Bispecific antibodies against CD3 and CD20 in combination therapy for treating diffuse large B-cell lymphoma

Abstract

Methods for the clinical treatment of diffuse large B-cell lymphoma (e.g., relapsed and / or refractory diffuse large B-cell lymphoma) in human subjects are provided using a bispecific antibody that binds CD3 and CD20, in combination with lenalidomide or ibrutinib and lenalidomide.

Description

[Technical field]

[0001] The present invention relates to bispecific antibodies targeting both CD3 and CD20 and the use of such antibodies in combination with lenalidomide, or in combination with lenalidomide and ibrutinib, for the treatment of diffuse large B-cell lymphoma (DLBCL), such as relapsed and / or refractory DLBCL. Advantageous treatment regimens are also provided. [Background technology]

[0002] DLBCL is the most common non-Hodgkin's lymphoma (NHL), and the standard first-line therapy is R-CHOP. The cure rate of this combination for the entire population of newly diagnosed DLBCL is 60%-70% (Sehn et al., Blood 2007;109:1867-61). Attempts to improve outcomes of first-line therapy, including dose escalation and the addition of other agents to enhance the regimen, have failed to provide sufficient evidence to change the standard of care.

[0003] Risk factors influencing the rate of CR to first-line treatment, disease recurrence, and OS are included in the International Prognostic Index (IPI) or revised IPI (R-IPI) and are as follows: age >60 years, ECOG >1 or KPS <60, LDH >ULN; extranodal disease >1 (≥2), and disease stage 3 or 4 (Project et al., N Engl J Med 1993;329:987-994; Sehn et al., supra). Patients in the good risk group (IPI factors of 1-2) have a 4-year PFS of 80% after standard first-line R-CHOP, whereas 45% of patients in the high risk group (IPI factors of 3-5) achieve a 4-year PFS and OS of only 55% (Sehn et al., supra). [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Sehn et al.,Blood 2007;109:1867-61 [Non-Patent Document 2] Project et al.,N Engl J Med 1993;329:987-994 Summary of the Invention [Problem to be solved by the invention]

[0005] Given the limited efficacy and long-term response of high-risk subjects to currently available treatments, new and effective treatments are needed. [Means for solving the problem]

[0006] Provided herein are methods of treating a human subject with diffuse large B-cell lymphoma (DLBCL), such as relapsed and / or refractory (R / R) DLBCL, by administering a bispecific antibody that binds CD3 and CD20 in combination with lenalidomide, or in combination with lenalidomide and ibrutinib, in a particularly advantageous clinical treatment regimen.

[0007] In one aspect, provided herein is a method of treating DLBCL, e.g., R / R DLBCL, in a human subject, comprising administering to the subject a combination of epcolitamab and lenalidomide, or epcolitamab, lenalidomide and ibrutinib, e.g., comprising administering to the subject an effective amount of lenalidomide, and epcolitamab, or ibrutinib, lenalidomide and epcolitamab.

[0008]

[0013] In one aspect, a method of treating DLBCL, e.g., R / R DLBCL, in a human subject, comprising administering to the subject a bispecific antibody (e.g., subcutaneously) and an effective amount of lenalidomide lenalidomide (e.g., orally), or an effective amount of lenalidomide (e.g., orally) and ibrutinib (e.g., orally), wherein the bispecific antibody (i) a first binding arm that binds to human CD3ε (epsilon) and comprises a first antigen-binding region comprising a variable heavy (VH) region and a variable light (VL) region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences found in the VH region sequence of SEQ ID NO: 6, and the VL region comprises the CDR1, CDR2 and CDR3 sequences found in the VL region sequence of SEQ ID NO: 7; (ii) a second binding arm that binds to human CD20 and comprises a second antigen-binding region comprising a VH region and a VL region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences found in the VH region sequence of SEQ ID NO: 13, and the VL region comprises the CDR1, CDR2 and CDR3 sequences found in the VL region sequence of SEQ ID NO: 14; Provided herein are methods of treating DLBCL, e.g., R / R DLBCL, in a human subject, where the bispecific antibody is administered at a dose of 24 mg or 48 mg, and lenalidomide, the bispecific antibody, and optionally ibrutinib, are administered in a 28 day cycle.

[0009] In some embodiments, the bispecific antibody is administered in a dose of 24 mg (or about a dose of 24 mg). In some embodiments, the bispecific antibody is administered in a dose of 48 mg (or about a dose of 48 mg).

[0010] In one embodiment, the bispecific antibody is administered at a dose of 24 mg or 48 mg once per week, for example, for 2.5 28-day cycles (weekly dosing). In some embodiments, the bispecific antibody is administered once every 4 weeks after the biweekly dosing, for example, for at least 8 28-day cycles, for example, until disease progression or unacceptable toxicity occurs. In further embodiments, a priming dose (e.g., 0.16 mg or about 0.16 mg) of the bispecific antibody is administered 2 weeks prior to administering the first weekly dose of 24 mg or 48 mg. In some embodiments, after administering the priming dose, an intermediate dose (e.g., 0.8 mg or about 0.8 mg) of the bispecific antibody is administered before administering the weekly dose of 24 mg or 48 mg. In some embodiments, the priming dose is administered 1 week prior to the intermediate dose, which is administered 1 week prior to the first weekly dose of 24 mg or 48 mg.

[0011] In some embodiments, ibrutinib is administered once a day (daily administration) in a 28 day cycle, e.g., in up to 24 28 day cycles, or in at least 24 28 day cycles, e.g., in 24 28 day cycles. In one embodiment, ibrutinib is administered at a dose of about 420 mg / day, e.g., 420 mg / day. In one embodiment, ibrutinib is administered at a dose of about 560 mg / day, e.g., 560 mg / day.

[0012] In some embodiments, lenalidomide is administered once daily on days 1 through 21 of a 28 day cycle, e.g., cycles 1 through 12 of a 28 day cycle. In some embodiments, lenalidomide is administered at a dose of about 25 mg, e.g., 25 mg, on cycles 1 through 12 of a 28 day cycle. In some embodiments, lenalidomide is administered at a dose of about 20 mg, e.g., 20 mg, on cycles 1 through 24 of a 28 day cycle.

[0013] In some embodiments, lenalidomide and optionally ibrutinib, and the bispecific antibody are administered on the same day (e.g., days 1, 8, 15, and 22 of cycles 1-3, and 1 of cycles 4-12, or days 1, 8, 15, and 22 of cycles 1-3, and day 1 of cycles 4-24).

[0014] In some embodiments, the administration occurs in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) in cycles 4 and beyond, a dose of 24 mg is administered on day 1, and so on; (b) lenalidomide was administered on days 1-21 of Cycle 1 or later; and (c) Ibrutinib is optionally administered on days 1-28 after Cycle 1.

[0015] In some embodiments, the administration occurs in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 12, a dose of 24 mg was administered subcutaneously on day 1, as follows: (b) Lenalidomide will be administered orally at a dose of 25 mg / day on days 1-21 of cycles 1-12.

[0016] In some embodiments, the administration occurs in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 24 mg is administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib will be administered orally at a dose of 560 mg / day on days 1-28 of cycles 1-24.

[0017] In some embodiments, the administration occurs in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 24 mg is administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib will be administered orally at a dose of 420 mg / day on days 1-28 of cycles 1-24.

[0018] In some embodiments, the administration occurs in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) in cycles 3 and beyond, a dose of 48 mg is administered on day 1, and so on; (b) lenalidomide was administered on days 1-21 of Cycle 1 or later; and (c) Ibrutinib is optionally administered on days 1-28 after Cycle 1.

[0019] In some embodiments, the administration occurs in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 12, a dose of 48 mg was administered subcutaneously on day 1, as follows: (b) Lenalidomide will be administered orally at a dose of 25 mg / day on days 1-21 of cycles 1-12.

[0020] In some embodiments, the administration occurs in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 48 mg was administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib will be administered orally at a dose of 560 mg / day on days 1-28 of cycles 1-24.

[0021] In some embodiments, the administration occurs in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 48 mg was administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib will be administered orally at a dose of 420 mg / day on days 1-28 of cycles 1-24.

[0022] In some embodiments, the bispecific antibody is administered subcutaneously. In some embodiments, ibrutinib is administered orally. In some embodiments, lenalidomide is administered orally.

[0023] In some embodiments, the DLBCL has histologically confirmed CD20+ disease. In some embodiments, the DLBCL is an aggressive B-cell lymphoma with MYC and BCL-2 and / or BCL-6 translocations (double hit or triple hit). In some embodiments, the DLBCL is follicular lymphoma grade 3B. In some embodiments, the DLBCL is relapsed and / or refractory DLBCL.

[0024] In some embodiments, the first antigen-binding region of the bispecific antibody comprises VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences set forth in SEQ ID NO:1, 2, and 3, respectively, and VLCDR1, VLCDR2, and VLCDR3 comprising the amino acid sequences set forth in SEQ ID NO:4, sequence GTN, and SEQ ID NO:5, respectively, and the second antigen-binding region comprises VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences set forth in SEQ ID NO:8, 9, and 10, respectively, and VLCDR1, VLCDR2, and VLCDR3 comprising the amino acid sequences set forth in SEQ ID NO:11, sequence DAS, and SEQ ID NO:12, respectively. In some embodiments, the first antigen-binding region of the bispecific antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO:6, and a VL region comprising the amino acid sequence of SEQ ID NO:7, and the second antigen-binding region comprises a VH region comprising the amino acid sequence of SEQ ID NO:13, and a VL region comprising the amino acid sequence of SEQ ID NO:14.

[0025] In some embodiments, the first binding arm of the bispecific antibody is derived from a humanized antibody, preferably a full-length IgG1, λ (lambda) antibody (e.g., SEQ ID NO: 22). In some embodiments, the second binding arm of the bispecific antibody is derived from a human antibody, preferably a full-length IgG1, κ (kappa) antibody (e.g., SEQ ID NO: 23). In some embodiments, the bispecific antibody is a full-length antibody with a human IgG1 constant region.

[0026] In some embodiments, a bispecific antibody comprises an inactive Fc region, e.g., an Fc region in which the amino acids at positions corresponding to positions L234, L235, and D265 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 are F, E, and A, respectively. In some embodiments, a bispecific antibody comprises substitutions that promote the formation of a bispecific antibody, e.g., in a first heavy chain, the amino acid at position corresponding to F405 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is L and in a second heavy chain, the amino acid at position corresponding to K409 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is R, or vice versa. In some embodiments, a bispecific antibody has both an inactive Fc region (e.g., substitutions at L234, L235, and D265 (e.g., L234F, L235E, and D265A)) and substitutions that promote bispecific antibody formation (e.g., F405L and K409R). In a further embodiment, the bispecific antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NOs: 19 and 20.

[0027] In some embodiments, the bispecific antibody comprises a first heavy chain and a first light chain comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 24 and 25, respectively, and a second heavy chain and a second light chain comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 26 and 27, respectively. In some embodiments, the bispecific antibody is epcolitamab, or a biosimilar thereof. [Brief description of the drawings]

[0028] [Figure 1] Figure 1 shows T cell activation in the presence of lenalidomide. T cells were pre-incubated for 3 days with assay medium or lenalidomide in the absence or presence of CD3 cross-linking with immobilized anti-CD3 (indicated on the x-axis), after which upregulation of CD69, CD25, PD-1 and LAMP-1 on CD4+ and CD8+ T cells was determined by flow cytometry. Data shown are geometric mean fluorescence intensity (FI) for four donors (each circle represents one condition for one donor). Pre-incubation conditions are indicated on the x-axis. [Diagram 2] Figure 1 shows the effect of lenalidomide on Duobody®-CD3xCD20-induced T cell-mediated cytotoxicity against CD20-expressing Daudi cells. T cells were incubated with lenalidomide (5 or 50 μM) or without lenalidomide for 3 days in the absence or presence of immobilized anti-CD3. T cells were then used in a cytotoxicity assay with DuoBody®-CD3xCD20 or DuoBody®-CD3xctrl (containing CD3 arms and non-binding control) and CD20-expressing Daudi cells as target cells (E:T ratio 2:1). Data shown are the mean percentage of cytotoxicity ± SD of replicates, normalized to media control (no antibody, no lenalidomide). [Diagram 3] FIG. 1 is a schematic diagram of the overall clinical trial design. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] definition The term "immunoglobulin" as used herein refers to a class of structurally related glycoproteins consisting of two pairs of polypeptide chains, one pair of light (L) low molecular weight chains and one pair of heavy (H) chains, all four interconnected by disulfide bonds. The structure of immunoglobulins is well characterized (see, e.g., Fundamental Immunology Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, NY (1989)). Briefly, each heavy chain typically comprises a heavy chain variable region (herein referred to as VH or VH), which is a nucleotide sequence that is linked to the heavy chain variable region by a ligand such as a nucleotide sequence. H and a heavy chain constant region (abbreviated as CH or C herein) H The heavy chain constant region is typically composed of three domains, CH1, CH2, and CH3. The hinge region is the region between the CH1 and CH2 domains of the heavy chain and is very flexible. Disulfide bonds in the hinge region are part of the interaction between the two heavy chains in an IgG molecule. Each light chain typically comprises a light chain variable region (herein VL or V Land a light chain constant region (abbreviated herein as CL or C L The light chain constant region is typically composed of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariable regions (or hypervariable regions that may be hypervariable in sequence and / or in the form of structurally defined loops), also called complementarity determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL is typically composed of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 (see also Chothia and Lesk J Mol Biol 1987;196:901-17). Unless otherwise indicated or inconsistent with the context, CDR sequences herein are identified according to the IMGT rules (Brochet X., Nucl Acids Res 2008; 36: W503-508; Lefranc MP., Nucl Acids Res 1999; 27: 209-12; www.imgt.org / ). Unless otherwise indicated or inconsistent with the context, references to amino acid positions within a constant region are according to EU numbering (Edelman et al., PNAS. 1969; 63: 78-85; Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition. 1991 NIH Publication No. 91-3242). For example, SEQ ID NO: 15 sets forth amino acids 118 to 447 of the IgG1 heavy chain constant region according to the EU numbering system.

[0030] The term "amino acid corresponding to position ..." as used herein refers to the amino acid position number of the human IgG1 heavy chain. Corresponding amino acid positions in other immunoglobulins can be found by alignment with human IgG1. Thus, an amino acid or segment in one sequence that "corresponds" to an amino acid or segment in another sequence is one that aligns with the other amino acid or segment, typically with default settings, using a standard sequence alignment program such as ALIGN, ClustalW or the like, and has at least 50%, at least 80%, at least 90%, or at least 95% identity to the human IgG1 heavy chain. It is within the capabilities of the skilled artisan to align sequences or segments in a sequence and thereby determine the corresponding positions in the sequence for the amino acid positions according to the invention.

[0031] The term "antibody" (Ab) as used herein in the context of the present invention refers to an immunoglobulin molecule that has the ability to specifically bind to an antigen under typical physiological conditions with a half-life of sufficient duration, such as at least about 30 minutes, at least about 45 minutes, at least about 1 hour, at least about 2 hours, at least about 4 hours, at least about 8 hours, at least about 12 hours, about 24 hours or more, about 48 hours or more, about 3, 4, 5, 6, 7 days or more, etc., or any other relevant functionally defined period (e.g., a sufficient time to induce, promote, enhance, and / or modulate a physiological response associated with antibody binding to the antigen and / or a sufficient time for the antibody to recruit effector activity). The variable regions of the heavy and light chains of the immunoglobulin molecule contain the binding domains that interact with the antigen. The term antibody also encompasses polyclonal antibodies, monoclonal antibodies (mAbs), antibody-like polypeptides, chimeric antibodies, and humanized antibodies, unless otherwise specified.

[0032] The term "antibody fragment" or "antigen-binding fragment" as used herein refers to a fragment of an immunoglobulin molecule that retains the ability to specifically bind to an antigen and can be produced by any known technique, such as enzymatic cleavage, peptide synthesis, and recombinant techniques. Examples of antibody fragments include: (i) Fab' or Fab fragments, monovalent fragments consisting of the VL, VH, CL and CH1 domains, or monovalent antibodies (Genmab) as described in WO2007059782; (ii) F(ab')2 fragments, bivalent fragments comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) Fd fragments consisting essentially of the VH and CH1 domains; (iv) Fv fragments consisting essentially of the VL and VH domains of a single arm of an antibody; (v) dAb fragments, also called domain antibodies, consisting essentially of the VH domain (Ward et al., Nature 1989;341:544-46) (Holt et al; Trends Biotechnol 2003;21:484-90); (vi) camelids or nanobodies (Revets et al; Expert Opin Biol Ther 2005;5:111-24) and (vii) isolated complementarity determining regions (CDRs). Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be linked by a synthetic linker that allows them to be produced using recombinant methods as a single protein chain in which the VL and VH regions pair to form a monovalent molecule (known as single chain antibodies or single chain Fvs (scFvs), see, for example, Bird et al., Science 1988;242:423-26 and Huston et al., PNAS 1988;85:5879-83). Such single chain antibodies are encompassed by the term antibody fragment, unless otherwise stated or clearly indicated by the context.

[0033] The term "antibody binding region" or "antigen binding region" as used herein refers to the region that interacts with an antigen and includes both the VH and VL regions. The term antibody as used herein refers to monospecific antibodies as well as multispecific antibodies that include multiple, e.g., two or more, e.g., three or more, different antigen binding regions. The term antigen binding region, unless otherwise specified or clearly contradicted by the context, includes antigen-binding fragments, i.e., fragments of antibodies that retain the ability to specifically bind to an antigen.

[0034] As used herein, the term "isotype" refers to an immunoglobulin class (e.g., IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM) encoded by heavy chain constant region genes. When a particular isotype, e.g., IgG1, is referred to, the term is not limited to a particular isotype sequence, e.g., a particular IgG1 sequence, but is used to indicate that the antibody is closer in sequence to that isotype, e.g., IgG1, than to other isotypes. Thus, for example, an IgG1 antibody may be a sequence variant of a naturally occurring IgG1 antibody, which may include constant region mutations.

[0035] The term "bispecific antibody" or "bs" or "bsAb" as used herein refers to an antibody that has two different antigen-binding regions defined by different antibody sequences. Bispecific antibodies can be of any format.

[0036] As used herein, the terms "half molecule," "Fab arm," and "arm" refer to one heavy-light chain pair.

[0037] When a bispecific antibody is described as comprising a half molecule antibody "derived from" a first parent antibody and a half molecule antibody "derived from" a second parent antibody, the term "derived from" indicates that the bispecific antibody was generated by recombining, by any known method, the half molecules from each of the first and second parent antibodies into the resulting bispecific antibody. In this context, "recombinant" is not intended to be limited by any particular recombination method and thus includes all of the methods for making bispecific antibodies described herein, including, for example, recombination by half molecule exchange (also known as "controlled Fab arm exchange"), as well as recombination at the nucleic acid level and / or by co-expression of two half molecules in the same cell.

[0038] The term "full length" as used herein in reference to an antibody indicates that the antibody is not a fragment, but contains all of the domains of a particular isotype that are normally found in that isotype in nature, e.g., the VH, CH1, CH2, CH3, hinge, VL and CL domains of an IgG1 antibody. Full length antibodies can be engineered. An example of a "full length" antibody is epcolitamab.

[0039] The term "Fc region" as used herein refers to the region of an antibody that consists of the Fc sequences of two immunoglobulin heavy chains, said Fc sequences including at least the hinge region, the CH2 domain, and the CH3 domain.

[0040] As used herein, the term "heterodimeric interaction between a first CH3 region and a second CH3 region" refers to an interaction between a first CH3 region and a second CH3 region in a first CH3 / second CH3 heterodimeric protein.

[0041] As used herein, the term "homodimeric interaction of a first CH3 region and a second CH3 region" refers to an interaction between a first CH3 region and another first CH3 region in a first CH3 / first CH3 homodimeric protein, and an interaction between a second CH3 region and another second CH3 region in a second CH3 / second CH3 homodimeric protein.

