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

Abstract

Methods for clinical treatment of diffuse large B-cell lymphoma (DLBCL) (e.g., previously untreated DLBCL) in human subjects are provided using a bispecific antibody that binds to CD3 and CD20 in combination with Pola-R-CHP (polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and prednisone).

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 polatuzumab vedotin and R-CHP rituximab, cyclophosphamide, doxorubicin and prednisone or equivalents, e.g., a prednisolone regimen, for treating, e.g., diffuse large B-cell lymphoma (DLBCL), e.g., previously untreated DLBCL, in subjects with an International Prognostic Index (IPI) score of 2 to 5. 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 having DLBCL, e.g., previously untreated DLBCL (e.g., DLBCL with an International Prognostic Index (IPI) score of 2 to 5), by administering a bispecific antibody that binds CD3 and CD20 in combination with polatuzumab vedotin, rituximab and cyclophosphamide, doxorubicin and prednisone or its equivalent, e.g., prednisolone (R-CHP), among other favorable clinical treatment regimens.

[0007] In one aspect, provided herein is a method of treating diffuse large B-cell lymphoma (DLBCL) in a human subject, the method comprising administering to the subject a combination of epcolitamab and polatuzumab vedotin and rituximab, cyclophosphamide, doxorubicin and prednisone or an equivalent thereof, e.g., prednisolone (R-CHP), e.g., a method comprising administering to the subject effective amounts of polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin and a glucocorticoid, e.g., effective amounts of polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin and prednisone or an equivalent thereof, e.g., prednisolone.

[0008] In one aspect, there is provided a method of treating diffuse large B-cell lymphoma (DLBCL) in a human subject, the method comprising administering to the subject a bispecific antibody and an effective amount of polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and prednisone or an equivalent thereof, e.g., prednisolone, 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; Here, methods are provided herein, where the bispecific antibody is administered at a dose of 24 mg or 48 mg, and polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin and prednisone or its equivalent, e.g., prednisolone, and the bispecific antibody are administered in a 21 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 (weekly administration / dose), e.g., once weekly for 3 and 1 / 3 21 day cycles (i.e., day 15 of cycle 1 and days 1, 8, and 15 of cycles 2-4). In some embodiments, the bispecific antibody is administered once every 3 weeks after the weekly administration, e.g., for at least 2 or 4 21 day cycles, e.g., for 4 21 day cycles. In further embodiments, a priming dose (e.g., at 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., at or about 0.8 mg) of the bispecific antibody is administered prior to administering the weekly dose of 24 mg or 48 mg. In some embodiments, the priming dose is administered one week before the intermediate dose, which is administered one week before the first weekly dose of 24 mg or 48 mg.

[0011] In some embodiments, polatuzumab vedotin is administered once every 3 weeks in a 21 day cycle, e.g., for at least six 21 day cycles, e.g., for six 21 day cycles. In some embodiments, polatuzumab vedotin is administered at a dose of 1.8 mg / kg.

[0012] In some embodiments, rituximab is administered in a 21 day cycle, once every 3 weeks, for example, 6 or 8 21 day cycles. In some embodiments, rituximab is administered at 375 mg / m 2 is administered at a dose of

[0013] In some embodiments, cyclophosphamide is administered in 21-day cycles, for example, 6 or 8 21-day cycles, every 3 weeks. In some embodiments, cyclophosphamide is administered at 750 mg / m 2 is administered at a dose of

[0014] In some embodiments, doxorubicin is administered in 21-day cycles, for example, 6 or 8 21-day cycles, every 3 weeks. In some embodiments, doxorubicin is administered at 50 mg / m 2 is administered at a dose of

[0015] In some embodiments, prednisone or its equivalent is administered once daily on days 1 through 5 of a 21 day cycle, for example, for 6 or 8 21 day cycles. In some embodiments, prednisone or its equivalent; e.g., prednisolone, is administered at a dose of 100 mg / day.

[0016] In some embodiments, polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, prednisone, and the bispecific antibody are administered on the same day (e.g., on day 1 of cycles 1-6).

[0017] In some embodiments, the administration occurs in a 21 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 a dose of 24 mg is administered on day 15; (ii) In cycles 2 to 4, a dose of 24 mg is administered on days 1, 8, and 15; (iii) In cycles 5 through 8, a dose of 24 mg is administered on day 1, and so on. (b) polatuzumab vedotin, rituximab, cyclophosphamide, and doxorubicin are administered on Day 1 of Cycles 1 to 6; and (c) Prednisone or its equivalent, e.g., prednisolone, is administered on days 1-5 of cycles 1-6.

[0018] In some embodiments, the administration occurs in a 21 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 a dose of 48 mg is administered on day 15; (ii) In cycles 2 to 4, a dose of 48 mg is administered on days 1, 8, and 15; (iii) In cycles 5 through 8, a dose of 48 mg was administered on day 1, and so on. (b) polatuzumab vedotin, rituximab, cyclophosphamide, and doxorubicin are administered on Day 1 of Cycles 1-6, and (c) Prednisone or its equivalent, e.g., prednisolone, is administered on days 1-5 of cycles 1-6.

[0019] In some embodiments, the bispecific antibody is administered subcutaneously. In some embodiments, polatuzumab vedotin is administered intravenously. In some embodiments, rituximab is administered intravenously. In some embodiments, cyclophosphamide is administered intravenously. In further embodiments, doxorubicin is administered intravenously. In some embodiments, prednisone is administered intravenously or orally.

[0020] In some embodiments, the bispecific antibody, polatuzumab vedotin, rituximab, cyclophosphamide and doxorubicin are administered sequentially.

[0021] In some embodiments, the DLBCL has histologically confirmed CD20+ disease. In some embodiments, the DLBCL is a B cell lymphoma with a MYC and Bcl-2 and / or Bcl-6 translocation (double-hit or triple-hit DLBCL). In some embodiments, the DLBCL is follicular lymphoma grade 3B. In some embodiments, the subject has an International Prognostic Index (IPI) score of 2-5 or a revised IPI score, e.g., a score of 2, 3, 4, or 5.

