Therapy Comprising an Anti-CD19 Antibody and an EZH2 Modulator

Abstract

The present disclosure provides methods, pharmaceutical compositions, and kits for the treatment of cancer in a patient in need thereof, the methods comprising administering to the patient in need thereof an EZH2 modulator, such as Compound I, or a pharmaceutically acceptable salt thereof in combination with one or more anti-CD19 antibodies. Also provided is a medicament for use in treating cancer.

Description

[Technical Field]

[0001] The present disclosure relates to combinations comprising an anti-CD19 antibody and an EZH2 modulator for use in treating various cancers. [Background technology]

[0002] Non-Hodgkin's lymphoma (NHL) is one of the most common cancers in the United States and Europe, with over 70,000 and 93,000 new cases diagnosed annually, respectively. (Siegel RL, et al., CA Cancer J. Clin. 68(1):7-30 (2018); Ferlay J., et al., Eur. J. Cancer 103:356-87 (2018)) NHL is a heterogeneous group of malignancies with diverse clinical features that are optimally managed through a variety of different treatment modalities. NHL ranges from more indolent variants, such as follicular and marginal zone lymphoma, to more aggressive subtypes, such as diffuse large B-cell lymphoma (DLBCL) and Burkitt lymphoma (BL). While systemic chemotherapy is the mainstay of treatment for most NHL variants, anti-tumor-directed monoclonal antibodies play an important role in the treatment of this disease. Oflazoglu E., et al., MAbs 2(1):14-9(2010). Monoclonal antibodies, such as rituximab, that target the B cell antigen CD20 are part of the standard treatment regimen for many B-cell NHLs. Keating GM, Drugs 70(11):1445-76(2010). However, when NHL becomes refractory to standard chemotherapy and antibody-based treatments, the overall prognosis is poor and long-term survival is limited. Therefore, novel and effective therapies are needed to address this particular unmet medical need.

[0003] Indolent NHL (iNHL) accounts for 40% of all NHL subtypes, with follicular lymphoma occurring most frequently. Harris NL, et al., Ann. Oncol. 10(12):1419-32 (1999). iNHL exhibits a wide range of disease characteristics. Patients often experience a chronic relapsing and remitting disease course, are exposed to several successive treatment regimens, and ultimately die from disease progression. Generally, treatment is reserved for patients who develop severe symptoms or are at sufficiently high risk to merit early treatment. Gribben JG, Blood 109(11):4617-26 (2007).

[0004] Patients with iNHL who initially responded to single-agent rituximab (complete response or partial response (PR) with a time to tumor growth inhibition of at least 6 months) and subsequently experienced relapse are frequently re-treated with rituximab alone or in combination with chemotherapy. Gribben JG, Blood 109(11):4617-26(2007); Kahl BS, et al., J. Clin. Oncol. 32(28):3096-102(2014); NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines): B-Cell Lymphomas (Version 3.2019), National Comprehensive Cancer Network, May 6, 2019. Patients who become refractory to rituximab alone or in combination with chemotherapy have few effective treatment options.

[0005] Aggressive non-Hodgkin lymphoma (aNHL) accounts for approximately 30-40% of all NHL cases (Project TN-HsLC, Blood 89(11):3909-18(1997)), and DLBCL is the most common histological subtype. Beham-Schmid C., Aggressive lymphoma 2016: revision of the WHO classification, Memo 10(4):248-54(2017). Combination chemotherapy with the addition of rituximab is the standard treatment for newly diagnosed DLBCL patients. However, approximately 40% of DLBCL patients relapse after initial immunochemotherapy. Vaidya R., et al., Ann. Oncol. 25(11):2124-33(2014). For eligible patients, a salvage chemotherapy regimen followed by autologous stem cell transplantation is the standard treatment. However, many patients are ineligible for transplant due to age and other medical comorbidities. Although various salvage regimens, including combination chemotherapy, are available for relapsed / refractory disease, there is currently no standard salvage regimen. There is an unmet need for NHL patients who have early relapse or who do not respond to anti-CD20 treatment regimens.

[0006] CD19 is a 95-kDa transmembrane glycoprotein of the immunoglobulin superfamily containing two extracellular immunoglobulin-like domains and an extensive cytoplasmic tail. This protein is a pan-B lymphocyte surface receptor and is ubiquitously expressed from early stages of pre-B cell development until it is downregulated during terminal differentiation into plasma cells. It is specific to the B lymphocyte lineage and is not expressed on hematopoietic stem cells or other immune cells, except for some follicular dendritic cells. CD19 functions as a positive regulator of B cell receptor (BCR) signaling and is important for B cell activation and proliferation and the development of humoral immune responses. It acts as a costimulatory molecule, in conjunction with CD21 and CD81, and is important for B cell responses to T cell-dependent antigens. The cytoplasmic tail of CD19 physically associates with a family of tyrosine kinases that trigger downstream signaling pathways via the Src family of protein tyrosine kinases. CD19 is an attractive target for cancers of lymphoid origin because it is highly expressed in nearly all chronic lymphocytic leukemias (CLL) and non-Hodgkin's lymphomas (NHL), as well as many other different types of leukemia, including acute lymphocytic leukemia (ALL) and hairy cell leukemia (HCL).

[0007] Tafasitamab (formerly MOR208 and XmAb® 5574) is a humanized monoclonal antibody targeting the antigen CD19, a transmembrane protein involved in B-cell receptor signaling. Tafasitamab has been engineered in the Fc region of IgG to enhance antibody-dependent cell-mediated cytotoxicity (ADCC), thereby improving a key mechanism for tumor cell killing and potentially resulting in enhanced efficacy compared to conventional, i.e., non-enhanced, antibodies. Tafasitamab has been or is currently being tested in several clinical trials, including for CLL, ALL, and NHL. In July 2020, tafasitamab received accelerated approval from the U.S. Food and Drug Administration (FDA) in combination with lenalidomide to treat adults with relapsed / relapsed DLBCL. In August 2021, the European Commission granted a conditional marketing authorization for tafasitamab in combination with lenalidomide, followed by tafasitamab monotherapy, for the treatment of adult patients with relapsed or refractory diffuse large B-cell lymphoma (DLBCL) who are ineligible for autologous stem cell transplantation (ASCT). Despite the recent discovery and development of several anti-cancer drugs, the poor prognosis of many types of cancer, including CD19-expressing tumors, means that improved methods or therapeutic approaches to treat such types of cancer remain needed.

[0008] In order to extend the lifespan of patients while maintaining a high quality of life, new combinations of therapeutic agents that have beneficial effects in cancer treatment are desired. New combinations can enhance the benefits compared to each of the agents alone. In particular, combination treatment regimens may be useful for patients with conditions, including proliferative diseases, and can potentially reduce the recurrence rate or even overcome the resistance to certain anticancer drugs that is sometimes seen in these patients. This is especially true when cancer may be resistant or refractory to currently available treatment regimens.

[0009] Therefore, new cancer treatment regimens, including combination therapies, are needed. The present inventors have discovered that the combined administration of a CD19-specific antibody or antibody fragment and an EZH2 modulator has excellent effects on the treatment of malignant lymphomas of B cell origin. Summary of the Invention

[0010] The present disclosure provides a novel combination comprising an anti-CD19 antibody and an EZH2 modulator for use in the treatment of cancer.

[0011] In one aspect, the present disclosure relates to a method of treating cancer comprising administering a combination of an EZH2 modulator and an anti-CD19 antibody to a human subject in need of cancer treatment.

[0012] In one aspect, the present disclosure relates to a method of treating cancer comprising administering to a patient in need thereof a combination of an EZH2 inhibitor and an anti-CD19 antibody or antibody fragment.

[0013] In some embodiments, the anti-CD19 antibody comprises a heavy chain variable region comprising an HCDR1 region comprising the sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising the sequence NPYNDG (SEQ ID NO: 2), and an HCDR3 region comprising the sequence GTYYYGTRVFDY (SEQ ID NO: 3), and a light chain variable region comprising an LCDR1 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising the sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6).

[0014] In some embodiments, the anti-CD19 antibody comprises a heavy chain variable region comprising an HCDR1 region of SYVMH (SEQ ID NO: 1), an HCDR2 region of NPYNDG (SEQ ID NO: 2), and an HCDR3 region of GTYYYGTRVFDY (SEQ ID NO: 3), and a light chain variable region comprising an LCDR1 region of RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region of RMSNLNS (SEQ ID NO: 5), and an LCDR3 region of MQHLEYPIT (SEQ ID NO: 6).

[0015] In some embodiments, the anti-CD19 antibody comprises a heavy chain variable region of EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSS (SEQ ID NO: 7), and a light chain variable region of DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIK (SEQ ID NO: 8).

[0016] In some embodiments, the anti-CD19 antibody has effector function. In another aspect, the CD19-specific antibody or antibody fragment has enhanced effector function. In one embodiment, the effector function is ADCC. In one embodiment, the CD19-specific antibody or antibody fragment has enhanced ADCC activity. In a further embodiment, the CD19-specific antibody or antibody fragment comprises an Fc domain comprising an amino acid substitution at positions S239 and / or I332, the numbering being according to the EU index in Kabat. In a further embodiment, the CD19-specific antibody or antibody fragment comprises an Fc domain comprising an amino acid substitution of S239D and an amino acid substitution of I332E, the numbering being according to the EU index in Kabat.

[0017] In some embodiments, the anti-CD19 antibody comprises the heavy chain constant region of ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 9).

[0018] In some embodiments, the anti-CD19 antibody comprises the light chain constant region of RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10).

