 wt.% to about 25 wt.% (e.g., about 15 wt.%, 16 wt.%, 17 wt.%, 18 wt.%, 19 wt.%, 20 wt.%, 21 wt.%, 22 wt.%, 23 wt.%, 24 wt.%, or 25 wt.%) of a disintegrant.

[0103] For example, disintegrants include low-substituted hydroxypropyl cellulose, sodium starch glycolate, or combinations thereof.

[0104] For example, the formulation contains about 1 wt.% to about 10 wt.% (e.g., about 1 wt.%, 2 wt.%, 3 wt.%, 4 wt.%, 5 wt.%, 6 wt.%, 7 wt.%, 8 wt.%, 9 wt.%, or 10 wt.%) of adhesive.

[0105] For example, the adhesive is hydroxypropyl cellulose.

[0106] For example, one or more pharmaceutically acceptable excipients further include lubricants.

[0107] For example, the formulation contains about 0.5 wt.% to about 5 wt.% (e.g., about 0.5 wt.%, 0.7 wt.%, 0.9 wt.%, 1 wt.%, 2 wt.%, 3 wt.%, 4 wt.%, or 5 wt.%) of a lubricant.

[0108] For example, a lubricant is magnesium stearate.

[0109] For example, one or more pharmaceutically acceptable excipients further include coating compositions.

[0110] For example, the formulation comprises about 1 wt.% to about 10 wt.% (e.g., about 1 wt.%, 2 wt.%, 3 wt.%, 4 wt.%, 5 wt.%, 6 wt.%, 7 wt.%, 8 wt.%, 9 wt.%, or 10 wt.%) of a coating composition.

[0111] For example, the coating composition is a water-soluble, immediate-release coating composition.

[0112] For example, the coating composition contains hydroxypropyl methylcellulose.

[0113] For example, the coating composition further comprises talc and polyethylene glycol.

[0114] For example, the coating composition further comprises a colorant, such as titanium dioxide, iron(III) oxide, or both.

[0115] For example, the coating composition comprises one or more of polyvinyl alcohol, hydroxypropyl methylcellulose, talc, and polyethylene glycol. For example, the coating composition further comprises titanium dioxide and / or iron(III) oxide. For example, the coating composition is Opadry03F45063 RED.

[0116] For example, the formulation comprises a therapeutic agent in an amount equal to about 40-60 wt.% of Compound 1, about 10-20 wt.% of a diluent, about 15-25 wt.% of a disintegrant, about 1-10 wt.% of a binder, about 0.5-5 wt.% of a lubricant, and about 1-10 wt.% of a coating composition. For example, the formulation comprises a therapeutic agent in an amount equal to about 40-60 wt.% of Compound 1, about 12-18 wt.% of a diluent, about 18-23 wt.% of a disintegrant, about 2-6 wt.% of a binder, about 1-3 wt.% of a lubricant, and about 2-6 wt.% of a coating composition.

[0117] For example, the formulation comprises a therapeutic agent, lactose monohydrate, low-substituted hydroxypropyl cellulose, sodium starch glycolate, hydroxypropyl cellulose, and magnesium stearate. For example, the formulation comprises a therapeutic agent in an amount equal to about 35-65 wt.%, or 40-60 wt.%, or 45-55 wt.% of Compound 1, about 10-20 wt.% of lactose monohydrate, about 11-19 wt.% of low-substituted hydroxypropyl cellulose, about 3-7 wt.% of sodium starch glycolate, about 1-10 wt.% of hydroxypropyl cellulose, and about 0.5-5 wt.% of magnesium stearate. For example, the formulation comprises a therapeutic agent in an amount equal to about 50 wt.% of Compound 1, about 17 wt.% of lactose monohydrate, about 15 wt.% of low-substituted hydroxypropyl cellulose, about 5 wt.% of sodium starch glycolate, about 4 wt.% of hydroxypropyl cellulose, and about 2 wt.% of magnesium stearate.

[0118] For example, the formulation comprises a therapeutic agent, lactose monohydrate, low-substituted hydroxypropyl cellulose, sodium starch glycolate, hydroxypropyl cellulose, magnesium stearate, and a coating composition. For example, the formulation comprises a therapeutic agent in an amount equal to about 40-60 wt.% of compound 1, about 10-20 wt.% of lactose monohydrate, about 11-19 wt.% of low-substituted hydroxypropyl cellulose, about 3-7 wt.% of sodium starch glycolate, about 1-10 wt.% of hydroxypropyl cellulose, about 0.5-5 wt.% of magnesium stearate, and about 1-10 wt.% of the coating composition. For example, the formulation consists of a therapeutic agent in an amount equal to about 47-48 wt.% of compound 1, about 16 wt.% of lactose monohydrate, about 14-15 wt.% of low-substituted hydroxypropyl cellulose, about 5 wt.% of sodium starch glycolate, about 4 wt.% of hydroxypropyl cellulose, about 2 wt.% of magnesium stearate, and about 4 wt.% of a coating composition.

[0119] For example, the formulation is an oral dosage form, each unit of which contains an amount of therapeutic agent equal to about 10 mg to about 1000 mg, or about 10 mg to about 800 mg, or about 10 mg to about 500 mg, or about 10 mg to about 400 mg of compound 1. For example, the oral dosage form is in tablet form. For example, the tablet contains an amount of therapeutic agent equal to about 25 mg to about 400 mg of compound 1. For example, the tablet contains an amount of therapeutic agent equal to about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, or about 400 mg of compound 1.

[0120] For example, the formulation is a solid formulation. For example, the formulation is substantially free of water. In this context, "substantially" free of water means that the water content of the formulation is less than 7%, less than 5%, less than 1%, or less than 0.5% of the total weight of the formulation at the time of packaging. In one embodiment, the amount of water is between 0.1% and 5% (e.g., 0.1%-1% or 0.1%-0.5%) of the total weight of the formulation. In one embodiment, the amount of water in the formulation of the present invention manufactured by spray coating is less than 0.5%.

[0121] This invention relates to oral formulations (e.g., in tablet form) as stable formulations. For example, the stable formulations of this invention maintain, over a period of time (e.g., 3 months, 12 months, 18 months, and 24 months), the amount of the active compound (e.g., compound 1 or a salt thereof) in the formulation at at least 90%, preferably at least 95%, and most preferably at least 99% of the amount of the active compound initially present in the formulation. Storage conditions may be 2-8 degrees Celsius (2°C-8°C), or 25 degrees Celsius (25°C) and 60% relative humidity, or 25°C and 75% relative humidity, or 40°C and 75% relative humidity.

[0122] This invention relates to pharmaceutical formulations, specifically solid pharmaceutical formulations comprising a therapeutic agent (e.g., compound 1, or a salt thereof, or a combination thereof) and one or more pharmaceutically acceptable excipients selected from sodium starch glycolate, carboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium, or low-substituted hydroxypropyl cellulose, and combinations thereof. In one embodiment, the excipients are selected from sodium starch glycolate, carboxymethyl cellulose, calcium carboxymethyl cellulose, or croscarmellose sodium, and combinations thereof. In another embodiment, the excipients are selected from sodium starch glycolate, or carboxymethyl cellulose, and combinations thereof. In yet another embodiment, the pharmaceutical formulation further comprises lactose, hydroxypropyl cellulose, or magnesium stearate, or combinations thereof.

[0123] In one embodiment, the pharmaceutical formulation comprises a therapeutic agent in an amount equal to about 25-75 wt.% of Compound 1, and about 5-35 wt.% of excipients selected from sodium starch glycolate, carboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium, or low-substituted hydroxypropyl cellulose, and combinations thereof. In one embodiment, the pharmaceutical formulation comprises a therapeutic agent in an amount equal to about 40-60 wt.% of Compound 1, about 10-30 wt.% of excipients, about 10-20 wt.% of diluent, about 2-6 wt.% of binder, and about 1-3 wt.% of lubricant selected from sodium starch glycolate, carboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium, or low-substituted hydroxypropyl cellulose, and combinations thereof.

[0124] In one embodiment, the pharmaceutical formulation comprises a therapeutic agent in an amount equal to about 50 wt.% of Compound 1, about 20 wt.% of excipients, about 10-20 wt.% of diluent, about 2-6 wt.% of binder, and about 1-3 wt.% of lubricant, wherein the excipients are selected from sodium starch glycolate, carboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium, or low-substituted hydroxypropyl cellulose, and combinations thereof. In another embodiment, the formulation comprises a therapeutic agent in an amount equal to about 50 wt.% of Compound 1, about 20 wt.% of excipients, about 10-20 wt.% of lactose monohydrate, about 2-6 wt.% of hydroxypropyl cellulose, and about 1-3 wt.% of magnesium stearate, wherein the excipients are selected from sodium starch glycolate, carboxymethyl cellulose, and combinations thereof.

[0125] In one embodiment of the invention, the composition of the preparation of the invention is provided in Table 3 of Example 2.

[0126] In one embodiment of the invention, the composition of the formulations of the invention is provided in Tables 5-6 of Example 3, such as one of formulation numbers 1-5.

[0127] In one embodiment, the formulation is a solid formulation. In one embodiment, the formulation is in the form of a powder, granules, capsules, lozenges, or tablets. In one embodiment, the unit dose is a powder or tablet. In one embodiment, the tablet is in a blister pack or strip. For example, the blister pack or strip may be made of a material impermeable to water vapor and oxygen. In one embodiment, the blister pack is made of metal foil. In one embodiment, the blister pack is a FOIL / FOIL blister pack. In one embodiment, the container of the blister pack is flushed with an inert gas (such as nitrogen or argon). In one embodiment, the container further includes a desiccant, such as a molecular sieve. In one embodiment, the unit dose is in a high-density polyethylene bottle with a seal. In one embodiment, the bottle further includes a desiccant. In one embodiment, the bottle further includes an oxygen scavenger and / or a molecular sieve. In one embodiment, the bottle is substantially impermeable to oxygen and water vapor (e.g., much more impermeable than an HDPE bottle), such as an OxyGuard bottle.