[0022] In some embodiments, a subject treated with the methods described herein achieves a complete response (CR), partial response (PR), or stable disease (SD), e.g., as defined by the Lugano criteria or LYRIC.

[0023] 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, the 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, the sequence DAS, and SEQ ID NO:12, respectively.

[0024] 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]A is a graph showing the minimal effect of CHP components on DuoBody® CD3xCD20-induced T cell-mediated cytotoxicity. B is a graph showing the minimal effect of CHP components on DuoBody® CD3xCD20-induced T cell-mediated cytotoxicity. C is a graph showing the minimal effect of CHP components on DuoBody® CD3xCD20-induced T cell-mediated cytotoxicity. Figure 1A: DuoBody® CD3xCD20 + cyclophosphamide; Figure 1B: DuoBody® CD3xCD20 + doxorubicin; Figure 1C: DuoBody® CD3xCD20 + prednisone. The left panel shows the DuoBody® CD3xCD20 dose response curve for one representative donor. The right panel shows results for four donors at 333 ng / mL DuoBody® CD3xCD20. [Diagram 2] FIG. 1 is a schematic diagram of the overall clinical trial design. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] 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) HThe 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 L and 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;196:90117). 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. The antibodies generated can have any isotype.

[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;341:54446) (Holt et al; Trends Biotechnol 2003;21:484-90); (vi) camelid 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;242:42326 and Huston et al., PNAS 1988;85:587983). 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 "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.

[0043] 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.

[0044] 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).

[0045] 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).

[0046] 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.

[0047] 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.

[0048] 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.

[0049] 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.

[0050] Conservative substitutions may be defined by substitutions within the classes of amino acids reflected in the table below. [Table 1]

[0051] 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*.

[0052] 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.

[0053] 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.

[0054] 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.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] The term "R-CHP" as used herein refers to a drug combination that includes rituximab, cyclophosphamide, doxorubicin, and prednisone. The term "R-CHP" is also intended to encompass regimens in which the rituximab component is replaced with a biosimilar thereof, and / or the branded or generic versions (generic equivalents) of cyclophosphamide, doxorubicin, and / or prednisone, as well as pharmacologic acceptable salts, isomers, racemates, solvates, complexes, and hydrates, anhydrous forms thereof, and any polymorphic or amorphous forms thereof, or combinations thereof, are used in the methods described herein.

[0059] The term "polatuzumab vedotin" (CAS number: 1313206-42-6; DrugBank accession number DB12240; Kyoto Encyclopedia of Genes and Genomes (KEGG) entry: D10761) as used herein refers to a disulfide with human Mus musculus monoclonal MCDS4409A heavy chain, human Mus musculus monoclonal MCDS4409A kappa chain, dimer, thioether with maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl monomethyl lauristatin E. Polatuzumab vedotin is commercially available as Polivy®. In certain embodiments of the methods described herein, polatuzumab vedotin can be replaced with its biosimilar. Thus, it will be understood that the term "polatuzumab vedotin" is intended to encompass a biosimilar of rituximab. The term "polatuzumab vedotin" also includes antibodies having the CDRs, variable regions, or heavy and light chains of polatuzumab vedotin. Biosimilars may be administered according to standard therapeutic dosages or at doses equivalent to the standard therapeutic dosages specified for polatuzumab vedotin.

[0060] The term "rituximab" (CAS number: 174722-31-7; DrugBank-DB00073; Kyoto Encyclopedia of Genes and Genomes (KEGG) entry D02994) as used herein refers to an engineered chimeric human gamma 1 mouse constant domain containing monoclonal antibody against human CD20. The chimeric antibody contains a human gamma 1 constant domain and is referred to as "C2B8" in U.S. Patent No. 5,736,137, the entire contents of which are incorporated herein by reference. Rituximab is commercially available, for example, as Rituxan®, MabThera®, or Zytux®. In certain embodiments of the methods described herein, rituximab may be replaced with its biosimilar. Thus, it will be understood that the term "rituximab" is intended to encompass biosimilars of rituximab. Antibodies having CDRs, variable regions, or heavy and light chains of rituximab are also encompassed by the term "rituximab." Non-limiting examples of biosimilars of rituximab include Truxima® (rituximab-abbs), Ruxience® (rituximab-pvvr), and Rixathon®. Biosimilars may be administered according to standard therapeutic dosages or at doses equivalent to the standard therapeutic dosages specified for rituximab.

[0061] As used herein, the term "cyclophosphamide" has the chemical name 2H-1,3,2-oxazaphosphorin-2-amine, N,N-bis(2-chloroethyl)tetrahydro-, 2-oxide (CAS number 50-18-0) and the chemical formula CH 15It refers to a nitrogen mustard alkylating agent having Cl2N2O2P. It is commercially available under trade names such as Endoxan®, Cytoxan®, Neosar®, Procytox®, and Revimmune®. The term "cyclophosphamide" is also intended to encompass branded and generic versions (generic equivalents) of cyclophosphamide, as well as pharma- ceutically acceptable salts, isomers, racemates, solvates, complexes, and hydrates, anhydrous forms thereof, and any polymorphic or amorphous forms thereof, or combinations thereof.

[0062] The term "doxorubicin" as used herein refers to an anthracycline antibiotic that is closely related to the natural product daunomycin and, like all anthracyclines, acts by intercalating DNA. Doxorubicin is commercially available under trade names such as Adriamycin PFS®, Adriamycin RDF®, or Rubex®. Typically, the drug is administered intravenously in the form of a hydrochloride salt (e.g., as doxorubicin hydrochloride). Doxorubicin hydrochloride has the chemical name 5,12-naphthacenedione, 10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-8-(hydroxyacetyl)-1-methoxy-, hydrochloride, (8S,10S)-(CAS number 25316-40-9) and has the chemical formula C 27 H 29 NO 11 The term "doxorubicin" is also intended to encompass branded and generic versions (generic equivalents) of doxorubicin, as well as its pharma- ceutically acceptable salts, isomers, racemates, solvates, complexes and hydrates, anhydrous forms, and any polymorphic or amorphous forms, or combinations thereof.