[0019] In some embodiments, the anti-CD19 antibody is ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQ It comprises a heavy chain constant region of PREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 9), and a light chain constant region of RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10).

[0020] In some embodiments, the anti-CD19 antibody is EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTI It comprises a heavy chain region of SKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 11), and a light chain region of DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12).

[0021] In some embodiments, the anti-CD19 antibody is tafasitamab.

[0022] In some embodiments, the EZH2 modulator is administered orally.

[0023] In some embodiments, the EZH2 modulator is administered intravenously. In some embodiments, the EZH2 modulator is administered subcutaneously.

[0024] In some embodiments, the EZH2 modulator is administered by intravenous infusion.

[0025] In some embodiments, the EZH2 modulator is an EZH2 inhibitor.

[0026] In some embodiments, the anti-CD19 antibody is administered intravenously.

[0027] In some embodiments, the anti-CD19 antibody is administered by intravenous infusion.

[0028] In some embodiments, the anti-CD19 antibody is administered by subcutaneous injection.

[0029] In some embodiments, the anti-CD19 antibody is administered subcutaneously.

[0030] In some embodiments, the cancer is a CD19-positive cancer.

[0031] In some embodiments, the cancer is a hematological malignancy.

[0032] In some embodiments, the cancer is lymphoma or leukemia.

[0033] In some embodiments, the cancer is chronic lymphocytic leukemia or non-Hodgkin's lymphoma.

[0034] In some embodiments, the cancer is CD19-positive chronic lymphocytic leukemia or CD19-positive non-Hodgkin's lymphoma.

[0035] In some embodiments, the cancer is non-Hodgkin's lymphoma.In some embodiments, the human subject suffers from relapsed or refractory non-Hodgkin's lymphoma.In some embodiments, the cancer is follicular lymphoma (FL), marginal zone lymphoma (MZL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL) or Burkitt's lymphoma.

[0036] In some embodiments, the EZH2 modulator is administered once every two weeks, once a week, twice a week, three times a week, or daily.

[0037] In some embodiments, the EZH2 modulator is administered twice weekly.

[0038] In some embodiments, the EZH2 modulator is administered once a week.

[0039] In some embodiments, the EZH2 modulator is administered on days 1, 4, 8, and 11 of a 21-day cycle.

[0040] In some embodiments, the anti-CD19 antibody is administered once every two weeks, once a week, twice a week, three times a week, or daily.

[0041] In some embodiments, the anti-CD19 antibody is administered once every two weeks.

[0042] In some embodiments, the anti-CD19 antibody is administered once a week.

[0043] In some embodiments, the anti-CD19 antibody is administered once every four weeks.

[0044] In some embodiments, the anti-CD19 antibody is administered once every eight weeks.

[0045] In some embodiments, the anti-CD19 antibody is administered on day 1 of a treatment cycle.

[0046] In some embodiments, the treatment cycle is 21 or 28 days.

[0047] In some embodiments, the EZH2 modulator and the anti-CD19 antibody are administered simultaneously once every 8 weeks, once every 4 weeks, once every 2 weeks, once a week, twice a week, three times a week, or daily.

[0048] In some embodiments, the EZH2 modulator and the anti-CD19 antibody are administered simultaneously on days 1, 4, 8, and 11 of a 21-day cycle.

[0049] In some embodiments, the EZH2 modulator is administered once every two weeks, once a week, twice a week, three times a week, daily, or on days 1, 4, 8, and 11 of a 21 day cycle, and the anti-CD19 antibody is separately administered once every eight weeks, once every four weeks, once every two weeks, once a week, twice a week, three times a week, or daily.

[0050] In one aspect, the present disclosure relates to a kit containing agents for use in treating cancer in a subject in need of such treatment. The kit includes an agent comprising an EZH2 modulator and instructions for administering the EZH2 modulator and one or more anti-CD19 antibodies or antibody fragments, or the kit includes an agent comprising the one or more anti-CD19 antibodies or antibody fragments and instructions for administering the one or more anti-CD19 antibodies or antibody fragments and an EZH2 modulator. The kit may include both an agent comprising an EZH2 modulator and an agent comprising one or more anti-CD19 antibodies, and instructions for administering the EZH2 modulator and the one or more anti-CD19 antibodies. The kit may also include one or more additional therapeutic agents.

[0051] In one aspect, the present disclosure relates to a medicament for use in treating cancer in a subject in need of such treatment. The medicament comprises an EZH2 modulator and one or more anti-CD19 antibodies or antibody fragments. The medicament may also include one or more additional therapeutic agents. [Brief explanation of the drawings]

[0052] [Figure 1] Viability assay. Effect of tafasitamab and compound 1 alone and in combination on viable cell counts in different lymphoma cell lines, as measured by CTG assay. [Figure 2] Estimation of combined effects using a linear mixed-effects model. Error bars indicate 95% confidence intervals. [Figure 3] Survival assay. Dose escalation curves of compound 1 and tazemetostat in the absence and presence of 0.1 nM tafasitamab in SU-DHL-4 (A) and SU-DHL-6 (B) cell lines. [Figure 4] Upregulation of cell surface CD19 levels by Compound 1 after 7 days of treatment. [Figure 5] Tumor growth kinetics (mean + / - SEM) after administration of different concentrations of Compound 1 and / or tafasitamab. [Figure 6] Mean tumor volumes for each treatment group on day 38 after tumor implantation (day 21 after treatment initiation). DETAILED DESCRIPTION OF THE INVENTION

[0053] definition To facilitate understanding of this disclosure, certain abbreviations, terms, and phrases are defined below.

[0054] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications cited herein are incorporated by reference in their entirety.

[0055] As used herein, the term "cancer" refers to a cellular disorder characterized by uncontrolled or disregulated cell proliferation, reduced cell differentiation, the ability to inappropriately invade surrounding tissues, and / or the ability to establish neoplasia in ectopic locations. The term "cancer" includes solid tumors and non-solid tumors, such as hematologic tumors. The term "cancer" encompasses diseases of the skin, tissues, organs, bone, cartilage, blood, and blood vessels. The term "cancer" further encompasses primary and metastatic cancers.

[0056] The term "CD19" refers to the protein known as CD19, and has the following synonyms: B4, B lymphocyte antigen CD19, B lymphocyte surface antigen B4, CVID3, differentiation antigen CD19, MGC12802, and T cell surface antigen Leu-12. The term also encompasses naturally occurring variants of CD19, such as splice variants, allelic variants, and isoforms. In embodiments, human CD19 has the amino acid sequence (SEQ ID NO: 13).

[0057] "MOR208" and "XmAb 5574" and "tafasitamab" are used synonymously for the anti-CD19 antibodies according to Table 1. Table 1 provides the amino acid sequence of MOR208 / tafasitamab. The MOR208 antibody is described in U.S. Pat. No. 8,524,867, which is incorporated by reference in its entirety.

[0058] The term "antibody" refers to an immunoglobulin molecule that recognizes and specifically binds to a target, such as a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or a combination of the above, via at least one antigen recognition site within the variable region of the immunoglobulin molecule. As used herein, the term "antibody" encompasses polyclonal antibodies, monoclonal antibodies, antibody fragments (e.g., Fab, Fab', F(ab')2, and Fv fragments), single-chain Fv (scFv) variants, multispecific antibodies, e.g., bispecific antibodies, e.g., those generated from at least two intact antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins containing an antigenic determinant of an antibody, and any other modified immunoglobulin molecule containing an antigen recognition site, so long as the antibody exhibits the desired biological activity. The antibody may be of any isotype (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass. Both the light and heavy chains are divided into regions of structural and functional homology. Different classes of immunoglobulins have different and well-known subunit structures and three-dimensional configurations. Antibodies can be naked or conjugated to other molecules, such as toxins, radioisotopes, etc.

[0059] The term "anti-CD19 antibody" or "antibody that binds to CD19" refers to an antibody that is capable of binding to CD19 with sufficient affinity that the antibody is useful as a diagnostic and / or therapeutic agent in targeting CD19.

[0060] A "monoclonal antibody" refers to a homogeneous or substantially homogeneous antibody population involved in highly specific recognition and binding of a single antigenic determinant, or epitope. This is in contrast to polyclonal antibodies, which typically contain different antibodies directed against different antigenic determinants. The term "monoclonal antibody" encompasses both intact and full-length monoclonal antibodies, as well as antibody fragments (e.g., Fab, Fab', F(ab')2, Fv), single-chain (scFv) variants, fusion proteins containing an antibody portion, and any other modified immunoglobulin molecule containing an antigen recognition site. Furthermore, "monoclonal antibody" refers to antibodies made in a variety of ways, including, but not limited to, by hybridoma, phage selection, recombinant expression, and transgenic animals.

[0061] The term "chimeric antibody" refers to an antibody in which the amino acid sequences of the immunoglobulin molecule are derived from more than one species. Typically, the variable regions of both the light and heavy chains correspond to the variable regions of antibodies from one species of mammal (e.g., mouse, rat, rabbit, etc.) having the desired specificity, affinity, and function, while the constant regions are homologous to sequences of antibodies from another species (usually human) to avoid eliciting an immune response in that species.