[0128] The present invention also relates to a solid pharmaceutical composition comprising a therapeutic agent and means for achieving immediate release of the therapeutic agent, wherein the therapeutic agent is selected from compound 1, its salts, and combinations thereof.

[0129] For example, a device for achieving rapid release of the therapeutic agent allows for the release of at least 90%, or at least 80%, or at least 70%, or at least 60% of the therapeutic agent after 60 minutes. For example, a device for achieving rapid release of the therapeutic agent allows for the release of at least 90%, or at least 80%, or at least 70%, or at least 60% of the therapeutic agent after 45 minutes. For example, a device for achieving rapid release of the therapeutic agent allows for the release of at least 80%, or at least 70%, or at least 60% of the therapeutic agent after 30 minutes. For example, according to the dissolution test 6.10 of JP16 or USP37... <711> The procedure for dissolving an immediate-release formulation, wherein the apparatus used to achieve immediate release of the therapeutic agent allows for a dissolution rate of at least approximately 90%, at least approximately 80%, or at least approximately 70% in the dissolution medium (pH 1.2, 900 mL, 37 ± 0.5 °C) within 60, 45, or 30 minutes from the start of a dissolution study using instrument 2 (paddle apparatus, paddle speed; 50 rpm). For example, according to the dissolution test 6.10 of JP16 or USP37. <711> The procedure for dissolving an immediate-release formulation, wherein the apparatus for achieving immediate release of the therapeutic agent allows for a dissolution rate of at least about 80%, or at least about 75%, or at least about 70%, or at least about 60% in the dissolution medium (pH 4.5 acetate buffer, 900 mL, 37 ± 0.5 °C) within 60, 45, or 30 minutes from the start of a dissolution study using instrument 2 (paddle apparatus, paddle speed; 50 rpm).

[0130] In another aspect, the present invention relates to a method for preparing the pharmaceutical formulations or compositions disclosed herein. The method comprises a) mixing a therapeutic agent, a diluent, a disintegrant, and optionally a lubricant to form a first mixture, wherein the therapeutic agent is selected from the group consisting of: compound 1, its salts, and combinations thereof. The method optionally includes one or more of the following steps:

[0131] b) Add an aqueous solution containing an adhesive, or an organic solvent-based solution (e.g., IPA, EtOH, etc.), or an organic / aqueous mixture (e.g., 1:1 EtOH:water) to the first mixture to form a second mixture;

[0132] c) Granulate the second mixture to form wet granules;

[0133] d) Dry these wet particles to form dried particles;

[0134] e) These dried particles are screened by size to obtain particles that meet the size requirements;

[0135] f) Mix these appropriately sized particles with a lubricant and a second disintegrant to form a third mixture;

[0136] g) Compress the third mixture to form tablets; and

[0137] h) Apply a coating suspension to these tablets to produce film-coated tablets.

[0138] In some embodiments, the present invention relates to a method for preparing the pharmaceutical formulation disclosed herein. The method includes a) mixing a therapeutic agent, a diluent, and a disintegrant to form a first mixture, wherein the therapeutic agent is selected from the group consisting of: compound 1, its salts, and combinations thereof.

[0139] The method optionally includes one or more of the following steps:

[0140] b) Granulate the first mixture into dry particles;

[0141] e) These dry particles are sorted by size;

[0142] f) Mix these appropriately sized particles with a lubricant and a second disintegrant to form a third mixture;

[0143] g) Compress the third mixture to form tablets; and

[0144] h) Apply a coating suspension to these tablets to produce film-coated tablets.

[0145] Where applicable, the method of the present invention may include one or more of the following features:

[0146] For example, this method does not include step e) size filtering.

[0147] For example, the method does not include step h) coating.

[0148] For example, the method does not include step f) mixing with the lubricant and the second disintegrant.

[0149] For example, the method does not include step g) compression.

[0150] For example, the diluent is lactose monohydrate.

[0151] For example, a disintegrant is low-substituted hydroxypropyl cellulose.

[0152] For example, the adhesive is hydroxypropyl cellulose.

[0153] For example, a lubricant is magnesium stearate.

[0154] For example, second disintegrants include low-substituted hydroxypropyl cellulose, sodium starch glycolate, or combinations thereof.

[0155] For example, the coating suspension contains hydroxypropyl methylcellulose. For example, the coating suspension further contains water, talc, and / or polyethylene glycol. For example, the coating suspension further contains one or more colorants.

[0156] In one embodiment of the present invention, a method for preparing the formulation of the present invention is provided. Figure 8 In the middle. For example Figure 8 As shown, the drug (e.g., compound 1, its salt, or a combination thereof), lactose monohydrate, and low-substituted hydroxypropyl cellulose are loaded into a high-shear mixer and mixed (Step 1: Mixing). Then, the hydroxypropyl cellulose is dissolved in purified water, and the solution is added to the mixer, and the mixture is granulated to produce wet granules (Step 2: Granulation). The wet granules are then dried using a fluidized bed dryer to produce dried granules (Step 3: Drying). Next, the dried granules are sieved for size screening (Step 4: Size Screening). Then, the size-fitted granules, low-substituted hydroxypropyl cellulose, sodium starch glycolate, and magnesium stearate are combined and lubricated in a rolling mixer (Step 5: Lubrication). The amounts of low-substituted hydroxypropyl cellulose, sodium starch glycolate, and magnesium stearate used in Step 5 are adjusted according to the yield of size-fitted granules. For example, the higher the yield of size-fitted granules, the higher the amount of disintegrant and lubricant used. Next, the lubricated granules are compressed into tablets using a tableting machine (Step 6: Tableting). Then, a pan coating machine is used to spray the coating suspension prepared by mixing OPADRY 03F45063 RED with purified water onto the tablets (step 7: film coating) to produce film-coated tablets.

[0157] In any of the methods or formulations described herein (e.g., oral dosage forms), in one embodiment, the cancer is advanced, refractory, or drug-resistant cancer. In any of the methods or formulations described herein (e.g., oral dosage forms), in one embodiment, the cancer is an INI1-deficient tumor.

[0158] In any of the methods or formulations described herein (e.g., oral dosage forms), in one embodiment, the subject is a human.

[0159] Where applicable, in any of the methods or formulations described herein (e.g., oral dosage forms), the cancer is a solid tumor. Examples of solid tumors described herein include, but are not limited to, colorectal adenocarcinoma, bile duct cancer, pancreatic cancer, Ewing's sarcoma, synovial sarcoma, alveolar sarcoma, alveolar soft tissue sarcoma, prostate cancer, rhabdomyosarcoma, malignant rhabdomyosarcoma, and urothelial carcinoma.

[0160] Where applicable, in any of the methods or formulations described herein (e.g., oral dosage forms), the cancer is B-cell lymphoma. Examples of B-cell lymphomas described herein include, but are not limited to, diffuse large B-cell lymphoma, follicular lymphoma, and marginal zone lymphoma.

[0161] Where applicable, in any of the methods or formulations described herein (e.g., oral dosage forms), the cancer is a cancer with abnormal H3-K27 methylation.

[0162] In any of the methods or formulations described herein (e.g., oral dosage forms), a compound having chemical formula (I) or a pharmaceutically acceptable salt thereof is administered orally for at least 7, 14, 21, 28, 35, 42, 47, 56, or 64 days. In some embodiments, administration is continuous administration without drug holidays. For example, a compound having chemical formula (I) or a pharmaceutically acceptable salt thereof is administered orally for 28 days in a 28-day cycle. In other embodiments, the compound is administered with drug holidays. For example, a compound having chemical formula (I) or a pharmaceutically acceptable salt thereof is administered orally for day 21 of a 28-day cycle, with a 7-day drug holiday for each cycle.

[0163] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, the range of a single dose is from about 100 mg to about 1600 mg.

[0164] In any of the methods or formulations described herein (e.g., oral dosage forms), a single dose of a compound having chemical formula (I) or a pharmaceutically acceptable salt thereof is 100, 200, 400, 800, or 1600 mg.

[0165] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, the therapeutically effective dose is a single 400 mg dose, wherein the single dose provides a bioequivalence of the mean AUC(0-12) from about 1720 ng*hr / ml to about 7798 ng*hr / ml (e.g., from about 1720 ng*hr / ml to about 3899 ng*hr / ml).

[0166] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, the therapeutically effective dose is a single 800 mg dose, wherein the single dose provides a bioequivalence of a mean AUC(0-12) from about 7798 ng*hr / ml to about 9441 ng*hr / ml.

[0167] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, the therapeutically effective dose is a single 1600 mg dose, wherein the single dose provides a bioequivalence of a mean AUC(0-12) from about 15596 ng*hr / ml to about 18882 ng*hr / ml.

[0168] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, the therapeutically effective dose is a single 400 mg dose, wherein the single dose provides a mean Cmax that is bioequivalent to a mean Cmax from about 476 ng / ml to about 1730 ng / ml (e.g., from about 476 ng / ml to about 865 ng / ml).

[0169] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, the therapeutically effective dose is a single 800 mg dose, wherein the single dose provides a bioequivalence of a mean Cmax from about 1730 ng / ml to about 2063 ng / ml.

[0170] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, the therapeutically effective dose is a single 1600 mg dose, wherein the single dose provides a bioequivalence of a mean Cmax from about 3460 ng / ml to about 4125 ng / ml.