[0042] The term "binding" as used herein in reference to the binding of an antibody to a given antigen typically refers to a binding rate of about 10, as determined by, for example, Biolayer Interferometry (BLI) technology on an Octet HTX instrument using the antibody as the ligand and the antigen as the analyte. -6 M or less, e.g. 10 -7 M or less, for example, about 10 -8 M or less, for example, about 10 -9 M or less, about 10 -10 M or less, or about 10 -11 K below M D and an antibody will bind with an affinity corresponding to a K for binding to a non-specific antigen other than the given antigen or a closely related antigen (e.g., BSA, casein). D at least 10 times lower, such as at least 100 times lower, such as at least 1,000 times lower, such as at least 10,000 times lower, such as at least 100,000 times lower, D binds to a given antigen with an affinity corresponding to the K D A lower amount of the antibody D Since it depends on the antibody's K D If the K of binding to the antigen is very low, D is the K for nonspecific antigen binding. D The amount lower may be at least 10,000 fold (ie, the antibody is highly specific).

[0043] The term "isolated antibody" as used herein refers to an antibody that is substantially free of other antibodies having different antigen specificities. In a preferred embodiment, an isolated bispecific antibody that specifically binds to CD20 and CD3 is further substantially free of monospecific antibodies that specifically bind to CD20 or CD3.

[0044] As used herein, the term "CD3" refers to the human cluster of differentiation 3 protein, which is part of the T cell coreceptor protein complex and is composed of four distinct chains. Because CD3 is found in other species, the term "CD3" is not limited to human CD3, unless consistent with the context. In mammals, the complex comprises the CD3 gamma (gamma) chain (human CD3 gamma chain UniProtKB / Swiss-Prot No P09693, or cynomolgus monkey CD3 gamma UniProtKB / Swiss-Prot No Q95LI7), the CD3 delta (delta) chain (human CD3 delta UniProtKB / Swiss-Prot No P04234, or cynomolgus monkey CD3 delta UniProtKB / Swiss-Prot No Q95LI8), two CD3 epsilon (epsilon) chains (human CD3 epsilon UniProtKB / Swiss-Prot No P07766, SEQ ID NO: 28; cynomolgus monkey CD3 epsilon UniProtKB / Swiss-Prot No Q95LI5; or rhesus monkey CD3 epsilon UniProtKB / Swiss-Prot No G7NCB9), and the CD3 zeta (zeta) chain (human CD3 These chains associate with molecules known as T cell receptors (TCRs) to generate activation signals in T lymphocytes. The TCR and CD3 molecules together comprise the TCR complex.

[0045] As used herein, the term "CD3 antibody" or "anti-CD3 antibody" refers to an antibody that specifically binds to the antigen CD3, in particular human CD3ε (epsilon).

[0046] The term "human CD20" or "CD20" refers to human CD20 (UniProtKB / Swiss-Prot No P11836, SEQ ID NO: 29) and includes any variants, isoforms, and species homologs of CD20 naturally expressed by cells, including tumor cells, or expressed on cells transfected with the CD20 gene or cDNA. Species homologs include rhesus monkey CD20 (macaca mulatta; UniProtKB / Swiss-Prot No H9YXP1) and cynomolgus monkey CD20 (macaca fascicularis; UniProtKB No G7PQ03).

[0047] As used herein, the term "CD20 antibody" or "anti-CD20 antibody" refers to an antibody that specifically binds to the antigen CD20, in particular human CD20.

[0048] As used herein, the term "CD3xCD20 antibody," "anti-CD3xCD20 antibody," "CD20xCD3 antibody" or "anti-CD20xCD3 antibody" refers to a bispecific antibody that contains two different antigen-binding regions, one of which specifically binds to the antigen CD20 and one of which specifically binds to CD3.

[0049] The term "DuoBody®-CD3xCD20" as used herein refers to an IgG1 bispecific CD3xCD20 antibody comprising a first heavy and light chain pair defined in SEQ ID NO: 24 and SEQ ID NO: 25, respectively, and a second heavy and light chain pair defined in SEQ ID NO: 26 and SEQ ID NO: 27. The first heavy and light chain pair comprises a region that binds to human CD3ε (epsilon), and the second heavy and light chain pair comprises a region that binds to human CD20. The first binding region comprises the VH and VL sequences defined by SEQ ID NO: 6 and 7, and the second binding region comprises the VH and VL sequences defined by SEQ ID NO: 13 and 14. This bispecific antibody can be prepared as described in WO2016 / 110576.

[0050] Antibodies comprising functional variants of the heavy chain, light chain, VL region, VH region, or one or more CDRs of the example antibodies are also provided herein. A functional variant of the heavy chain, light chain, VL, VH, or CDR used in the context of an antibody still allows the antibody to retain at least a significant proportion (at least about 90%, 95% or more) of the functional characteristics of the "reference" and / or "parent" antibody, including affinity and / or specificity / selectivity for a particular epitope of CD20 and / or CD3, Fc inactivation, and PK parameters such as half-life, Tmax, Cmax, etc. Such functional variants typically retain significant sequence identity with the parent antibody and / or have heavy and light chains of substantially similar length. The percent identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., % homology = number of identical positions / total number of positions x 100), taking into account the number of gaps that need to be introduced for optimal alignment of the two sequences and the length of each gap. Percent identity between two nucleotide or amino acid sequences can be determined using, for example, the algorithm of E. Meyers and W. Miller, Comput. Appl. Biosci 4,11-17 (1988), as incorporated into the ALIGN program (version 2.0), using a PAM120 weight residual table, a gap length penalty of 12, and a gap penalty of 4. Additionally, the algorithm of Needleman and Wunsch, J Mol Biol 1970;48:444-453 can be used to determine percent identity between two amino acid sequences. Exemplary variants include those that differ from the heavy and / or light chain, VH and / or VL, and / or CDR regions of a parent antibody sequence primarily by conservative substitutions, for example, 10, for example 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the substitutions in the variant can be conservative amino acid residue substitutions.

[0051] Conservative substitutions may be defined by substitutions within the classes of amino acids reflected in the table below. [Table 1]

[0052] Unless otherwise indicated, the following nomenclature is used to describe the mutations: i) substitution of an amino acid at a given position is written, for example, K409R, which means substitution of lysine at position 409 with arginine. ii) For certain variants, specific three-letter or one-letter codes are used, including the codes Xaa and X to indicate any amino acid residue. Thus, substitution of lysine with arginine at position 409 is named K409R, and substitution of lysine with any amino acid residue at position 409 is named K409X. In case of deletion of lysine at position 409, it is indicated by K409*.

[0053] The term "humanized antibody" as used herein refers to a genetically engineered non-human antibody comprising a human antibody constant domain and a non-human variable domain modified to contain a high level of sequence homology to the human variable domain. This can be achieved by grafting the six non-human antibody CDRs that together form the antigen binding site into a homologous human acceptor framework region (FR) (see WO 92 / 22653 and EP 0629240). Substitution (backmutation) of framework residues from the parent antibody (i.e., non-human antibody) into the human framework region may be required to fully reconstitute the binding affinity and binding specificity of the parent antibody. Structural homology modeling can help to identify amino acid residues in the framework region that are important for the binding properties of the antibody. Thus, a humanized antibody may comprise non-human CDR sequences, primarily human framework regions, optionally containing one or more amino acid backmutations to the non-human amino acid sequences, and a fully human constant region. As used herein, the VH and VL of the CD3 arms in DuoBody®-CD3xCD20 represent humanized antigen-binding regions. Optionally, further amino acid modifications, not necessarily back mutations, can be applied to obtain a humanized antibody with favorable characteristics such as affinity and biochemical properties.

[0054] The term "human antibody" as used herein refers to an antibody having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody" as used herein is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences. The VH and VL of the CD20 arms used in DuoBody®-CD3xCD20 represent human antigen-binding regions. The human monoclonal antibodies of the invention may be produced by a variety of techniques, including conventional monoclonal antibody methodologies, such as the standard somatic cell hybridization technique of Kohler and Milstein, Nature 256:495 (1975). Although somatic cell hybridization procedures are in principle preferred, other techniques for producing monoclonal antibodies can be used, for example viral or oncogenic transformation of B lymphocytes or phage display techniques using libraries of human antibody genes. A suitable animal system for preparing hybridomas secreting human monoclonal antibodies is the murine system. Hybridoma production in mice is a very well established procedure. Immunization protocols and techniques for isolating immunized splenocytes for fusion are known in the art. Fusion partners (e.g. mouse myeloma cells) and fusion procedures are also known. Thus, human monoclonal antibodies can be made, for example, using transgenic or transchromosomal mice or rats carrying parts of the human immune system rather than the mouse or rat system. Thus, in one embodiment, human antibodies are obtained from transgenic animals (e.g. mice or rats) that have human germline immunoglobulin sequences instead of animal immunoglobulin sequences.In such embodiments, the antibody is derived from human germline immunoglobulin sequences introduced into the animal, with the final antibody sequence being the result of further modification of said human germline immunoglobulin sequences by somatic hypermutation and affinity maturation by the endogenous animal antibody machinery (see, e.g., Mendez et al. Nat Genet 1997;15:146-56). The VH and VL regions of the CD20 arms used in DuoBody®-CD3xCD20 represent human antigen-binding regions.

[0055] The term "biosimilar" (e.g., an approved reference product / biological drug) as used herein refers to a biological product that is similar to the reference product based on (a) analytical studies showing that the biological product is highly similar to the reference product, despite minor differences in clinically inactive ingredients; (b) animal testing (including evaluation of toxicity); and / or (c) data from one or more clinical trials (including evaluation of immunogenicity and pharmacokinetics or pharmacodynamics) sufficient to demonstrate safety, purity, and potency (e.g., no clinically significant differences in product safety, purity, and potency between the biological product and the reference product) for one or more appropriate conditions of use for which the reference product has been approved, intended for use, and for which approval is sought. In some embodiments, the biosimilar biological product and the reference product utilize the same mechanism(s) of action for one or more conditions of use prescribed, recommended, or suggested in the proposed labeling, but only to the extent that the mechanism(s) of action are known for the reference product. In some embodiments, one or more conditions of use prescribed, recommended, or suggested in the proposed labeling for the biological product have been previously approved for the reference product. In some embodiments, the route of administration, dosage form, and / or strength of the biological product is the same as that of the reference product. A biosimilar can be, for example, a currently known antibody that has the same primary amino acid sequence as a commercially available antibody, but may be made in a different cell type or by a different production, purification, or formulation method.

[0056] The term "reducing conditions" or "reducing environment" as used herein refers to a state or environment in which a substrate, here a cysteine ​​residue in the hinge region of an antibody, is more likely to be reduced than oxidized.

[0057] The term "recombinant host cell" (or simply "host cell"), as used herein, is intended to refer to a cell into which an expression vector has been introduced, such as an expression vector encoding an antibody described herein. Recombinant host cells include, for example, transfectomas, such as CHO, CHO-S, HEK, HEK293, HEK-293F, Expi293F, PER.C6 or NS0 cells, and lymphocytic cells.

[0058] As used herein, the term "diffuse large B-cell lymphoma" or "DLBCL" refers to a neoplasm of germinal center B lymphocytes with a diffuse growth pattern and a high intermediate proliferation index. DLBCL represents approximately 30% of all lymphomas. Subtypes of DLBCL appear to have different outlooks (prognoses) and responses to treatment. DLBCL can affect any age group, but occurs primarily in older adults (the average age is in the mid-60s). "Double hit" and "triple hit" DLBCL refer to DLBCL with MYC and BCL2 and / or BCL6 translocations, which belong to the category of high-grade B-cell lymphoma (HGBCL) with MYC and BCL2 and / or BCL6 translocations according to the WHO 2016 classification (Swerdlow SH, Campo E, Harris NL, et al. WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues (Revised ed.4th). Lyon, France: IARC Press (2017)). Follicular lymphoma grade 3B is also often considered equivalent to DLBCL and is therefore treated as described above.

[0059] As used herein, the term "recurrent diffuse large B-cell lymphoma" or "recurrent DLBCL" refers to diffuse large B-cell lymphoma that previously responded to treatment but has progressed 6 months or more after completing treatment.

[0060] As used herein, the term "refractory diffuse large B-cell lymphoma" or "refractory DLBCL" refers to diffuse large B-cell lymphoma that has progressed during treatment, failed to achieve an objective response to previous treatment, or progressed within 6 months of completing treatment (including maintenance therapy). As used herein, the term "R / R diffuse large B-cell lymphoma" or "R / R DLBCL" is intended to refer to relapsed and / or refractory diffuse large B-cell lymphoma, unless otherwise specified.

[0061] As used herein, the term "ibrutinib" refers to a compound of the chemical formula C 25 -H 24 -N6-O2 and the chemical name: 1-((3R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo(3,4-d)pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (Chemical Abstracts Service No. 936563-96-1). Ibrutinib is available, for example, under the trade name Imbruvica®. The term "ibrutinib" is also intended to encompass branded and generic versions (generic equivalents) of ibrutinib, as well as pharmaceutically acceptable salts, isomers, racemates, solvates, complexes and hydrates, anhydrous forms, and any polymorphic or amorphous forms thereof, or combinations thereof.

[0062] As used herein, the term "lenalidomide" has the chemical formula C 13 H 13It refers to a thalidomide derivative having N3O3 and the chemical name: 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione (Chemical Abstracts Service No. 191732-72-6). Lenalidomide is available, for example, under the trade name Revlimid®. The term "lenalidomide" is also intended to encompass branded and generic versions (generic equivalents) of lenalidomide, as well as pharmaceutically acceptable salts, isomers, racemates, solvates, complexes and hydrates, anhydrous forms, and any polymorphic or amorphous forms thereof, or combinations thereof.

[0063] The term "treatment" refers to the administration of an effective amount of a therapeutically active antibody as described herein for the purpose of alleviating, ameliorating, arresting or eradicating (curing) a symptom or condition, such as DLBCL. Treatment may result in a complete response (CR), partial response (PR), or stable disease (SD), e.g., as defined by the Lugano criteria and / or LYRIC. Treatment may be continued, e.g., until disease progression or unacceptable toxicity.

[0064] The term "administering" or "administration" as used herein refers to the physical introduction of a composition (or formulation) containing a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those of skill in the art. Preferred routes of administration for the antibodies described herein include intravenous, intraperitoneal, intramuscular, subcutaneous, spinal or other parenteral routes of administration, for example by injection or infusion. The phrase "parenteral administration" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion, and in vivo electroporation. Alternatively, the therapeutic agents described herein can be administered via a parenteral route, such as a topical, epidermal or mucosal route of administration, for example, intranasal, oral, intravaginal, rectal, sublingual or topical. Administration can also be, for example, once, multiple times, and / or over one or more extended periods of time. In the methods described herein, the bispecific antibody (e.g., epcolitamab) is administered subcutaneously. Other agents used in combination with the bispecific antibody, such as for cytokine release syndrome prophylaxis and / or tumor lysis syndrome (TLS) prophylaxis, can be administered via other routes, such as intravenously or orally.

[0065] The term "effective amount" or "therapeutically effective amount" refers to an amount effective at the dosage and duration required to achieve the desired therapeutic result. For example, the dosages defined herein for a bispecific antibody (e.g., epcolitamab) administered subcutaneously, i.e., 24 mg or 48 mg, can be defined as such an "effective amount" or "therapeutically effective amount". The therapeutically effective amount of an antibody can vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the antibody to elicit a desired response in the individual. A therapeutically effective amount is also an amount in which any toxic or adverse effects of the antibody or antibody portion are outweighed by therapeutically beneficial effects. In some embodiments, patients treated with the methods described herein show improved ECOG performance status. A therapeutically effective amount or dosage of a drug includes a "prophylactically effective amount" or "prophylactically effective dosage", which is any amount of a drug that inhibits the onset or recurrence of a disease when administered alone or in combination with another therapeutic agent to a subject at risk of developing a disease or disorder (e.g., cytokine release syndrome) or at risk of suffering from a recurrence of the disease.

[0066] As used herein, the term "inhibiting the growth" of a tumor includes any measurable reduction in tumor growth, such as an inhibition of tumor growth of at least about 10%, e.g., at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 99%, or 100%.

[0067] As used herein, the term "subject" refers to a human patient, e.g., a human patient with diffuse large B-cell lymphoma. The terms "subject" and "patient" are used interchangeably herein.

[0068] The term "buffering agent" as used herein refers to a pharma- ceutically acceptable buffering agent. The term "buffering agent" encompasses agents that maintain the pH value of a solution within an acceptable range, including, but not limited to, acetate, histidine, TRIS (registered trademark) (tris(hydroxymethyl)aminomethane), citrate, succinate, glycolate, and the like. In general, a "buffering agent" as used herein has a pKa and buffering capacity suitable for a pH range of about 5 to about 6, preferably about 5.5.

[0069] As used herein, "progressive disease" or "PD" refers to a situation in which one or more indicators of diffuse large B-cell lymphoma indicate that the disease is progressing despite treatment. In one embodiment, progressive disease is defined based on the Lugano Response Criteria for Malignant Lymphoma ("Lugano Criteria") and / or the Lymphoma Response to Immunomodulatory Therapy Criteria (LYRIC). Details regarding the Lugano criteria / classification system, including definitions of complete response (CR), partial response (PR), no response / stable disease (NR.SD), and progressive disease (PD), are provided in Cheson et al. J Clin Oncol 2014;32:3059-68, the contents of which are incorporated herein by reference (see in particular Table 3 of Cheson et al., 2014). Further details regarding the Lugano criteria / classification system are provided in Table 3.

[0070] As used herein, a "surfactant" is a compound typically used in pharmaceutical formulations to prevent drug adsorption and / or aggregation on surfaces. Additionally, surfactants reduce the surface tension (or interfacial tension) between two liquids or between a liquid and a solid. For example, exemplary surfactants can significantly reduce surface tension when present at very low concentrations (e.g., 5% w / v or less, e.g., 3% w / v or less, e.g., 1% w / v or less, e.g., 0.4% w / v or less, e.g., less than 0.1% w / v, e.g., 0.04% w / v). Surfactants are amphiphilic, which means that they are usually composed of both hydrophilic and hydrophobic or lipophilic groups and thus can form micelles or similar self-assembled structures in aqueous solutions. Known surfactants for pharmaceutical use include glycerol monooleate, benzethonium chloride, docusate sodium, phospholipids, polyethylene alkyl ethers, sodium lauryl sulfate and tricaprylin (anionic surfactants); benzalkonium chloride, cytrimide, cetylpyridinium chloride and phospholipids (cationic surfactants); and alpha tocopherol, glycerol monooleate, myristyl alcohol, phospholipids, poloxamers, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stellarate, polyoxyl hydroxystearates, polyoxyl glycerides, polysorbates such as polysorbate 20 or polysorbate 80, propylene glycol dilaurate, propylene glycol monolaurate, sorbitan esters sucrose palmitate, sucrose stearate, tricaprylin and TPGS (nonionic and zwitterionic surfactants).

[0071] As used herein, a "diluent" is one that is pharma- ceutically acceptable (safe and non-toxic for administration to humans) and useful for preparing a diluent of a pharmaceutical composition or pharmaceutical formulation (the terms "composition" and "formulation" are used interchangeably herein). Preferably, such dilution of the composition dilutes only the antibody concentration and not the buffer and stabilizer. Thus, in one embodiment, the diluent contains the same concentrations of buffer and stabilizer as are present in the pharmaceutical composition of the present invention. Further exemplary diluents include sterile water, bacteriostatic water for injection (BWFI), a pH buffer, preferably an acetate buffer, sterile saline, such as water for injection, Ringer's solution, or dextrose solution. In one embodiment, the diluent comprises or consists essentially of an acetate buffer and sorbitol.

[0072] As used herein, the term "about" refers to a value that is 10% or more and 10% or less of a particular value.