[0063] "Prednisone" is a synthetic glucocorticoid with anti-inflammatory and immunosuppressant properties. It is a prodrug that is metabolized in the liver to prednisolone, the active form of the drug. Prednisone is commercially available under trade names such as Deltasone®, Liquid Pred®, Rayos®, and Orasone®, among others. Prednisone has the chemical name 17,21-dihydroxypregna-1,4-diene-3,11,20-trione (CAS number 53-03-2). The term "prednisone" is also intended to encompass branded and generic versions (generic equivalents) of prednisone, as well as its pharma- ceutically acceptable salts, isomers, racemates, solvates, complexes, and hydrates, anhydrous forms, and any polymorphic or amorphous forms thereof, or combinations thereof.

[0064] 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), as defined, for example, by the Lugano criteria and / or LYRIC.

[0065] 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 occur, for example, once, multiple times, and / or over one or more extended periods. In the methods described herein, the bispecific antibody (e.g., epcolitamab) is administered subcutaneously.

[0066] 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.

[0067] 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%.

[0068] As used herein, the term "subject" refers to a human patient, e.g., a human patient with DLBCL. The terms "subject" and "patient" are used interchangeably herein.

[0069] 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.

[0070] As used herein, "progressive disease" or "PD" refers to a situation in which one or more indices of DLBCL 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 the 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).

[0071] 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).

[0072] 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.

[0073] As used herein, the term "about" refers to a value that is 10% or more and 10% or less of a particular value.

[0074] DLBCL treatment regimens Provided herein is a method of treating DLBCL in a human subject using a bispecific antibody that binds CD3 and CD20 ("anti-CD3xCD20 antibody"), e.g., an isolated anti-CD3xCD20 antibody, such as epcolitamab, that binds human CD3 and human CD20, in combination with polatuzumab vedotin and R-CHP (i.e., rituximab, cyclophosphamide, doxorubicin, and prednisone or its equivalent, e.g., prednisolone). The method is useful, for example, for treating DLBCL (e.g., DLBCL in which the subject has an International Prognostic Index (IPI) score of 2-5, e.g., a score of 2, 3, 4, or 5) with histologically confirmed CD20+ disease (e.g., previously untreated DLBCL, etc.). It is understood that the methods of treating DLBCL (e.g., newly diagnosed, previously untreated (IPI 2-5) DLBCL) with a bispecific antibody that binds both CD3 and CD20 described herein also encompass the corresponding use of the bispecific antibody to treat DLBCL (e.g., newly diagnosed, previously untreated (IPI 2-5) DLBCL) in a human subject. In some embodiments, the subject receiving therapy according to the invention has not received previous therapy for DLBCL or follicular lymphoma grade 3B.

[0075] Thus, in one aspect, there is provided a method of treating DLBCL in a human subject, the method comprising administering a bispecific antibody and an effective amount of polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and prednisone or an equivalent thereof, e.g., prednisolone, 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; Here, methods are provided herein, where the bispecific antibody is administered at a dose of 24 mg or 48 mg, and the polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, prednisone or an equivalent thereof, e.g., prednisolone, and the bispecific antibody are administered in a 21 day cycle.

[0076] 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.

[0077] 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).

[0078] 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.

[0079] In some embodiments, the bispecific antibody dose is administered once a week in a 21-day cycle (weekly dosing). In one embodiment, a weekly dose of 24 mg or 48 mg is administered in 3 and 1 / 3 21-day cycles (i.e., 10 times; day 15 of cycle 1 and days 1, 8, and 15 of cycles 2-4). In one embodiment, a weekly dose of 24 mg is administered in 3 and 1 / 3 21-day cycles (i.e., 10 times; day 15 of cycle 1 and days 1, 8, and 15 of cycles 2-4). In one embodiment, a weekly dose of 48 mg is administered in 3 and 1 / 3 21-day cycles (i.e., 10 times; day 15 of cycle 1 and days 1, 8, and 15 of cycle 2). In some embodiments, after weekly dosing, the dosing interval can be reduced to once every 3 weeks. In one embodiment, administration once every three weeks is administered for at least four 21 day cycles, such as four 21 day cycles (i.e., at least four times, such as four times). In one embodiment, administration once every three weeks is administered for four 21 day cycles (i.e., four times).

[0080] In one embodiment, weekly doses of the bispecific antibody are administered in a 21 day cycle in cycles 5-8 (which may include priming and intermediate doses, as described below), and once every 3 weeks doses of the bispecific antibody are administered in a 21 day cycle in cycles 5-8.

[0081] 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 doses are administered once every three weeks and / or doses are administered at the same level. Thus, if a dose of 48 mg is selected, the same dose of 48 mg is preferably administered for each weekly administration and each administration once every three weeks. 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.

[0082] Thus, in some embodiments, a priming dose of the bispecific antibody can be administered prior to administration of the 24 mg or 48 mg weekly dose. 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. In one embodiment, the 0.16 mg priming dose is administered two weeks prior to administration of the first weekly dose of 24 mg in cycle 1. In one embodiment, the 0.16 mg priming dose is administered two weeks prior to administration of the first weekly dose of 48 mg in cycle 1.