[0171] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, the administration includes orally administering the dosage form to the subject twice or three times daily.

[0172] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, the single dose provides a median Tmax from about 1 hour to about 2 hours.

[0173] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, each unit dose of the oral dosage form or formulation contains a therapeutic agent in an amount equal to about 25 mg to about 400 mg (e.g., equal to about 25 mg to about 200 mg) of EPZ-6438.

[0174] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, dissolution testing is performed according to JP16 6.10 or USP37. <711> The procedure for dissolving intermediate-release dosage forms, wherein the oral dosage form or formulation provides at least about 90%, or at least about 80%, or at least about 70% dissolution in the dissolution medium (pH 1.2, 37±0.5°C) within 60 minutes from the start of a dissolution study using instrument 2 (paddle apparatus, paddle speed; 50 rpm).

[0175] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, dissolution testing is performed according to JP16 6.10 or USP37. <711> The procedure for dissolving intermediate-release dosage forms, wherein the oral dosage form or formulation provides at least about 90%, or at least about 80%, or at least about 70% dissolution in the dissolution medium (pH 1.2, 37±0.5°C) within 45 minutes from the start of a dissolution study using instrument 2 (paddle apparatus, paddle speed; 50 rpm).

[0176] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, dissolution testing is performed according to JP16 6.10 or USP37. <711> The procedure for dissolving intermediate-release dosage forms, wherein the oral dosage form or formulation provides at least about 90%, or at least about 80%, or at least about 70% dissolution in the dissolution medium (pH 1.2, 37±0.5°C) within 30 minutes from the start of a dissolution study using instrument 2 (paddle apparatus, paddle speed; 50 rpm).

[0177] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, dissolution testing is performed according to JP16 6.10 or USP37. <711> The procedure for dissolving intermediate-release dosage forms, wherein the oral dosage form or formulation provides at least about 80%, or at least about 75%, or at least about 70%, or at least about 60% dissolution in the dissolution medium (pH 4.5 acetate buffer, 37 ± 0.5 °C) within 60 minutes from the start of a dissolution study using instrument 2 (paddle apparatus, paddle speed; 50 rpm).

[0178] In any of the methods or formulations described herein (e.g., oral dosage forms), in some embodiments, the oral dosage form or formulation comprises sodium starch glycolate or carboxymethyl cellulose or combinations thereof as a pharmaceutically acceptable carrier or excipient.

[0179] Other compounds suitable for the methods of the present invention are described in U.S. Publication No. 20120264734, the contents of which are incorporated herein by reference in their entirety. Additionally, compound 1 is suitable as part of a combination therapy administered with one or more other therapeutic agents or modalities suitable for administration together, sequentially, or alternately.

[0180] In one embodiment, the subject is given compound 1 or a pharmaceutically acceptable salt thereof at a dose of about 100 mg to about 3200 mg per day, such as about 100 mg BID to about 1600 mg BID (e.g., 100 mg BID, 200 mg BID, 400 mg BID, 800 mg BID, or 1600 mg BID), for the treatment of cancer (e.g., solid tumors, B-cell lymphomas, or cancers with aberrant H3-K27 methylation).

[0181] In one embodiment, the subject is given compound 1 or a pharmaceutically acceptable salt thereof at a dose of about 100 mg to about 3200 mg per day, such as about 100 mg BID to about 1600 mg BID (e.g., 100 mg BID, 200 mg BID, 400 mg BID, 800 mg BID, or 1600 mg BID), for the treatment of INI1-deficient tumors.

[0182] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof is administered in combination (simultaneously or sequentially) with a standard of care (such as one or more components of R-CHOP, a BCL inhibitor, or a BCR inhibitor). For example, compound 1 (or EPZ-6438) or a pharmaceutically acceptable salt thereof has an additive or synergistic effect when combined with a drug targeting the BCR / PI3K pathway in cell lines containing mutant EZH2 or WT EZH2 germinal center lymphoma cell lines. No effect was observed in ABC lymphoma cell lines when they were exposed to a combination of EPZ-6438 and a drug targeting the BCR / PI3K pathway. Importantly, EPZ-6438 in combination with a drug targeting the BCR / PI3K pathway showed a synergistic effect in germinal center B-cell lymphoma (GCB lymphoma) cell lines, regardless of whether the GCB-lymphoma cell lines contain WT or mutant EZH2 protein.

[0183] Other embodiments or examples of the combination therapy are described in a co-pending application, International Application PCT / US2014 / 069167, which claims priority and benefit to USSN 61 / 913,063, filed December 6, 2013; USSN 61 / 934,338, filed January 31, 2014; and USSN 61 / 992,881, filed May 13, 2014, the contents of which are hereby incorporated by reference in their entirety.

[0184] In one embodiment, the compound of the present invention is the compound itself, i.e., a free base or a "naked" molecule. In another embodiment, the compound is its salt, such as a mono-HCl salt or tri-HCl salt, mono-HBr salt or tri-HBr salt of a naked molecule.

[0185] Unless otherwise indicated herein or clearly contradicted by the context, the use of the articles “a / an” and “the” herein shall be interpreted to include both the singular and plural. For example, the term “disintegrant” refers to one or more disintegrants included in or suitable for use in the formulations described herein. Similarly, the term “therapeutic agent” refers to one or more therapeutic agents included in or suitable for use in the formulations described herein. For example, the formulations described herein may include only Compound 1 as a therapeutic agent or active ingredient, or may include a mixture of Compound 1 and another compound (e.g., an HBr salt of Compound 1 or another anticancer drug). Unless otherwise indicated, the terms “comprising,” “having,” “including,” and “containing” shall be interpreted as open-ended terms (i.e., meaning “including but not limited to”). Furthermore, whenever “comprising” or another open-ended term is used in an embodiment, it should be understood that the same embodiment may be required more narrowly using the intermediate term “consistently composed of” or the closed term “composed of.”

[0186] The concentration of the therapeutic agent in the formulation is expressed as an amount equal to a certain amount of Compound 1. As used herein, the term "equivalent" or weight percentage refers to the amount of drug adjusted according to a potency correction factor (a value obtained from a assay value derived from Compound 1). Methods for determining equivalent amounts are well known in the art (see, for example, http: / / www.fda.gov / downloads / Drugs / ... / Guidances / ucm070246.pdf).

[0187] When used in conjunction with numerical values, the terms "about," "approximately," or "approximately" mean including a set of values ​​or a series of values. For example, "about X" includes a series of values ​​of ±10%, ±5%, ±2%, ±1%, ±0.5%, ±0.2%, or ±0.1% of X, where X is a numerical value. Additionally, "about X" may also include a series of X ±0.5, X ±0.4, X ±0.3, X ±0.2, or X ±0.1, where X is a numerical value. In one embodiment, the term "about" refers to a series of values ​​that are more than or less than 5% of a specified value. In another embodiment, the term "about" refers to a series of values ​​that are more than or less than 2% of a specified value. In yet another embodiment, the term "about" refers to a series of values ​​that are more than or less than 1% of a specified value.

[0188] In this specification, for convenience, the structural formula of a compound may represent a particular isomer in some cases; however, this invention includes all isomers, such as geometric isomers, optical isomers based on asymmetric carbon, stereoisomers, tautomers, etc. Furthermore, compounds represented by chemical formulas may exist in polymorphic forms. It should be noted that any crystalline form, mixture of crystalline forms, or its anhydrous form or hydrate is included within the scope of this invention. Moreover, so-called metabolites generated by the in vivo degradation of the compounds of this invention are included within the scope of this invention.

[0189] Furthermore, the structures and other compounds discussed in this invention include all their transisomers. A “transisomer” is a class of stereoisomers in which the atoms of the two isomers have different spatial arrangements. Transisomers attribute their existence to rotational restriction caused by the impeded rotation of a large group around a central bond. Such transisomers typically exist as mixtures; however, due to recent advances in chromatographic techniques, mixtures of two transisomers can now be separated under selected conditions.

[0190] A tautomer is one of two or more structural isomers that exist in equilibrium and readily transform from one isomeric form to another. This transformation results in the migration of hydrogen atoms, accompanied by the conversion of adjacent conjugated double bonds. Tautomers exist in solution as a mixture of tautomer groups. In solutions where tautomerization is possible, tautomers will reach chemical equilibrium. The exact ratio of tautomers depends on several factors, including temperature, solvent, and pH. The concept of tautomers that can interconvert through tautomerism is called the phenomenon of tautomerism.

[0191] Of the many possible types of tautomerism, two are commonly observed. In keto-enol tautomerism, electrons and hydrogen atoms are transferred simultaneously. Ring-chain tautomerism occurs when the aldehyde group (-CHO) in the sugar molecule reacts with a hydroxyl group (-OH) in the same molecule, forming a cyclic (ring-shaped) form, as seen in glucose.

[0192] Common tautomer pairs include: keto-enol, amide-nitrile, lactam-lactamimide, amide-imine tautomerism in heterocycles (e.g., nucleobases such as guanine, thymine, and cytosine), amine-enamine, and enamine-enamine. An example of a keto-enol equilibrium, as shown below, is between pyridin-2(1H)-one and the corresponding pyridin-2-ol.

[0193]

[0194] It should be understood that the compounds used in the formulations of this invention can be described as different tautomers. It should also be understood that when a compound has a tautomer form, all tautomer forms are intended to be included within the scope of this invention, and the naming of the compound does not exclude any tautomer form.