[0073] Diffuse large B-cell lymphoma treatment regimens Provided herein is a method of treating diffuse large B-cell lymphoma (DLBCL) in a human subject using a bispecific antibody that binds both CD3 and CD20 ("anti-CD3xCD20 antibody"), e.g., an isolated anti-CD3xCD20 antibody that binds human CD3 and human CD20, in combination with lenalidomide or in combination with lenalidomide and ibrutinib. The method is also useful, for example, for treating relapsed and / or refractory diffuse large B-cell lymphoma (R / R diffuse large B-cell lymphoma). It is understood that the method of treating diffuse large B-cell lymphoma (e.g., R / R diffuse large B-cell lymphoma) with a bispecific antibody that binds both CD3 and CD20 described herein also encompasses the corresponding use of the bispecific antibody to treat diffuse large B-cell lymphoma (e.g., R / R diffuse large B-cell lymphoma) in a human subject.

[0074] Thus, in one aspect, a method of treating diffuse large B-cell lymphoma in a human subject comprising administering a bispecific antibody and an effective amount of lenalidomide (e.g., orally), a bispecific antibody and an effective amount of ibrutinib (e.g., orally) and lenalidomide (e.g., orally), wherein the bispecific antibody: (i) a first binding arm that binds to human CD3ε (epsilon) and comprises a first antigen-binding region comprising a variable heavy (VH) region and a variable light (VL) region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences found in the VH region sequence of SEQ ID NO: 6, and the VL region comprises the CDR1, CDR2 and CDR3 sequences found in the VL region sequence of SEQ ID NO: 7; (ii) a second binding arm that binds to human CD20 and comprises a second antigen-binding region comprising a VH region and a VL region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences found in the VH region sequence of SEQ ID NO: 13, and the VL region comprises the CDR1, CDR2 and CDR3 sequences found in the VL region sequence of SEQ ID NO: 14; Provided herein are methods, wherein the bispecific antibody is administered at a dose of 24 mg or 48 mg (or at a dose of about 24 mg or about 48 mg), and the lenalidomide, or lenalidomide and ibrutinib, and the bispecific antibody are administered in a 28 day cycle.

[0075] In some embodiments, the bispecific antibody is a full-length antibody. In some embodiments, the bispecific antibody is an antibody with an inactive Fc region. In some embodiments, the bispecific antibody is a full-length antibody with an inactive Fc region.

[0076] In some embodiments, the bispecific antibody is administered in a dose of 24 mg (or about a dose of 24 mg). In some embodiments, the bispecific antibody is administered in a dose of 48 mg (or about a dose of 48 mg).

[0077] With respect to a 24 mg or 48 mg dose (or a dose of about 24 mg or about 48 mg) of a bispecific antibody being administered, or any other specific dose, it is understood that this amount refers to the amount of the bispecific antibody presenting the full-length antibody, for example, epcolitamab, as defined in the Examples section. Thus, a 24 mg dose of the bispecific antibody may be referred to as the administration of a dose of the bispecific antibody described herein, where the dose corresponds to a 24 mg dose of epcolitamab. A person skilled in the art can easily determine the amount of the antibody to be administered, for example, when the molecular weight of the antibody used is substantially different from the molecular weight of the full-length antibody, such as epcolitamab. For example, the amount of the antibody can be calculated by dividing the molecular weight of the antibody by the weight of the full-length antibody, such as epcolitamab, and multiplying the result by the specific dose described herein. To the extent that a bispecific antibody (e.g. a functional variant of DuoBody® CD3xCD20) has characteristics highly similar to DuoBody® CD3xCD20 in terms of plasma half-life, Fc inactivation, and / or binding properties to CD3 and CD20, i.e. in terms of CDR and epitope binding characteristics, such an antibody is suitable for use in the methods provided herein at the doses described for full-length antibodies such as epcolitamab.

[0078] In some embodiments, the bispecific antibody dose is administered once a week in a 28 day cycle (weekly dosing). In one embodiment, a weekly dose of 24 or 48 mg is administered in 2.5 28 day cycles (i.e., 10 times). In one embodiment, a weekly dose of 24 mg or 48 mg is administered in 2.5 28 day cycles on days 15 and 22 of cycle 1, and days 1, 8, 15, and 22 of cycles 2 and 3. In some embodiments, after the weekly dosing, the dosing interval can be reduced to once every 4 weeks. In one embodiment, administration once every four weeks may be performed over an extended period of time, for example, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least fifteen, at least twenty, or from one to twenty-five, one to fifteen, one to ten, one to five, five to twenty-five, five to fifteen, or five to ten, 28-day cycles. In a preferred embodiment, administration once every four weeks is performed for up to eight 28-day cycles, for example eight 28-day cycles, or nine 28-day cycles. In another preferred embodiment, administration once every four weeks is performed for up to twenty 28-day cycles, for example twenty 28-day cycles, or twenty-one 28-day cycles.

[0079] In one embodiment, weekly doses of the bispecific antibody are administered in 28 day cycles in cycles 1-3 (which may include priming and intermediate doses, as described below), and one dose every 4 weeks is administered from cycle 4 onwards, e.g., cycles 4-12, or cycles 4-24, or until disease progression or unacceptable toxicity is observed in the subject.

[0080] It is understood that the doses referred to herein may also be referred to as full or flat doses, for example in the above scenarios where weekly and / or every 4 weeks doses are administered at the same level. Thus, if a dose of 48 mg is selected, preferably the same dose of 48 mg is administered in each weekly administration and each every 4 weeks administration. Before administering the dose, a priming or priming and then intermediate (second priming) dose can be administered. This may be advantageous as it may help to reduce the risk and severity of cytokine release syndrome (CRS), a side effect that may occur during treatment with the bispecific anti-CD3xCD20 antibodies described herein. Such priming, or priming and intermediate doses are lower doses compared to flat or full doses.

[0081] Thus, in some embodiments, a priming dose of the bispecific antibody can be administered in cycle 1 of a 28 day cycle prior to administration of the weekly dose of 24 mg or 48 mg. In one embodiment, the priming dose is administered two weeks prior to administration of the first weekly dose of 24 mg or 48 mg in cycle 1. In one embodiment, the priming dose is 0.16 mg (or about 0.16 mg) of the full length bispecific antibody.

[0082] In some embodiments, an intermediate dose of the bispecific antibody is administered after the priming dose and before the 24 mg or 48 mg weekly doses. In one embodiment, the priming dose is administered one week before the intermediate dose (i.e., on day 1 of cycle 1), and the intermediate dose is administered one week before the first of the 24 mg or 48 mg weekly doses (i.e., on day 8 of cycle 1). In one embodiment, the intermediate dose is at or about 800 μg (0.8 mg) of the full length bispecific antibody.

[0083] The methods described herein include treating a human subject having diffuse large B-cell lymphoma (e.g., R / R diffuse large B-cell lymphoma) with a bispecific antibody that binds CD3 and CD20 in combination with a regimen of lenalidomide or lenalidomide or ibrutinib.

[0084] In some embodiments, lenalidomide, or ibrutinib and lenalidomide, are administered at doses supported by clinical studies, according to local guidelines, and / or in accordance with the relevant local labeling.

[0085] In some embodiments, ibrutinib is administered according to the product label or summary of product characteristics (see, e.g., IMBRUVICA® (ibrutinib) prescribing information available at https: / / www.accessdata.fda.gov / drugsatfda_docs / label / 2016 / 205552s007lbl.pdf). In some embodiments, ibrutinib is administered at a dose of 420 mg (or at a dose of about 420 mg). In other embodiments, ibrutinib is administered at a dose of 560 mg (or at a dose of about 560 mg). In some embodiments, a biosimilar of ibrutinib is used in place of ibrutinib in the methods described herein.

[0086] In some embodiments, lenalidomide is administered according to the product label or summary of product characteristics (see, e.g., REVLIMID® Prescribing Information available at www.accessdata.fda.gov / drugsatfda_docs / label / 2013 / 021880s034lbl.pdf).

[0087] In one embodiment, ibrutinib is administered once a day in a 28-day cycle (daily administration; 7QW). In one embodiment, the daily administration of ibrutinib is performed in at least one 28-day cycle (i.e., 4 times), such as at least 10 28-day cycles, such as at least 20 28-day cycles, such as 24 28-day cycles. In one embodiment, lenalidomide is administered according to local guidelines and local labeling. In some embodiments, lenalidomide is administered at a dose of 10 mg to 25 mg (or at a dose of about 10 mg to 25 mg). In some embodiments, lenalidomide is administered at a dose of 20 mg to 30 mg (or at a dose of about 20 mg to 30 mg). In one embodiment, lenalidomide is administered at a dose of 20 mg (or at a dose of about 20 mg). In one embodiment, lenalidomide is administered at a dose of 25 mg (or at a dose of about 25 mg). In one embodiment, lenalidomide is administered as an oral dose. In one embodiment, lenalidomide is administered as a capsule for oral administration.

[0088] In one embodiment, lenalidomide is administered once a day for 21 consecutive days (i.e., days 1-21) in a 28-day cycle, i.e., from day 1 to day 21 of the 28-day cycle. In one embodiment, lenalidomide is administered in at least one 28-day cycle, such as at least five 28-day cycles, at least ten 28-day cycles, at least fifteen 28-day cycles, at least twenty 28-day cycles, or at least twenty-four 28-day cycles. In one embodiment, lenalidomide is administered up to twelve, such as twelve 28-day cycles (i.e., days 1-21 of cycles 1-12 of a 28-day cycle). In one embodiment, lenalidomide is administered up to twenty-four 28-day cycles, such as twenty-four 28-day cycles (i.e., days 1-21 of cycles 1-24 of a 28-day cycle). In one embodiment, lenalidomide is administered at a dose of 25 mg (or at a dose of about 25 mg) on ​​days 1-21 of cycles 1-12 of a 28 day cycle. In one embodiment, lenalidomide is administered at a dose of 25 mg (or at a dose of about 25 mg) on ​​days 1-21 of cycles 1-24 of a 28 day cycle.

[0089] In certain embodiments, the bispecific antibody, ibrutinib and / or lenalidomide are administered simultaneously, hi some embodiments, the lenalidomide and the bispecific antibody are administered on the same day (e.g., days 1, 8, and 15 of cycles 1-12).

[0090] In some embodiments, ibrutinib, lenalidomide and the bispecific antibody are administered on the same day (e.g., days 1, 8, and 15 of cycles 1-21).

[0091] In some embodiments, the bispecific antibody, ibrutinib, and / or lenalidomide are administered sequentially.

[0092] In some embodiments, ibrutinib (e.g., orally), lenalidomide (e.g., orally), and the bispecific antibody (e.g., subcutaneously) are administered in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) in cycles 4 and beyond, a dose of 24 mg is administered on day 1, and so on; (b) lenalidomide is administered on days 1-21 of cycle 1 or later; (c) Ibrutinib is optionally administered on days 1-28 after Cycle 1.

[0093] In some embodiments, lenalidomide (e.g., orally) and the bispecific antibody (e.g., subcutaneously) are administered in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 12, a dose of 24 mg was administered subcutaneously on day 1, as follows: (b) Lenalidomide will be administered orally at a dose of 25 mg / day on days 1-21 of cycles 1-12.

[0094] In some embodiments, ibrutinib (e.g., orally), lenalidomide (e.g., orally), and the bispecific antibody (e.g., subcutaneously) are administered in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 24 mg is administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; (c) Ibrutinib will be administered orally at a dose of 560 mg / day on days 1-28 of cycles 1-24.

[0095] In some embodiments, ibrutinib (e.g., orally), lenalidomide (e.g., orally), and the bispecific antibody (e.g., subcutaneously) are administered in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 24 mg is administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; (c) Ibrutinib will be administered orally at a dose of 420 mg / day on days 1-28 of cycles 1-24.

[0096] In some embodiments, ibrutinib (e.g., oral), lenalidomide (e.g., oral), and the bispecific antibody (e.g., subcutaneous) are administered in a 28 day cycle. (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) in cycles 3 and beyond, a dose of 48 mg is administered on day 1, and so on; (b) lenalidomide is administered on days 1-21 of cycle 1 or later; (c) Ibrutinib is optionally administered on days 1-28 after Cycle 1.

[0097] In some embodiments, the lenalidomide (e.g., orally) and the bispecific antibody (e.g., subcutaneously) are administered in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 12, a dose of 48 mg was administered subcutaneously on day 1, as follows: (b) Lenalidomide will be administered orally at a dose of 25 mg / day on days 1-21 of cycles 1-12.

[0098] In some embodiments, ibrutinib (e.g., orally), lenalidomide (e.g., orally), and the bispecific antibody (e.g., subcutaneously) are administered in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 48 mg was administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib will be administered orally at a dose of 560 mg / day on days 1-28 of cycles 1-24.

[0099] In some embodiments, ibrutinib (e.g., orally), lenalidomide (e.g., orally), and the bispecific antibody (e.g., subcutaneously) are administered in a 28 day cycle; (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 48 mg was administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib will be administered orally at a dose of 420 mg / day on days 1-28 of cycles 1-24.

[0100] In one embodiment, the dosing of the bispecific antibody and lenalidomide in a 28 day cycle is as follows:

[0101] Bispecific antibody (subcutaneous): Cycle 1, Day 1: Priming dose (0.16 mg) Cycle 1, Day 8: Intermediate dose (0.8 mg) Cycle Days 1, 15, and 22: All doses (24 or 48 mg) Cycles 2-3, 1, 8, 15, and 22: All doses (24 or 48 mg) Cycles 4-12, Day 1: All doses (24 or 48 mg) Lenalidomide (oral): Cycles 1-12, days 1-21: 25 mg / day

[0102] In a further embodiment, the dosing of the bispecific antibody, ibrutinib and lenalidomide in a 28 day cycle is as follows:

[0103] Bispecific antibody (subcutaneous): Cycle 1, Day 1: Priming dose (0.16 mg) Cycle 1, Day 8: Intermediate dose (0.8 mg) Cycle Days 1, 15, and 22: All doses (24 or 48 mg) Cycles 2-3, 1, 8, 15, and 22: All doses (24 or 48 mg) Cycles 4-24, Day 1: All doses (24 or 48 mg) Ibrutinib (oral): Cycle days 1-24 and 1-28: 420 mg / day or 560 mg / day Lenalidomide (oral): Cycle 1-24, days 1-21: 20 mg / day

[0104] In some embodiments, the subject has DLBCL with histologically confirmed CD20+ disease.

[0105] In some embodiments, the DLBCL is an aggressive B-cell lymphoma with a MYC and BCL-2 and / or BCL-6 translocation (double hit or triple hit).

[0106] In some embodiments, the DLBCL is follicular lymphoma grade 3B.

[0107] In some embodiments, the DLBCL is relapsed and / or refractory DLBCL.

[0108] In some embodiments, the DLBCL has relapsed, i.e., it previously responded to a previous treatment but has progressed after the previous treatment, with progression beginning 6 months or more after completion of the previous treatment.

[0109] In some embodiments, the DLBCL is refractory, i.e., has either progressed during previous therapy, failed to achieve an objective response to previous therapy, or progressed within 6 months after completion of previous therapy, including maintenance therapy.

[0110] In some embodiments, the subject has disease that is relapsed or refractory to at least one prior systemic anti-lymphoma therapy containing an anti-CD20 monoclonal antibody.

[0111] In some embodiments, the DLBCL is not refractory to previous chimeric antigen receptor T cell (CAR-T) therapy.

[0112] In some embodiments, the subject has failed autologous stem cell transplantation (ASCT) or is ineligible for ASCT.

[0113] In some embodiments, the subject is not refractory to lenalidomide or ibrutinib. In the context of this embodiment, refractory is defined as follows:

[0114] Best response to previous regimen(s) with stable disease (SD) or progressive disease (PD), or Progressive disease within 6 months of completion of previous regimen(s)

[0115] In one embodiment, the subject has previously received at least one treatment with an anti-CD20 monoclonal antibody in combination with another systemic therapy.

[0116] In one embodiment, the subject has undergone prior CAR-T therapy or is ineligible for or unable to receive CAR-T therapy.

[0117] In a further embodiment, the subject has not received prior treatment with ibrutinib.

[0118] In some embodiments, the subject has an Eastern Cooperative Oncology Group (ECOG) performance status (ECOG PS) of 0, 1, or 2. Information regarding ECOG PS scores can be found, for example, in Oken et al, Am J Clin Oncol 1982 Dec;5(6):649-55).

[0119] In some embodiments, subjects have measurable disease, defined as (a) ≧1 measurable nodal disease (long axis >1.5 cm and short axis >1.0 cm) or ≧1 measurable extranodal disease (long axis >1 cm) on CT or MRI.

[0120] In one embodiment, the subject has one or more measurable sites of disease, defined as at least one measurable nodal lesion (long axis ≧1.5 cm and short axis >1.0 cm) or ≧1 measurable extranodal lesion (long axis ≧1.0 cm) on a positron emission tomography / computed tomography (PET / CT) scan and CT scan or MRI showing (one or more) PET positive lesions.

[0121] In some embodiments, the subject has laboratory values ​​that meet the following criteria prior to receiving the first dose of the bispecific antibody:

[0122] - Absolute neutrophil count (ANC) ≥ 1.0 x 109 / L (use of growth factors is permitted if evidence of bone marrow involvement is present, but subjects must not have received growth factors within 14 days prior to screening) - Hemoglobin ≥ 8.0 g / dL (RBC transfusion is permitted, but subjects must not have received a blood transfusion within 7 days prior to screening) - Platelet count ≥ 75 x 109 / L, or ≥ 50 x 109 / L if bone marrow infiltration or splenomegaly is present (platelet transfusions are permitted, but subjects must not have received a transfusion within 7 days prior to screening). - Serum aspartate transaminase (AST) or alanine transaminase (ALT) levels ≤ 3 x ULN - Total bilirubin value ≦1.5×ULN or ≦5×ULN (for subjects with disease or liver involvement of non-hepatic origin). Subjects with Gilbert's syndrome may have total bilirubin values ​​above 1.5×ULN, but direct bilirubin must be less than 2×ULN. - Estimated creatinine clearance (CrCl) ≥ 50 mL / min (calculated by the Cockcroft-Gault formula, corrected as necessary for factors such as body weight) Prothrombin time (PT) / international normalized ratio (INR) / activated partial thromboplastin time (aPTT) ≦1.5×ULN (excluding patients receiving anticoagulant therapy)

[0123] In further embodiments, the subject is: Must have a diagnosis of DLBCL (de novo or histologically transformed from follicular lymphoma or nodal marginal zone lymphoma) with histologically confirmed CD20+ disease, documented in the pathology report, including the following according to the WHO 2016 classification: Must have DLBCL, not otherwise specified (NOS) Patients must have aggressive B-cell lymphoma with MYC and BCL-2 and / or BCL-6 translocations for WHO 2016 ("double hit" or "triple hit") Note: High-grade B-cell lymphoma NOS or other double / triple-hit lymphomas (with histology inconsistent with DLBCL) are not eligible Must have follicular lymphoma grade 3B -No prior treatment with bispecific antibodies targeting CD3 and CD20 Must have one or more measurable sites of disease: Must have a positron emission tomography / computed tomography (PET / CT) scan demonstrating (one or more) PET-positive lesions and at least one measurable nodal lesion (long axis ≥ 1.5 cm and short axis > 1.0 cm) or ≥ 1 measurable extranodal lesion (long axis ≥ 1.0 cm) on a CT scan or MRI Must be eligible and have a need for treatment initiation based on symptoms and / or disease burden as assessed by the investigator. Eastern Cooperative Oncology Group (ECOG) performance status 0-2 No unresolved toxicity from prior anticancer therapy defined as unresolved Common Terminology Criteria for Adverse Events (CTCAE, v5.0), Grade 1, except for alopecia. No current evidence of primary central nervous system (CNS) tumor or known CNS involvement (including leptomeningeal disease) at the time of screening. No history of severe allergic or anaphylactic reactions to anti-CD20 mAb therapy, or known significant allergy or intolerance to any component or excipient of epcolitamab or any component of the study drug combination (e.g., lenalidomide, ibrutinib, etc.) - Patients cannot have undergone an autologous stem cell transplant within 3 months prior to screening. Patients may not have received any chemotherapy, non-investigational, or investigational antineoplastic agent (except for CD20 mAbs) within 4 weeks or 5 half-lives (whichever is shorter) prior to the first dose of epcolitamab. No clinically significant cardiovascular disease, including: Myocardial infarction or stroke within 6 months prior to enrollment or Any of the following conditions within 3 months prior to enrollment: diseases / conditions related to or affecting unstable or uncontrolled cardiac function (e.g., unstable angina, congestive heart failure, New York Heart Association class III-IV), uncontrolled cardiac arrhythmias or Other clinically significant electrocardiogram (ECG) abnormalities within 6 months prior to enrollment unless deemed stable and appropriately treated. - Absence of hepatitis, current alcohol abuse, or clinically significant liver disease, including cirrhosis. - Not having active Hepatitis B virus (HBV) or Hepatitis C virus (HCV) infection.