[0062] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a compound, or a combination of one or more compounds, that, when administered (sequentially or simultaneously), elicits a desired biological or pharmaceutical response, e.g., destroys, slows or stops the growth of, or slows or stops the progression of cancer in a patient. A therapeutically effective amount may vary depending on the intended use (e.g., in vitro or in vivo) or the patient and condition being treated, and may depend on factors such as the patient's weight and age, the severity of the condition, and the method of administration, and can be readily determined by one of ordinary skill in the art. The term "effective amount" or "therapeutically effective amount" also applies to an amount, such as one or more doses, that induces a specific response in a target cell, e.g., reduced platelet adhesion and / or cell migration. For example, in some embodiments, a "therapeutically effective amount" as used herein refers to an amount of an EZH2 modulator and an amount of an anti-CD19 antibody that exert a beneficial effect when administered separately or in combination. In some embodiments, the combined effect is additive. In some embodiments, the combined effect is synergistic. Furthermore, one skilled in the art will recognize that in combination therapy, the amount of EZH2 modulator and / or the amount of the anti-CD19 antibody may be used in a "sub-therapeutic amount," i.e., less than the therapeutically effective amount of the EZH2 modulator or anti-CD19 antibody alone.

[0063] In any form or composition, the administered dose(s) or therapeutically effective (total) amount may be: (i) BSA, e.g., mg / m 2 or (ii) the amount, e.g., as mg, of therapeutic agent(s) per patient.

[0064] The term "about" refers to approximately, in the region of, roughly, or approximately. When the term "about" is used in conjunction with a numerical value or numerical range, it means that the stated numerical value or numerical range is an approximation within experimental variability (or within statistical experimental error), and thus the numerical value or numerical range may vary, for example, by 1% to 15% of the stated numerical value or numerical range. Generally, the term "about" is used herein to modify numerical values ​​above and below the stated value with a variance of ±10%.

[0065] As used herein, "patient" generally refers to a mammal (e.g., a human) who has been diagnosed with, is exhibiting symptoms of, or is believed to be afflicted with a disease, disorder, or condition (e.g., cancer). The term "patient" is used interchangeably with the term "subject" herein.

[0066] As used herein, "body surface area" (BSA) is calculated using a standard nomogram. For example,

number

[0067] The terms "combination," "pharmaceutical combination," "combined administration," "administered in combination," and "administering a combination" refer to the administration of one treatment in addition to another. Each component may be administered simultaneously or sequentially at different times in any order. Thus, "in combination with" refers to the administration of more than one pharmaceutically active component (including, but not limited to, an EZH2 modulator and an anti-CD19 antibody disclosed herein) to a patient. Combined administration can refer to simultaneous or sequential administration of an EZH2 modulator and an anti-CD19 antibody disclosed herein.

[0068] The terms "concurrently" and "simultaneously" refer to administration of an EZH2 modulator and an anti-CD19 antibody disclosed herein to a patient at the same time or at two different times that are about 2 hours or less apart. The concurrent administration of the EZH2 modulator and the anti-CD19 antibody can be in a single dosage form or in separate dosage forms.

[0069] The terms "continuous" and "sequentially" refer to the administration of an EZH2 modulator and an anti-CD19 antibody disclosed herein to a patient at two different times that are separated by more than about 2 hours, e.g., about 3 hours, about 4 hours, about 5 hours, about 8 hours, about 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, or even longer.

[0070] The term "intermission" refers to the period after one or more specific pharmaceutically active ingredients are administered to a patient in an intermittent regimen. Intermission refers to a rest period during which the specific pharmaceutically active ingredient is not administered for at least one day.

[0071] As used interchangeably herein, the terms "synergy," "synergistic," "synergistic," and "synergistic effect" refer to a situation in which the combination of two or more drugs produces an effect that is greater than the sum of the effects of each of the individual drugs. The terms encompass not only the alleviation of symptoms of the disorder being treated, but also improved side effect profile, improved tolerability, improved patient compliance, improved efficacy, or any other improvement in clinical outcome.

[0072] As used herein, the illustrative terms "include," "such as," "for example," "eg," etc. (and variations thereof, e.g., "includes," "including," "examples") are intended to be open-ended unless otherwise specified. That is, unless expressly stated otherwise, such terms are intended to imply "including but not limited to," e.g., "including" means "including, but not limited to."

[0073] When used in context to describe a chemical group that may have multiple points of attachment, a hyphen (-) indicates the point of attachment of that group to the variable for which it is defined. For example, -S(C3-C7)cycloalkyl and -S[halo(C1-C4)alkyl] mean that the point of attachment of the group occurs on the sulfur atom.

[0074] As used herein, the terms "halo" and "halogen" refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), and iodine (iodo, -I).

[0075] As used herein, unless otherwise specified, the term "alkyl" refers to a monovalent saturated straight- or branched-chain hydrocarbon radical having 1 to 10 carbon atoms. Examples of alkyl radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, sec-pentyl, isopentyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, sec-hexyl, and the like.

[0076] The term "haloalkyl" includes mono-, poly-, and perhaloalkyl groups, where the halogens are independently selected from fluorine, chlorine, bromine, and iodine.

[0077] An "alkoxy" is an alkyl group attached to another moiety via an oxygen linker (-O(alkyl)). Non-limiting examples include methoxy, ethoxy, propoxy, and butoxy.

[0078] A "haloalkoxy" is a haloalkyl group that is attached to another moiety via an oxygen atom, such as, but not limited to, -CHCF2 or -OCF3.

[0079] The term "cycloalkyl" refers to a fully saturated 3- to 12-membered (e.g., 3- to 7-membered) monocyclic, bicyclic (e.g., bridged or spiro bicyclic ring), or polycyclic (e.g., tricyclic) hydrocarbon ring system. Monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Bridged bicyclic cycloalkyl groups include, but are not limited to, bicyclo[3.2.1]octane, bicyclo[2.2.1]heptane, bicyclo[3.1.0]hexane, bicyclo[1.1.1]pentane, and the like. Spiro bicyclic cycloalkyl groups include, for example, spiro[3.6]decane, spiro[4.5]decane, and the like. Fused cycloalkyl rings include, for example, decahydronaphthalene, octahydropentalene, and the like. When specified as optionally substituted or substituted, a substituent on a cycloalkyl (e.g., in the case of an optionally substituted cycloalkyl) may be located at any substitutable position, including, for example, the position at which the cycloalkyl group is attached.

[0080] The term "heterocyclyl" refers to a 3- to 12-membered (e.g., 4-, 5-, 6-, and 7-membered) saturated or partially unsaturated heterocyclic ring containing 1 to 4 heteroatoms independently selected from N, O, and S. It can be monocyclic, bicyclic (e.g., bridged, fused, or spiro bicyclic rings), or tricyclic. The terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," "heterocyclic moiety," and "heterocyclic radical" are used interchangeably herein. A heterocyclyl ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, pyrrolidinyl, pyridinonyl, pyrrolidonyl, piperidinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyrimidinyl, oxetanyl, azetidinyl, and tetrahydropyrimidinyl. Heterocyclyl groups can be monocyclic or bicyclic. The term "heterocyclyl" also includes unsaturated heterocyclic radicals fused to another unsaturated heterocyclic radical or to an aryl or heteroaryl ring, such as tetrahydronaphthyridine, indolinone, dihydropyrrolotriazole, imidazopyrimidine, quinolinone, dioxaspirodecane. When specified as optionally substituted or substituted, the substituent on the heterocyclyl (e.g., in the case of an optionally substituted heterocyclyl) may be at any substitutable position, including, for example, the position at which the heterocyclyl group is attached.

[0081] The term "spiro" refers to two rings that share one ring atom (eg, carbon).

[0082] The term "fused" refers to two rings that share two adjacent ring atoms with each other.

[0083] The term "bridged" refers to two rings which share three or more ring atoms with one another.

[0084] Unless otherwise stated, structures depicted herein are also meant to include chemical compounds that differ only in the presence of one or more isotopically enriched atoms, for example, the replacement of a hydrogen atom by deuterium or tritium, or 13 C or 14 Chemical compounds having the present structure, except for the replacement of a carbon atom with a C-enriched carbon, are within the scope of this invention. In one embodiment, all atoms on the compounds described herein (e.g., the disclosed EZH2 modulators) are present at natural abundance.

[0085] Unless a stereochemical configuration is indicated, structures depicted herein are intended to include all stereochemical forms of the structure, i.e., the R and S configurations for each asymmetric center. Accordingly, unless otherwise indicated, single stereochemical isomers as well as enantiomeric, racemic, and diastereomeric mixtures of the present chemical compounds are within the scope of the present invention. When a stereochemical configuration is indicated for a compound, the diastereomeric or enantiomeric excess of the compound is, in some embodiments, at least 99.0%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9%.

[0086] Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic / anionic or basic / cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include, for example, salts of inorganic acids (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, and sulfuric acid) and salts of organic acids (e.g., acetic acid, benzenesulfonic acid, benzoic acid, methanesulfonic acid, and p-toluenesulfonic acid).

[0087] The present disclosure provides a combination therapy for cancer patients, comprising administering to a subject in need thereof a therapeutically effective amount of at least one EZH2 modulator, e.g., an EZH2 inhibitor.

[0088] EZH2 modulators described herein include, for example, small molecules capable of modulating EZH2 activity, such as small molecule inhibitors of EZH2. Modulation can be measured in vitro, in vivo, or a combination thereof. In one embodiment, EZH2 modulators described herein include: [ka] and those described in WO 2022 / 0135582, WO 2022 / 035303, WO 2020 / 171606, WO 2019 / 094552, WO 2019 / 204490, WO 2019 / 226491, and WO 2021 / 016414, each of which is incorporated herein by reference.