[0083] In some embodiments, after administration of the priming dose, an intermediate dose of the bispecific antibody is administered before administration of the 24 mg or 48 mg weekly dose. 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). The priming dose is administered on day 1, the intermediate dose on day 8, followed by the first weekly dose of 24 mg or 48 mg on day 15 of cycle 1. In one embodiment, the intermediate dose is at or about 800 μg (0.8 mg) of the full length bispecific antibody. In one embodiment, the 0.16 mg priming dose is administered one week before the 0.8 mg intermediate dose (i.e., on day 1 of cycle 1), which is administered one week before the first weekly dose of 24 mg (i.e., on day 8 of cycle 1). In one embodiment, the 0.16 mg priming dose is administered one week before the 0.8 mg intermediate dose (i.e., on day 1 of cycle 1), which is administered one week before the first weekly dose of 48 mg (i.e., on day 8 of cycle 1).

[0084] The methods described herein involve treating a human subject with DLBCL with a bispecific antibody that binds CD3 and CD20 in combination with polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin and prednisone or its equivalent, e.g., prednisolone.

[0085] In some embodiments, rituximab, cyclophosphamide, doxorubicin and prednisone or its equivalents, e.g., prednisolone, are administered at standard therapeutic doses for R-CHP, e.g., as supported by clinical trials, in accordance with local guidelines, and / or in accordance with relevant local labeling.

[0086] For example, in some embodiments, polatuzumab vedotin is administered according to the relevant local product label or Summary of Product Characteristics (see, e.g., POLIVY® (polatuzumab vedotin-piiq) Prescribing Information available at https: / / www.accessdata.fda.gov / drugsatfda_docs / label / 2019 / 761121s000lbl.pdf). In some embodiments, a biosimilar of polatuzumab vedotin is used in place of polatuzumab vedotin in the methods described herein.

[0087] In some embodiments, rituximab is administered according to the relevant local product label or summary of product characteristics (see, e.g., RITUXAN® (rituximab) prescribing information available at www.accessdata.fda.gov / drugsatfda_docs / label / 2013 / 103705s5414lbl.pdf). In some embodiments, a biosimilar of rituximab is used in place of rituximab in the methods described herein.

[0088] In some embodiments, cyclophosphamide is administered according to the relevant local product label or Summary of Product Characteristics (see, e.g., CYCLOPHOSPHAMIDE INJECTION Prescribing Information available at www.accessdata.fda.gov / drugsatfda_docs / label / 2013 / 012141s090,012142s112lbl.pdf).

[0089] In some embodiments, doxorubicin is administered according to the relevant local product label or Summary of Product Characteristics (see, e.g., ADRIAMYCIN (Doxorubicin HCl) For Injection (Lyophilized) and ADRIAMYCIN (Doxorubicin HCL) Injection (0.9% Sodium Chloride and Water) Prescribing Information available at www.accessdata.fda.gov / drugsatfda_docs / label / 2012 / 062921s022lbl.pdf; and Doxorubicin Hydrochloride For Injection and Doxorubicin Hydrochloride Injection Prescribing Information available at www.accessdata.fda.gov / drugsatfda_docs / label / 2010 / 050467s070lbl.pdf).

[0090] In some embodiments, prednisolone is administered instead of prednisone in the R-CHP regimen.

[0091] In one embodiment, polatuzumab vedotin is administered according to local guidelines and local labeling. In some embodiments, polatuzumab vedotin is administered at a dose of 1.8 mg / kg (or at a dose of about 1.8 mg / kg). In some embodiments, polatuzumab vedotin is administered intravenously.

[0092] In one embodiment, polatuzumab vedotin is administered once every three weeks. In some embodiments, polatuzumab vedotin is administered once every three weeks (Q3W) in a 21-day cycle. In some embodiments, administration of polatuzumab vedotin once every three weeks is performed for six 21-day cycles (i.e., six times). In a preferred embodiment, polatuzumab vedotin is administered intravenously at a dose of 1.8 mg / kg once every three weeks for six 21-day cycles (i.e., six times).

[0093] In one embodiment, rituximab is administered according to local guidelines and local labeling. In some embodiments, rituximab is administered at a dose of 375 mg / m 2 (or about 375 mg / m 2In some embodiments, rituximab is administered intravenously.

[0094] In one embodiment, rituximab is administered once every three weeks. In some embodiments, rituximab is administered once every three weeks (Q3W) in a 21-day cycle. In some embodiments, rituximab is administered once every three weeks for six 21-day cycles (i.e., six times). In a preferred embodiment, rituximab is administered intravenously at a dose of 375 mg / m2 once every three weeks for six 21-day cycles (i.e., six times).

[0095] In some embodiments, cyclophosphamide is administered according to local guidelines and local labeling. In some embodiments, cyclophosphamide is administered at 750 mg / m 2 (or about 750 mg / m 2 In some embodiments, cyclophosphamide is administered intravenously. In some embodiments, cyclophosphamide is administered once every three weeks. In some embodiments, rituximab is administered once every three weeks (Q3W) in a 21-day cycle. In some embodiments, cyclophosphamide is administered once every three weeks for six 21-day cycles (i.e., six times). In a preferred embodiment, cyclophosphamide is administered intravenously at a dose of 750 mg / m2 once every three weeks for six 21-day cycles (i.e., six times).

[0096] In some embodiments, doxorubicin is administered according to local guidelines and local labeling. In some embodiments, doxorubicin is administered at a dose of 50 mg / m 2 at a dose of about 50 mg / m 2In some embodiments, doxorubicin is administered intravenously. In some embodiments, doxorubicin is administered once every three weeks. In some embodiments, doxorubicin is administered once every three weeks (Q3W) in a 21-day cycle. In some embodiments, doxorubicin is administered once every three weeks for six 21-day cycles (i.e., six times). In a preferred embodiment, doxorubicin is administered intravenously at a dose of 50 mg / m2 once every three weeks for six 21-day cycles (i.e., six times).

[0097] In a preferred embodiment, polatuzumab vedotin is preferably administered intravenously at a dose of 1.8 mg / kg, rituximab is preferably administered at a dose of 375 mg / m2, cyclophosphamide is preferably administered at a dose of 750 mg / m2, and doxorubicin is preferably administered at a dose of 50 mg / m2, and rituximab, cyclophosphamide and doxorubicin are preferably administered intravenously once every three weeks for six 21-day cycles.