[0124] Subjects who are positive for hepatitis B core antibody (HBcAb), hepatitis B surface antigen (HBsAg), or hepatitis C antibody must have a negative polymerase chain reaction (PCR) result prior to enrollment. Those with a positive PCR result will be excluded.

[0125] No known history of Human Immunodeficiency Virus (HIV) infection. Note: HIV testing does not need to be done at the time of screening unless required according to local guidelines or institutional standards. No known active bacterial, viral, fungal, mycobacterial, parasitic, or other infection (except fungal infections of the nail bed) requiring intravenous (IV) therapy or IV antibiotics within 2 weeks prior to enrollment. No evidence of significant uncontrolled complications that could affect protocol compliance or interpretation of the results. · No history of other previous malignancies, except for: Malignancies that have been treated with curative intent, have had no known active disease for at least 3 years prior to the first dose of study drug, and are considered by the treating physician to be at low risk of recurrence Non-melanoma skin cancer or lentigo maligna that has been adequately treated without evidence of disease Successfully treated carcinoma in situ with no evidence of disease Localized prostate cancer, post-radical prostatectomy with no elevated prostate-specific antigen (PSA) level <0.1ng / mL - Not have undergone lesion-directed radiation therapy or major surgery within 4 weeks of enrollment. - No neurological deficits greater than Grade 1. · You must not have a history of active tuberculosis (TB) or completed treatment for active TB within the past 12 months.

[0126] NOTE: Interferon-gamma release assay (IGRA) testing does not need to be done at screening unless active or latent TB is suspected. Subjects with a positive IGRA and active pulmonary TB must be ruled out with clinical evaluation and radiological imaging. Subjects with a positive IGRA and no evidence of active disease may be enrolled after treatment for latent TB infection (recommended total of 6 months of isoniazid monotherapy) has been initiated.

[0127] · No evidence of CMV viremia at screening (defined as a positive level above the lower limit of detection). · Not have a current autoimmune disease requiring immunosuppressive therapy, except for up to 20 mg of prednisone (or equivalent) daily. Absence of any life-threatening illness, medical condition, or organ system dysfunction that, in the opinion of the investigator, may compromise the subject's safety or place the results of the study at undue risk. · No current seizure disorder requiring treatment. No known active SARS-CoV-2 infection. If a subject has signs / symptoms suggestive of SARS-CoV-2 infection or is known to have had recent exposure to a SARS-CoV-infected individual, they should receive molecular (e.g., PCR) testing or two negative antigen test results, at least 24 hours apart, to rule out SARS-CoV-2 infection.

[0128] Subjects who do not meet the SARS-CoV-2 infection eligibility criteria must fail screening and may be rescreened only after meeting the following SARS-CoV-2 infection viral clearance criteria: At least 10 days have elapsed since the first positive test result in an asymptomatic patient, or at least 10 days have elapsed since recovery (defined as resolution of fever and improvement of symptoms without the use of antipyretic medications). Patients must not have had major surgery within 4 weeks of receiving the first dose of study drug.

[0129] In one embodiment, the subject has no current evidence of primary central nervous system (CNS) tumors or known CNS involvement (including leptomeningeal disease) at the time of screening.

[0130] Subjects may not have a history of severe allergic or anaphylactic reactions to anti-CD20 monoclonal antibody therapy or known significant allergies or intolerances to any components or excipients of epcolitamab or to any components of the study drug combination (e.g., lenalidomide, ibrutinib, etc.).

[0131] In one embodiment, subjects must not have undergone an autologous stem cell transplant within 3 months prior to screening.

[0132] In one embodiment, subjects must not have received a chemotherapeutic agent, non-investigational agent, or investigational anti-neoplastic agent (except for CD20 monoclonal antibodies) within 4 weeks or 5 half-lives (whichever is shorter) prior to the first dose of epcolitamab.

[0133] In one embodiment, the subject is free of clinically significant cardiovascular disease, including: Myocardial infarction or stroke within 6 months prior to enrollment or Any of the following conditions within 3 months prior to enrollment: diseases / conditions related to or affecting unstable or uncontrolled cardiac function (e.g., unstable angina, congestive heart failure, New York Heart Association class III-IV), uncontrolled cardiac arrhythmias or Other clinically significant electrocardiogram (ECG) abnormalities within 6 months prior to enrollment unless deemed stable and appropriately treated.

[0134] Left ventricular ejection fraction (LVEF) must be within the institutional normal range by multiple gated acquisition (MUGA) or transthoracic echocardiography at screening.

[0135] In one embodiment, the subject has no history of other prior malignancies, except for: Malignant tumors that have been treated with curative intent, have had no known active disease for at least 3 years prior to the first dose of study drug, and are felt by the treating physician to be at low risk of recurrence Non-melanoma skin cancer or lentigo maligna that has been adequately treated without evidence of disease Adequately treated carcinoma with no evidence of disease Localized prostate cancer, post-radical prostatectomy with no elevated prostate-specific antigen (PSA) level <0.1ng / mL

[0136] In one embodiment, subjects have not undergone lesion-directed radiation therapy or major surgery within 4 weeks of enrollment.

[0137] In one embodiment, the subject does not have a grade >1 neuropathy.

[0138] Human subjects receiving the treatments described herein may be patients who meet one or more of the inclusion criteria described in Example 3 or who do not meet one or more of the exclusion criteria described in Example 3.

[0139] The methods described herein are advantageous for treating diffuse large B-cell lymphoma, such as relapsed and / or refractory diffuse large B-cell lymphoma. Treatment is maintained continuously, for example, using the treatment regimen described herein. However, treatment can be terminated if progressive disease develops or unacceptable toxicity occurs.

[0140] The response of subjects with diffuse large B-cell lymphoma to treatment using the methods described herein can be evaluated according to the Lugano Response Criteria for Malignant Lymphoma (also referred to herein as "Lugano Criteria") and / or the Lymphoma Response to Immunomodulatory Therapy Criteria (also referred to herein as "LYRIC"), as described in Example 3. In one embodiment, complete response (CR), partial response (PR), and stable disease (SD) are evaluated using the Lugano criteria. In some embodiments, patients who show disease progression, also referred to as progressive disease (PD), according to the Lugano criteria are further evaluated according to LYRIC. Details regarding the Lugano criteria / classification system, including the definitions of complete response, partial response, no response / stable disease, and progressive disease, are provided in Cheson et al. J Clin Oncol 2014;32:3059-68 (see especially Table 3 in Cheson et al., 2014). Details regarding Lugano are provided in Example 2 herein.

[0141] In some embodiments, a subject is treated with the methods described herein until the subject exhibits progressive disease (PD), e.g., as defined by the Lugano criteria and / or LYRIC. In one embodiment, a subject is treated with the methods described herein until the subject exhibits progressive disease (PD), as defined by both the Lugano criteria and LYRIC.

[0142] Subjects treated according to the methods described herein preferably experience an improvement in at least one symptom of diffuse large B-cell lymphoma. In one embodiment, the improvement is measured by a reduction in the amount and / or size of measurable tumor lesions. In some embodiments, the lesions can be measured by CT (computed tomography), PET-CT (positron emission tomography-computed tomography), or MRI (magnetic resonance imaging) films. In some embodiments, cytology or histology can be used to assess response to treatment. In some embodiments, bone marrow aspirate, bone marrow biopsy, tumor biopsy, physical examination, and / or laboratory tests (e.g., tumor cells in peritoneal or pleural fluid) can be used to assess response to treatment.

[0143] In one embodiment, the subject being treated exhibits a complete response (CR), partial response (PR), or stable disease (SD) as defined by the Lugano criteria or LYRIC (see, e.g., Example 2 herein). In some embodiments, the methods described herein optionally result in at least one therapeutic benefit selected from prolonged survival (e.g., progression-free survival or overall survival, etc.) compared to another therapy (e.g., treatment with lenalidomide or lenalidomide and ibrutinib alone, etc.).

[0144] In one embodiment, the bispecific antibodies used in the methods described herein are administered subcutaneously and thus have a formulation and / or concentration that allows for subcutaneous (sc) administration, i.e., pharma- ceutically acceptable sc administration at the doses described herein. In some embodiments, the subcutaneous administration is by injection. For example, formulations for Duobody® CD3xCD20 that are compatible with subcutaneous formulations and may be used in the methods described herein have been previously described (see, e.g., WO2019155008, incorporated herein by reference). In some embodiments, the bispecific antibodies may be formulated with sodium acetate trihydrate, acetic acid, sodium hydroxide, sorbitol, polysorbate 80, and water for injection and may have a pH of 5.5 or about 5.5. In some embodiments, the bispecific antibodies are provided as a 5 mg / mL or 60 mg / mL concentrate. In other embodiments, the desired dose of the bispecific antibody is reconstituted to a volume of about 1 mL for subcutaneous injection.

[0145] In one embodiment, a pharmaceutical composition suitable for a bispecific antibody may comprise the bispecific antibody, 20-40 mM acetate, 140-160 mM sorbitol, and a surfactant, such as polysorbate 80, and a pH of 5.3-5.6. In another embodiment, a pharmaceutical formulation may comprise an antibody concentration in the range of 5-100 mg / mL, e.g., 48 or 60 mg / mL of the bispecific antibody, 30 mM acetate, 150 mM sorbitol, 0.04% w / v polysorbate 80, and may have a pH of 5.5. Such formulations may be diluted, for example with a formulation buffer, to allow for proper dosing and subcutaneous administration.

[0146] The volume of the pharmaceutical composition is appropriately selected to allow subcutaneous administration of the antibody. For example, the administered volume is in the range of about 0.3 mL to about 3 mL, for example, 0.3 mL to 3 mL. The administered volume may be 0.5 mL, 0.8 mL, 1 mL, 1.2 mL, 1.5 ml, 1.7 mL, 2 mL, or 2.5 mL, or about 0.5 mL, about 0.8 mL, about 1 mL, about 1.2 mL, about 1.5 ml, about 1.7 mL, about 2 mL, or about 2.5 mL. Thus, in one embodiment, the administered volume is 0.5 mL or about 0.5 mL. In some embodiments, the administered volume is 0.8 mL or about 0.8 mL. In some embodiments, the administered volume is 1 mL or about 1 mL. In some embodiments, the administered volume is 1.2 mL or about 1.2 mL. In some embodiments, the administered volume is 1.5 mL or about 1.5 mL. In some embodiments, the administered volume is at or about 1.7 mL. In some embodiments, the administered volume is at or about 2 mL. In some embodiments, the administered volume is at or about 2.5 mL.

[0147] In one embodiment, ibrutinib is formulated in a pharmaceutical composition comprising pharma- ceutically acceptable excipients for administration (e.g., orally) in accordance with local standard of care practices, e.g., as specified by local guidelines or local product labeling. For example, in some embodiments, ibrutinib is provided in an oral dosage form, e.g., a capsule.

[0148] In one embodiment, lenalidomide is formulated into a pharmaceutical composition comprising pharma- ceutically acceptable excipients suitable for administration (e.g., oral administration), e.g., in accordance with local standard of care practices, e.g., as specified by local guidelines or local product labeling. In some embodiments, lenalidomide is formulated in an oral dosage form, e.g., a capsule. In some embodiments, lenalidomide is formulated as a capsule comprising lenalidomide, anhydrous lactose, microcrystalline cellulose, croscarmellose sodium, and magnesium stearate.

[0149] In one embodiment, the bispecific antibody used in the methods described herein comprises: (i) a first binding arm that binds to human CD3ε (epsilon) and comprises a first antigen-binding region comprising a variable heavy (VH) region and a variable light (VL) region, wherein the VH region comprises CDR1, CDR2 and CDR3 sequences within the amino acid sequence of SEQ ID NO: 6, and the VL region comprises CDR1, CDR2 and CDR3 sequences within the amino acid sequence of SEQ ID NO: 7; (ii) a second binding arm that binds to human CD20 and comprises a second antigen-binding region comprising a VH region and a VL region, wherein the VH region comprises CDR1, CDR2 and CDR3 sequences within the amino acid sequence of SEQ ID NO: 13, and the VL region comprises CDR1, CDR2 and CDR3 sequences within the amino acid sequence of SEQ ID NO: 14.

[0150] CDR1, CDR2 and CDR3 regions can be identified from the variable heavy and light chain regions using methods known in the art. The CDR regions from the variable heavy and light chain regions can be annotated according to IMGT (see Lefranc et al., Nucleic Acids Research 1999;27:209-12,1999] and Brochet. Nucl Acids Res 2008;36:W503-8).

[0151] In some embodiments, the bispecific antibody comprises: (i) a first binding arm comprising a first antigen-binding region that binds to human CD3ε (epsilon) and comprises the amino acid sequences VHCDR1, VHCDR2, and VHCDR3 shown in SEQ ID NOs: 1, 2, and 3, respectively, and VLCDR1, VLCDR2, and VLCDR3 that comprise the amino acid sequences shown in SEQ ID NO: 4, sequence GTN, and SEQ ID NO: 5, respectively; (ii) a second binding arm that binds to human CD20 and comprises a second antigen-binding region comprising VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively, and VLCDR1, VLCDR2, and VLCDR3 comprising the amino acid sequences set forth in SEQ ID NO: 11, sequence DAS, and SEQ ID NO: 12, respectively.

[0152] In some embodiments, the bispecific antibody comprises: (i) a first binding arm comprising a first antigen-binding region that binds to human CD3ε (epsilon) and comprises a VH region comprising the amino acid sequence of SEQ ID NO:6 and a VL region comprising the amino acid sequence of SEQ ID NO:7; (ii) a second binding arm that binds to human CD20 and comprises a second antigen-binding region comprising a VH region comprising the amino acid sequence of SEQ ID NO: 13 and a VL region comprising the amino acid sequence of SEQ ID NO: 14.

[0153] In one embodiment, the bispecific antibody is a full-length antibody. In some embodiments, the bispecific antibody has an inactive Fc region. In some embodiments, the bispecific antibody is a full-length antibody and has an inactive Fc region. In some embodiments, the first binding arm to CD3 is derived from a humanized antibody, such as a full-length IgG1, λ (lambda) antibody, such as H1L1 described in WO2015001085, which is incorporated herein by reference, and / or the second binding arm to CD20 is derived from a human antibody, such as a full-length IgG1, κ (kappa) antibody, such as clone 7D8 described in WO2004035607, which is incorporated herein by reference. Bispecific antibodies can be produced from two half-molecule antibodies, including the respective first and second binding arms shown in, for example, SEQ ID NOs: 24 and 25, and SEQ ID NOs: 26 and 27. The half antibodies can be produced in CHO cells, and the bispecific antibodies can be generated, for example, by Fab arm exchange. In one embodiment, the bispecific antibody is a functional variant of DuoBody® CD3xCD20.

[0154] Thus, in some embodiments, the bispecific antibody comprises: (i) a first binding arm comprising a first antigen-binding region that binds human CD3ε (epsilon) and comprises a VH region comprising an amino acid sequence that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:6, or a VH region comprising the amino acid sequence of SEQ ID NO:6 but with one, two or three mutations (e.g., amino acid substitutions), and a VL region comprising an amino acid sequence that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:7, or a VL region comprising the amino acid sequence of SEQ ID NO:7 but with one, two or three mutations (e.g., amino acid substitutions); (ii) a second binding arm that binds human CD20 and comprises a second antigen-binding region comprising a VH region comprising an amino acid sequence at least 85%, 90%, 95%, 98% or 99% identical to SEQ ID NO: 13, or a VH region comprising the amino acid sequence of SEQ ID NO: 13 but with one, two or three mutations (e.g., amino acid substitutions), and a VL region comprising an amino acid sequence at least 85%, 90%, 95%, 98% or 99% identical to SEQ ID NO: 14, or a VL region comprising the amino acid sequence of SEQ ID NO: 14 but with one, two or three mutations (e.g., amino acid substitutions).

[0155] In one embodiment, the bispecific antibody comprises: (i) a first binding arm comprising a first antigen-binding region that binds to human CD3ε (epsilon) and comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 24 and a light chain comprising the amino acid sequence of SEQ ID NO: 25; (ii) a second binding arm that binds to human CD20 and comprises a second antigen-binding region comprising a VH region comprising the amino acid sequence of SEQ ID NO:26 and a VL region comprising the amino acid sequence of SEQ ID NO:27.

[0156] In some embodiments, the bispecific antibody comprises (i) a first binding arm comprising a first antigen-binding region that binds human CD3ε (epsilon) and comprises a heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO:24, or a heavy chain comprising the amino acid sequence of SEQ ID NO:24 but with one, two or three mutations (e.g., amino acid substitutions), and a light chain comprising an amino acid sequence at least 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO:25, or a light chain region comprising the amino acid sequence of SEQ ID NO:25 but with one, two or three mutations (e.g., amino acid substitutions); (ii) a second binding arm comprising a heavy chain that binds human CD20 and comprises an amino acid sequence at least 85%, 90%, 95%, 98% or 99% identical to SEQ ID NO: 26, or a heavy chain comprising the amino acid sequence of SEQ ID NO: 26 but with one, two or three mutations (e.g., amino acid substitutions), and a light chain comprising an amino acid sequence at least 85%, 90%, 95%, 98% or 99% identical to SEQ ID NO: 27, or a light chain region comprising the amino acid sequence of SEQ ID NO: 27 but with one, two or three mutations (e.g., amino acid substitutions).

[0157] Various constant regions or variants thereof may be used in the bispecific antibody. In one embodiment, the antibody comprises an IgG constant region, such as a human IgG1 constant region, such as the human IgG1 constant region defined in SEQ ID NO: 15, or any other suitable IgG1 allotype. In some embodiments, the bispecific antibody is a full-length antibody having a human IgG1 constant region. In some embodiments, the first binding arm of the bispecific antibody is derived from a humanized antibody, preferably a full-length IgG1, λ (lambda) antibody. In one embodiment, the first binding arm of the bispecific antibody is derived from a humanized antibody, such as a full-length IgG1, λ (lambda) antibody, and thus comprises a λ light chain constant region. In some embodiments, the first binding arm comprises a λ light chain constant region defined in SEQ ID NO: 22. In some embodiments, the second binding arm of the bispecific antibody is derived from a human antibody, preferably a full-length IgG1, κ (kappa) antibody. In some embodiments, the second binding arm of the bispecific antibody may be derived from a human antibody, preferably a full-length IgG1, κ (kappa) antibody, and thus comprises a κ light chain constant region. In some embodiments, the second binding arm comprises a kappa light chain constant region defined in SEQ ID NO: 23. In preferred embodiments, the first binding arm comprises a lambda light chain constant region defined in SEQ ID NO: 22 and the second binding arm comprises a kappa light chain constant region defined in SEQ ID NO: 23.