[0089] In some embodiments, the EZH2 modulator is of Formula I: [ka] or a pharmaceutically acceptable salt thereof, wherein: R 1 is halo, -S(C1-C4)alkyl, -S(C3-C7)cycloalkyl, or -S[halo(C1-C4)alkyl]; X is CH or N; R 2 is hydrogen, halo, (C1-C4)alkyl, or halo(C1-C4)alkyl; R 3 is halo, (C1-C4)alkyl, or halo(C1-C4)alkyl; R 4 is (C3-C7)cycloalkyl or 4- to 7-membered heterocyclyl, each of which is optionally selected from halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, and -NR a R b and R ais hydrogen, (C1-C4) alkyl, or halo(C1-C4) alkyl; R b is (C1-C4)alkyl, halo(C1-C4)alkyl, or a 4- to 7-membered heterocyclyl, which heterocyclyl is optionally substituted with 1 to 3 groups selected from halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, and halo(C1-C4)alkoxy; or R a and R b together with the nitrogen atom to which they are attached, form halo, (C1-C4)alkyl, halo(C1-C4)alkyl, and -OR c and forming a 4- to 7-membered heterocyclyl optionally substituted with 1 to 3 groups selected from R c is (C1-C4)alkyl, halo(C1-C4)alkyl, or (C3-C7)cycloalkyl; R 5 is halo, (C1-C4)alkyl, or halo(C1-C4)alkyl.

[0090] In some embodiments, the EZH2 modulator is of Formula I, or a pharmaceutically acceptable salt thereof, wherein: R 1 is halo or -S(C1-C4)alkyl; R 2 is the halo and R 3 is (C1-C4) alkyl, R 4 is (C3-C7)cycloalkyl or (C4-C7)heterocyclyl, each of which is optionally halo(C1-C4)alkyl and -NR a R b and is substituted with 1 to 2 groups selected from R a is hydrogen or (C1-C4) alkyl, R b is (C1-C4) alkyl or (C4-C7) heterocyclyl, wherein the heterocyclyl is optionally substituted with halo(C1-C4) alkyl, or R a and R b together with the nitrogen atom to which they are attached, form halo and -OR c and forming a 4- to 7-membered nitrogen-containing heterocyclyl optionally substituted with 1 to 3 groups selected from R c is (C1-C4)alkyl, halo(C1-C4)alkyl, or (C3-C7)cycloalkyl; R 5 is halo or —S(C1-C4)alkyl.

[0091] In some embodiments, the EZH2 modulator is of Formula II: [ka] or a pharmaceutically acceptable salt thereof, wherein the variables are as described in any of the above embodiments.

[0092] In some embodiments, R in the EZH2 modulator of Formula I or II, or a pharmaceutically acceptable salt thereof, 1 is chloro, and the remaining variables are as described in any of the above embodiments.

[0093] In some embodiments, R in the EZH2 modulator of Formula I or II, or a pharmaceutically acceptable salt thereof, 1 is -SCH3, and the remainder of the variables are as described in any of the above embodiments.

[0094] In some embodiments, R in the EZH2 modulator of Formula I or II, or a pharmaceutically acceptable salt thereof, 4 is cyclohexyl or piperidinyl, each of which is optionally selected from halo(C-C)alkyl and -NR a R b and the remaining variables are as described in any of the above embodiments.

[0095] In some embodiments, the EZH2 modulator is of Formula III: [ka] or a pharmaceutically acceptable salt thereof, wherein R 6 is halo(C1-C4)alkyl, and the remaining variables are as described in any of the above embodiments.

[0096] In some embodiments, the EZH2 modulator is of formula IV: [ka]

[0097] or a pharmaceutically acceptable salt thereof, wherein the variables are as described in any of the above embodiments.

[0098] In some embodiments, the EZH2 modulator is of Formula V: [ka]

[0099] or a pharmaceutically acceptable salt thereof, wherein the variables are as described in any of the above embodiments.

[0100] In some embodiments, the EZH2 modulator is of Formula VI or VII: [ka] or [ka] or a pharmaceutically acceptable salt thereof, wherein the variables are as described in any of the above embodiments.

[0101] In some embodiments, R in the above EZH2 modulators and pharmaceutically acceptable salts thereofb is (C1-C4) alkyl or oxetanyl, wherein the oxetanyl is optionally substituted with halo(C1-C4) alkyl, or R a and R b together with the nitrogen atom to which they are attached form halo or -OR c and the remaining variables are as described in any of the above embodiments.

[0102] In some embodiments, R in the above EZH2 modulators and pharmaceutically acceptable salts thereof a is hydrogen or methyl, and R b is methyl or oxetanyl, wherein the oxetanyl is optionally substituted with —CH 2 F or —CF 3 , and the remaining variables are as described in any of the above embodiments.

[0103] In some embodiments, the EZH2 modulators and pharmaceutically acceptable salts thereof a and R b together with the nitrogen atom to which they are attached, form one or two fluoro or -OR c and forming an azetidinyl optionally substituted with R c is —CH 3 , —CHF 2 , or cyclopropyl, and the remainder of the variables are as described in any of the above embodiments.

[0104] In some embodiments, the EZH2 modulator has the following chemical structure: [ka] or a pharmaceutically acceptable salt thereof.

[0105] In some embodiments, the EZH2 modulator has the following chemical structure: [ka] or a pharmaceutically acceptable salt thereof.

[0106] In some embodiments, the EZH2 modulator has the following chemical structure: [ka] The crystalline form is crystalline form 1 of 10.0 o , 13.3 o , 14.9 o , 20.2 o , 20.8 o , 22.2 o , and 22.5 o X-ray powder diffraction peaks at at least three 2θ angles selected from 10.0 o , 13.3 o , 14.9 o , 20.2 o , 20.8 o , 22.2 o , and 22.5 o X-ray powder diffraction peaks at at least four 2θ angles selected from 10.0 o , 13.3 o , 14.9 o , 20.2 o , 20.8 o , 22.2 o , and 22.5 o X-ray powder diffraction peaks at at least five 2θ angles selected from 10.0 o , 13.3 o , 14.9 o , 20.2 o , 20.8 o , 22.2 o , and 22.5 o X-ray powder diffraction peaks at at least six 2θ angles selected from 10.0 o , 13.3 o , 14.9 o , 20.2 o , 20.8 o , 22.2 o , and 22.5 o X-ray powder diffraction peaks at 2θ angles selected from: 10.0 o , 10.2 o , 12.3 o , 12.7 o , 13.3o , 14.9 o , 15.3 o , 20.2 o , 20.8 o , 21.3 o , 22.2 o , 22.5 o , and 23.8 o or 10.0 o , 10.2 o , 11.0 o , 11.4 o , 11.8 o , 12.3 o , 12.7 o , 13.3 o , 14.9 o , 15.3 o , 16.1 o , 17.4 o , 20.2 o , 20.8 o , 21.3 o , 22.2 o , 22.5 o , and 23.8 o The compound is characterized by an x-ray powder diffraction peak at a 2θ angle selected from:

[0107] In some embodiments, the EZH2 modulator has the following chemical structure: [ka] The crystalline form is crystalline form 1 of 14.9 o , 20.2 o , and 20.8 o X-ray powder diffraction peaks at 2θ angles selected from: 10.0 o , 14.9 o , 20.2 o , and 20.8 o X-ray powder diffraction peaks at 2θ angles selected from: 10.0 o , 14.9 o , 20.2 o , 20.8 o , and 22.2 o or 10.0 o , 13.3o , 14.9 o , 20.2 o , 20.8 o , and 22.2 o The compound is characterized by an x-ray powder diffraction peak at a 2θ angle selected from:

[0108] The present disclosure provides, inter alia, combination treatments comprising administering a therapeutically effective amount of at least one anti-CD19 antibody (e.g., tafasitamab) to a subject in need thereof.

[0109] The use of anti-CD19 antibodies in non-specific B-cell lymphoma is discussed in WO2007076950 (US2007154473), both of which are incorporated by reference in their entireties. The use of CD19 antibodies in CLL, NHL, and ALL is described in Scheuermann et al., CD19 Antigen in Leukemia and Lymphoma Diagnosis and Immunotherapy, Leukemia and Lymphoma, Vol. 18, 385-397 (1995), which is incorporated by reference in its entirety.

[0110] Further antibodies specific for CD19 are disclosed in WO2005012493 (US7109304), WO2010053716 (US12 / 266,999) (Immunomedics), WO2007002223 (US8097703) (Medarex), WO2008022152 (US12 / 377,251) and WO2008150494 (Xencor), WO2008031056 (US11 / 852,106) (Medimmune), WO 2007076950 (US11 / 648,505) (Merck Patent GmbH), WO 2009 / 052431 (US12 / 253,895) (Seattle Genetics), and WO2010095031(12 / 710,442) (Glenmark Pharmaceuticals), WO2012010562 and WO2012010561 (International Drug Development), WO2011147834 (Roche Glycart), and WO2012156455 (Sanofi), all of which are incorporated by reference in their entireties.

[0111] The dose of the antibody contained in the pharmaceutical composition containing the combination according to the present disclosure administered to a patient may vary depending on the patient's age and size, symptoms, condition, route of administration, etc. The dose is usually calculated according to body weight or body surface area, age, or per person. The frequency and duration of treatment may be adjusted depending on the severity of the condition. Effective dosages and schedules for administering pharmaceutical compositions containing antibodies or antibody fragments specific for CD19 may be empirically determined. For example, the patient's progress can be monitored by periodic evaluation, and the dosage adjusted accordingly. Furthermore, interspecies scaling of dosages may be performed using methods well known in the art (e.g., Mordenti et al., 1991, Pharmaceut. Res. 8:1351).

[0112] The anti-CD19 antibody can be administered as a fixed dose or in mg / kg patient weight dose.The dose can also be selected to reduce or avoid the production of antibodies against the anti-CD19 antibody.The dosage regimen is adjusted to achieve a desired response, for example, a therapeutic response or a combined therapeutic effect.Generally, the dose of the anti-CD19 antibody can be used to provide the subject with a bioavailable amount of the drug.