[0195] The terms "crystalline polymorph," "polymorph," or "crystal form" refer to crystalline structures in which a compound (or its salt or solvate) can crystallize in different crystalline stacks, all of which have the same elemental composition. Different crystal forms typically exhibit different XRPD patterns, infrared spectra, melting points, densities, hardness, crystal shapes, optical and electrical properties, stability, and solubility. Recrystallization solvents, crystallization rates, storage temperatures, and other factors can cause one crystal form to become dominant. Crystalline polymorphs of a compound can be prepared by crystallization under different conditions.

[0196] The compounds of the present invention can be crystalline, semi-crystalline, amorphous, mesocrystalline, etc.

[0197] The compounds of the present invention include the compounds themselves, as well as their N-oxides, salts, solvates, and prodrugs (if applicable). For example, salts can be formed between an anion and a positively charged group (e.g., amino) on a substituted purine or 7-denitropurine compound. Suitable anions include chloride, bromide, iodide, sulfate, hydrogen sulfate, aminosulfonate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, glutamate, glucuronate, glutarate, malate, maleate, succinate, fumarate, tartrate, toluenesulfonate, salicylate, lactate, naphthalenesulfonate, and acetate. Similarly, salts can also be formed between a cation and a negatively charged group (e.g., carboxylate) on a substituted purine or 7-denitropurine compound. Suitable cations include sodium, potassium, magnesium, calcium, and ammonium cations (such as tetramethylammonium ions). Substituted purines or 7-denitropurines also include salts containing a quaternary nitrogen atom. Examples of prodrugs include esters and other pharmaceutically acceptable derivatives that, when administered to a subject, provide the active substituted purine or 7-denitropurine compound.

[0198] Furthermore, the compounds or crystal forms of the present invention (e.g., salts of the compounds or crystal forms) may exist in hydrated or dehydrated (anhydrous) forms or as solvates with other solvent molecules. Non-limiting examples of hydrates include hemihydrates, monohydrates, dihydrates, trihydrates, etc. Non-limiting examples of solvates include ethanol solvates, acetone solvates, etc.

[0199] A solvate is a solvent addition form containing stoichiometric or non-stoichiometric amounts of solvent. Some compounds tend to capture solvent molecules in a fixed molar ratio in a crystalline solid state, thus forming a solvate. If the solvent is water, the resulting solvate is a hydrate; and if the solvent is an alcohol, the resulting solvate is an alcohol. A hydrate is formed by the combination of one or more water molecules with one molecule of the substance, where the water retains its molecular state as H₂O. A hemihydrate is formed by the combination of one water molecule with more than one molecule of the substance, where the water retains its molecular state as H₂O.

[0200] This invention aims to include all isotopes of atoms present in the compounds of this invention. Isotopes include atoms that have the same number of atoms but different mass numbers. By way of general example and without limitation, hydrogen isotopes include tritium and deuterium, and carbon isotopes include C-13 and C-14.

[0201] As used herein, “subject” and “subject in need of” are interchangeable, both referring to a subject with a disorder in which EZH2-mediated protein methylation plays a role, or a subject with an increased risk of developing such a disorder relative to the majority population. “Subject” includes mammals. Mammals can be, for example, humans or suitable non-human mammals such as primates, mice, rats, dogs, cats, cows, horses, goats, camels, sheep, or pigs. Subjects can also be birds or poultry. In one embodiment, the mammal is a human. Subjects in need of “their” can be individuals who have been previously diagnosed or identified with cancer or a precancerous condition. Subjects in need of “their” can also be individuals who have (e.g., are currently suffering from) cancer or a precancerous condition. Alternatively, subjects in need of “their” can be individuals with an increased risk of developing such a disorder relative to the majority population (i.e., subjects who are predisposed to developing such a disorder relative to the majority population). Subjects in need of “their” can have a precancerous condition. Subjects in need of “their” have an INI1-deficient tumor.

[0202] INI1 is a regulatory complex that antagonizes the enzymatic function of EZH2. Due to various genetic alterations, INI1 has lost its regulatory function. As a result, EZH2 activity is misregulated, leading to EZH2 playing a driving oncogenic role in a group of genetically determined cancers, including synovial sarcoma and malignant rhabdomyosarcoma.

[0203] Synovial sarcoma is a malignant soft tissue tumor and one of the most common soft tissue tumors in adolescents and young patients. The average age of patients at diagnosis is approximately 30 years old.

[0204] Malignant rhabdomyosarcoma, or MRT, is a rare and deadly form of childhood cancer caused by specific genetic alterations that lead to dysfunction of EZH2. MRT typically occurs in the kidneys or brain and in children under two years of age.

[0205] Subjects in need may have refractory or drug-resistant cancer (i.e., cancer that is unresponsive or has not yet responded to treatment). Subjects may be resistant at the start of treatment or may become resistant during treatment. In some embodiments, the cancer of the subject in need has relapsed after a recent therapeutic remission. In some embodiments, the subject in need has received all known effective therapies for cancer treatment but has failed. In some embodiments, the subject in need has received at least one prior therapy. In a preferred embodiment, the subject has cancer or a cancerous condition.

[0206] As used herein, “treating” describes the management and care of a patient for the purpose of combating a disease, symptom, or disorder, and includes administering the compounds of the present invention, or pharmaceutically acceptable salts, prodrugs, metabolites, polymorphs, or solvates thereof, to alleviate or eliminate symptoms or complications of the disease, symptom, or disorder. The term “treating” may also include treatment in in vitro cell or animal models.

[0207] The compounds of the present invention, or pharmaceutically acceptable salts, prodrugs, metabolites, polymorphs, or solvates thereof, may or may also be used to prevent associated diseases, conditions, or disorders, or to identify suitable candidates for this purpose. As used herein, “preventing” or “protecting from” describes reducing or eliminating the onset of symptoms or complications of such disease, condition, or disorder.

[0208] The methods and uses described herein may include the step of detecting the presence or absence of one or more EZH2 mutations in a sample from a subject before and / or after administering the compound or composition described herein to a subject in need of it. The term "sample" means any biological sample derived from a subject, including but not limited to cell, tissue, bodily fluids (including but not limited to mucus, blood, plasma, serum, urine, saliva, and semen), tumor cells, and tumor tissue. Preferably, the sample is selected from bone marrow, peripheral blood cells, blood, plasma, and serum. The sample may be provided by the subject being treated or tested. Alternatively, the sample may be obtained by a physician according to standard practice in the art.

[0209] Point mutations in the EZH2 gene at single amino acid residues (e.g., Y641, A677, and A687) have been reported to be associated with lymphoma. Further examples of EZH2 mutants, as well as methods for mutation detection and treatments for mutation-related disorders, are described, for example, in U.S. Patent Application Publication No. 20130040906, the entire contents of which are incorporated herein by reference.

[0210] Those skilled in the art can obtain detailed descriptions of the known or equivalent techniques discussed herein by referring to general references. These publications include Ausubel et al., Current Protocols in Molecular Biology, John Wiley and Sons, Inc. (2005); Sambrook et al., Molecular Cloning, A Laboratory Manual (3rd edition), Cold Spring Harbor Press, Cold Spring Harbor, New York (2000); Coligan et al., Current Protocols in Immunology, John Wiley and Sons, New York; Enna et al., Current Protocols in Pharmacology, John Wiley and Sons, New York; and Fingl et al., The Pharmacological Basis of Therapeutics (1975), Remington's Pharmaceutical Sciences, Mack Publishing. Co., Easton, Pennsylvania, 18th edition (1990). These documents may, of course, be consulted in matters concerning the preparation or use of the invention.

[0211] The present invention also provides pharmaceutical compositions comprising one or more active compounds (e.g., compound 1 or a salt thereof) in combination with at least one pharmaceutically acceptable excipient or carrier.

[0212] A “pharmaceutical composition” is a formulation containing a compound of the invention in a form suitable for administration to a subject. In one embodiment, the pharmaceutical composition is in bulk or in unit dosage form. As used herein, the term “unit dosage form” refers to a physically discrete unit suitable as a single dose for a subject to be treated; each unit contains a predetermined amount of active compound bound to a desired drug carrier to produce the desired therapeutic effect. Unit dosage forms are any of a variety of forms, including, for example, IV bags, tablets, single pumps or vials on aerosol inhalers. The amount of active ingredient (e.g., a formulation of the disclosed compound or its salts, hydrates, solvates or isomers) in a unit dosage composition is an effective amount and varies depending on the specific treatment involved. Those skilled in the art will recognize that it is sometimes necessary to routinely change the dosage based on the patient’s age and condition. The dosage will also depend on the route of administration. A variety of routes are considered, including oral, pulmonary, rectal, parenteral, percutaneous, subcutaneous, intravenous, intramuscular, intraperitoneal, inhalation, buccal, sublingual, intrapleural, intrathecal, intranasal, etc. Dosage forms for topical or transdermal administration of the compounds of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. In one embodiment, the active compound is mixed under aseptic conditions with a pharmaceutically acceptable carrier and any desired preservatives, buffers, or propellants.

[0213] In one embodiment, the unit dosage form is an oral dosage form. In another embodiment, the unit dosage form is a tablet.

[0214] As used herein, the phrase “pharmaceutically acceptable” means compounds, materials, compositions, carriers, and / or dosage forms that, to the extent of reasonable medical judgment, are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0215] "Pharmaceutically acceptable excipients" means excipients used in the preparation of pharmaceutical compositions that are generally safe, non-toxic, and not biologically or otherwise undesirable, and includes excipients acceptable for veterinary and human pharmaceutical use. As used in this specification and claims, "pharmaceutically acceptable excipients" includes one or more such excipients. For example, pharmaceutically acceptable excipients for formulations of the present invention may be diluents or inert carriers, lubricants, binders, or combinations thereof. Pharmaceutically acceptable excipients for formulations of the present invention may further include fillers, antimicrobial agents, antioxidants, anti-caking agents, coating agents, or mixtures thereof.