[0158] It is understood that the constant region portion of the bispecific antibody may contain modifications that allow for efficient formation / production of the bispecific antibody and / or provide an inactive Fc region. Such modifications are well known in the art.

[0159] Different formats of bispecific antibodies are known in the art (reviewed in Kontermann, Drug Discov Today 2015;20:838-47; MAbs, 2012;4:182-97). Thus, the bispecific antibodies used in the methods and uses described herein are not limited to any particular bispecific format or method of making them. For example, bispecific antibodies can include, but are not limited to, bispecific antibodies with complementary CH3 domains that force heterodimerization, knobs-into-hole molecules (Genentech, WO 9850431), CrossMAbs (Roche, WO 2011117329), or electrostatically matched molecules (Amgen, EP 1870459 and WO 2009089004; Chugai, US 201000155133; Oncomed, WO 2010129304).

[0160] Preferably, the bispecific antibody comprises an Fc region comprising a first heavy chain having a first Fc sequence comprising a first CH3 region and a second heavy chain having a second Fc sequence comprising a second CH3 region, the sequences of the first and second CH3 regions being different such that a heterodimeric interaction between the first and second CH3 regions is stronger than a homodimeric interaction of the first and second CH3 regions, respectively. Further details regarding these interactions and how they may be achieved are provided, for example, in WO2011131746 and WO2013060867 (Genmab), which are incorporated herein by reference. In one embodiment, the bispecific antibody comprises in the first heavy chain (i) an amino acid L at a position corresponding to F405 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 and in the second heavy chain an amino acid R at a position corresponding to K409 of the human IgG1 heavy chain constant region of SEQ ID NO: 15, or vice versa.

[0161] Bispecific antibodies may contain modifications in the Fc region to render the Fc region inactive or inactive. Thus, in the bispecific antibodies disclosed herein, one or both heavy chains may be modified such that the antibody induces Fc-mediated effector function to a lesser extent compared to a bispecific antibody without the modification. Fc-mediated effector function can be measured by determining Fc-mediated CD69 expression on T cells (i.e., CD69 expression as a result of CD3 antibody-mediated Fcγ receptor-dependent CD3 cross-linking), by binding to Fcγ receptors, by binding to C1q, or by induction of Fc-mediated cross-linking of FcγR. In particular, the heavy chain constant region sequence may be modified such that Fc-mediated CD69 expression is reduced by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 99% or 100% when compared to a wild-type (unmodified) antibody, as determined, for example, in a PBMC-based functional assay as described in Example 3 of WO2015001085. Modification of the heavy and light chain constant region sequences may also result in a reduction in binding of C1q to the antibody. Compared to the unmodified antibody, the reduction may be at least 70%, at least 80%, at least 90%, at least 95%, at least 97% or 100%, and C1q binding may be determined, for example, by ELISA. Furthermore, the Fc region may be modified such that the antibody mediates at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 99% or 100% reduced Fc-mediated T cell proliferation compared to an unmodified antibody, as measured in a PBMC-based functional assay. Examples of amino acid positions that may be modified, for example in an IgG1 isotype antibody, include positions L234 and L235. Thus, in one embodiment, the bispecific antibody may comprise a first heavy chain and a second heavy chain, in both of which the amino acid residues at positions corresponding to positions L234 and L235 of the human IgG1 heavy chain according to Eu numbering are F and E, respectively.Furthermore, the D265A amino acid substitution can reduce binding to all Fcγ receptors and prevent ADCC (Shields et al., JBC 2001;276:6591-604). Thus, a bispecific antibody can comprise a first heavy chain and a second heavy chain, in which the amino acid residue at the position corresponding to position D265 of the human IgG1 heavy chain according to Eu numbering is A in both the first heavy chain and the second heavy chain.

[0162] In one embodiment, in the first and second heavy chains of the bispecific antibody, the amino acids at positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain are, respectively, F, E, and A. An antibody with these amino acids at these positions is an example of an antibody with an inactive or non-active Fc region.

[0163] In some embodiments, the bispecific antibody comprises a first heavy chain and a second heavy chain, and in both the first heavy chain and the second heavy chain, the amino acids at positions corresponding to positions L234, L235, and D265 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 are F, E, and A, respectively. In some embodiments, the bispecific antibody comprises a first heavy chain and a second heavy chain, and in the first heavy chain, the amino acid at position corresponding to F405 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is L and in the second heavy chain, the amino acid at position corresponding to K409 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is R, or vice versa. In a preferred embodiment, the bispecific antibody comprises a first heavy chain and a second heavy chain, (i) in both the first and second heavy chains, the amino acids at positions corresponding to positions L234, L235, and D265 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 are F, E, and A, respectively; (ii) in the first heavy chain, the amino acid at the position corresponding to F405 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is L and in the second heavy chain, the amino acid at the position corresponding to K409 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is R, or vice versa.

[0164] With respect to the bispecific antibodies described herein, those having a combination of the three amino acid substitutions L234F, L235E and D265A, and additionally having the K409R or F405L mutations as described above, may be designated with the suffix "FEAR" or "FEAL", respectively.

[0165] The amino acid sequence of the wild type IgG1 heavy chain constant region may be identified herein as SEQ ID NO: 15. Consistent with the embodiments disclosed above, the bispecific antibody may comprise an IgG1 heavy chain constant region carrying an F405L substitution, may have the amino acid sequence shown in SEQ ID NO: 17 and / or an IgG1 heavy chain constant region carrying a K409R substitution, may have the amino acid sequence shown in SEQ ID NO: 18, and may have further substitutions that render the Fc region inactive or inactive. Thus, in one embodiment, the bispecific antibody comprises an IgG1 heavy chain constant region in combination with one amino acid sequence of an IgG1 heavy chain constant region with L234F, L235E, D265A and F405L substitutions (e.g., as shown in SEQ ID NO: 19) and the other amino acid sequence of an IgG1 heavy chain constant region with L234F, L235E, D265A and K409R substitutions (e.g., as shown in SEQ ID NO: 20). Thus, in some embodiments, the bispecific antibody comprises heavy chain constant regions comprising the amino acid sequences of SEQ ID NOs: 19 and 20.

[0166] In a preferred embodiment, the bispecific antibody used in the methods and uses described herein comprises a first binding arm comprising a heavy chain and a light chain as defined in SEQ ID NOs: 24 and 25, respectively, and a second binding arm comprising a heavy chain and a light chain as defined in SEQ ID NOs: 26 and 27, respectively. Such an antibody is referred to herein as DuoBody® CD3xCD20. Variants of such antibodies are also contemplated for use in the methods and uses described herein. In some embodiments, the bispecific antibody comprises a heavy chain and a light chain consisting of the amino acid sequences set forth in SEQ ID NOs: 24 and 25, respectively, and a heavy chain and a light chain consisting of the amino acid sequences set forth in SEQ ID NOs: 26 and 27, respectively. In some embodiments, the bispecific antibody is epcolitamab (CAS 2134641-34-0) or a biosimilar thereof.

[0167] Medical Use Further provided herein is a bispecific antibody for use in the above disclosed methods.

[0168] In certain embodiments, the bispecific antibody is for use in a method of treating diffuse large B-cell lymphoma (DLBCL) in a human subject, wherein the bispecific antibody is administered to the subject in combination with an effective amount of lenalidomide, and optionally, an effective amount of ibrutinib, wherein the bispecific antibody is (i) a first binding arm that binds to human CD3ε (epsilon) and comprises a first antigen-binding region comprising a variable heavy (VH) region and a variable light (VL) region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences found in the VH region sequence of SEQ ID NO: 6, and the VL region comprises the CDR1, CDR2 and CDR3 sequences found in the VL region sequence of SEQ ID NO: 7; (ii) a second binding arm that binds to human CD20 and comprises a second antigen-binding region comprising a VH region and a VL region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences found in the VH region sequence of SEQ ID NO: 13, and the VL region comprises the CDR1, CDR2 and CDR3 sequences found in the VL region sequence of SEQ ID NO: 14; The bispecific antibody is administered at a dose of 24 mg or 48 mg, and the lenalidomide, bispecific antibody and optionally ibrutinib are administered in a 28-day cycle.

[0169] Also provided herein are bispecific antibodies for the manufacture of a medicament for use in the methods disclosed above.

[0170] In particular, the bispecific antibody is for the manufacture of a medicament for use in the treatment of diffuse large B-cell lymphoma (DLBCL) in a human subject, wherein the bispecific antibody is administered to the subject in combination with an effective amount of lenalidomide and optionally an effective amount of ibrutinib, wherein the bispecific antibody is (i) a first binding arm that binds to human CD3ε (epsilon) and comprises a first antigen-binding region comprising a variable heavy (VH) region and a variable light (VL) region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences found in the VH region sequence of SEQ ID NO: 6, and the VL region comprises the CDR1, CDR2 and CDR3 sequences found in the VL region sequence of SEQ ID NO: 7; (ii) a second binding arm that binds to human CD20 and comprises a second antigen-binding region comprising a VH region and a VL region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences found in the VH region sequence of SEQ ID NO: 13, and the VL region comprises the CDR1, CDR2 and CDR3 sequences found in the VL region sequence of SEQ ID NO: 14; The bispecific antibody is administered at a dose of 24 mg or 48 mg, and the lenalidomide, bispecific antibody and optionally ibrutinib are administered in a 28-day cycle.

[0171] kit Also provided herein is a kit comprising a pharmaceutical composition comprising a bispecific antibody that binds CD3 and CD20 according to the present invention, such as DuoBody® CD3xCD20 or epcolitamab, and a pharma- ceutically acceptable carrier, in a therapeutically effective amount adapted for use in the methods described herein. The kit may also comprise a pharmaceutical composition comprising ibrutinib (e.g., for oral administration) and / or lenalidomide (e.g., for oral administration). The kit may further comprise a pharmaceutical composition comprising lenalidomide (e.g., for oral administration). The kit may also optionally comprise instructions, including, for example, an administration schedule, to enable a practitioner (e.g., a physician, a nurse, or a patient) to administer one or more compositions contained therein to a patient with diffuse large B-cell lymphoma. The kit may also comprise one or more syringes.

[0172] Optionally, the kits contain multiple packages of single-dose pharmaceutical compositions each containing an effective amount of a bispecific antibody for a single administration according to the methods described herein. They may also contain multiple packages of single-dose pharmaceutical compositions containing doses of ibrutinib and / or lenalidomide according to standard practice regimens. Apparatus or devices necessary for administering the pharmaceutical composition(s) may also be included in the kits.

[0173] Further embodiments 1. A method of treating diffuse large B-cell lymphoma (DLBCL) in a human subject, the method comprising administering to the subject a bispecific antibody, an effective amount of lenalidomide, and optionally an effective amount of ibrutinib, wherein the bispecific antibody is (i) a first binding arm that binds to human CD3ε (epsilon) and comprises a first antigen-binding region comprising a variable heavy (VH) region and a variable light (VL) region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences found in the VH region sequence of SEQ ID NO: 6, and the VL region comprises the CDR1, CDR2 and CDR3 sequences found in the VL region sequence of SEQ ID NO: 7; (ii) a second binding arm that binds to human CD20 and comprises a second antigen-binding region comprising a VH region and a VL region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences found in the VH region sequence of SEQ ID NO: 13, and the VL region comprises the CDR1, CDR2 and CDR3 sequences found in the VL region sequence of SEQ ID NO: 14; The bispecific antibody is administered at a dose of 24 mg or 48 mg, and the lenalidomide, bispecific antibody and optionally ibrutinib are administered in a 28 day cycle.

[0174] 2. The method of embodiment 1, wherein the bispecific antibody is administered at a dose of 24 mg.

[0175] 3. The method of embodiment 1, wherein the bispecific antibody is administered at a dose of 48 mg.

[0176] 4. The method of any one of embodiments 1-3, wherein the bispecific antibody is administered once a week (administered every week).

[0177] The method of embodiment 4, wherein weekly doses of 5.24 mg or 48 mg are administered for 2.5 28 day cycles.

[0178] 6. The method of embodiment 4 or 5, wherein after the weekly administration, the bispecific antibody is administered once every 4 weeks, for example, for a 28 day cycle, on day 1 of each 28 day cycle.

[0179] 7. The method of embodiment 6, wherein one administration every 4 weeks is administered for at least eight 28-day cycles, such as eight 28-day cycles.

[0180] 8. The method of embodiment 6, wherein one administration every 4 weeks is administered for at least 20 28-day cycles, such as 20 28-day cycles.

[0181] The method of any one of embodiments 4-8, wherein a priming dose of the bispecific antibody is administered in cycle 1 of a 28 day cycle prior to the weekly administration of 9.24 mg or 48 mg.

[0182] 10. The method of embodiment 9, wherein the priming dose is administered 2 weeks prior to administration of the first weekly dose of 24 mg or 48 mg.

[0183] 11. The method of embodiment 9 or 10, wherein the priming dose is 0.16 mg.

[0184] 12. The method of any one of embodiments 9-11, wherein an intermediate dose of the bispecific antibody is administered after the priming dose and before the first weekly dose of 24 mg or 48 mg.

[0185] 13. The method of embodiment 12, wherein a priming dose is administered on day 1, an intermediate dose is administered on day 8, followed by first weekly doses of 24 mg or 48 mg on days 15 and 22 of cycle 1.

[0186] 14. The method of embodiment 12 or 13, wherein the intermediate dose is 0.8 mg.

[0187] 15. The method of any one of embodiments 1-14, wherein lenalidomide is administered once daily on days 1 through 21 of a 28-day cycle.

[0188] 16. The method of any one of embodiments 1-15, wherein lenalidomide is administered from cycle 1 to cycle 12 of a 28 day cycle.

[0189] 17. The method of any one of embodiments 1-15, wherein lenalidomide is administered from cycle 1 to cycle 24 of a 28 day cycle.

[0190] 18. The method according to any one of embodiments 1-17, wherein lenalidomide is administered at a dose of 20-30 mg, such as 25 mg.

[0191] 19. The method of any one of embodiments 1-17, wherein lenalidomide is administered at a dose of 20-30 mg in cycles 1 to 12 of a 28 day cycle.

[0192] 20. The method of any one of embodiments 1-17, wherein lenalidomide is administered at a dose of 25 mg in cycles 1 to 12 of a 28 day cycle.

[0193] 21. The method according to any one of embodiments 1-14, wherein lenalidomide is administered in a dose of 10 to 25 mg, such as 25 mg.

[0194] 22. The method of any one of embodiments 1-14 and 21, wherein lenalidomide is administered at a dose of 10-25 mg in cycles 1 to 24 of a 28 day cycle.

[0195] 23. The method of any one of embodiments 1-14 and 21-22, wherein lenalidomide is administered at a dose of 20 mg in cycles 1 to 24 of a 28 day cycle.

[0196] 24. The method of any one of embodiments 1-14 and 21-23, wherein ibrutinib is administered once daily from day 1 to day 28 of a 28-day cycle.

[0197] 25. The method of any one of embodiments 1-14 and 21-24, wherein ibrutinib is administered from cycle 1 to cycle 24 of a 28 day cycle.

[0198] 26. The method of any one of embodiments 1-14 and 21-25, wherein ibrutinib is administered at a dose of 280 mg to 560 mg, for example 280, 420 mg or 560 mg.

[0199] 27. The method of any one of embodiments 1-14 and 21-25, wherein ibrutinib is administered at a dose of 560 mg in cycles 1 to 24 of a 28 day cycle, or at a dose of 420 mg in cycles 1 to 24 of a 28 day cycle.

[0200] 28. Treatment is on a 28-day cycle. (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) in cycles 4 and beyond, a dose of 24 mg is administered on day 1, and so on; (b) lenalidomide is administered on days 1-21 of Cycle 1 or later; and (c) ibrutinib is optionally administered on days 1-28 after Cycle 1; The method according to any one of embodiments 1, 2, and 4 to 27.

[0201] 29. Treatment is on a 28-day cycle. (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 12, a dose of 24 mg was administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 25 mg / day on days 1-21 of cycles 1-12; The method according to any one of embodiments 1, 2, and 4 to 28.

[0202] 30. Treatment is in a 28-day cycle. (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 24 mg is administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib will be administered orally at a dose of 560 mg / day on days 1-28 of cycles 1-24. The method according to any one of embodiments 1, 2, and 4 to 29.

[0203] 31. Treatment is in a 28-day cycle. (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a dose of 24 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 24 mg is administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib will be administered orally at a dose of 420 mg / day on days 1-28 of cycles 1-24. The method according to any one of embodiments 1, 2, and 4 to 29.

[0204] 32. Treatment is in a 28-day cycle. (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) in cycles 3 and beyond, a dose of 48 mg is administered on day 1, and so on; (b) lenalidomide is administered on days 1-21 of Cycle 1 or later; and (c) ibrutinib is optionally administered on days 1-28 after Cycle 1; The method according to any one of embodiments 1 and 3 to 27.

[0205] 33. Treatment is in a 28-day cycle. (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 12, a dose of 48 mg was administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 25 mg / day on days 1-21 of cycles 1-12; The method according to any one of embodiments 1, 3 to 27 and 31.

[0206] 34. Treatment is in a 28-day cycle. (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 48 mg was administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib will be administered orally at a dose of 560 mg / day on days 1-28 of cycles 1-24. The method according to any one of embodiments 1, 3-27 and 31-33.

[0207] 35. Treatment is in a 28-day cycle. (a) a bispecific antibody comprising: (i) in cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) in cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In cycles 4 through 24, a dose of 48 mg was administered subcutaneously on day 1, as follows: (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib will be administered orally at a dose of 420 mg / day on days 1-28 of cycles 1-24. The method according to any one of embodiments 1, 3-27 and 31-32.

[0208] 36. The method of any one of embodiments 1 to 35, wherein the bispecific antibody is administered subcutaneously.

[0209] 37. The method of any one of embodiments 1-36, wherein ibrutinib is administered orally.

[0210] 38. The method of any one of embodiments 1-37, wherein lenalidomide is administered orally.

[0211] 39. The method of any one of embodiments 1-38, wherein the bispecific antibody, lenalidomide and optionally ibrutinib are administered sequentially.

[0212] 40. The method of any one of embodiments 1 to 39, wherein the DLBCL has histologically confirmed CD20+ disease.

[0213] 41. The method according to any one of the preceding embodiments, wherein DLBCL is an aggressive B-cell lymphoma with MYC and BCL-2 and / or BCL-6 translocations (double or triple hit).

[0214] 42. The method according to any one of embodiments 1 to 41, wherein the DLBCL is follicular lymphoma grade 3B.

[0215] 43. The method according to any one of embodiments 1 to 42, wherein the DLBCL is relapsed and / or refractory DLBCL.

[0216] 44. The method of any one of embodiments 1 to 43, wherein the DLBCL has relapsed, i.e., previously responded to a previous treatment, but has progressed after the previous treatment, with progression beginning 6 months or more after completion of the previous treatment.

[0217] 45. The method according to any one of embodiments 1 to 44, wherein the DLBCL is refractory, i.e., has either progressed during a previous treatment, failed to achieve an objective response to a previous treatment, or progressed within 6 months after completion of a previous treatment, including maintenance therapy.

[0218] 46. ​​The method of any one of embodiments 1-45, wherein the subject has disease that is relapsed or refractory to at least one prior systemic anti-lymphoma therapy containing an anti-CD20 monoclonal antibody.

[0219] 47. The method of any one of embodiments 1 to 46, wherein the DLBCL is not refractory to previous chimeric antigen receptor T cell (CAR-T) therapy.

[0220] 48. The method of any one of embodiments 1-46, wherein the subject is not refractory to lenalidomide or ibrutinib.

[0221] 49. The method of any one of embodiments 1-48, wherein the subject has previously received at least one treatment with an anti-CD20 monoclonal antibody in combination with another systemic therapy.

[0222] 50. The method of any one of embodiments 1-49, wherein the subject has previously undergone CAR-T therapy or is ineligible for or unable to undergo CAR-T therapy.