[0113] The anti-CD19 antibody can be administered once a week, once every two weeks, or once every four weeks.

[0114] For example, a dose ranging from about 9 mg / kg to about 30 mg / kg may be administered. In certain embodiments, a subject in need of treatment with an anti-CD19 antibody is administered the antibody at a dose of about 9 mg / kg or about 30 mg / kg. With respect to dose or dosage, the term "about" is intended to indicate a range of ±10% of the recited dose; for example, a dose of about 3 mg / kg is 2.7 mg / kg to 3.3 mg / kg of patient body weight.

[0115] The anti-CD19 antibody may be administered at a dose of at least 9 mg / kg. In another embodiment, the anti-CD19 antibody may be administered at a dose of 9 mg / kg to 12 mg / kg. In another embodiment, the anti-CD19 antibody may be administered at a dose of 9 mg / kg.

[0116] The anti-CD19 antibody may be administered at a dose of 24 mg / kg or more. In another embodiment, the anti-CD19 antibody may be administered at a dose of 24 mg / kg to 30 mg / kg. In another embodiment, the anti-CD19 antibody may be administered at a dose of 24 mg / kg. In another embodiment, the anti-CD19 antibody may be administered at a dose of 30 mg / kg. The pharmaceutical composition may include dosage forms for intravenous, subcutaneous, intradermal, and intramuscular injection. These injectable formulations may be prepared by known methods. For example, the injectable formulation may be prepared by dissolving, suspending, or emulsifying the above-mentioned antibody or its salt in a sterile aqueous or oily medium commonly used for injections. Exemplary pharmaceutical compositions comprising an antibody specific for CD19 that can be used in connection with the present disclosure are disclosed, for example, in WO2008 / 022152 or WO2018 / 002031, both of which are incorporated by reference in their entireties.

[0117] For certain administration methods used in embodiments of the present disclosure, such as intravenous administration, it is preferable to administer the drug according to the patient's weight. For other administration methods used in embodiments of the present disclosure, such as subcutaneous administration, it is preferable to administer the drug at a fixed, constant dose. Those skilled in the art will understand when a dose in one administration method is equivalent or substantially equivalent to another dose in another administration method. For example, the pharmacodynamics of a particular drug are usually taken into account in making a rational decision to administer the drug in the required form and at the required effective dose. Antibodies administered according to the present disclosure can be administered to patients in therapeutically effective amounts.

[0118] In some embodiments, the present disclosure relates to methods of treating cancer in a patient in need thereof by administering to the patient a combination of an EZH2 modulator and one or more anti-CD19 antibodies.

[0119] In some embodiments, the present disclosure relates to methods of treating cancer by administering to a patient in need thereof a combination of an EZH2 modulator and an anti-CD19 antibody.

[0120] In some embodiments, the present disclosure relates to the use of a combination of an EZH2 modulator and an anti-CD19 antibody for the treatment of cancer in a patient.

[0121] In some embodiments, the present disclosure relates to a composition comprising an EZH2 modulator for use in treating cancer in a patient, wherein the patient is also treated with an anti-CD19 antibody. In some aspects, the present disclosure relates to a composition comprising an EZH2 modulator for use in treating a disease in a patient, wherein the disease is cancer, wherein the EZH2 modulator is in combination with an anti-CD19 antibody. In some embodiments, the EZH2 modulator can be administered simultaneously or sequentially with the anti-CD19 antibody.

[0122] In some embodiments, the present disclosure relates to methods of treating cancer, comprising administering to a patient in need thereof a therapeutically effective amount of a combination of an EZH2 modulator and an anti-CD19 antibody.

[0123] In some embodiments, the present disclosure relates to methods of treating cancer by administering to a patient a combination of an EZH2 modulator and an anti-CD19 antibody.

[0124] In another aspect, the present disclosure relates to the use of a combination of an EZH2 modulator and an anti-CD19 antibody for the treatment of cancer.

[0125] In some embodiments, the methods of treating cancer described herein may include a combination of an EZH2 modulator, an anti-CD19 antibody, and one or more additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents may be chemotherapeutic agents. In some embodiments, the one or more additional therapeutic agents may include, but are not limited to, lenalidomide, fludarabine, cyclophosphamide, doxorubicin, vincristine, methotrexate, anthracycline chemotherapy agents, prednisone, methylprednisolone, glucocorticoids, ibritumomab tiuxetan, acetaminophen, antihistamines, and combinations thereof. In some embodiments, the one or more additional therapeutic agents may include R-CHOP.

[0126] In embodiments, the one or more additional therapeutic agents is lenalidomide.

[0127] In some embodiments, the cancer is a CD19-positive cancer.

[0128] In some embodiments, the cancer is a hematological cancer. Non-limiting examples of hematological cancers include acute myeloid leukemia (AML), chronic myeloid leukemia (CML), including accelerated phase CML and blastic phase of CML (CML-BP), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), Hodgkin's lymphoma (HL), B-cell lymphoma, T-cell lymphoma, follicular lymphoma (FL), marginal zone lymphoma (MZL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), and non-Hodgkin's lymphoma (NHL), including Burkitt's lymphoma, multiple myeloma (MM), amyloidosis, Waldenstrom's disease, and leukemia. Myelodysplastic syndromes (MDS), including Ström's macroglobulinemia, refractory anemia (RA), refractory anemia with ringed sideroblasts (RARS), refractory anemia with excess blasts (RAEB), and accelerated RAEB (RAEB-T), and myeloproliferative disorders. In some embodiments, the cancer is chronic lymphocytic leukemia (CLL), Hodgkin's lymphoma, or non-Hodgkin's lymphoma, including follicular lymphoma (FL), marginal zone lymphoma (MZL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), and Burkitt's lymphoma.

[0129] In some embodiments, the cancer is chronic lymphocytic leukemia. In some embodiments, the cancer is CD19-positive chronic lymphocytic leukemia.

[0130] In some embodiments, the cancer is a non-Hodgkin's lymphoma, including follicular lymphoma (FL), marginal zone lymphoma (MZL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), and Burkitt's lymphoma. In some embodiments, the cancer is a B-cell lymphoma.

[0131] In one embodiment, the lymphoma is non-Hodgkin's lymphoma. In another embodiment, the non-Hodgkin's lymphoma is DLBCL. In another embodiment, the DLBCL is relapsed, refractory, or relapsed and refractory DLBCL. In one embodiment, the DLBCL is relapsed DLBCL. In one embodiment, the DLBCL is refractory DLBCL. In one embodiment, the DLBCL is relapsed and refractory DLBCL. In another embodiment, the DLBCL is relapsed and refractory DLBCL, and patients with relapsed and refractory DLBCL are not eligible for autologous stem cell transplantation (ASCT).

[0132] In another embodiment, the DLBCL is primary refractory DLBCL.

[0133] In one embodiment, the DLBCL is primary DLBCL. In another embodiment, the DLBCL is newly diagnosed DLBCL. In another embodiment, the DLBCL is newly diagnosed DLBCL, and the patient with the newly diagnosed DLBCL is at least 80 years old and is not eligible for autologous stem cell transplant (ASCT). In another embodiment, the DLBCL is newly diagnosed frail / unfit DLBCL. In another embodiment, the DLBCL is newly diagnosed DLBCL, and the patient with the newly diagnosed DLBCL is frail and 80 years old or older. In another embodiment, the DLBCL is newly diagnosed DLBCL, and the patient with the newly diagnosed DLBCL is frail, 80 years old or older, has an ADL score of 6, and an IADL score of 8. In another embodiment, the DLBCL is newly diagnosed DLBCL, and the patient with the newly diagnosed DLBCL is unfit, aged 65 years or older, and has at least one of the following comorbidities: (i) (LVEF) ≥ 30% to < 50%, (ii) history of myocardial infarction within 6 months prior to screening, (iii) ischemic heart disease, or (iv) history of stroke within 12 months prior to screening.

[0134] In another embodiment, the DLBCL is newly diagnosed DLBCL, and the patient with the newly diagnosed DLBCL is (a) frail, aged 80 years or older, with an ADL score of 6 and an IADL score of 8, or (b) unfit, aged 65 years or older, and has at least one of the following comorbidities: (i) (LVEF) ≥ 30% to < 50%, (ii) a history of myocardial infarction within 6 months prior to screening, (iii) ischemic heart disease, or (iv) a history of stroke within 12 months prior to screening.

[0135] In another embodiment, the DLBCL is newly diagnosed DLBCL. In another embodiment, the DLBCL is newly diagnosed DLBCL and the patient with the newly diagnosed DLBCL is at least 80 years old. In another embodiment, the DLBCL is newly diagnosed DLBCL and the newly diagnosed patient is at least 80 years old and has an ADL score of 6 and an IADL score of 8. In another embodiment, the DLBCL is newly diagnosed DLBCL and the patient with the newly diagnosed DLBCL is at least 65 years old and has at least one of the following comorbidities: (i) (LVEF) ≥ 30 to < 50%, (ii) a history of myocardial infarction within 6 months prior to screening, (iii) ischemic heart disease, or (iv) a history of stroke within 12 months prior to screening.

[0136] In another embodiment, the DLBCL is newly diagnosed DLBCL, and the patient with the newly diagnosed DLBCL is (a) at least 80 years old and has an ADL score of 6 and an IADL score of 8, or (b) at least 65 years old and has at least one of the following comorbidities: (i) (LVEF) ≥ 30 to < 50%, (ii) a history of myocardial infarction within 6 months prior to screening, (iii) ischemic heart disease, or (iv) a history of stroke within 12 months prior to screening.