[0216] Examples of pharmaceutically acceptable excipients include, but are not limited to, binders, fillers, disintegrants, lubricants, antimicrobial agents, antioxidants, and coating agents.

[0217] Exemplary adhesives include, but are not limited to, corn starch, potato starch, other starches, gelatin, natural and synthetic gums such as gum arabic, xanthan gum, sodium alginate, alginic acid, other alginates, powdered tragacanth gum, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose), polyvinylpyrrolidone (e.g., povidone, cropovidone, copovidone, etc.), methylcellulose, Methocel, and pregelatinized starches (e.g., STARCH sold by Colorcon Ltd.). and STARCH 1500 Hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose (FMC Corporation, Marcus Hook, Pennsylvania, USA), Emdex, Plasdone, or mixtures thereof; fillers: talc, calcium carbonate (e.g., granules or powder), dicalcium phosphate, tricalcium phosphate, calcium sulfate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrin, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, glucose, fructose, honey, anhydrous lactose, lactose monohydrate, lactose and aspartame, lactose and cellulose, lactose and microcrystalline cellulose, maltodextrin, maltose, mannitol, microcrystalline cellulose & guar gum, molasses, sucrose, or mixtures thereof.

[0218] Exemplary disintegrants include, but are not limited to: agar, alginate, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacolin potassium, sodium starch glycolate (such as Explotab), potato or cassava starch, other starches, pregelatinized starches, clay, other alginates, other celluloses, gums (such as gellan gum), low-substituted hydroxypropyl cellulose, ployplasdone, or mixtures thereof.

[0219] Exemplary lubricants include, but are not limited to: calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, glyceryl behenate, stearic acid, sodium lauryl sulfate, sodium stearoyl fumarate (such as Pruv), vegetable fatty acid lubricants, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, syloid silica (AEROSIL 200, WRGrace Co., Baltimore, MD, USA), condensed aerosols of synthetic silica (Deaussa, Piano, TD, USA), coking silica (CAB-O-SIL, Cabot, Boston, MD, USA), or mixtures thereof.

[0220] Exemplary anti-caking agents include, but are not limited to: calcium silicate, magnesium silicate, silica, colloidal silica, talc, or mixtures thereof.

[0221] Exemplary antimicrobial agents include, but are not limited to: benzalkonium chloride, benzyl chloride, benzoic acid, benzyl alcohol, butylparaben, hexadecylpyridine chloride, cresol, chlorobutanol, dehydroacetic acid, ethylparaben, methylparaben, phenol, phenethyl alcohol, phenoxyethanol, phenylmercuric acetate, phenylmercuric nitrate, potassium sorbate, propylparaben, sodium benzoate, sodium dehydroacetate, sodium propionate, sorbic acid, thimersol, thymo, or mixtures thereof.

[0222] Exemplary antioxidants include, but are not limited to: ascorbic acid, BHA, BHT, EDTA, or mixtures thereof.

[0223] Exemplary coating agents include, but are not limited to: sodium carboxymethyl cellulose, cellulose acetate phthalate, ethyl cellulose, gelatin, pharmaceutical glaze, hydroxypropyl cellulose, hydroxypropyl methyl cellulose (hydroxypropyl methyl cellulose), hydroxypropyl methyl cellulose phthalate, methyl cellulose, polyethylene glycol, polyvinyl acetate phthalate, shellac, sucrose, titanium dioxide, carnauba wax, microcrystalline wax, gellan gum, maltodextrin, methacrylates, microcrystalline cellulose, and carrageenan, or mixtures thereof.

[0224] The formulations described herein may also include other excipients and their categories, including but not limited to those described herein. Polosham (e.g.) And Poloxamer 188), ascorbic acid, glutathione, protease inhibitors (e.g., soybean trypsin inhibitor, organic acids), pH lowering agents, creams and lotions (like maltodextrin and carrageenan); materials for chewable tablets (like glucose, fructose, lactose monohydrate, lactose and aspartame, lactose and cellulose, maltodextrin, maltose, mannitol, microcrystalline cellulose and guar gum, sorbitol crystals); parenteral agents (like mannitol and povidone); plasticizers (like dibutyl sebate, plasticizers for coating, polyvinyl acetate phthalates); powder lubricants (like glyceryl behenate); for packaging Gel spheres (like sugar balls); rounding agents (like glyceryl behenate and microcrystalline cellulose); suspending / gelling agents (like carrageenan, gellan gum, mannitol, microcrystalline cellulose, povidone, sodium starch glycolate, xanthan gum); sweeteners (like aspartame, aspartame and lactose, glucose, fructose, honey, maltodextrin, maltose, mannitol, molasses, sorbitol crystals, sorbitol-specific solution, sucrose); wet granulation agents (like calcium carbonate, anhydrous lactose, lactose monohydrate, maltodextrin, mannitol, microcrystalline cellulose, povidone, starch), caramel, sodium carboxymethyl cellulose, cherry flavoring. Cream flavor and cherry flavor), anhydrous citric acid, citric acid, powdered sugar, D&C Red No. 33, D&C Yellow #10 Aluminum Lake, disodium EDTA, 15% ethanol, FD&C Yellow No. 6 Aluminum Lake, FD&C Blue #1 Aluminum Lake, FD&C Blue No. 1, FD&C Blue No. 2 Aluminum Lake, FD&C Green No. 3, FD&C Red No. 40, FD&C Yellow No. 6 Aluminum Lake, FD&C Yellow No. 6, FD&C Yellow No. 10, Glyceryl palmitate, Glyceryl monostearate, Indigo carmine, Lecithin, Mannitol, Methylparaben and Propylparaben, Monoammonium glycyrrhizate, Natural and artificial orange flavor, Medicinal glaze, Poloxamer 188, Polydextrose, Polysorbate 20, Polysorbate 80, Povidone, Pregelatinized corn starch, Pregelatinized starch, Red iron oxide, Sodium saccharin, Sodium carboxymethyl ether, Sodium chloride, Sodium citrate, Sodium phosphate, Strawberry flavor, Synthetic black iron oxide, Synthetic red iron oxide, Titanium dioxide, and White wax.

[0225] In some embodiments, the formulation of the present invention is a solid oral dosage form, which may optionally be used with, for example, [other ingredients]. Blue (OY-LS-20921) White (YS-2-7063) Coating systems for white (YS-1-7040) and black ink (S-1-8106) coatings (e.g.) The process involves using the fx film coating system.

[0226] As used herein, the term "therapeutic effective amount" refers to the amount of a pharmaceutical agent used to treat, improve, or prevent an identified disease or symptom, or to demonstrate a detectable therapeutic or inhibitory effect. This effect can be detected by any assay method known in the art. The precise effective amount used on a subject depends on the subject's weight, build, and health status; the nature and severity of the symptom; and the chosen therapeutic agent. The therapeutic effective amount for a given situation can be determined by routine testing within the scope of a clinician's technique and judgment. In one preferred aspect, the disease or symptom to be treated is cancer. In another aspect, the disease or symptom to be treated is a cell proliferation disorder.

[0227] The pharmaceutical compositions containing the active compound of the present invention can be manufactured in a manner generally known (e.g., by conventional methods such as mixing, dissolving, granulation, forming sugar-coated tablets, grinding, emulsifying, encapsulating, embedding, or lyophilizing). The pharmaceutical compositions can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers (including excipients and / or adjuvants) that facilitate the processing of the active compound into a pharmaceutically usable formulation. Of course, the appropriate formulation depends on the chosen route of administration.

[0228] Particularly advantageous is the formulation of oral compositions in dosage units to facilitate administration and dosage uniformity. As used herein, a unit dosage form refers to a physically discrete unit suitable as a single dose for a subject to be treated; each unit contains a predetermined amount of active compound bound to a desired drug carrier to produce the desired therapeutic effect. The specifications of the unit dosage forms of the present invention are indicated and directly depend on the unique characteristics of the active compound and the specific therapeutic effect to be achieved.

[0229] In therapeutic applications, among the factors influencing the selected dosage, the dosage of the pharmaceutical composition used according to the invention varies particularly based on the pharmaceutical agent, the age, weight, and clinical condition of the receiving patient, and the experience and judgment of the clinician or physician administering the treatment. Generally, the dosage should be sufficient to cause a slowing of tumor growth, and preferably regression, and even more preferably complete regression. An effective amount of a pharmaceutical agent is an amount that provides an objectively identifiable improvement as noted by a clinician or other qualified observer. For example, regression of a patient's tumor can be measured with reference to the diameter of the tumor. A decrease in tumor diameter indicates regression. The inability of the tumor to reappear after cessation of treatment also indicates regression. As used herein, the term "dosage-effective manner" refers to the amount of active compound that produces the desired biological effect in a subject or cell.

[0230] The pharmaceutical composition may be included in a container, package, or dispenser along with the instructions for use.

[0231] The compounds in the formulations of this invention can further form salts. All of these forms are also considered within the scope of the claimed invention.