[0223] 51. The method of any one of embodiments 1-50, wherein the subject has not received prior treatment with ibrutinib.

[0224] 52. (i) a first antigen-binding region of a bispecific antibody comprises VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences shown in SEQ ID NO:1, 2, and 3, respectively, and VLCDR1, VLCDR2, and VLCDR3 comprising the amino acid sequences shown in SEQ ID NO:4, sequence GTN, and SEQ ID NO:5, respectively; (ii) The method according to any one of embodiments 1 to 51, wherein the second antigen-binding region of the bispecific antibody comprises VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences shown in SEQ ID NOs: 8, 9, and 10, respectively, and VLCDR1, VLCDR2, and VLCDR3 comprising the amino acid sequences shown in SEQ ID NO: 11, sequence DAS, and SEQ ID NO: 12, respectively.

[0225] 53. (i) The first antigen-binding region of the bispecific antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO:6 and a VL region comprising the amino acid sequence of SEQ ID NO:7; (ii) The method according to any one of embodiments 1 to 52, wherein the second antigen-binding region of the bispecific antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 13 and a VL region comprising the amino acid sequence of SEQ ID NO: 14.

[0226] 54. The method according to any one of embodiments 1 to 53, wherein the first binding arm of the bispecific antibody is derived from a humanized antibody, preferably a full-length IgG1, λ (lambda) antibody.

[0227] 55. The method of embodiment 54, wherein the first binding arm of the bispecific antibody comprises a lambda light chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 22.

[0228] 56. The method according to any one of the preceding embodiments, wherein the second binding arm of the bispecific antibody is derived from a human antibody, preferably a full-length IgG1, κ (kappa) antibody.

[0229] 57. The method of embodiment 56, wherein the second binding arm comprises a kappa light chain constant region comprising the amino acid sequence set forth in SEQ ID NO:23.

[0230] 58. The method of any one of embodiments 1 to 57, wherein the bispecific antibody is a full-length antibody having a human IgG1 constant region.

[0231] 59. The method of any one of embodiments 1 to 58, wherein the bispecific antibody comprises an inactive Fc region.

[0232] 60. The method of any one of embodiments 1 to 59, wherein the bispecific antibody comprises a first heavy chain and a second heavy chain, and in both the first heavy chain and the second heavy chain, the amino acids at positions corresponding to positions L234, L235, and D265 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 are F, E, and A, respectively.

[0233] 61. The method of any one of embodiments 1 to 60, wherein the bispecific antibody comprises a first heavy chain and a second heavy chain, wherein in the first heavy chain, the amino acid at the position corresponding to F405 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is L and in the second heavy chain, the amino acid at the position corresponding to K409 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is R, or vice versa.

[0234] 62. The bispecific antibody comprises a first heavy chain and a second heavy chain; (i) in both the first and second heavy chains, the amino acids at positions corresponding to positions L234, L235, and D265 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 are F, E, and A, respectively; (ii) The method according to any one of embodiments 1 to 61, wherein in the first heavy chain, the amino acid at the position corresponding to F405 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is L and in the second heavy chain, the amino acid at the position corresponding to K409 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is R, or vice versa.

[0235] 63. The method of embodiment 62, wherein the bispecific antibody comprises a heavy chain constant region comprising the amino acid sequences of SEQ ID NOs: 19 and 20.

[0236] 64. The method according to any one of embodiments 1 to 63, wherein the bispecific antibody comprises a heavy chain and a light chain comprising the amino acid sequences set forth in SEQ ID NOs: 24 and 25, respectively, and a heavy chain and a light chain comprising the amino acid sequences set forth in SEQ ID NOs: 26 and 27, respectively.

[0237] 65. The method according to any one of embodiments 1 to 64, wherein the bispecific antibody comprises a heavy chain and a light chain consisting of the amino acid sequences of SEQ ID NOs: 24 and 25, respectively, and a heavy chain and a light chain consisting of the amino acid sequences of SEQ ID NOs: 26 and 27, respectively.

[0238] 66. The method of any one of embodiments 1 to 65, wherein the bispecific antibody is epcolitamab, or a biosimilar thereof.

[0239] [Example] DuoBody®-CD3xCD20 DuoBody®-CD3xCD20 is a bsAb that recognizes the T cell antigen CD3 and the B cell antigen CD20. DuoBody®-CD3xCD20 triggers potent T cell-mediated killing of CD20 expressing cells. DuoBody®-CD3xCD20 has a regular IgG1 structure.

[0240] Two parent antibodies, IgG1-CD3-FEAL, a humanized IgG1 lambda, CD3ε specific antibody with heavy and light chain sequences as set forth in SEQ ID NOs: 24 and 25, respectively, and IgG1-CD20-FEAR derived from human IgG1 kappa CD20 specific antibody 7D8 with heavy and light chain sequences as set forth in SEQ ID NOs: 26 and 27, respectively, were produced as separate biological intermediates. Each parent antibody contains one of the complementary mutations in the CH3 domain required for the generation of DuoBody® molecules (F405L and K409R, respectively). The parent antibodies contained three additional mutations in the Fc region (L234F, L235E and D265A; FEA). The parent antibodies were produced in mammalian Chinese Hamster Ovary (CHO) cell lines using standard suspension cell culture and purification techniques. DuoBody®-CD3xCD20 was then produced by a controlled Fab arm exchange (cFAE) process (Labrijn et al. 2013, Labrijn et al. 2014, Gramer et al. 2013). The parent antibodies are mixed and subjected to controlled reducing conditions. This results in the separation of the parent antibodies which reassemble under reoxidation. In this way, a highly pure preparation of DuoBody®-CD3xCD20 (approximately 93-95%) was obtained. After further polishing / purification, a final product close to 100% purity was obtained. The DuoBody®-CD3xCD20 concentration was calculated based on the theoretical extinction coefficient ε = 1.597 mL mg -1 cm -1 The final product was stored at 4° C. The product has the international trade name epcolitamab.

[0241] Epcolitamab is prepared as a colorless to slightly yellow, sterile, clear solution (5 mg / mL or 60 mg / mL) supplied as a concentrate for subcutaneous (SC) injection. Epcolitamab contains a buffer and a tonicity agent. All excipients and their amounts in the formulated product are pharma- ceutically acceptable for subcutaneous injection products. The appropriate dose is reconstituted to a volume of approximately 1 mL for subcutaneous injection.

[0242] [Example 1] Effect of lenalidomide on epcolitamab-induced T cell-mediated cytotoxicity in vitro This experiment was performed to determine the effect of lenalidomide on T cell activation and T cell-mediated cytotoxicity induced by Duobody®-CD3xCD20.

[0243] Briefly, T cells were activated with immobilized anti-CD3 in the presence or absence of lenalidomide (5 or 50 μM) for 3 days. Cross-linking of CD3 on T cells in the presence of lenalidomide resulted in increased T cell activation measured by upregulation of activation markers CD69, CD25 on the T cell surface, and release of granzyme B and IFNγ, compared to the absence of lenalidomide (see FIG. 1). T cells activated in the presence or absence of lenalidomide were then tested for their cytotoxicity in response to epcolitamab. In T cells pretreated with lenalidomide and anti-CD3, higher maximum cytotoxicity percentages and higher activity were observed at lower epcolitamab concentrations in a typical dose-response curve, while none of the controls caused further target cell lysis (see FIG. 2).

[0244] This indicates that lenalidomide may enhance the epcolitamab-induced T cell activation observed in patients, leading to more efficient T cell-mediated cytotoxicity against target cells.

[0245] [Example 2] A Phase 1b / 2 Open-Label Study to Evaluate the Safety and Tolerability of Epcolitamab in Combination with Lenalidomide and Ibrutinib in Subjects with Diffuse Large B-Cell Lymphoma A Phase 1b / 2, open-label, multicenter, interventional study evaluating the safety, tolerability, and preliminary efficacy of epcolitamab in combination with lenalidomide, or in combination with lenalidomide and ibrutinib, in subjects diagnosed with DLBCL. The study will include a dose-escalation phase followed by an expansion phase.

[0246] Overview of ongoing clinical trials with epcolitamab Epcolitamab as monotherapy is currently in clinical trials for the treatment of R / R B-NHL (ClinicalTrials.gov Identifier: NCT03625037).

[0247] A phase 1 study evaluating SC epcolitamab monotherapy included subjects with R / R NHL, including DLBCL. The dose escalation portion of the study evaluated a range of doses (12-60 mg). After a weekly priming dose of 0.16 mg and a weekly intermediate dose of 0.8 mg, a total dose of 48 mg was selected as the RP2D.

[0248] The phase 2 study included subjects with R / R NHL, including DLBCL, as well as treatment-naïve DLBCL, evaluated at doses of 24 mg and 48 mg in the escalation phase. Clinically significant and compelling efficacy with epcolitamab was observed among patients with relapsed or refractory (R / R) B-NHL in a phase 1 / 2 study (NCT03625037), including deep and durable responses (overall response rate (ORR), 63%; complete response (CR) rate, 39%; median duration of response (DOR), 12 months) in a population with highly refractory large B-cell lymphoma (n=157) with a manageable safety profile (J. Clin. Oncol. December 22, 2022: DOI https: / / doi.org / 10.1200 / JCO.22.01725).

[0249] Combination therapy with epcolitamab and rituximab plus lenalidomide (R2) is being investigated in an ongoing Phase 1 / 2 study (NCT04663347). 2 Combination treatment with rituximab resulted in a high CMR rate in patients with R / R FL, with 73% (30 / 41) of patients achieving a complete metabolic response (CMR). All CRS events were grade 1 (27%) or grade 2 (10%), most were related to the first full dose, were low grade, and resolved. No new safety signals were detected (Falchi et al, Blood (2022) 140 (Supplement 1): 1464-1466).

[0250] In patients with newly diagnosed high-risk DLBCL, those with previously untreated FL grade 1-3A who met GELF criteria were treated with epcolitamab 48 mg + R 2 Patients received 28 days of 12C. Epcolitamab was administered QW at C1-2 and Q4W+ for up to 2 years. Step-down dosing and corticosteroid prophylaxis was required during C1 to mitigate cytokine release syndrome (CRS). As of June 10, 2022, 41 patients were receiving treatment. Median age was 57 years (range, 39-78), median time from initial diagnosis to first dose was 12 weeks (range, 2-352), most patients (85%) had grade 2 / 3A FL, 90% had stage III / IV disease, and 34% had FLIPI 3-5. At a median follow-up of 4.4 months (range, 0.7-7.5), 88% of patients continued treatment. The most common treatment-emergent adverse events (TEAEs) were CRS (51%; 34% grade 1, 17% grade 2), neutropenia, pyrexia, injection site reactions, fatigue, headache, constipation, and rash. Most CRS events occurred after the first full dose and all resolved in a median of 4 days. No cases of ICANS or clinical tumor lysis syndrome were observed. There was one fatal TEAE: COVID-19 pneumonia (not related to epcolitamab). In efficacy-evaluable patients (n=29), the overall response rate and complete metabolic response rate were 90% and 69%. All responses were ongoing at the time of data cutoff.

[0251] From this study, epcolitamab + R 2 concluded that EGFR demonstrated a manageable safety profile in patients with newly diagnosed DLBCL, similar to patients with R / R disease. CRS events were low grade and occurred near the time of the first full dose.

[0252] Objectives and Endpoints Main purpose To characterize the safety and toxicity profile of epcolitamab when co-administered with lenalidomide or lenalidomide and ibrutinib in subjects with DLBCL. To determine the recommended dose for further investigation of epcolitamab when coadministered with lenalidomide or lenalidomide and ibrutinib in subjects with DLBCL.

[0253] Secondary Objectives To evaluate the anti-NHL activity of epcolitamab when administered in combination with lenalidomide or lenalidomide and ibrutinib in subjects with DLBCL. To characterize the pharmacokinetics of epcolitamab when administered in combination with lenalidomide or lenalidomide and ibrutinib in subjects with DLBCL.

[0254] exploratory purpose Evaluating potential mechanisms of response or resistance to treatment Evaluation of the immunogenicity of epcolitamab To assess the impact on patients' quality of life (QOL) via patient-reported outcome measures (PROs), Functional Assessment of Cancer Therapy-Lymphoma (FACT-Lym) and EuroQol 5 Dimensions 5 Levels (EQ-5D-5L).

[0255] Primary Endpoint The primary endpoint was dose-limiting toxicity (DLT) of epcolitamab in combination with lenalidomide or in combination with lenalidomide and ibrutinib.

[0256] Secondary Endpoints Overall response rate (ORR) by Lugano 2014 criteria as assessed by investigator for epcolitamab in combination with lenalidomide or lenalidomide and ibrutinib. Anti-lymphoma activity of epcolitamab in combination with lenalidomide or lenalidomide and ibrutinib: Duration of response (DOR) determined according to the Lugano 2014 criteria as assessed by the investigator Progression-free survival (PFS) determined according to the Lugano 2014 criteria as assessed by the investigator Complete response (CR) rate determined according to the Lugano 2014 criteria as assessed by the investigator Time to response (TTR) determined according to the Lugano 2014 criteria as assessed by the investigator Time to next anti-lymphoma treatment (TTNT) Rate and duration of minimal residual disease (MRD) negativity · Overall survival (OS)

[0257] Safety Endpoints Safety and tolerability evaluations during the study may include, but are not limited to, the following:

[0258] Monitoring the severity and incidence of adverse events (AEs), including adverse events of special interest (AESIs) Cytokine release syndrome (CRS), immune cell-associated neurotoxicity syndrome (ICANS), and clinical tumor lysis syndrome (CTLS) Clinical Laboratory Tests (Haematology, Chemistry, and Urinalysis) Monitor the incidence and severity of laboratory changes Physical Examination Measurement of vital signs Electrocardiogram (ECG) variables

[0259] Pharmacokinetic endpoints Maximum observed plasma concentration (C max ), C max Time to max ), and pharmacokinetic (PK) parameter values ​​including area under the plasma concentration versus time curve (AUC) will be determined using non-compartmental methods for epcolitamab in combination with lenalidomide or lenalidomide and ibrutinib. Epcolitamab anti-drug antibodies (ADA) and neutralizing ADA in combination with lenalidomide or lenalidomide and ibrutinib.

[0260] Overview of study design A schematic of the overall study design is shown in Figure 3.

[0261] Test Arm The following regimens will be evaluated initially in the appropriate populations:

[0262] Arm 1: Epcolitamab in combination with lenalidomide in subjects with R / R DLBCL Arm 2: Epcolitamab in combination with ibrutinib and lenalidomide in subjects with R / R DLBCL

[0263] Study treatment Arm 1: 12 cycles of epcolitamab in combination with lenalidomide Lenalidomide 25 mg orally (PO) will be administered on days 1-21 (except days 22-28) of cycles 1-12. Epcolitamab will be administered in 28-day cycles for a total of 12 cycles as described below.

[0264] Arm 2: 24 cycles of epcolitamab in combination with lenalidomide and ibrutinib Ibrutinib (420 mg or 560 mg) will be administered orally on days 1-28 of cycles 1-24. Lenalidomide 20 mg will be administered orally on days 1-21 of cycles 1-24. Epcolitamab will be administered as above for a total of 24 28-day cycles.

[0265] Arm 1 and Arm 2: Epcolitamab in combination with study drug(s) will be administered using a step-up dosing regimen: a priming dose of 0.16 mg (Day 1 of Cycle 1), followed by an intermediate dose of 0.8 mg (Day 8 of Cycle 1), and a total dose of 24 or 48 mg at the assigned dose level (Day 15 of Cycle 1 onwards). Epcolitamab will be administered as a SC injection once every week (QW) in cycles 2-3, followed by once every 4 weeks (Q4W) in cycles 4 through 12 (Arm 1) or cycles 4 through 24 (Arm 2).

[0266] Each arm consists of two phases: dose escalation (n-12 subjects for each dose level) and expansion (n=20 subjects). Within each arm, subjects can only participate in one phase. The dose escalation and expansion phases of each arm consist of a screening period, a treatment period, a post-treatment follow-up period, a safety follow-up period, and a survival follow-up period.

[0267] Dose Escalation Phase The dose escalation phase is designed to evaluate the initial safety and tolerability of lenalidomide or epcolitamab in combination with lenalidomide or ibrutinib.

[0268] Dose escalation will be guided by a Bayesian optimal interval (BOIN) design. For each arm, initial enrollment in the dose escalation cohort will consist of at least three DLT-evaluable subjects. For each arm, epcolitamab will be administered initially in combination with the corresponding antineoplastic agent. Arm 1 will begin with an epcolitamab dose level of 48 mg. Arm 2 will begin with an epcolitamab dose level of 24 mg, and if acceptable safety and tolerability are observed during the DLT period, the epcolitamab dose will be escalated to the next dose level of 48 mg. Decisions to de-escalate or escalate to a higher dose of epcolitamab will be made according to the BOIN design and based on the cumulative number of subjects who experience dose-limiting toxicity (DLT).

[0269] For arm 2, the initial dose level of ibrutinib will be 420 mg. Escalation to a 560 mg ibrutinib dose may be considered if permitted by the escalation decision rules. In a single cohort, only one agent (either epcolitamab or ibrutinib) may be escalated (i.e., a single cohort may not escalate both agents simultaneously).

[0270] Table 2 below provides titration decision rules for the BOIN design with a target toxicity rate of 0.25 and an optimal interval of (0.204, 0.304). [Table 2]

[0271] Dose-limiting toxicities (DLTs) will be evaluated during each dose-escalation cohort to define the recommended phase 2 dose (RP2D). In this study, the DLT evaluation period is defined as the first 4 weeks, i.e., 28 days after the first dose of epcolitamab.

[0272] After all subjects at a dose level have completed the DLT evaluation period, all available data will be evaluated to recommend the next dose level.

[0273] After completion of the dose escalation phase, the Sponsors will review the cumulative study data and recommend the declared dose of epcolitamab to be used in the dose expansion phase. The overall data, including safety (i.e., AEs and safety laboratory values, as well as observations made after the end of the DLT evaluation period), pharmacokinetics, pharmacodynamics, and preliminary efficacy, will be evaluated to guide further development of the expansion phase.

[0274] Expansion Stage The objective of the expansion phase is to evaluate the safety, tolerability, and preliminary clinical activity of epcolitamab at recommended doses in combination with antineoplastic agents.

[0275] The expansion phase of the study will enroll a total of approximately 20 subjects in each arm. Epcolitamab will be administered at the recommended phase 2 dose determined (RP2D) in combination with lenalidomide, or lenalidomide and ibrutinib, in the same manner as was done in dose escalation.

[0276] A toxicity monitoring rule will be implemented in each expansion cohort after six subjects have been enrolled. The rule will monitor the occurrence of DLT in each expansion cohort and will pause enrollment into the cohort if the posterior probability of a DLT rate exceeding 0.25 is greater than 80%. The prior distribution of DLT rates in each expansion cohort is assumed to follow a beta (1.5,4.5) distribution, reflecting a prior mean DLT rate of 0.25 and an effective sample size of 6. This corresponds to the target toxicity rate defined in the dose escalation portion (0.25) and the preliminary recommended dose and the minimum number of subjects to be enrolled in the schedule specified for further investigation during dose escalation (6).