[0137] In one embodiment, the lymphoma is a CD20-expressing lymphoma. In another embodiment, the lymphoma is a CD19-expressing lymphoma. In another embodiment, the lymphoma is a CD20-expressing CD19-expressing lymphoma.

[0138] In one embodiment, the subject has previously received CAR-T therapy.

[0139] In some embodiments, the cancer is a CD19-positive non-Hodgkin's lymphoma. In some embodiments, the cancer is a CD19-positive aggressive non-Hodgkin's lymphoma. In some embodiments, the cancer is a CD19-positive indolent non-Hodgkin's lymphoma. In some embodiments, the cancer is a relapsed or refractory non-Hodgkin's lymphoma. In some embodiments, the cancer is a relapsed or refractory CD19-positive aggressive non-Hodgkin's lymphoma. In some embodiments, the cancer is a relapsed or refractory CD19-positive indolent non-Hodgkin's lymphoma.

[0140] In some embodiments, the cancer is recurrent. In some embodiments, a recurrent cancer is one that has recurred after a period of time during which the cancer could not be detected.

[0141] In some embodiments, the cancer is refractory. In some embodiments, refractory cancers do not respond to cancer treatment and are also known as resistant cancers. In some embodiments, the cancer is resistant to rituximab. In some embodiments, the cancer does not respond to treatment with rituximab. In some embodiments, the cancer is a rituximab-resistant recurrent cancer. In some embodiments, the patient has become refractory to a rituximab-containing regimen. In some embodiments, the tumor is unresectable. In some embodiments, the cancer has not been previously treated. In some embodiments, the cancer is locally advanced. In some embodiments, "locally advanced" refers to a cancer that is somewhat widespread but still limited to one area. In some cases, "locally advanced" can refer to a small tumor that has not spread but has invaded nearby organs or tissues, making it difficult to remove by surgery alone. In some embodiments, the cancer is metastatic.

[0142] In some embodiments, the patient has relapsed or refractory CD19-positive non-Hodgkin's lymphoma. In some embodiments, the patient has both CD19-positive non-Hodgkin's lymphoma and relapsed or refractory non-Hodgkin's lymphoma.

[0143] In some embodiments, the patient has relapsed or refractory CD19-positive, aggressive non-Hodgkin's lymphoma. In some embodiments, the patient has relapsed or refractory CD19-positive, aggressive non-Hodgkin's lymphoma and has progressed on at least one prior treatment regimen.

[0144] In some embodiments, the patient has relapsed or refractory CD19-positive indolent non-Hodgkin's lymphoma. In some embodiments, the patient has relapsed or refractory CD19-positive indolent non-Hodgkin's lymphoma and has progressed on at least two previous treatment regimens. In some embodiments, the patient has relapsed or refractory CD19-positive indolent non-Hodgkin's lymphoma and is refractory to any anti-CD20 monoclonal antibody or any anti-CD19 monoclonal antibody. In some embodiments, the patient has relapsed or refractory CD19-positive indolent non-Hodgkin's lymphoma and has progressed on at least two previous treatment regimens and is refractory to anti-CD20 monoclonal antibodies.

[0145] In some embodiments, the present disclosure relates to a medicament for use in treating cancer in a patient in need of such treatment, the medicament comprising an EZH2 modulator and an anti-CD19 antibody, in a single dosage form or in separate dosage forms.

[0146] In some embodiments, the medicaments described herein may comprise a combination of an EZH2 modulator, an anti-CD19 antibody, and optionally one or more additional therapeutic agents.

[0147] In some embodiments, the disclosure relates to the use of an EZH2 modulator in the manufacture of a medicament for the treatment of cancer, wherein the EZH2 modulator is administered together with an anti-CD19 antibody, wherein the medicaments are in a single dosage form or in separate dosage forms. In some embodiments, the EZH2 modulator is administered together with the anti-CD19 antibody and one or more additional therapeutic agents.

[0148] In some embodiments, the present disclosure relates to the use of an EZH2 modulator for the manufacture of a medicament for treating cancer in a patient, wherein the patient is also treated with an anti-CD19 antibody, and optionally one or more additional therapeutic agents. In some embodiments, the EZH2 modulator can be administered simultaneously or sequentially with the anti-CD19 antibody. In some embodiments, the EZH2 modulator is included in the same composition as the anti-CD19 antibody. In some embodiments, the EZH2 modulator is included in a separate composition from the anti-CD19 antibody. In some embodiments, the EZH2 modulator is included in the same composition as the one or more additional therapeutic agents. In some embodiments, the EZH2 modulator is included in the same composition as the anti-CD19 antibody, and optionally one or more additional therapeutic agents. In some embodiments, the EZH2 modulator is included in a separate composition from the one or more additional therapeutic agents. In some embodiments, the EZH2 modulator is included in a separate composition from the anti-CD19 antibody, and optionally one or more additional therapeutic agents.

[0149] In another aspect, the disclosure relates to the use of a combination of an EZH2 modulator and an anti-CD19 antibody in the manufacture of a medicament for use in treating cancer. In some embodiments, the disclosure relates to the use of a combination of an EZH2 modulator and an anti-CD19 antibody, and optionally one or more additional therapeutic agents, in the manufacture of a medicament for use in treating cancer.

[0150] In another aspect, the disclosure relates to the use of an EZH2 modulator in the manufacture of a medicament for the treatment of cancer, wherein the EZH2 modulator is administered together with an anti-CD19 antibody, and optionally one or more additional therapeutic agents.

[0151] In one aspect, the present disclosure relates to an anti-CD19 antibody for use in treating cancer, wherein the anti-CD19 antibody is administered in combination with an EZH2 modulator to a subject in need of such treatment.

[0152] In one aspect, the present disclosure relates to an anti-CD19 antibody for use in treating cancer, wherein the anti-CD19 antibody is administered in combination with an EZH2 modulator.

[0153] In some embodiments, the anti-CD19 antibody comprises a heavy chain variable region comprising an HCDR1 region comprising the sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising the sequence NPYNDG (SEQ ID NO: 2), and an HCDR3 region comprising the sequence GTYYYGTRVFDY (SEQ ID NO: 3), and a light chain variable region comprising an LCDR1 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising the sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6).

[0154] In some embodiments, the anti-CD19 antibody comprises a heavy chain variable region comprising an HCDR1 region of SYVMH (SEQ ID NO: 1), an HCDR2 region of NPYNDG (SEQ ID NO: 2), and an HCDR3 region of GTYYYGTRVFDY (SEQ ID NO: 3), and a light chain variable region comprising an LCDR1 region of RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region of RMSNLNS (SEQ ID NO: 5), and an LCDR3 region of MQHLEYPIT (SEQ ID NO: 6).

[0155] In some embodiments, the anti-CD19 antibody comprises a heavy chain variable region of EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSS (SEQ ID NO: 7), and a light chain variable region of DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIK (SEQ ID NO: 8).

[0156] In some embodiments, the anti-CD19 antibody has effector function. In another aspect, the CD19-specific antibody or antibody fragment has enhanced effector function. In one embodiment, the effector function is ADCC. In one embodiment, the CD19-specific antibody or antibody fragment has enhanced ADCC activity. In a further embodiment, the CD19-specific antibody or antibody fragment comprises an Fc domain containing amino acid substitutions at positions S239 and / or I332, the numbering being according to the EU index in Kabat.

[0157] In some embodiments, the anti-CD19 antibody comprises the heavy chain constant region of ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 9).

[0158] In some embodiments, the anti-CD19 antibody comprises the light chain constant region of RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10).

[0159] In some embodiments, the anti-CD19 antibody is ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQ It comprises a heavy chain constant region of PREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 9), and a light chain constant region of RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10).

[0160] In some embodiments, the anti-CD19 antibody is EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTI It comprises a heavy chain region of SKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 11), and a light chain region of DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12).

[0161] In some embodiments, the anti-CD19 antibody is tafasitamab.

[0162] The EZH2 modulator may be administered in combination with the anti-CD19 antibody, and optionally one or more additional therapeutic agents, in a single dosage form or in separate dosage forms. In some embodiments, when administered in separate dosage forms, the anti-CD19 antibody may be administered before, simultaneously with, or after administration of the EZH2 modulator. In some embodiments, when administered in separate dosage forms, one or more doses of the EZH2 modulator may be administered before the anti-CD19 antibody. In some embodiments, the anti-CD19 antibody is administered before administration of the EZH2 modulator. As used herein, administration of an EZH2 modulator, an anti-CD19 antibody, and optionally one or more additional therapeutic agents in "combination" refers not only to simultaneous or sequential administration of the agents, but also to administration of the agents in a single treatment cycle, as will be understood by those skilled in the art. When an EZH2 modulator is administered in combination with the anti-CD19 antibody and, optionally, one or more additional therapeutic agents, a therapeutically effective amount of the combination may be administered.

[0163] The EZH2 modulator may be administered by any method known to those of skill in the art. For example, in some embodiments, the EZH2 modulator may be administered in the form of a pharmaceutical composition comprising the EZH2 modulator and a pharmaceutically acceptable carrier, such as those described herein. In some embodiments, the pharmaceutical composition is suitable for oral administration. In some embodiments, the pharmaceutical composition is a tablet or capsule suitable for oral administration. In some other embodiments, the pharmaceutical composition is a liquid dosage form suitable for oral administration. In some embodiments, the pharmaceutical composition is suitable for intravenous administration. In some embodiments, the pharmaceutical composition is suitable for subcutaneous administration. In some embodiments, these compositions optionally further comprise one or more additional therapeutic agents.