[0098] In some embodiments, prednisone or its equivalent, such as prednisolone, is administered according to local guidelines and local labeling. In some embodiments, prednisone or its equivalent, such as prednisolone, is administered at a dose of 100 mg (or at a dose of about 100 mg). In some embodiments, prednisone or its equivalent, such as prednisolone, is administered intravenously and / or orally. In some embodiments, prednisone or its equivalent, such as prednisolone, is administered intravenously. In some embodiments, prednisone or its equivalent, such as prednisolone, is administered orally.

[0099] In one embodiment, prednisone or its equivalent, e.g., prednisolone, is administered once a day for five consecutive days (i.e., days 1-5) in a 21-day cycle. In one embodiment, prednisone or its equivalent, e.g., prednisolone, is administered for six 21-day cycles (e.g., days 1-5 of cycles 1-6 of a 21-day cycle). In one embodiment, prednisone or its equivalent, e.g., prednisolone, is administered once a day for days 1-5 of a 21-day cycle. In some embodiments, prednisone or its equivalent, e.g., prednisolone, is administered once a day for five consecutive days (i.e., days 1-5) in six 21-day cycles (e.g., days 1-5 of cycles 1-6 of a 21-day cycle). In a preferred embodiment, prednisone or its equivalent, e.g., prednisolone, is administered intravenously at a dose of 100 mg / day once daily for five consecutive days (i.e., days 1-5) for six 21-day cycles (e.g., days 1-5 of cycles 1-6 of a 21-day cycle). In another preferred embodiment, prednisone or its equivalent, e.g., prednisolone, is administered orally at a dose of 100 mg / day once daily for five consecutive days (i.e., days 1-5) for six 21-day cycles (e.g., days 1-5 of cycles 1-6 of a 21-day cycle). In another preferred embodiment, prednisone or its equivalent, e.g., prednisolone, is administered intravenously and / or orally at a dose of 100 mg / day once daily for five consecutive days (i.e., days 1-5) for six 21-day cycles (e.g., days 1-5 of cycles 1-6 of a 21-day cycle).

[0100] In one embodiment, polatuzumab vedotin (e.g., intravenous), rituximab (e.g., intravenous), cyclophosphamide (e.g., intravenous), doxorubicin (e.g., intravenous), prednisone or its equivalent, e.g., prednisolone (e.g., intravenous or oral), and a bispecific antibody (e.g., subcutaneous) are administered in a 21 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 a dose of 24 mg is administered on day 15; (ii) In cycles 2 to 4, a dose of 24 mg is administered on days 1, 8, and 15; (iii) In cycles 5 through 8, a dose of 24 mg is administered on day 1, and so on. (b) polatuzumab vedotin, rituximab, cyclophosphamide, and doxorubicin are administered on Day 1 of Cycles 1-6, and (c) Prednisone or its equivalent, e.g., prednisolone, is administered on days 1-5 of cycles 1-6.

[0101] In one embodiment, polatuzumab vedotin, rituximab (e.g., intravenously), cyclophosphamide (e.g., intravenously), doxorubicin (e.g., intravenously), prednisone or its equivalent, e.g., prednisolone (e.g., intravenously or orally), and a bispecific antibody (e.g., subcutaneously) are administered in a 21 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 a dose of 48 mg is administered on day 15; (ii) In cycles 2 to 4, a dose of 48 mg is administered on days 1, 8, and 15; (iii) In cycles 5 through 8, a dose of 48 mg was administered on day 1, and so on. (b) polatuzumab vedotin, rituximab, cyclophosphamide, and doxorubicin are administered on Day 1 of Cycles 1 to 6; and (c) Prednisone or its equivalent, e.g., prednisolone, is administered on days 1-5 of cycles 1-6.

[0102] In the above two embodiments, polatuzumab vedotin is preferably administered at a dose of 1.8 mg / kg, rituximab is preferably administered at a dose of 375 mg / m2, cyclophosphamide is preferably administered at a dose of 750 mg / m2, doxorubicin is preferably administered at a dose of 50 mg / m2, and prednisone or its equivalent, e.g., prednisolone, is preferably administered at a dose of 100 mg / day.

[0103] In one embodiment, polatuzumab vedotin (e.g., intravenous), rituximab (e.g., intravenous), cyclophosphamide (e.g., intravenous), doxorubicin (e.g., intravenous), prednisone or its equivalent, e.g., prednisolone (e.g., intravenous or oral), and the bispecific antibody epcolitamab (e.g., subcutaneous) are administered in a 21 day cycle; (a) The bispecific antibody epcolitamab is (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 a dose of 24 mg is administered on day 15; (ii) In cycles 2 to 4, a dose of 24 mg is administered on days 1, 8, and 15; (iii) In cycles 5 through 8, a dose of 24 mg is administered on day 1, and so on. (b) polatuzumab vedotin, rituximab, cyclophosphamide, and doxorubicin are administered on Day 1 of Cycles 1-6, and (c) Prednisone or its equivalent, e.g., prednisolone, is administered on days 1-5 of cycles 1-6.

[0104] In one embodiment, polatuzumab vedotin (e.g., intravenous), rituximab (e.g., intravenous), cyclophosphamide (e.g., intravenous), doxorubicin (e.g., intravenous), prednisone or its equivalent, e.g., prednisolone (e.g., intravenous or oral), and the bispecific antibody epcolitamab (e.g., subcutaneous) are administered in a 21 day cycle; (a) The bispecific antibody epcolitamab is (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 a dose of 48 mg is administered on day 15; (ii) In cycles 2-4, a dose of 48 mg is administered on days 1, 8, and 15; (iii) In cycles 5 through 8, a dose of 48 mg was administered on day 1, and so on. (b) polatuzumab vedotin, rituximab, cyclophosphamide, and doxorubicin are administered on Day 1 of Cycles 1-6, and (c) Prednisone or its equivalent, e.g., prednisolone, is administered on days 1-5 of cycles 1-6.