[0232] As used herein, "pharmaceutically acceptable salt" refers to a derivative of the compounds of the present invention, wherein the parent compound is modified by preparing a salt of its acid or base. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral salts or organic acid salts of basic residues, such as amines, and basic or organic salts of acidic residues (such as carboxylic acids). Pharmaceutically acceptable salts include conventional non-toxic salts or, for example, quaternary ammonium salts of parent compounds formed from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include, but are not limited to, those derived from inorganic and organic acids selected from 2-acetoxybenzoic acid, 2-hydroxyethanesulfonic acid, acetic acid, ascorbic acid, benzenesulfonic acid, benzoic acid, bicarbonate, carbonic acid, citric acid, edemaic acid, ethanedisulfonic acid, 1,2-ethanesulfonic acid, fumaric acid, glucoheponic acid, gluconic acid, glutamic acid, glycolic acid, ethylene glycol arsanonic acid, hexylresorcinic acid, and hydrabamic acid. Hydrobromic acid, hydrochloric acid, hydroiodic acid, hydroxymaleic acid, hydroxynaphthenic acid, hydroxyethanesulfonic acid, lactic acid, lactobionic acid, lauryl sulfonic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, naphthalenesulfonic acid, nitric acid, oxalic acid, primordial acid, pantothenic acid, phenylacetic acid, phosphoric acid, polygalacturonic acid, propionic acid, salicylic acid, stearic acid, subacetic acid, succinic acid, aminosulfonic acid, p-aminobenzenesulfonic acid, sulfuric acid, tannic acid, tartaric acid, toluenesulfonic acid, and common amino acids such as glycine, alanine, phenylalanine, and arginine.

[0233] Other pharmaceutically acceptable examples of salts include hexanoic acid, cyclopentanepropionic acid, pyruvic acid, malonic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo-[2.2.2]-oct-2-en-1-carboxylic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, and kojic acid. The invention also covers salts formed when: an acidic proton present in the parent compound is replaced by a metal ion (e.g., an alkali metal ion, an alkaline earth metal ion, or an ammonium ion); or coordinated with an organic base (e.g., ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucosamine, etc.). In salt form, it should be understood that the ratio of the cation or anion of the compound to the salt can be 1:1, or any ratio other than 1:1, such as 3:1, 2:1, 1:2, or 1:3.

[0234] It should be understood that all references to pharmaceutically acceptable salts include the same salt in its solvation form (solvent) or crystalline form (polymorph) as defined herein.

[0235] These compounds, or their pharmaceutically acceptable salts, esters, or prodrugs, are administered orally, nasally, dermally, pulmonaryly, by inhalation, buccally, sublingually, intraperitoneally, subcutaneously, intramuscularly, intravenously, rectally, intrapleurally, intrathecally, or parenterally. In one embodiment, the compound is administered orally. Those skilled in the art will recognize the advantages of certain routes of administration.

[0236] The dosage regimen for these compounds is chosen based on a variety of factors, including the patient's type, species, age, weight, sex, and medical condition; the severity of the condition to be treated; the route of administration; the patient's renal and hepatic function; and the specific compound or its salt used. A physician or veterinarian with general skills can easily determine and prescribe the effective amount of the required medication to prevent, combat, or halt the progression of the condition.

[0237] The formulation and administration techniques of the compounds disclosed herein can be found in Remington: The Science and Practice of Pharmacy, 19th Edition, Mack Publishing, Easton, Pennsylvania (1995). In one embodiment, the compounds described herein and their pharmaceutically acceptable salts are used in combination with a pharmaceutically acceptable carrier or diluent in pharmaceutical formulation. Suitable pharmaceutically acceptable carriers include inert solid fillers or diluents and sterile water or organic solutions. The compounds will be present in such pharmaceutical compositions in an amount sufficient to provide the desired dose within the range described herein.

[0238] Unless otherwise stated, all percentages and ratios used herein are by weight. Other features and advantages of the invention will be apparent from various examples. The examples provided illustrate different components and methods useful in carrying out the invention. These examples do not limit the claimed invention. Based on this disclosure, those skilled in the art can identify and utilize other components and methods that can be used to carry out the invention.

[0239] Unless explicitly stated or obvious in the context, the term “or” as used herein should be understood as inclusive.

[0240] Unless the context otherwise clarifies, all numerical values ​​provided in this article are modified by the term “about”.

[0241] The list of abbreviations used in this disclosure and the accompanying drawings is provided below.

[0242] AE: Adverse Events

[0243] AUC: Area under the plasma concentration-time curve

[0244] AUC(0-x): Area under the plasma concentration-time curve from time zero to x hours after administration.

[0245] AUC(0-t): Area under the plasma concentration-time curve from time zero to the final quantifiable concentration.

[0246] AUC(0-inf): Area under the plasma concentration-time curve from zero to infinity.

[0247] ANCOVA: Analysis of Covariance

[0248] BID: Twice a day

[0249] CI: Confidence Interval

[0250] Cmax: Maximum drug concentration

[0251] Cx: Plasma concentration x hours after administration

[0252] CV: Coefficient of Variation

[0253] DLT: Dose-limiting toxicity

[0254] MTD: Maximum tolerated dose

[0255] PO: Oral administration

[0256] PD: Pharmacodynamics

[0257] PK: Pharmacokinetics

[0258] T1 / 2: Terminal elimination half-life

[0259] Tmax: Time to reach maximum (peak) concentration after drug administration

[0260] Example

[0261] Example 1: PK / PD Study

[0262] The first-in-human phase 1 study of EPZ-6438, a histone methyltransferase inhibitor of zeste enhancer homolog 2 (EZH2), as a single agent in patients with advanced solid tumors (“ST”) or B-cell lymphoma.

[0263] Phase 1 dose escalation will be conducted to determine the safety, PK, PD, and preliminary antitumor activity of the maximum tolerated dose (MTD) in patients with B-cell lymphoma or advanced solid tumors (“pt”).

[0264] Methods: EPZ-6438 was administered sequentially at PO BID up to the maximum feasible dose of 1600 mg BID to age-matched individuals at 3 to 6 points. Blood samples were collected for PK analysis and skin biopsies for PD analysis. PD samples were stained with an H3K27me3-specific antibody, and the percentage change in H3K27Me3-positive cells from baseline was determined. PK / PD relationships were analyzed. Tumor assessment was performed every 8 weeks.

[0265] As of August 14, 2014, 21 patients had been recruited and treated at five dose levels: 100, 200, 400, 800, and 1600 mg BID. Diagnoses of B-cell NHL included follicular lymphoma (FL, n=4), DLBCL (n=4, including one patient with primary mediastinal lymphoma [PMBL] and marginal zone lymphoma (“pt”) (n=1)). ST pts included one pt with MRT. Eleven pts (7 with ST and 4 with B-cell NHL) had at least one filed post-treatment evaluation and were evaluable for efficacy. The median age was 59 years (range 23–83 years). One case of thrombocytopenic DLT was reported at the 1600 mg BID. Frequent adverse events (AEs) unrelated to causation were asthenia (8 pts), anemia (4 pts), decreased appetite and diarrhea (3 pts each), pulmonary embolism, insomnia, muscle cramps, thrombocytopenia, nausea and vomiting (2 pts each). Five out of 21 pts experienced grade 3 / 4 AEs. EPZ-6438PK exhibited rapid absorption (Tmax = 1–2 hr), dose-related increase in exposure, and rapid elimination (half-life approximately 4 hr). Lower EPZ-6438 exposure with multiple dosings was associated with higher metabolite exposure. Exposure-related reductions were observed in H3K27Me3-positive cells of the skin. Partial response was shown in 2 out of 4 evaluable NHL pts (1 with converted DLBCL, 100 mg BID, and 1 with PMBL, 200 mg BID) and in 1 pt with INI1-deficient MRT (800 mg BID). Further age-group expansion is underway.

[0266] This study showed that EPZ-6438 remained well tolerated up to a BID of 1600 mg, with preliminary evidence of activity in NHL and MRT and exposure-related inhibition of H3K27Me3. MTD was not yet reached. These data support the continued progress of EPZ-6438 in B-cell lymphoma and INI1-deficient tumors in a phase 2 study.

[0267] Pharmacokinetics

[0268] In dose-escalation studies, the pharmacokinetics of EPZ-6438 and its major deethylated metabolite ER-897387 were characterized after single (day 1) and multiple (day 15) administrations to subjects with staged solid tumors or B-cell lymphomas. The administered doses were 100 mg BID as a suspension (n=3) or tablet (n=3) formulation, and 200, 400, 800, and 1600 mg as tablet formulations. On day 1, EPZ-6438 was rapidly absorbed, and peak plasma concentrations of EPZ-6438 were observed approximately 1 to 2 hours after administration (Table 1). Plasma concentrations decreased in a biexponential manner, and quantifiable levels of EPZ-6438 and its metabolite ER-897387 remained measurable until 12 hours after administration (Figure 1). The mean terminal half-life (t) of EPZ-6438... 1 / 2 The exposure time is approximately 3 to 6 hours. EPZ-6438 exposure is slightly above dose-proportion, highly variable (%CV = 32%-95%), and comparable between tablet and suspension formulations. On day 1, regarding C... max and AUC (0-12h) In terms of conversion, the ranges from approximately 39% to 104% for EPZ-6438 and from 58% to 156% for ER-897387.

[0269] After multiple administrations (day 15), the median time (t) to reach maximum plasma concentration was... max The half-life is 1 to 2 hours. The half-life across the dose range remains unchanged on day 15. 1 / 2 = Approximately 3-6 hours). Following multiple doses, EPZ-6438 exposure showed a dose-dependent decrease. The cumulative rate of EPZ-6438 (R = AUC) after administration of 100 mg and 1600 mg doses was [not specified]. D15 / AUC D1 The figures are approximately 74% and 44%, respectively.

[0270] The maximum concentration of ER-897387 was observed 0.5 to 2 hours after administration, and its elimination was parallel to that of EPZ-6438 (t). 1 / 2 =3-6 hours). After multiple administrations, the amount of metabolites formed increases ( Figure 2 The increase in ER-897387 exposure on day 15 was associated with the decrease in EPZ-6438 exposure, thus indicating metabolic induction.