[0164] The anti-CD19 antibody can be administered by any method known to those of skill in the art. In some embodiments, the anti-CD19 antibody is administered intravenously (IV). In some embodiments, the anti-CD19 antibody is administered subcutaneously (SC). In some embodiments, the anti-CD19 antibody is administered orally. For example, the anti-CD19 antibody can be administered in the form of a second composition, in some embodiments, a pharmaceutical composition comprising the anti-CD19 antibody and a pharmaceutically acceptable carrier, such as those described herein. In some aspects, the pharmaceutical composition is suitable for oral administration. In some embodiments, the pharmaceutical composition is a tablet or capsule suitable for oral administration. In some other embodiments, the pharmaceutical composition is a liquid dosage form suitable for oral administration. In some embodiments, these compositions optionally further comprise one or more additional therapeutic agents.

[0165] The amount or appropriate dose of the disclosed methods depends on several factors, including the nature of the severity of the condition being treated, the particular inhibitor, the route of administration, and the age, weight, general condition, and response of the individual patient. In some embodiments, an appropriate dose level is one that achieves a therapeutic response as measured by tumor regression or other standard criteria such as disease progression, progression-free survival, or overall survival. In some embodiments, an appropriate dose level is one that achieves this therapeutic response while also minimizing any side effects associated with the administration of the therapeutic agent. An appropriate dose level may prolong the therapeutic response and / or prolong life.

[0166] It will be appreciated that an appropriate dose of the EZH2 modulator, e.g., Compound I, that elicits a biological or medical response in a subject can be administered, for example, at a dose of 0.01 to 100 mg / kg body weight / day. In one embodiment, the dose of the EZH2 modulator, e.g., Compound I, ranges from about 10 mg / kg body weight / day to about 150 mg / kg body weight / day. In another embodiment, the dose of the EZH2 modulator, e.g., Compound I, ranges from about 50 mg to about 375 mg per day. In another embodiment, the dose of the EZH2 modulator, e.g., Compound I, is about 375 mg per day. In another embodiment, the dose of the EZH2 modulator, e.g., Compound I, is 375 mg per day.

[0167] It will be understood that the appropriate doses of the EZH2 modulator, the anti-CD19 antibody, and optionally one or more additional therapeutic agents may be taken at any time during the day or night. In some embodiments, an appropriate dose of each agent is taken in the morning. In some other embodiments, an appropriate dose of each agent is taken in the evening. In some embodiments, an appropriate dose of each of the agents is taken both in the morning and in the evening. It will be understood that the appropriate doses of each agent may be taken with or without food. In some embodiments, an appropriate dose of an agent is taken with a meal. In some embodiments, an appropriate dose of an agent is taken on an empty stomach.

[0168] In some embodiments, the EZH2 modulator is administered on a daily schedule. In some embodiments, the EZH2 modulator is administered every other day. In some embodiments, the EZH2 modulator is administered every three days. In some embodiments, the EZH2 modulator is administered on a twice-weekly schedule. In some embodiments, the EZH2 modulator is administered on a three-times-weekly schedule. In some embodiments, the EZH2 modulator is administered on a weekly schedule. In some embodiments, the EZH2 modulator is administered on a biweekly schedule.

[0169] In some embodiments, the EZH2 modulator is administered at least three times, every other day, within a 7-day cycle. In some embodiments, the EZH2 modulator is administered on days 1 and 4 of the 7-day cycle. In some embodiments, the EZH2 modulator is administered every day in a 7-day cycle, followed by an intermission. In some embodiments, the EZH2 modulator is administered for two consecutive days in at least one 7-day cycle, followed by a five-day intermission. In some embodiments, the EZH2 modulator is administered for three consecutive days in at least one 7-day cycle, followed by a four-day intermission. In some embodiments, the EZH2 modulator is administered for four consecutive days in at least one 7-day cycle, followed by a three-day intermission. In some embodiments, the EZH2 modulator is administered for five consecutive days in at least one 7-day cycle, followed by a two-day intermission. In some embodiments, there is a rest period between one or more 7-day treatment cycles. In some embodiments, there is a 7-day rest period between one or more 7-day treatment cycles.

[0170] The present description contemplates administering the drug for one or more treatment cycles, e.g., 1, 2, 3, 4, 5, 6, or more treatment cycles. In some embodiments, the treatment cycle is about 7 days to about 56 days, or longer. In some embodiments, the treatment cycle is 7 days, 14 days, 21 days, 28 days, 35 days, 42 days, 49 days, or 56 days. In some embodiments, the treatment cycle is 21 days or 28 days. In some embodiments, there is a rest period between one or more treatment cycles. For example, in some embodiments, there is a rest period at the end of the treatment cycle. In some embodiments, there is a rest period between the second and third treatment cycles, but not between the first and second treatment cycles. In other embodiments, there may be a rest period between the first and second treatment cycles, but not between the second and third treatment cycles. Dosing schedules include, for example, administering the EZH2 modulator once per treatment cycle, e.g., day 1 of a 21-day cycle, twice per treatment cycle, e.g., days 1 and 15 of a 21-day cycle, or days 1 and 15 of a 28-day cycle, three times per treatment cycle, e.g., days 1, 8, and 15 of a 21-day cycle, or days 1, 8, and 15 of a 28-day cycle, and four times per treatment cycle, e.g., days 1, 4, 8, and 11 of a 21-day cycle, or days 1, 4, 8, and 11 of a 28-day cycle. Other dosing schedules are encompassed by the invention.

[0171] In some embodiments, the EZH2 modulator is administered within a 21 day cycle. In some embodiments, the EZH2 modulator is administered at least twice within a 21 day cycle. In some embodiments, the EZH2 modulator is administered on day 1 within a 21 day cycle. In some embodiments, the EZH2 modulator is administered on day 8 within a 21 day cycle. In some embodiments, the EZH2 modulator is administered on days 1 and 8 within a 21 day cycle.

[0172] In some embodiments, the EZH2 modulator is administered for a period of one year or less. In some embodiments, the EZH2 modulator is administered for a period of one year or more.

[0173] In some embodiments, the EZH2 modulators or anti-CD19 antibodies described herein may be manufactured for inclusion in a kit. A "kit" is any article of manufacture (e.g., a package or container) containing at least one reagent or chemotherapeutic agent. Kits for use in the methods herein may include an EZH2 modulator, such as a compound of Formula I or a pharmaceutically acceptable salt thereof. In some embodiments, the kit may further include an anti-CD19 antibody, and optionally one or more additional therapeutic agents. In some embodiments, the kit may include a compound of Formula I or a pharmaceutically acceptable salt thereof, an anti-CD19 antibody, and optionally one or more additional therapeutic agents. In some embodiments, the kit may include one or more EZH2 modulators or pharmaceutically acceptable salts thereof. In some embodiments, the kit may include one or more anti-CD19 antibodies.

[0174] In some embodiments, the present disclosure relates to a kit comprising agents for use in treating cancer in a patient in need of such treatment. The kit may comprise an agent comprising an EZH2 modulator and, optionally, instructions for administering the EZH2 modulator and an anti-CD19 antibody, or the kit may comprise an agent comprising an anti-CD19 antibody and, optionally, instructions for administering the anti-CD19 antibody and the EZH2 modulator. The kit may comprise an agent comprising an EZH2 modulator and an anti-CD19 antibody and, optionally, instructions for administering the EZH2 modulator and the anti-CD19 antibody, wherein the agents are in a single dosage form or in separate dosage forms. In some embodiments, the kit optionally comprises one or more additional therapeutic agents.

[0175] In some embodiments, a kit comprising an EZH2 modulator and an anti-CD19 antibody may further comprise another component or reagent. In some embodiments, a reagent included in the kit may be a diluent for preparing the EZH2 modulator for administration. In some embodiments, a reagent included in the kit may be a diluent for preparing the anti-CD19 antibody for administration. In some embodiments, a component included in the kit may be a container for mixing the combination of the EZH2 modulator and the anti-CD19 antibody.

[0176] In another aspect, the present disclosure relates to a kit for treating cancer comprising at least one agent comprising at least one dose of an EZH2 modulator and at least one agent comprising at least one dose of an anti-CD19 antibody, wherein the kit for treating cancer further comprises administration instructions for administering the agents for treatment of a patient identified in need of such treatment.

[0177] In order that this disclosure may be more fully understood, the following examples are set forth, which are illustrative only and are not intended to limit the scope of the disclosure in any way. [Table 1-1] [Table 1-2] [Example]

[0178] Example 1: In vitro methods cell line A panel of six lymphoma cell lines was included in the analysis (five DLBCL and one Burkitt's lymphoma). All cell lines were cultured in RPMI 1640 medium (Gibco) supplemented with 10%–20% FCS (Sigma) at 37°C in 5% CO.

[0179] compound Compound 1 was prepared according to the methods described in WO 2021 / 016409, the contents of which are incorporated herein by reference.

[0180] Viability assay To analyze direct cytotoxic and antiproliferative effects on target cells, cell viability assays were performed using the CellTiter-Glo kit (Promega), a luminescent assay for measuring viable cell number based on quantification of adenosine triphosphate (ATP) levels. Cells were treated with Compound 1 or tazemetostat for 7 days (SU-DHL-4, SU-DHL-6, Ramos, U-2932) or 11 days (SU-DHL-8, NU-DUL-1), with tafasitamab added for only the last day (SU-DHL-6, NU-DUL-1) or 4 days (all other cell lines). To avoid overgrowth, cell density was adjusted on days 3 and 7 of the assay. The assays were performed in clear 48-well and 96F-well bottom plates in a final volume of 1 mL or 200 μL, respectively, in a cell incubator at 37°C and 5% CO2. All test conditions were performed in triplicate in each experiment.