[0105] In the above two embodiments, polatuzumab vedotin is preferably administered at a dose of 1.8 mg / kg, rituximab is preferably administered at a dose of 375 mg / m2, cyclophosphamide is preferably administered at a dose of 750 mg / m2, doxorubicin is preferably administered at a dose of 50 mg / m2, and prednisone or its equivalent, e.g., prednisolone, is preferably administered at a dose of 100 mg / day.

[0106] In one embodiment, the dosing of the bispecific antibody, polatuzumab and R-CHP in a 21 day cycle is as follows:

[0107] Bispecific antibody (subcutaneous): Cycle 1, Day 1: Priming dose (0.16 mg) Cycle 1, Day 8: Intermediate dose (0.8 mg) Cycle 1, Day 15: All doses (24 or 48 mg) Cycles 2-4, days 1, 8, and 15: all doses (24 or 48 mg) Cycle 5-8, Day 1: Full dose (24 or 48 mg) every 3 weeks Polatuzumab (intravenous): Cycles 1-6, day 1: 1.8 mg / kg R-CHP: Rituximab: Cycles 1-6, Day 1: 375 mg / m 2 (intravenous) Cyclophosphamide: Cycles 1-6, Day 1: 750 mg / m 2 (intravenous) Doxorubicin: Cycles 1-6, Day 1: 50 mg / m 2 (intravenous) Prednisone or equivalent: Cycles 1-6, days 1-6: 100 mg (orally)

[0108] In one embodiment, the subject treated with the methods described herein is an adult male or female who is at least 18 years of age.

[0109] In one embodiment, the subject undergoing treatment with the methods described herein has documented 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: Thus, in one embodiment, the subject has DLBCL, NOS (not otherwise specified). In some embodiments, the subject has "double hit" or "triple hit" DLBCL, classified in the WHO 2016 as HGBCL with MYC and BCL2 and / or BCL6 translocations. In some embodiments, the subject has follicular lymphoma grade 3B.

[0110] In one embodiment, a subject with DLBCL has an International Prognostic Index (IPI) score of 2-5, such as an IPI score of 2, 3, 4, or 5. IPI risk factors include: (1) Ann Arbor stage III or IV, (2) age >60 years, (3) elevated levels of lactate dehydrogenase, (4) ECOG performance score >2, and (5) more than one extranodal site.

[0111] In one embodiment, the subject with DLBCL has not had prior treatment for DLBCL or follicular lymphoma grade 3B.

[0112] In a further embodiment, the subject has not received prior treatment with a bispecific antibody targeting CD3 and CD20.

[0113] In one embodiment, the subject has an Eastern Cooperative Oncology Group (ECOG) performance status (ECOG PS) of 0 to 2, such as 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).

[0114] 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.

[0115] In some embodiments, the subject has laboratory values ​​that meet the following criteria prior to receiving the first dose of the bispecific antibody:

[0116] - 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 hepatic 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 (except in patients receiving anticoagulant therapy)

[0117] 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, including the following according to the WHO 2016 classification, documented in the pathology report: - 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 - Cannot have had prior treatment with a bispecific antibody 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 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 Common Terminology Criteria for Adverse Events (CTCAE, v5.0), not resolved to Grade 1, except for alopecia. - No current evidence of primary central nervous system (CNS) tumors 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 concomitant (e.g., lenalidomide, rituximab, etc.) - Cannot have undergone an autologous stem cell transplant within 3 months prior to screening. - Patients 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) prior to the first dose of epcolitamab. - Absence 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.

[0118] - 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.

[0119] 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. - 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 compliance with the protocol 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 - No lesion-directed radiation therapy or major surgery within 4 weeks of enrollment. - No neurological deficits greater than Grade 1. - Must not have a history of active tuberculosis (TB) or completed treatment for active TB within the past 12 months.

[0120] 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.

[0121] - No evidence of CMV viremia at screening (defined as a positive level above the lower limit of detection). - No 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 study results 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.

[0122] 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 undergone major surgery within 4 weeks of the first dose of study drug.

[0123] 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.

[0124] 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, rituximab, etc.).

[0125] In one embodiment, subjects must not have undergone an autologous stem cell transplant within 3 months prior to screening.

[0126] 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.

[0127] 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 associated with 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.

[0128] Left ventricular ejection fraction (LVEF) must be within the institutional normal range by multiple gated echocardiography (MUGA) or transthoracic echocardiography at screening.

[0129] 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

[0130] In one embodiment, subjects have not undergone lesion-directed radiation therapy or major surgery within 4 weeks of enrollment.

[0131] In one embodiment, the subject does not have a grade >1 neuropathy.

[0132] 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.

[0133] The methods described herein are advantageous for treating DLBCL with an International Prognostic Index (IPI) score of 2 to 5, such as 2, 3, 4, or 5. Treatment is maintained continuously, for example, using a treatment regimen described herein. However, treatment can be terminated if progressive disease develops or unacceptable toxicity occurs.

[0134] In one embodiment, the method of the invention is for first line treatment of DLBCL.

[0135] The response of subjects with DLBCL 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 LYRIC are provided in Example 3 herein.

[0136] 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.

[0137] Subjects treated according to the methods described herein preferably experience an improvement in at least one symptom of DLBCL. 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.

[0138] In one embodiment, the treated subject exhibits a complete response (CR), partial response (PR), or stable disease (SD) as defined by the Lugano criteria and / or LYRIC (see, e.g., Example 3 herein). In some embodiments, the methods described herein result in at least one therapeutic benefit selected from prolonged survival (e.g., progression-free survival or overall survival, etc.), optionally compared to another treatment, such as treatment with R-CHP alone.

[0139] 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.

[0140] 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 some embodiments, 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.

[0141] 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.