[0277] If the number of expansion cohort subjects experiencing DLTs exceeds the toxicity boundary at any time after six subjects have been enrolled, further enrollment into that cohort will be paused and a comprehensive safety review of all available data will be conducted. Based on toxicity monitoring rules, enrollment into an expansion cohort will be paused if the number of subjects experiencing DLTs meets any of the following boundaries:

[0278] Three or more of the six registered subjects 4 or more of the 7-9 registered subjects 5 or more of the 10-12 registered subjects More than 6 of the 13-16 registered subjects More than 7 of the 17-19 registered subjects 8 or more of the 20 registered subjects

[0279] Inclusion criteria Subjects must meet all of the following criteria to be included in the study: · An adult male or female, at least 18 years of age. Laboratory tests that meet the following criteria during the screening period prior to the first dose of study drug: Absolute neutrophil count (ANC) ≥ 1.0 x 109 / L (growth factor use is permitted if evidence of bone marrow involvement is present, but subjects must not have received growth factors within 14 days prior to screening) Hemoglobin ≥ 8.0 g / dL (RBC transfusions are permitted, but subjects must not have received a blood transfusion within 7 days prior to screening) Platelet count ≥ 75 x 109 / L, or ≥ 50 x 109 / L if bone marrow infiltration or splenomegaly is present (platelet transfusions are permitted, but subjects must not have received a transfusion within 7 days prior to screening). Serum aspartate transaminase (AST) or alanine transaminase (ALT) levels ≤ 3 × ULN Total bilirubin level ≦1.5×ULN or ≦5×ULN (for subjects with disease or liver involvement of non-hepatic origin). Subjects with Gilbert's syndrome may have total bilirubin levels above 1.5×ULN, but direct bilirubin must be less than 2×ULN. Estimated creatinine clearance (CrCl) ≥ 50 mL / min (calculated by the Cockcroft-Gault formula, corrected as necessary for factors such as body weight) Prothrombin time (PT) / international normalized ratio (INR) / activated partial thromboplastin time (aPTT) ≦1.5×ULN (except in patients receiving anticoagulant therapy) Subject must be able to tolerate subcutaneous injections Subjects must have sufficient fresh or paraffin-embedded tissue available at the time of screening.

[0280] Disease / Condition Activity Diagnosis of DLBCL (de novo or histologically transformed from follicular lymphoma or nodal marginal zone lymphoma) with histologically confirmed CD20+ disease, including the following according to the WHO 2016 classification, as recorded in the pathology report: DLBCL, Not Otherwise Specified (NOS) High-grade B-cell lymphoma with MYC and BCL-2 and / or BCL-6 translocations according to WHO 2016 ("double hit" or "triple hit") Note: High-grade B-cell lymphoma NOS or other double / triple-hit lymphomas (with histology inconsistent with DLBCL) are not eligible Follicular lymphoma grade 3B Subjects must not have received prior treatment with a bispecific antibody targeting CD3 and CD20. Subjects must have one or more measurable sites of disease:

[0281] Positron emission tomography / computed tomography (PET / CT) scan demonstrating PET-positive lesion(s) and At least one measurable nodal lesion (long axis ≥ 1.5 cm and short axis > 1.0 cm) or one or more measurable extranodal lesions (long axis ≥ 1.0 cm) on CT or MRI Subjects must be eligible and require treatment initiation based on symptoms and / or disease burden as assessed by the investigator. Subjects must have Eastern Cooperative Oncology Group (ECOG) performance status 0-2. Subjects will have no unresolved toxicity from prior anticancer therapy, defined as unresolved Common Terminology Criteria for Adverse Events (CTCAE, v5.0), Grade 1, except for alopecia. Other eligibility criteria (e.g., laboratory, cardiac criteria) must also be met. Subjects have no current evidence of primary central nervous system (CNS) tumors or known CNS involvement (including leptomeningeal disease) at the time of screening. Subjects have no history of severe allergic or anaphylactic reactions to anti-CD20 mAb therapy, or known significant allergies or intolerances to any components or excipients of epcolitamab or to any components of the study drug combination (e.g., lenalidomide, ibrutinib, etc.). Subjects must not have undergone an autologous stem cell transplant within 3 months prior to screening. Subjects must not have received any chemotherapy, non-investigational, or investigational antineoplastic agent (except for CD20 mAbs) within 4 weeks or 5 half-lives (whichever is shorter) of the first dose of epcolitamab. Subjects were free of clinically significant cardiovascular disease, including: Myocardial infarction or stroke within 6 months prior to enrollment or Any of the following conditions within 3 months prior to enrollment: diseases / conditions related to or affecting unstable or uncontrolled cardiac function (e.g., unstable angina, congestive heart failure, New York Heart Association class III-IV), uncontrolled cardiac arrhythmias or Other clinically significant electrocardiogram (ECG) abnormalities within 6 months prior to enrollment unless deemed stable and appropriately treated or Left ventricular ejection fraction <45%.

[0282] Subjects are free of clinically significant liver disease, including hepatitis, current alcohol abuse, or cirrhosis. Subjects do not have active Hepatitis B virus (HBV) or Hepatitis C virus (HCV) infection.

[0283] Subjects who are positive for hepatitis B core antibody (HBcAb), hepatitis B surface antigen (HBsAg), or hepatitis C antibody must have a negative polymerase chain reaction (PCR) result prior to enrollment. Those with a positive PCR result will be excluded.

[0284] Subjects have no known history of Human Immunodeficiency Virus (HIV) infection. Note: HIV testing does not need to be done at the time of screening unless required according to local guidelines or institutional standards. Subjects must not have any known active bacterial, viral, fungal, mycobacterial, parasitic, or other infection (except fungal infections of the nail bed) requiring intravenous (IV) therapy or IV antibiotics within 2 weeks prior to enrollment. Subjects have no evidence of significant uncontrolled comorbidities that may affect protocol compliance or interpretation of results. Subjects must have no history of other previous malignancies, except for: Malignancies that have been treated with curative intent, have had no known active disease for at least 3 years prior to the first dose of study drug, and are considered by the treating physician to be at low risk of recurrence Non-melanoma skin cancer or lentigo maligna that has been adequately treated without evidence of disease Successfully treated carcinoma in situ with no evidence of disease Localized prostate cancer, post-radical prostatectomy with no elevated prostate-specific antigen (PSA) level <0.1ng / mL Subjects did not receive lesion-directed radiation therapy if only one target lesion was involved and there was no way to follow up on other target lesions that did not receive radiation therapy, or if they underwent major surgery within 4 weeks of enrollment. Subjects do not have grade >1 neuropathy. Subjects must not have a history of active tuberculosis (TB) or completed treatment for active TB within the past 12 months.

[0285] NOTE: Interferon-gamma release assay (IGRA) testing does not need to be done at screening unless active or latent TB is suspected. Subjects with a positive IGRA and active pulmonary TB must be ruled out with clinical evaluation and radiological imaging. Subjects with a positive IGRA and no evidence of active disease may be enrolled after treatment for latent TB infection (recommended total of 6 months of isoniazid monotherapy) has been initiated.

[0286] Subjects have no evidence of cytomegalovirus (CMV) viremia (defined as a positive level above the lower limit of detection) at screening. Subjects have no current autoimmune disease requiring immunosuppressive therapy, except for up to 20 mg of prednisone (or equivalent) daily. The subject is free of any life-threatening illness, medical condition, or organ system dysfunction that, in the opinion of the investigator, may compromise the subject's safety or place the study results at undue risk. Subjects have no current seizure disorder requiring treatment. Subjects do not have known active SARS-CoV-2 infection. If a subject has signs / symptoms suggestive of SARS-CoV-2 infection or is known to have had recent exposure to a SARS-CoV-infected individual, they should undergo molecular (e.g., PCR) testing or two negative antigen test results, at least 24 hours apart, to rule out SARS-CoV-2 infection.

[0287] Subjects who do not meet the SARS-CoV-2 infection eligibility criteria must fail screening and may only be rescreened after meeting the following SARS-CoV-2 infection viral clearance criteria:

[0288] At least 10 days have elapsed since the first positive test result in an asymptomatic patient, or at least 10 days have elapsed since recovery (defined as resolution of fever and improvement of symptoms without the use of antipyretic medications).

[0289] Subjects must not have undergone major surgery within 4 weeks of the first dose of study drug.

[0290] Further eligibility criteria specific to Arm 1 Subjects must have relapsed / refractory DLBCL Note: Relapsed disease is defined as disease that previously responded to treatment but progressed 6 months or more after completion of treatment. Refractory disease is defined as disease that has progressed during treatment, failed to achieve an objective response to previous treatment, or progressed within 6 months after completion of treatment (including maintenance therapy). Subjects must have R / R disease to at least one prior systemic anti-lymphoma therapy (radiotherapy is not considered systemic therapy) containing an anti-CD20 monoclonal antibody. Subjects who have previously received only anti-CD20 monoclonal antibody monotherapy are not eligible. Subjects must not be refractory (defined as best response of SD or PD) to prior CAR-T therapy. Subjects must have either failed a previous autologous stem cell transplant (ASCT), been deemed not eligible for ASCT due to age, performance status, comorbidities and / or inadequate response, or refused ASCT. Subjects must not have documented refractory disease to lenalidomide and, in the opinion of the investigator, be suitable for treatment with lenalidomide. Note: Refractoriness is defined as follows: o Best response to prior regimen(s) with stable disease (SD) or progressive disease (PD); or o Progressive disease within 6 months of completion of previous regimen(s) Subjects must be willing to receive aspirin prophylaxis or prophylactic anticoagulation therapy for thromboembolic events (or as per local guidelines for lenalidomide administration). Female subjects of childbearing potential must be on at least two protocol-specified methods of birth control that are effective from 30 days prior to enrollment until at least 12 months after the last dose of study drug. Non-childbearing female subjects are not required to use birth control. Subject is willing to comply with a pregnancy risk minimization plan associated with lenalidomide treatment. Subjects must not have had exposure to lenalidomide within 12 months prior to screening.

[0291] Additional eligibility criteria specific to Arm 2: Subjects must have R / R DLBCL (see above for definition). Subjects must have received at least one prior treatment that must have included an anti-CD20 monoclonal antibody in combination with another systemic therapy. Subjects must have received a prior CAR-T cell therapy, with the exception of those who achieved a response to a prior CAR-T ≥ 90 days prior to the first dose of epcolitamab or those who were refractory to CAR-T ≥ 60 days prior to the first dose of epcolitamab. Note: Refractoriness is defined as follows: o Best response to prior regimen(s) with stable disease (SD) or progressive disease (PD); or o Progressive disease within 6 months of completion of previous regimen(s) Subjects must have either failed a previous ASCT, were not considered eligible for ASCT due to age, performance status, comorbidities and / or inadequate response, or declined ASCT. Subjects must not have documented refractory disease to lenalidomide and, in the opinion of the investigator, be suitable for treatment with lenalidomide. Subjects must not have received prior treatment with ibrutinib and, in the opinion of the investigator, must be suitable for treatment with ibrutinib. Subjects must not have a known bleeding diathesis (e.g., von Willebrand disease) or hemophilia. Subjects must not require treatment with strong cytochrome P450 (CYP) 3A inhibitors. Subjects must be willing to receive aspirin prophylaxis or prophylactic anticoagulation therapy for thromboembolic events (or as per local guidelines for lenalidomide administration). Female subjects of childbearing potential must be on at least two protocol-specified methods of birth control that are effective from 30 days prior to enrollment until at least 12 months after the last dose of study drug. Non-childbearing female subjects are not required to use birth control. Subject is willing to comply with a pregnancy risk minimization plan associated with lenalidomide treatment. Subjects must be able to swallow capsules and must not have any disease that significantly affects gastrointestinal function (e.g., stomach or small intestine resection, symptomatic inflammatory bowel disease, or partial or complete ileus).

[0292] Dose-limiting toxicity DLT-evaluable subjects in the dose escalation phase were defined as those who received at least 3 doses of epcolitamab at the assigned dose level in the first cycle or experienced a DLT during the 28-day period following the first dose of epcolitamab.

[0293] The DLT evaluation period was defined as the first 4 weeks, i.e., 28 days after the first dose of epcolitamab, if the subject received at least 3 doses of epcolitamab during this period.

[0294] The following qualify as DLTs unless the investigator can attribute the event to a clearly identifiable cause, e.g., underlying disease, disease progression / recurrence, other comorbidities, or concomitant therapy:

[0295] Grade 5 toxicity CRS classification according to the American Society for Transplantation and Cellular Therapy (ASTCT) criteria and DLT criteria for CRS o Grade 4 CRS by ASTCT criteria or ICANS o Grade 3 CRS by ASTCT criteria or ICANS that has not improved to Grade 2 or less or has not resolved within 48 hours (Grade 0) Neutropenia Grade 4 lasting more than 7 days as determined by CTCAE. Febrile neutropenia of CTCAE grade ≥ 3 lasting more than 2 days. CTCAE grade 4 thrombocytopenia lasting more than 7 days. Non-hematologic toxicity of grade 3 or greater as determined by CTCAE, except for: Grade 3 fever (>40.0°C for ≤24 hours) Grade 3 hypotension (resolved within 24 hours) o Laboratory values ​​outside the normal range that have no clinical consequences, are clinically transient, are essentially isolated, and resolve within 7 days (this includes electrolyte abnormalities that respond to medical intervention) o AST and / or ALT Grade 3 with return to Grade 1 or baseline within 7 days. Grade 3 nausea that responds to optimal antiemetic therapy within 3 days. o Grade 3 vomiting that responds to optimal antiemetic therapy within 3 days. o Grade 3 diarrhea that responds to optimal antidiarrheal treatment within 3 days. Grade 3 fatigue / asthenia if present at baseline or lasting <14 days after the last dose of epcolitamab. o Other grade 3 toxicity related to prior chemotherapy that was present at baseline (grade 1 or 2) and returned to baseline within 7 days.

[0296] Alopecia (no grade classification) Frequent laboratory monitoring of complete blood counts with differential should be initiated to document the onset and resolution of hematologic AEs. All AEs occurring during the defined DLT evaluation period will be evaluated according to the criteria above. All AEs, including those not eligible for DLTs, will be monitored and included in the evaluation of the toxicity profile of epcolitamab, unless the event is clearly determined to be unrelated to epcolitamab.

[0297] Adverse events of special interest The following adverse events of particular interest will be monitored during the study. Cytokine Release Syndrome (CRS) · Clinical tumor lysis syndrome (CTLS) Immune Cell-Associated Neurotoxicity Syndrome (ICANS)

[0298] CRS prophylaxis and premedication Premedication with corticosteroids, antihistamines, and antipyretics is mandatory as described in section 3.4 of the Operations Manual. For the first four doses of epcolitamab, premedication with antihistamines, antipyretics, and corticosteroids is mandatory, and an additional 3 days of corticosteroids are required after each of these first four doses to prevent / reduce the severity of symptoms from potential CRS. For the first four doses of epcolitamab, subjects must perform self-administered oral temperature monitoring three times per day (approximately every 6-8 hours during waking hours) for the first four days after epcolitamab administration. These temperature checks are to ensure that fever, an early sign of CRS, has not developed. If epcolitamab is administered beyond the fourth dose (i.e., the second full dose), CRS prophylaxis with corticosteroids is optional unless CRS grade 2 or higher occurs, in which case CRS prophylaxis should be continued until a dose of epcolitamab has been given without subsequent CRS. Pre-medication corticosteroid administration can be either IV or PO at the recommended dose or an equivalent dose.

[0299] Test evaluation Assessment of disease response and progressive disease On-treatment evaluation: Response at on-treatment time points should be interpreted according to the Lugano classification for patients showing CR, PR, and SD. For patients showing PD by Lugano classification, further evaluation should be performed to see if the subject can be considered to have IR (by LYRIC).

[0300] Lugano response criteria for malignant lymphoma Target and non-target lesions Target lesions consist of up to six of the largest dominant nodes, nodal masses, or other lymphomatous lesions measurable in two diameters, preferably from different body regions that represent the subject's overall disease burden, including mediastinal and retroperitoneal disease, if applicable. At baseline, measurable nodes must be greater than 15 mm in longest diameter (longest transverse diameter of lesion; LDi). Measurable extranodal disease may be included in six representative target lesions. At baseline, measurable extranodal lesions should be greater than 10 mm in LDi.

[0301] All other lesions (including nodal, extranodal, and evaluable disease) should be followed as non-target lesions (e.g., skin, GI, bone, spleen, liver, kidney, pleural or pericardial effusion, ascites, bone, bone marrow).

[0302] Mitotic and confluent lesions Lesions may split or become confluent over time. In the case of splitting lesions, the individual products of the nodule perpendicular diameter (PPD) should be summed together to represent the PPD of the splitting lesion, and this PPD is added to the sum of the PPD of the remaining lesions to measure the response. If subsequent growth of any or all of these individual nodules occurs, the nadir of each individual nodule is used to determine progression. In the case of confluent lesions, the PPD of the confluent mass should be compared to the sum of the individual nodules, and a greater than 50% increase in the PPD of the confluent mass compared to the sum of the individual nodules is required to show PD. LDi and minimum diameter (shortest axis perpendicular to LDi; SDi) are no longer necessary to determine progression.

[0303] [Table 3] TIFF2025503176000004.tif208154TIFF2025503176000005.tif93154

[0304] 1. A score of 3 in many subjects indicates a good prognosis with standard therapy, especially at interim scans. However, in studies including PET where deprogression is investigated, it may be preferable to consider a score of 3 as an inadequate response (to avoid undertreatment). · Measured dominant (target) lesions: up to the six largest dominant nodules, nodal masses, and extranodal lesions selected to be clearly measurable in two diameters. o The nodes should preferably be from different regions of the body and should include the mediastinal and retroperitoneal regions, if applicable. o Non-nodular lesions include those in parenchymal organs (e.g., liver, spleen, kidneys, lungs), gastrointestinal lesions, skin lesions, or visible on palpation. · Non-measured lesions: Any disease not selected as measured, dominant disease and truly evaluable disease should be considered as not measured. o These sites include any nodules, nodal masses, and extranodal sites that are not selected as dominant or measurable or that do not meet the requirements for measurability but are still considered abnormal, as well as truly evaluable disease (this is any site of suspicion of disease where it would be difficult to quantitatively follow up measurements, including pleural effusions, ascites, bone lesions, leptomeningeal disease, abdominal masses, and other lesions that cannot be confirmed and subsequently imaged). In Waldeyer's ring or extranodal sites (e.g., GI tract, liver, bone marrow), FDG uptake may be greater than in the mediastinum with a complete metabolic response, but should not be higher than the surrounding normal physiologic uptake (e.g., with bone marrow activation as a result of chemotherapy or myeloid growth factors).

[0305] 2. PET 5PS: 1=no uptake above background; 2=uptake ≦ mediastinum; 3=uptake > mediastinum but ≦ liver; 4=moderate uptake > liver; 5=uptake significantly higher than liver and / or new lesions; ×=areas of new uptake unlikely to be associated with lymphoma.

[0306] Source: Cheson BD, Fisher R, Barrington SF, et al. Recommendations for initial evaluation, staging, and response assessment of Hodgkin and non-Hodgkin lymphoma: the Lugano classification.J Clin Oncol.2014;32:3059-68.

[0307] Lymphoma Response to Immunomodulatory Therapy Criteria (LYRIC) Clinical trials have shown that cancer immunotherapy can result in early apparent radiological progression (including the appearance of new lesions) followed by a delayed response. Because this initial increase in tumor size can be caused by immune cell infiltration in the context of a T-cell response, this progression may not represent true disease progression and is therefore termed "pseudoprogression."

[0308] The association of epcolitamab (GEN3013; DuoBody®-CD3xCD20) with pseudoprogression is currently unknown, but its mechanism of action means that pseudoprogression would be expected.

[0309] Current Lugano response assessment criteria do not take into account pseudoprogression, and there is a significant risk of premature discontinuation of potentially effective immunomodulatory agents after the observation of an atypical response, which is characterized by either early progression of existing lesions, a subsequent response, or the development of new lesions with or without tumor shrinkage elsewhere.

[0310] LYRIC is a modification of the Lugano response evaluation criteria that is adapted for immune-based therapies and implements a new palliative response category called "indeterminate response" (IR). 5 This IR designation was introduced to potentially identify "atypical response" cases until confirmed as flare / pseudoprogression or true PD by either biopsy or subsequent imaging. The LYRIC and Lugano criteria will be evaluated in this study.