[0271] Table 1. Mean (standard deviation) pharmacokinetic parameters of EPZ-6438 and ER-897387 after single-dose (day 1) twice-daily administration of EPZ-6438

[0272]

[0273] *n=5 for t 1 / 2

[0274] The statistics for #Tmax are represented as the range of median and minimum values ​​minus maximum value.

[0275] Table 2. Mean (standard deviation) pharmacokinetic parameters of EPZ-6438 and ER-897387 after multiple-dose (day 15) twice-daily administration of EPZ-6438.

[0276]

[0277]

[0278] Pharmacodynamics

[0279] There was no direct correlation between the maximum percentage change in tumor size and EPZ-6438 dose (Figure 3). The maximum changes in tumor size observed in the 100 mg and 200 mg dose groups were -78% and -87%, respectively. Higher EPZ-6438 exposure was not associated with greater tumor reduction.

[0280] Skin biopsies were collected prior to administration (day 1 of cycle 1) and day 1 of cycle 2 for immunohistochemical analysis. Skin samples were stained with an H3K27Me3-specific antibody, and the percentage change in H3K27Me3-positive cells in the epidermis from baseline was determined. During day 1 of cycle 2 of EPZ-6438 administration, a consistent decrease in the number of H3K27Me3-positive cells was observed, confirming inhibition of histone H3 lysine 27 trimethylation in the skin. Figure 4 This effect is dose-dependent. Figure 5 And it was not correlated with the maximum changes in the tumor. Figure 6 ).

[0281] Decreased histone H3 lysine 27 trimethylation is associated with EPZ-6438 exposure. Figure 7 An inhibitory EmaxPK / PD model was fitted to describe this relationship.

[0282]

[0283] The model is parameterized in terms of maximum inhibition (Imax) and exposure associated with 50% of maximum inhibition (IC50). The model estimates Imax = -44.4% and IC50 = 487 ng·hr / mL. The model predicts EPZ-6438 exposure associated with 90% of maximum inhibition (IC90) to be 4421 ng·hr / mL. This estimate is comparable to the EPZ-6438 exposure maintained after an 800 mg dose (mean day 15 AUC = 4553 ng·hr / mL).

[0284] The results show that:

[0285] EPZ-6438 is rapidly absorbed and eliminated (t1 / 2 approximately 3-6 hours).

[0286] EPZ-6438 exposure is greater than dose-proportional and highly variable (%CV = 32%-95%).

[0287] EPZ-6438 is extensively metabolized (AUC metabolite / parent = 58%-156%).

[0288] • The significant decrease in EPZ-6438 exposure with repeated dosing was associated with increased metabolite formation.

[0289] No direct correlation was observed between the maximum reduction in tumor size and EPZ-6438 or exposure. The maximum reduction in tumor size was observed at the lowest doses (100 and 200 mg).

[0290] • There is a correlation between the bioactivity in the skin and EPZ-6438 exposure.

[0291] The model predicts that at a sustained EPZ-6438 exposure at a dose of 800 mg, exposure leading to near-maximal inhibition (90%) will be observed.

[0292] Example 2: Film-coated tablets

[0293] Formulations of EPZ-6438 were prepared according to the methods disclosed herein. Table 3 below provides the components and dosages for 50 mg, 100 mg, and 200 mg strength tablets:

[0294] Table 3. Components and Composition of EPZ-6438 Film-Coated Tablets NF = National Formulary (United States), USP = United States Pharmacopeia, qs = Quantity.

[0295] a: Adjust the amount of EPZ-6438 drug according to the potency correction factor (derived value of the free form measurement).

[0296] b: Adjust the amount of lactose monohydrate compound based on the amount of EPZ-6438 drug to maintain the weight of the core tablet.

[0297] c: Removed during the drying process.

[0298] d: Adjust according to the yield of qualified particles.

[0299] Dissolution tests were conducted on tablets containing compound 1HBr with strengths of 50 mg, 100 mg, and 200 mg EPZ-6438, according to industry standards, under standard testing conditions. Figure 9 and Figure 10 Dissolution curves under different conditions are provided.

[0300] Preparation of formulations in Examples 31-5 and Comparative Examples 1-8

[0301] The components used for sample preparation are listed in Table 4 below.

[0302] Table 4 Component Table

[0303]

[0304]

[0305] Tables 5 and 6 below list the components of formulations 1-5 (referred to as “Ex.1” to “Ex.5” in Tables 5-6) and the components of comparative examples 1-8 (referred to as “C.Ex.1” to “C.Ex.8” in Tables 5-6), as well as the amount of each component in each formulation sample.

[0306] Table 5 (50 tablets)

[0307]

[0308]

[0309] a: As is % (as purity of free base), which is 86.3%. This is equivalent to 200 mg of EPZ-6438 free base.

[0310] Table 6 (12.5 tablets)

[0311]

[0312]

[0313] a: As is % (as purity of free base), which is 86.3%. This is equivalent to 200 mg of EPZ-6438 free base.

[0314] Preparation No. 1 was prepared according to the following method. Other samples were prepared using a method similar to that used for Preparation No. 1, except for differences in wet granulation scale and / or the different components and their amounts. The wet granulation method shown in Table 5 was performed for samples in a 50-tablet scale. The granulation method shown in Table 6 was performed for samples in a 12.5-tablet scale. The weighed amounts of components were calculated based on the wet granulation scale and the preparations shown in Tables 5 and 6.

[0315] Preparation of Formulation No. 1

[0316] 11.59 g of EPZ-6438 drug, 3.23 g of lactose monohydrate, 0.80 g of hydroxypropyl cellulose, and 2.00 g of sodium starch glycolate were mixed using a mortar and pestle. The mixture was wet-granulated using a mortar and pestle while gradually adding an appropriate amount of purified water. The wet granules were dried using a constant-temperature oven set at 70°C. The dried granules were then passed through a sieve with a 710 μm opening. 40.0 mg of sodium starch glycolate and 7.6 mg of magnesium stearate (per 352.4 mg of sieved granules) were added to the sieved granules, and the components were lubricated by shaking in a glass vial. The lubricated granules, equivalent to one tablet, were compressed at 1450 kgf using a single-punch tablet press equipped with a 10.0 mm diameter punch, yielding tablets containing 200 mg of EPZ-6438 drug as a free base.

[0317] Example 4 Dissolution Test

[0318] According to the 37th edition of the USP <711> Dissolution was performed using Instrument 2 (paddle method). A 0.1 mol / L HCl solution and a pH 4.5 50 mmol / L acetate buffer were selected as the dissolution medium and prepared according to USP, version 37. Dissolution test conditions are summarized in Table 7. Samples were periodically collected from the container at predetermined times after the start of the dissolution test and filtered through a UHE-1400 filter with approximately 20 μm openings. Standard solutions were prepared at concentrations close to 100% dissolution by dissolving EPZ-6438 in the dissolution medium. The absorbance of the filtered samples and standard solutions was measured using a spectrophotometer, and the dissolution rate was calculated based on the absorbance and concentration of the standard solutions.

[0319] Table 7

[0320]

[0321] Dissolution test results

[0322] Dissolution tests were performed on each sample using two containers, and the average dissolution rates are shown in Tables 8-9 and 9-10. Figure 11-14 The leaching test results in 0.1 mol / L HCl solution are shown in Table 8 and... Figure 11-12 The dissolution test results in pH 4.5 acetate buffer are shown in Table 9 and 1. Figure 13-14 middle.

[0323] Table 8

[0324]

[0325] Table 9

[0326]

[0327] Preparations 1-5 dissolved rapidly in 0.1 mol / L HCl solution, with an average dissolution rate exceeding 80% at 30 minutes. Furthermore, preparations 1-4 achieved an average dissolution rate of 40% or higher at pH 4.5 acetate buffer at 30 minutes. In particular, preparations 1 and 5 dissolved 80% or more at pH 4.5 acetate buffer at 30 minutes.

[0328] All publications and patent documents cited herein are incorporated herein by reference, as each such publication or document is specifically and individually indicated to be incorporated herein by reference. The citation of publications and patent documents is not intended to acknowledge any publication or patent document as applicable prior art, nor does it constitute any admission of its content or date. The invention has now been described in writing, and those skilled in the art will recognize that the invention can be practiced in various embodiments, and that the foregoing description and the examples below are for illustrative purposes and do not limit the subsequent claims.

[0329] The invention may be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the foregoing embodiments should be considered illustrative rather than limiting of the invention described herein in all respects. Thus, the scope of the invention is indicated by the appended claims rather than the foregoing description, and all variations intended to fall within the meaning and equivalence of the claims are included therein.

Claims

1. Use of a solid pharmaceutical formulation comprising: N-((4,6-dimethyl-2-oxo-l,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4- yl)amino)-4-methyl-4'-(morpholinomethyl)-[l,l'-biphenyl]-3-carboxamide: at a content of 50 wt% ± 2.5 wt%; or a salt of Compound 1 at a content equivalent to 50 wt% ± 2.5 wt% of Compound 1, or a combination of Compound 1 and a salt of Compound 1, the combination being present at a content equivalent to 50 wt% ± 2.5 wt% of Compound 1; lactose monohydrate at a content of 16 wt% ± 0.8 wt%; a disintegrant at a content of 19 wt% ± 0.95 wt%, the disintegrant comprising low-substituted hydroxypropylcellulose and sodium starch glycolate; hydroxypropylcellulose at a content of 4 wt% ± 0.2 wt%; and magnesium stearate at a content of 2 wt% ± 0.2 wt%; in the manufacture of a medicament for the treatment of follicular lymphoma (FL), diffuse large B-cell lymphoma (DLBCL), primary mediastinal large B-cell lymphoma (PMBCL), or marginal zone lymphoma (MZL).