[0142] In one embodiment, rituximab is formulated in a pharmaceutical composition comprising pharma- ceutically acceptable excipients for administration (e.g., intravenous administration) according to local standard of care practices, e.g., as specified by local guidelines or local product labeling. For example, in some embodiments, rituximab is provided as a sterile, clear, preservative-free liquid concentrate for intravenous administration. In one embodiment, rituximab is supplied at a concentration of 10 mg / mL in either 100 mg / 10 mL or 500 mg / 50 mL single-use vials. In some embodiments, rituximab is formulated for injection in polysorbate 80 (0.7 mg / mL), sodium citrate dihydrate (7.35 mg / mL), sodium chloride (9 mg / mL), and water at pH 6.5.

[0143] In one embodiment, polatuzumab vedotin, cyclophosphamide, doxorubicin, and prednisone are formulated into a pharmaceutical composition containing pharma- ceutically acceptable excipients for administration (e.g., intravenous administration) according to local standard therapeutic practice, e.g., as specified by local guidelines or local product labeling, or as directed by the manufacturer. In some embodiments, polatuzumab vedotin, cyclophosphamide, doxorubicin, and prednisone are diluted from a stock solution or reconstituted if in lyophilized form, e.g., according to instructions on the product label (e.g., 0.9% saline). In some embodiments, prednisone is formulated into a pharmaceutical composition for oral administration.

[0144] 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.

[0145] 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).

[0146] 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 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.

[0147] 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 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.

[0148] 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.

[0149] 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).

[0150] 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.

[0151] 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).

[0152] 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 one embodiment, the bispecific antibody is a full-length antibody with a human IgG1 constant region. In one embodiment, 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 one embodiment, the second binding arm of the bispecific antibody is derived from a human antibody, preferably a full-length IgG1, κ (kappa) antibody. In one embodiment, the second binding arm of the bispecific antibody is derived from a human antibody, preferably a full-length IgG1, κ (kappa) antibody, and thus may comprise 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.

[0153] 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.

[0154] 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).

[0155] 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.

[0156] 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.

[0157] 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 of the human IgG1 heavy chain are F, E, and A, respectively. An antibody with these amino acids at these positions is an example of an antibody with an inactive or non-active Fc region. In one embodiment, the bispecific antibody comprises a first heavy chain and a second heavy chain, and 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. In one embodiment, 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.

[0158] 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.

[0159] 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 one embodiment, the bispecific antibody comprises heavy chain constant regions comprising the amino acid sequences of SEQ ID NOs: 19 and 20.

[0160] 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.

[0161] Medical Use Further provided herein is a bispecific antibody for use in the above disclosed methods.

[0162] 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 effective amounts of (a) polatuzumab vedotin, (b) rituximab, (c) cyclophosphamide, (d) doxorubicin, and (e) prednisone or an equivalent thereof, 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 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; and Including, The bispecific antibody is administered at a dose of 24 mg or 48 mg, and the polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, prednisone or equivalent, and the bispecific antibody are administered in a 21-day cycle.

[0163] Also provided herein are bispecific antibodies for the manufacture of a medicament for use in the methods disclosed above.

[0164] In certain embodiments, the bispecific antibody is for the manufacture of a medicament for use in treating diffuse large B-cell lymphoma (DLBCL) in a human subject, wherein the bispecific antibody is administered to the subject in combination with effective amounts of (a) polatuzumab vedotin, (b) rituximab, (c) cyclophosphamide, (d) doxorubicin, and (e) prednisone or an equivalent thereof, 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 polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, prednisone or equivalent, and the bispecific antibody are administered in a 21-day cycle.

[0165] 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 polatuzumab vedotin (e.g., for intravenous administration), rituximab (e.g., for intravenous administration), cyclophosphamide (e.g., for intravenous administration), doxorubicin (e.g., for intravenous administration), and / or prednisone or its equivalent (e.g., for intravenous or oral administration). The kit may also optionally comprise instructions, including, for example, a dosing schedule, to enable a practitioner (e.g., a doctor, a nurse, or a patient) to administer one or more compositions contained therein to a patient with DLBCL. The kit may also comprise one or more syringes.

[0166] Optionally, the kits include 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 include multiple packages of single-dose pharmaceutical compositions containing doses of polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and / or prednisone or equivalents according to standard treatment regimens. Apparatus or devices necessary for administering the pharmaceutical composition(s) may also be included in the kits.

[0167] 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 and an effective amount of (a) polatuzumab vedotin, (b) rituximab, (c) cyclophosphamide, (d) doxorubicin, and (e) prednisone or an equivalent thereof, 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 polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, prednisone or equivalents, and the bispecific antibody are administered in a 21 day cycle.

[0168] 2. The method of embodiment 1, wherein the bispecific antibody is administered at a dose of 24 mg.

[0169] 3. The method of embodiment 1, wherein the bispecific antibody is administered at a dose of 48 mg.

[0170] 4. The method of any one of embodiments 1-3, wherein the bispecific antibody is administered once a week (administered every week).

[0171] The method of embodiment 4, wherein weekly administration of 5.24 mg or 48 mg is administered over 3 and 1 / 3 21 day cycles.

[0172] 6. The method of embodiment 4 or 5, wherein after the weekly administration, the bispecific antibody is administered once every three weeks, for example, on day 1 of each 21 day cycle.

[0173] 7. The method of embodiment 6, wherein one administration every 3 weeks is administered for at least four 21-day cycles.

[0174] 8. The method of embodiment 7, wherein one administration every three weeks is administered for four 21-day cycles.

[0175] The method of any one of embodiments 4-8, wherein a priming dose of the bispecific antibody is administered in cycle 1 of a 21 day cycle prior to the weekly administration of 9.24 mg or 48 mg.

[0176] 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.

[0177] 11. The method of embodiment 9 or 10, wherein the priming dose is 0.16 mg.

[0178] 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.

[0179] 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 a first weekly dose of 24 mg or 48 mg on day 15 of cycle 1.