[0311] Indeterminate Response (IR) Category A subject exhibiting PD according to the Lugano classification is considered to have IR in one or more of the following three situations:

[0312] IR(1): An increase in total tumor burden (assessed by sum of products of diameters [SPD]) of ≥50% for up to six target lesions during the first 12 weeks of treatment without clinical progression.

[0313] IR(2): Appearance of a new lesion or growth of one or more existing lesions ≧50% at any time during treatment; occurs in the absence of an overall progression of overall tumor burden (increase in SPD<50%) as measured by SPD of up to six lesions at any time during treatment.

[0314] IR(3): Increased FDG uptake in one or more lesions without a concomitant increase in lesion size or number.

[0315] Screening evaluation: At screening, FDG-PET / CT and diagnostic CT or MRI scans should be read according to the Lugano classification, as described above.

[0316] On-treatment evaluation: Response at on-treatment time points should be interpreted according to the Lugano classification for patients showing CR, PR, and SD. For patients showing PD by Lugano classification, further evaluation should be performed to see if the subject can be considered to have IR (by LYRIC).

[0317] Statistical analysis of efficacy Descriptive statistics and subject listing will be used to summarize data for each epcolitamab dose level (24 mg and 48 mg). For continuous variables, number of observations, mean, standard deviation, median, and range will be used. For categorical variables, frequencies and percentages will be summarized. For time-to-event endpoints, Kaplan-Meier estimates will be provided.

[0318] Summary and analysis of key secondary efficacy endpoints Overall response rate (ORR) is defined as the proportion of subjects achieving a best overall response of CR or PR as determined by the Lugano 2014 criteria as assessed by the investigator. Point estimates are provided for each arm with 95% exact confidence intervals (CI).

[0319] Duration of response (DOR) is defined for subjects who achieved a best overall response of CR or PR ("responders") as the time in months from the initial CR / PR to the earliest occurrence of disease progression or death from any cause as determined by the Lugano 2014 criteria as assessed by the investigator. Responders alive without radiological disease progression are censored at the time of the last adequate disease assessment.

[0320] The number of responders, number of DOR events and earliest attributable event (disease progression or death) will be summarized by arm. The Kaplan-Meier method will be used to estimate the distribution of DoR in each arm.

[0321] Progression-free survival (PFS) is defined for subjects in all arms as the time in months from first dose of study drug to the earliest occurrence of disease progression as determined by the Lugano 2014 criteria as assessed by the investigator or death from any cause. Subjects surviving without disease progression are censored at the time of the last adequate disease assessment. Subjects surviving without post-baseline disease assessment are censored at the date of first dose of study drug.

[0322] The number of PFS events and the earliest attributable event (disease progression or death) are presented by arm. The distribution of PFS is estimated using the Kaplan-Meier method.

[0323] Complete response rate is defined as the proportion of subjects achieving a best overall response of CR as determined by the Lugano 2014 criteria as assessed by the investigator. Point estimates are provided for each arm with 95% exact confidence intervals (CI).

[0324] Time to response (TTR) is defined as the time in months from the first dose of study drug to first CR / PR for subjects who achieved a best overall response of CR or PR as determined by the Lugano 2014 criteria as assessed by the investigator ("responders").

[0325] The number of responders along with a narrative summary of TTR will be provided for each arm.

[0326] Overall survival (OS) is defined for subjects in all arms as the time in months from first dose of epcolitamab to death from any cause. Subjects still alive at the end of the study or at the time of analysis will be censored at their last known survival date.

[0327] Kaplan-Meier estimates of the distribution of deaths and OS are provided.

[0328] Statistical Analysis of Safety The safety and tolerability of epcolitamab in combination with other agents will be evaluated by assessment of study drug exposure, incidence of dose interruptions, attenuations, delays and discontinuations, AEs including AESIs, SAEs, deaths, and changes in adverse events and vital sign parameters.

[0329] Treatment-emergent AEs will be summarized by Preferred Terms within System Organ Class in the Medical Dictionary for Regulatory Activities. The number and percentage of subjects experiencing DLTs will be summarized. Further details will be provided in the SAP.

[0330] Where applicable, blood chemistry and hematology laboratory determinations will be categorized and summarized according to NCI CTCAE. Further details will be provided in the SAP.

[0331] Statistical analysis of pharmacokinetics Plasma concentrations of epcolitamab will be tabulated along with PK parameter values ​​for drug within each cohort. Summary statistics will be calculated by sampling time for PK concentrations and by cycle and / or visit for PK parameters. Results of epcolitamab ADA (and nAb, if applicable) will be summarized. Additional exploratory analyses may be performed if deemed appropriate.

[0332] Preliminary results Arm 1: Dose escalation (epcolitamab 24 mg + Len 25 mg): Number of subjects registered: 5 Number of subjects expected to have at least one post-baseline efficacy assessment: 5 Number of subjects with available post-baseline efficacy assessments: 3 ORR=100%(3 / 3) CRR=100%(3 / 3) Arm 1: Dose expansion (epcolitamab 48 mg + Len 25 mg): Number of subjects registered: 17 Number of subjects expected to have at least one post-baseline efficacy assessment: 3 Number of subjects with available post-baseline efficacy assessments: 0

[0333] [Table 4] TIFF2025503176000007.tif234154TIFF2025503176000008.tif224153

[0334] Bold and underlined are F; E; A; L and R, corresponding to positions 234, 235; 265; 405 and 409, respectively, according to EU numbering. In the variable region, the CDR regions, annotated according to the IMGT definition, are underlined.

Claims

1. (i) a first binding arm that binds to human CD3ε (epsilon) and comprises a first antigen-binding region comprising a variable heavy (VH) region and a variable light (VL) region, wherein the VH region comprises the CDR1, CDR2, and CDR3 sequences found in the VH region sequence of SEQ ID NO: 6, and the VL region comprises the CDR1, CDR2, and CDR3 sequences found in the VL region sequence of SEQ ID NO: 7; and (ii) a second binding arm that binds to human CD20 and comprises a second antigen-binding region comprising a VH region and a VL region, wherein the VH region comprises the CDR1, CDR2, and CDR3 sequences found in the VH region sequence of SEQ ID NO: 13, and the VL region comprises the CDR1, CDR2, and CDR3 sequences found in the VL region sequence of SEQ ID NO: 14; A therapeutic amount for diffuse large B-cell lymphoma (DLBCL) in a human subject, The method includes administering to the subject a bispecific antibody, an effective amount of lenalidomide, and optionally an effective amount of ibrutinib; wherein said bispecific antibody is administered at a dose of 24 mg or 48 mg, and wherein lenalidomide, said bispecific antibody and optionally ibrutinib are administered in a 28 day cycle.

2. 10. The pharmaceutical composition of claim 1, wherein the bispecific antibody is administered at a dose of 24 mg.

3. 10. The pharmaceutical composition of claim 1, wherein the bispecific antibody is administered at a dose of 48 mg.

4. 10. The pharmaceutical composition of claim 1, wherein the bispecific antibody is administered once a week (administered weekly).

5. 5. The pharmaceutical composition of claim 4, wherein weekly administration of 24 mg or 48 mg is administered over 2.5 28-day cycles.

6. 6. The pharmaceutical composition of claim 4 or 5, wherein after said weekly administration, the bispecific antibody is administered once every four weeks, for example in a 28 day cycle, on day 1 of each 28 day cycle.

7. 7. The pharmaceutical composition of claim 6, wherein the once every four weeks administration occurs over at least eight 28-day cycles, such as eight 28-day cycles or nine 28-day cycles.

8. 7. The pharmaceutical composition of claim 6, wherein the once every four weeks administration occurs over at least 20 28-day cycles, such as 20 28-day cycles or 21 28-day cycles.

9. 5. The pharmaceutical composition of claim 4, wherein a priming dose of the bispecific antibody is administered in cycle 1 of a 28 day cycle prior to the weekly administration of 24 mg or 48 mg.

10. 10. The pharmaceutical composition of claim 9, wherein the priming dose is administered two weeks before the first weekly dose of 24 mg or 48 mg.

11. 11. The pharmaceutical composition of claim 9 or 10, wherein the priming dose is 0.16 mg.

12. 10. The pharmaceutical composition of claim 9, wherein an intermediate dose of the bispecific antibody is administered after the priming dose and before the first weekly dose of 24 mg or 48 mg.

13. 13. The pharmaceutical composition of claim 12, wherein the priming dose is administered on day 1, the intermediate dose is administered on day 8, followed by first weekly doses of 24 mg or 48 mg on days 15 and 22 of cycle 1.

14. 14. The pharmaceutical composition of claim 12 or 13, wherein the intermediate dose is 0.8 mg.

15. 2. The pharmaceutical composition of claim 1, wherein lenalidomide is administered once daily on days 1 through 21 of a 28-day cycle.

16. 2. The pharmaceutical composition of claim 1, wherein lenalidomide is administered from cycle 1 to cycle 12 of a 28-day cycle.

17. 2. The pharmaceutical composition of claim 1, wherein lenalidomide is administered from cycle 1 to cycle 24 of a 28-day cycle.

18. 2. The pharmaceutical composition of claim 1, wherein lenalidomide is administered at a dose of 20 to 30 mg, such as 25 mg.

19. 2. The pharmaceutical composition of claim 1, wherein lenalidomide is administered at a dose of 20 to 30 mg in cycles 1 to 12 of a 28-day cycle.

20. 2. The pharmaceutical composition of claim 1, wherein lenalidomide is administered at a dose of 25 mg in cycles 1 to 12 of a 28-day cycle.

21. 2. The pharmaceutical composition of claim 1, wherein lenalidomide is administered at a dose of 10 to 25 mg, for example 25 mg.

22. 22. The pharmaceutical composition of claim 1 or 21, wherein lenalidomide is administered at a dose of 10 to 25 mg in cycles 1 to 24 of a 28-day cycle.

23. 22. The pharmaceutical composition of claim 1 or 21, wherein lenalidomide is administered at a dose of 20 mg in cycles 1 to 24 of a 28-day cycle.

24. 22. The pharmaceutical composition of claim 1 or 21, wherein ibrutinib is administered once daily on days 1 through 28 of a 28-day cycle.

25. 22. The pharmaceutical composition of claim 1 or 21, wherein ibrutinib is administered from cycle 1 to cycle 24 of a 28-day cycle.

26. 22. The pharmaceutical composition of claim 1 or 21, wherein ibrutinib is administered at a dose of 280 mg to 560 mg, such as 280, 420 mg or 560 mg.

27. 22. The pharmaceutical composition of claim 1 or 21, wherein ibrutinib is administered at a dose of 560 mg on cycles 1 to 24 of a 28-day cycle, or at a dose of 420 mg on cycles 1 to 24 of a 28-day cycle.

28. Administration is given in 28-day cycles, (a) the bispecific antibody is administered as follows: (i) In Cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In cycles 2 and 3, a 24 mg dose is administered on days 1, 8, 15, and 22; (iii) In Cycle 4 and beyond, a dose of 24 mg is administered on Day 1; (b) lenalidomide is administered on Days 1-21 of Cycle 1 onwards; and (c) ibrutinib is optionally administered on days 1-28 from Cycle 1 onwards; The pharmaceutical composition of claim 1.

29. Administration is given in 28-day cycles, (a) the bispecific antibody is administered subcutaneously as follows: (i) In Cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a 24 mg dose is administered on days 1, 8, 15, and 22; (iii) In Cycles 4-12, a dose of 24 mg is administered on Day 1; (b) lenalidomide is administered orally at a dose of 25 mg / day on days 1-21 of cycles 1-12; and The pharmaceutical composition of claim 1.

30. Administration is given in 28-day cycles, (a) the bispecific antibody is administered subcutaneously as follows: (i) In Cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a 24 mg dose is administered on days 1, 8, 15, and 22; (iii) In Cycles 4-24, a 24 mg dose is administered on Day 1; (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib is administered orally at a dose of 560 mg / day on days 1-28 of cycles 1-24. The pharmaceutical composition of claim 1.

31. Administration is given in 28-day cycles, (a) the bispecific antibody is administered subcutaneously as follows: (i) In Cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 24 mg are administered on days 15 and 22; (ii) In Cycles 2-3, a 24 mg dose is administered on days 1, 8, 15, and 22; (iii) In Cycles 4-24, a 24 mg dose is administered on Day 1; (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib is administered orally at a dose of 420 mg / day on days 1-28 of cycles 1-24. The pharmaceutical composition of claim 1.

32. Administration is given in 28-day cycles, (a) the bispecific antibody is administered as follows: (i) In Cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) In cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In Cycle 3 and beyond, a dose of 48 mg is administered on Day 1; (b) lenalidomide is administered on Days 1-21 of Cycle 1 or later; and (c) ibrutinib is optionally administered on days 1-28 from Cycle 1 onwards; The pharmaceutical composition of claim 1.

33. Administration is given in 28-day cycles, (a) the bispecific antibody is administered subcutaneously as follows: (i) In cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) In cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In Cycles 4-12, a dose of 48 mg is administered on Day 1; (b) lenalidomide is administered orally at a dose of 25 mg / day on days 1-21 of cycles 1-12; and The pharmaceutical composition of claim 1.

34. Administration is given in 28-day cycles, (a) the bispecific antibody is administered subcutaneously as follows: (i) In Cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) In cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In Cycles 4-24, a dose of 48 mg was administered on day 1; (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib is administered orally at a dose of 560 mg / day on days 1-28 of cycles 1-24. The pharmaceutical composition of claim 1.

35. Administration is given in 28-day cycles, (a) the bispecific antibody is administered subcutaneously as follows: (i) In Cycle 1, a priming dose of 0.16 mg is administered on day 1, an intermediate dose of 0.8 mg is administered on day 8, and doses of 48 mg are administered on days 15 and 22; (ii) In cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15, and 22; (iii) In Cycles 4-24, a dose of 48 mg is administered on Day 1; (b) lenalidomide is administered orally at a dose of 20 mg / day on days 1-21 of cycles 1-24; and (c) Ibrutinib is administered orally at a dose of 420 mg / day on days 1-28 of cycles 1-24. The pharmaceutical composition of claim 1.

36. 10. The pharmaceutical composition of claim 1, wherein the bispecific antibody is administered subcutaneously.

37. 10. The pharmaceutical composition of claim 1, wherein ibrutinib is administered orally.

38. 10. The pharmaceutical composition of claim 1, wherein lenalidomide is administered orally.

39. 10. The pharmaceutical composition of claim 1, wherein the bispecific antibody, ibrutinib, and lenalidomide are administered sequentially.

40. 2. The pharmaceutical composition of claim 1, wherein the DLBCL has histologically confirmed CD20+ disease.

41. 2. The pharmaceutical composition according to claim 1, wherein the DLBCL is an aggressive B-cell lymphoma with MYC and BCL-2 and / or BCL-6 translocation (double-hit or triple-hit).

42. The pharmaceutical composition of claim 1, wherein the DLBCL is follicular lymphoma grade 3B.

43. The pharmaceutical composition of claim 1, wherein the DLBCL is relapsed and / or refractory DLBCL.

44. 2. The pharmaceutical composition of claim 1, wherein the DLBCL has relapsed, i.e., previously responded to a previous treatment but has progressed after the previous treatment, with progression starting 6 months or more after completion of the previous treatment.

45. 2. The pharmaceutical composition of claim 1, wherein the DLBCL is refractory, i.e., has either progressed during previous therapy, failed to achieve an objective response to previous therapy, or progressed within 6 months of completion of previous therapy, including maintenance therapy.

46. 10. The pharmaceutical composition of claim 1, wherein the subject has disease that is relapsed or refractory to at least one prior systemic anti-lymphoma therapy containing an anti-CD20 monoclonal antibody.

47. 2. The pharmaceutical composition of claim 1, wherein the DLBCL is not refractory to a previous chimeric antigen receptor T cell (CAR-T) therapy.

48. 2. The pharmaceutical composition of claim 1, wherein the subject is not refractory to lenalidomide or ibrutinib.

49. 10. The pharmaceutical composition of claim 1, wherein the subject has previously received at least one treatment with an anti-CD20 monoclonal antibody in combination with another systemic therapy.

50. 10. The pharmaceutical composition of claim 1, wherein the subject has previously received CAR-T therapy or is ineligible for or unable to receive CAR-T therapy.

51. 10. The pharmaceutical composition of claim 1, wherein the subject has not received prior treatment with ibrutinib.

52. (i) the first antigen-binding region of the bispecific antibody comprises VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and VLCDR1, VLCDR2, and VLCDR3 comprising the amino acid sequences set forth in SEQ ID NO: 4, SEQ ID NO: GTN, and SEQ ID NO: 5, respectively; and (ii) the second antigen-binding region of the bispecific antibody comprises VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively, and VLCDR1, VLCDR2, and VLCDR3 comprising the amino acid sequences set forth in SEQ ID NO: 11, SEQ ID NO: DAS, and SEQ ID NO: 12, respectively; The pharmaceutical composition of claim 1.

53. (i) the first antigen-binding region of the bispecific antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 6 and a VL region comprising the amino acid sequence of SEQ ID NO: 7; and (ii) the second antigen-binding region of the bispecific antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 13 and a VL region comprising the amino acid sequence of SEQ ID NO: 14; The pharmaceutical composition of claim 1.

54. 2. The pharmaceutical composition of claim 1, wherein the first binding arm of the bispecific antibody is derived from a humanized antibody, preferably a full-length IgG1, λ (lambda) antibody.

55. 55. The pharmaceutical composition of claim 54, wherein the first binding arm of the bispecific antibody comprises a lambda light chain constant region comprising the amino acid sequence set forth in SEQ ID NO:

22.

56. 2. The pharmaceutical composition of claim 1, wherein the second binding arm of the bispecific antibody is derived from a human antibody, preferably a full-length IgG1, κ (kappa) antibody.

57. 57. The pharmaceutical composition of claim 56, wherein the second binding arm comprises a kappa light chain constant region comprising the amino acid sequence set forth in SEQ ID NO:

23.

58. 2. The pharmaceutical composition of claim 1, wherein the bispecific antibody is a full-length antibody having a human IgG1 constant region.

59. The pharmaceutical composition of claim 1 , wherein the bispecific antibody comprises an inactive Fc region.

60. 2. The pharmaceutical composition of claim 1, wherein the bispecific antibody comprises a first heavy chain and a second heavy chain, wherein in both the first heavy chain and the second heavy chain, the amino acids at positions corresponding to L234, L235, and D265 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 are F, E, and A, respectively.

61. the bispecific antibody comprises a first heavy chain and a second heavy chain, wherein in the first heavy chain, the amino acid at the position corresponding to F405 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is L, and wherein in the second heavy chain, the amino acid at the position corresponding to K409 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is R, or vice versa; The pharmaceutical composition of claim 1.

62. the bispecific antibody comprises a first heavy chain and a second heavy chain, (i) in both the first and second heavy chains, the amino acids at positions corresponding to L234, L235, and D265 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 are F, E, and A, respectively; and (ii) in the first heavy chain, the amino acid at the position corresponding to F405 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is L, and in the second heavy chain, the amino acid at the position corresponding to K409 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is R, or vice versa; The pharmaceutical composition of claim 1.

63. 63. The pharmaceutical composition of claim 62, wherein the bispecific antibody comprises a heavy chain constant region comprising the amino acid sequences of SEQ ID NOs: 19 and 20.

64. 2. The pharmaceutical composition of claim 1, wherein the bispecific antibody comprises heavy and light chains comprising the amino acid sequences set forth in SEQ ID NOs: 24 and 25, respectively, and heavy and light chains comprising the amino acid sequences set forth in SEQ ID NOs: 26 and 27, respectively.

65. 2. The pharmaceutical composition of claim 1, wherein the bispecific antibody comprises heavy and light chains consisting of the amino acid sequences of SEQ ID NOs: 24 and 25, respectively, and heavy and light chains consisting of the amino acid sequences of SEQ ID NOs: 26 and 27, respectively.

66. 2. The pharmaceutical composition of claim 1, wherein the bispecific antibody is epcolitamab or a biosimilar thereof.

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