[0181] Cell viability was analyzed using a Tecan Infinite F500 plate reader according to the manufacturer's protocol. The relative viable cell number was calculated from the measured relative luminescence units (RLU) using the following formula:

number

[0182] CD19 expression The cell surface CD19 expression of SU-DHL-4 and SU-DHL-6 cells was analyzed after 7 days of treatment with Compound 1. The remaining cells after CTG readout were stained in duplicate with a live / dead staining kit (L / D fixable yellow stain, Invitrogen) and a PE-labeled anti-CD19 detection antibody (clone HIB-19, Biolegend), and CD19 expression on the surface of live cells was measured using a flow cytometer (FACS Verse). Relative CD19 expression was calculated from the measured median fluorescence intensity (MFI) using the following formula:

number

[0183] Data analysis For graphical representation and IC50 determination, the mean values ​​of triplicates (survival assay) or duplicates (CD19 expression) from individual experiments were imported into GraphPad Prism software (version 8). The mean values ​​- / + standard deviation of the relative viable cell counts and relative CD19 expression were calculated from 2 to 11 individual experiments and displayed as scatter bar graphs. IC50 curves were determined by applying a nonlinear regression curve-fit model (sigmoidal dose-response, variable slope). The combined effect of both compounds on cell survival was estimated by a linear mixed-effects model of the following form:

number

[0184] Total mean RLU values ​​were calculated for: ·e μ Untreated samples by ·e (μ+α) Tafasitamab-treated samples ·e (μ+β) Compound 1 treated samples with ·e (μ+α+β+αβ) Tafasitamab + Compound 1 treated samples The percent inhibition of tafasitamab, compound 1, and the combined effect of both compounds were calculated for the following: ·(e α Tafasitamab by -1)*-100 ·(e β Compound 1 by -1)*-100 ·(e αβ Combined effect of -1)*-100

[0185] In vitro results Tafasitamab mediates B-cell lysis through direct cytotoxic effects and immune effector mechanisms such as ADCC and ADCP, while compound 1 has cytotoxic and antiproliferative effects on tumor cells. The combined effects of both drugs were analyzed in samples to determine their common mechanism of action: their direct effect on viable target cell numbers. The lymphoma cell lines used in the in vitro experiments included cell lines derived from different NHL subtypes (ABC DLBCL, GCB DLBCL, and BL), with different EHZ2 status (wild-type and variants with gain-of-function mutations) and sensitivity. Figure 1 shows the effects of tafasitamab, compound 1, and the combination of both on tumor cell survival at predetermined suboptimal dose levels. In seven of the seven cell lines examined, this combination of compounds reduced relative viable cell numbers better than either compound individually. The combined effects of tafasitamab and compound 1 were estimated using a linear mixed-effects model. An estimated combined effect less than 0 indicates antagonism, 0 indicates an additive effect, and values ​​greater than 0 indicate an additive effect (synergism) greater than the sum of the individual effects. The individual effects of both compounds estimated by the applied model (Figures 2A and 2B) correlated well with the results shown in Figure 1. Evaluation of the combined effect resulted in a significant synergistic decrease in survival of SU-DHL-4, SU-DHL-6, and SU-DHL-8, a trend toward a synergistic decrease in survival of NU-DUL-1, and an additive, but not synergistic, decrease in survival of U-2932 and Ramos cells (Figure 2C).

[0186] The ADCC mechanism of action of tafasitamab was not adversely affected by compound 1 when effector cells were pretreated with compound 1 (data not shown).

[0187] The effects of compound 1 alone and in combination with tafasitamab were compared with another EZH2 inhibitor, tazemetostat. In both tested cell lines, SU-DHL-4 and SU-DHL-6, compound 1 inhibited cell viability at lower concentrations than tazemetostat. The median inhibitory concentration (IC50) of the compound 1 / tafasitamab combination was 23-fold (14.3 nM vs. 332 nM in SU-DHL-4) or 35-fold (2.3 nM vs. 79.4 nM in SU-DHL-6) improved compared with the tazemetostat / tafasitamab combination. Overall, tafasitamab can generally improve the activity of EZH2 inhibitors, as demonstrated by the combined effect with tazemetostat in CTG assays using SU-DHL-4 and SU-DHL-6 cells (Figures 3A and 3B, Table 1). [Table 2]

[0188] In parallel with the survival readout, SU-DHL-4 and SU-DHL-6 cells treated with compound 1 were analyzed for expression of the tafasitamab target molecule, CD19. As determined by flow cytometry analysis after 7 days of treatment, compound 1 led to increased surface levels of CD19, which could explain the improved cytotoxic effect of tafasitamab (Figure 4).

[0189] Example 2: In vivo methods method In vivo studies were performed in CB17-SCID mice bearing WSU-DLCL2 tumors. 4 x 10^6 WSU-DLCL2 cells (a DLBCL cell line) were implanted sc into the left flank of 6- to 8-week-old female CB17-SCID mice with Matrigel (1:1) per animal. On study day 17, when the mean tumor volume reached approximately 100 mm^3, animals were randomly assigned to treatment groups (8 mice per group) and treatment began. Compound 1 was administered daily po at 35 mg / kg or 50 mg / kg (administration volume: 10 mL / kg, formulation buffer: 20% propylene glycol, 10% Solutol HS15, 70% phosphate buffer) and tafasitamab was administered biweekly iv at 3 mg / kg or 10 mg / kg (administration volume: 5 mL / kg, formulation buffer: PBS). Tumor volume and body weight were measured twice weekly. Mice were sacrificed when tumor volumes reached >1500 mm^3 or at the latest at the end of the study, 69 days after tumor cell implantation. Mean tumor growth curves were calculated using GraphPad Prism software (version 8.4.3).

[0190] result Proof of concept in vivo studies were performed in WSU-DLCL2 tumor-bearing CB17-SCID mice. Table 2 summarizes the nine different treatment groups. [Table 3]

[0191] Figures 5A and 5B show the tumor growth kinetics (mean + / - SEM) of the in vivo model performed. The mean tumor volumes of the individual treatment groups at 38 days after tumor implantation (21 days after treatment initiation) are shown in Figure 6.

[0192] Monotherapy with compound 1 (50 and 35 mg / kg) or tafasitamab (10 and 3 mg / kg), and combination treatment of 35 mg / kg compound 1 with either of the two doses of tafasitamab tested, did not show any tumor growth inhibition in this model. However, treatment with the higher dose of compound 1 (50 mg / kg) plus both doses of tafasitamab (10 and 3 mg / kg) showed a combination effect. Compared to vehicle-treated controls, the combination of 50 mg / kg compound 1 and 10 mg / kg tafasitamab resulted in 26% tumor growth inhibition, and the combination of 50 mg / kg compound 1 and 3 mg / kg tafasitamab resulted in 36% tumor growth inhibition. Body weight progressed normally in all animals in this study (data not shown).

Claims

1. A method for treating cancer, comprising administering to a human subject in need thereof a combination of an EZH2 modulator and an anti-CD19 antibody.

2. The EZH2 modulator has the formula: 【Chemical 1】 or a pharmaceutically acceptable salt thereof.

3. The method of any one of claims 1 to 2, wherein the anti-CD19 antibody comprises a heavy chain variable region comprising an HCDR1 region comprising the sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising the sequence NPYNDG (SEQ ID NO: 2), and an HCDR3 region comprising the sequence GTYYYGTRVFDY (SEQ ID NO: 3), and a light chain variable region comprising an LCDR1 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising the sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6).

4. the anti-CD19 antibody comprises a heavy chain variable region of EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSS (SEQ ID NO: 7); and The method of any one of claims 1 to 3, comprising the light chain variable region of DIVMTQSPATHLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIK (SEQ ID NO: 8).

5. 6. The method of claim 1, wherein the anti-CD19 antibody comprises an Fc domain comprising an amino acid substitution at positions S239 and / or I332, and wherein the numbering is according to the EU index in Kabat.

6. 5. The method of claim 1, wherein the anti-CD19 antibody comprises an Fc domain comprising an amino acid substitution of S239D and an amino acid substitution of I332E, and the numbering is according to the EU index in Kabat.

7. The anti-CD19 antibody is EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPDVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQPREPQVYTLP The heavy chain region of PSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 11), and DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGS 7. The method of any one of claims 1 to 6, comprising the light chain region of GTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12).

8. The method of any one of claims 1 to 7, wherein the EZH2 modulator is administered orally.

9. The method of any one of claims 1 to 8, wherein the EZH2 modulator is administered intravenously or subcutaneously.

10. 10. The method of any one of claims 1 to 9, wherein the EZH2 modulator is administered by intravenous infusion.

11. The method according to any one of claims 1 to 10, wherein the cancer is a CD19-positive cancer.

12. The method according to any one of claims 1 to 11, wherein the cancer is a hematological tumor.

13. The method of any one of claims 1 to 12, wherein the cancer is lymphoma or leukemia.

14. The method of any one of claims 1 to 13, wherein the cancer is chronic lymphocytic leukemia, acute lymphoblastic leukemia, or non-Hodgkin's lymphoma.

15. 15. The method of any one of claims 1 to 14, wherein the cancer is a non-Hodgkin's lymphoma selected from the group consisting of follicular lymphoma (FL), small lymphocytic lymphoma, mucosa-associated lymphoid tissue lymphoma, marginal zone lymphoma (MZL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), and Burkitt's lymphoma.

Images