2. The use of claim 1, wherein the solid pharmaceutical formulation comprises a hydrobromide salt of Compound 1 at a content equivalent to 50 wt% ± 2.5 wt% of Compound 1; and a coating composition at a content of 3 wt% ± 0.06 wt%. ("Compound 1"), 3. Use of a solid pharmaceutical formulation comprising: N-((4,6-dimethyl-2-oxo-l,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4- yl)amino)-4-methyl-4'-(morpholinomethyl)-[l,l'-biphenyl]-3-carboxamide: at a content of 50 wt% ± 2.5 wt%; or a salt of Compound 1 at a content equivalent to 50 wt% ± 2.5 wt% of Compound 1, or a combination of Compound 1 and a salt of Compound 1, the combination being present at a content equivalent to 50 wt% ± 2.5 wt% of Compound 1; lactose monohydrate at a content of 16 wt% ± 0.8 wt%; a disintegrant at a content of 19 wt% ± 0.95 wt%, the disintegrant comprising low-substituted hydroxypropylcellulose and sodium starch glycolate; hydroxypropylcellulose at a content of 4 wt% ± 0.2 wt%; and magnesium stearate at a content of 2 wt% ± 0.2 wt%; in the manufacture of a medicament for the treatment of follicular lymphoma (FL), diffuse large B-cell lymphoma (DLBCL), primary mediastinal large B-cell lymphoma (PMBCL), or marginal zone lymphoma (MZL).

4. The use of claim 3, wherein the solid pharmaceutical formulation comprises: a hydrobromide salt of Compound 1 at a content of 55 wt% ± 0.55 wt%; lactose monohydrate at a content of 16 wt% ± 0.8 wt%; a disintegrant at a content of 19 wt% ± 0.95 wt%, the disintegrant comprising low-substituted hydroxypropylcellulose and sodium starch glycolate; hydroxypropylcellulose at a content of 4 wt% ± 0.2 wt%; magnesium stearate at a content of 2 wt% ± 0.2 wt%; and a coating composition at a content of 3 wt% ± 0.06 wt%.

5. Use of a solid pharmaceutical formulation comprising: N-((4,6-dimethyl-2-oxo-l,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4- yl)amino)-4-methyl-4'-(morpholinomethyl)-[l,l'-biphenyl]-3-carboxamide: at a content of 50 wt% ± 2.5 wt%; or a salt of Compound 1 at a content equivalent to 50 wt% ± 2.5 wt% of Compound 1, or a combination of Compound 1 and a salt of Compound 1, the combination being present at a content equivalent to 50 wt% ± 2.5 wt% of Compound 1; lactose monohydrate at a content of 16 wt% ± 0.8 wt%; a disintegrant at a content of 19 wt% ± 0.95 wt%, the disintegrant comprising low-substituted hydroxypropylcellulose and sodium starch glycolate; hydroxypropylcellulose at a content of 4 wt% ± 0.2 wt%; and magnesium stearate at a content of 2 wt% ± 0.2 wt%; in the manufacture of a medicament for the treatment of follicular lymphoma (FL), diffuse large B-cell lymphoma (DLBCL), primary mediastinal large B-cell lymphoma (PMBCL), or marginal zone lymphoma (MZL). ​ ​ ​ ​ ​ ​ ("Compound 1"), wherein ​ ​ ​ ​ ​ ​ ​ ​ N-((4,6-dimethyl-2-oxo-l,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4- yl)amino)-4-methyl-4'-(morpholinomethyl)-[l,l'-biphenyl]-3-carboxamide: ("Compound 1"), wherein The solid pharmaceutical formulation comprises: hydrobromide salt of Compound 1 at a content of 50 wt% ± 2.5 wt%; lactose monohydrate; low-substituted hydroxypropylcellulose at a content of 15 wt% ± 0.75 wt%; sodium starch glycolate at a content of 5 wt% ± 0.5 wt%; hydroxypropylcellulose at a content of 4 wt% ± 0.2 wt%; magnesium stearate at a content of 2 wt% ± 0.2 wt%; and coating composition at a content of 3 wt% ± 0.06 wt%.

5. Use of a solid pharmaceutical formulation comprising: N-((4,6-dimethyl-2-oxo-l,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4- yl)amino)-4-methyl-4'-(morpholinomethyl)-[l,l'-biphenyl]-3-carboxamide: ("Compound 1"), wherein The solid pharmaceutical formulation comprises: hydrobromide salt of Compound 1 at a content of 40-60 wt%; lactose monohydrate; low-substituted hydroxypropylcellulose at a content of 11-19 wt%; sodium starch glycolate at a content of 3-7 wt%; hydroxypropylcellulose at a content of 1-10 wt%; magnesium stearate at a content of 0.5-5 wt%; and coating composition at a content of 1-10 wt%.

6. The use of claim 4 or 5, wherein the solid pharmaceutical formulation comprises lactose monohydrate at a content of 20 wt% ± 2 wt%.

7. Use of a solid pharmaceutical formulation consisting of: N-((4,6-dimethyl-2-oxo-l,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4- yl)amino)-4-methyl-4'-(morpholinomethyl)-[l,l'-biphenyl]-3-carboxamide at a content of 40-60 wt%: ("Compound 1"), or a salt of Compound 1 at a content equivalent to 40-60 wt% of Compound 1, or a combination of Compound 1 and a salt of Compound 1, the combination being present at a content equivalent to 40-60 wt% of Compound 1; lactose monohydrate; low-substituted hydroxypropylcellulose at a content of 11-19 wt%; sodium starch glycolate at a content of 3-7 wt%; hydroxypropylcellulose at a content of 1-10 wt%; magnesium stearate in an amount of 2 wt% ± 0.1 wt%; and a coating composition in an amount of 3 wt% ± 0.06 wt%.

8. The use of claim 4, 5, or 7, wherein the lactose monohydrate is in an amount of 10-20 wt%.

9. The use of any one of claims 2-5 and 7, wherein the coating composition comprises hypromellose, talc, polyethylene glycol, titanium dioxide, and iron (III) oxide.

10. The use of claim 7, wherein the pharmaceutical formulation comprises a salt of Compound 1 in an amount equivalent to 50 wt% ± 2.5 wt% of Compound 1.

11. The use of claim 10, wherein the salt of Compound 1 is a hydrobromide salt of Compound 1.

12. The use of claim 1, wherein the pharmaceutical formulation comprises a combination of Compound 1 and a salt of Compound 1, the combination being present in an amount equivalent to 47-50 wt% of Compound 1.

13. The use of claim 12, wherein the salt of Compound 1 is a hydrobromide salt of Compound 1.

14. Use of a solid pharmaceutical formulation in the manufacture of a medicament for the treatment of follicular lymphoma (FL), diffuse large B-cell lymphoma (DLBCL), primary mediastinal large B-cell lymphoma (PMBCL), or marginal zone lymphoma (MZL), the solid pharmaceutical formulation comprising: N-((4,6-dimethyl-2-oxo-l,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4- yl)amino)-4-methyl-4'- (morpholinomethyl)-[l,l'-biphenyl]-3-carboxamide: ("Compound 1"), wherein the solid pharmaceutical formulation comprising: a hydrobromide salt of Compound 1 in an amount of 55 wt% ± 0.55 wt%; lactose monohydrate in an amount of 16 wt% ± 0.8 wt%; low-substituted hydroxypropylcellulose in an amount of 15 wt% ± 0.75 wt%; sodium starch glycolate in an amount of 5 wt% ± 0.5 wt%; hydroxypropylcellulose in an amount of 4 wt% ± 0.2 wt%; magnesium stearate in an amount of 2 wt% ± 0.1 wt%; and a coating composition in an amount of 3 wt% ± 0.06 wt%.

15. The use of claim 14, wherein the coating composition comprises hypromellose, talc, polyethylene glycol, titanium dioxide, and iron (III) oxide.

16. The use of any one of claims 1-5, 7, and 14, wherein the medicament is for the treatment of follicular lymphoma (FL).

17. The use of any one of claims 1-5, 7, and 14, wherein the medicament is for the treatment of diffuse large B-cell lymphoma (DLBCL).

18. The use of any one of claims 1-5, 7, and 14, wherein the medicament is for the treatment of primary mediastinal large B-cell lymphoma (PMBCL).

19. The use of any one of claims 1-5, 7, and 14, wherein the medicament is for the treatment of marginal zone lymphoma (MZL).

20. The use of any one of claims 1-5, 7, and 14, wherein the solid pharmaceutical formulation is in the form of a tablet.

21. The use of claim 20, wherein the tablet consists of an inner phase and an outer phase.

22. The use of claim 21, wherein the tablet comprises Compound 1 equivalent to 100 mg, 200 mg, or 400 mg.

23. The use of any one of claims 1-5, 7, and 14, wherein the solid pharmaceutical formulation is made by a process comprising: a) mixing a therapeutic agent, a diluent, a disintegrant, and / or a lubricant to form a first mixture, wherein the therapeutic agent is a salt thereof, or a combination thereof, and optionally one or more steps selected from the group consisting of: b) adding to the first mixture an aqueous, organic solvent-based, or organic / aqueous mixture solution comprising a binder to form a second mixture; c) granulating the second mixture to form wet granules; d) drying the wet granules to form dried granules; e) size sieving the dried granules to obtain size-qualified granules; f) mixing the size-qualified granules with a glidant and a second disintegrant to form a third mixture; g) compressing the third mixture to form tablets; and h) applying a coating suspension to the tablets to produce film-coated tablets.

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