[0180] 14. The method of embodiment 12 or 13, wherein the intermediate dose is 0.8 mg.

[0181] 15. The method of any one of embodiments 1 to 14, wherein the bispecific antibody is administered subcutaneously.

[0182] 16. The method of any one of embodiments 1-15, wherein polatuzumab vedotin is administered once every three weeks.

[0183] 17. The method of any one of embodiments 1-16, wherein administration of polatuzumab vedotin once every 3 weeks occurs in six 21-day cycles.

[0184] 18. The method of any one of embodiments 1-17, wherein polatuzumab vedotin is administered at a dose of 1.8 mg / kg.

[0185] 19. The method of any one of embodiments 1-18, wherein polatuzumab vedotin is administered on day 1 of each 21-day cycle.

[0186] 20. The method of any one of embodiments 1-18, wherein rituximab is administered once every three weeks.

[0187] 21. The method of embodiment 20, wherein one administration of rituximab every 3 weeks is administered for six 21-day cycles.

[0188] 22. Rituximab at 375 mg / m 2 The method of any one of embodiments 1-21, wherein the dose is administered.

[0189] 23. The method of any one of embodiments 1-22, wherein rituximab is administered on day 1 of each 21-day cycle.

[0190] 24. The method of any one of embodiments 1-23, wherein cyclophosphamide is administered once every three weeks.

[0191] 25. The method of embodiment 24, wherein one administration of cyclophosphamide is administered every 3 weeks for six 21-day cycles.

[0192] 26. Cyclophosphamide 750 mg / m 2The method of any one of embodiments 1-25, wherein the dose is

[0193] 27. The method of any one of embodiments 1-26, wherein cyclophosphamide is administered on day 1 of each 21-day cycle.

[0194] 28. The method of any one of embodiments 1-27, wherein doxorubicin is administered once every three weeks.

[0195] 29. The method of embodiment 28, wherein one administration of doxorubicin is administered every 3 weeks for six 21-day cycles.

[0196] 30. Doxorubicin 50 mg / m 2 The method of any one of embodiments 1-29, wherein the dose is administered.

[0197] 31. The method of any one of embodiments 1-30, wherein doxorubicin is administered on day 1 of each 21-day cycle.

[0198] 32. The method of any one of embodiments 1-31, wherein the equivalent amount of prednisone is prednisolone.

[0199] 33. The method of any one of embodiments 1-32, wherein prednisone or prednisolone is administered once daily on days 1 through 5 of a 21-day cycle.

[0200] 34. The method of embodiment 33, wherein prednisone or prednisolone is administered in six 21-day cycles.

[0201] 35. The method of any one of embodiments 1-34, wherein prednisone or prednisolone is administered at a dose of 100 mg / day.

[0202] 36. The method of any one of embodiments 1-35, wherein prednisone or prednisolone is administered on days 1-5 of each 21-day cycle.

[0203] 37. The method of any one of embodiments 1-36, wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, prednisone or equivalents thereof, and the bispecific antibody are administered on the same day (e.g., days 1 of cycles 1-6 or cycles 1-8 of a 21-day cycle).

[0204] 38. Treatment is in a 21-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 a dose of 24 mg is administered on day 15; (ii) In cycles 2 to 4, a dose of 24 mg is administered on days 1, 8, and 15; (iii) In cycles 5 through 8, a dose of 24 mg is administered on day 1, and so on. (b) polatuzumab vedotin, rituximab, cyclophosphamide, and doxorubicin are administered on Day 1 of Cycles 1 to 6; and (c) prednisone or its equivalent is administered on days 1-5 of cycles 1-6; The method according to any one of embodiments 1, 2, and 4 to 37.

[0205] 39. Treatment is in a 21-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 a dose of 48 mg is administered on day 15; (ii) In cycles 2-4, a dose of 48 mg is administered on days 1, 8, and 15; (iii) In cycles 5 through 8, a dose of 48 mg was administered on day 1, and so on. (b) polatuzumab vedotin, rituximab, cyclophosphamide, and doxorubicin are administered on Day 1 of Cycles 1 to 6; and (c) prednisone or its equivalent is administered on days 1-5 of cycles 1-6; The method according to any one of embodiments 1 and 3 to 37.

[0206] 40. The method of any one of embodiments 1 to 39, wherein the bispecific antibody is administered subcutaneously.

[0207] 41. The method of any one of embodiments 1-40, wherein rituximab is administered intravenously.

[0208] 42. The method of any one of embodiments 1-41, wherein cyclophosphamide is administered intravenously.

[0209] 43. The method of any one of embodiments 1-42, wherein doxorubicin is administered intravenously.

[0210] 44. The method of any one of embodiments 1-43, wherein prednisone or prednisolone is administered intravenously or orally.

[0211] 45. The method of any one of embodiments 1 to 44, wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, prednisone and the bispecific antibody are administered sequentially.

[0212] 46. ​​The method of any one of embodiments 1 to 45, wherein the DLBCL has histologically confirmed CD20+ disease.

[0213] 47. 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] 48. The method of any one of embodiments 1 to 47, wherein the DLBCL is follicular lymphoma grade 3B.

[0215] 49. The method of any one of embodiments 1-48, wherein the subject has an International Prognostic Index (IPI) score of 2 to 5.

[0216] 50. The method of any one of embodiments 1-49, wherein the subject has not received prior treatment for DLBCL or follicular lymphoma grade 3B.

[0217] 51. (i) a first antigen-binding region of a 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; (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, sequence DAS, and SEQ ID NO: 12, respectively; The method according to any one of embodiments 1 to 50.

[0218] 52. (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 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 method according to any one of embodiments 1 to 51.

[0219] 53. The method according to any one of embodiments 1 to 52, wherein the first binding arm of the bispecific antibody is derived from a humanized antibody, preferably a full-length IgG1, λ (lambda) antibody.

[0220] 54. The method of embodiment 53, 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.

[0221] 55. The method according to any one of embodiments 1 to 54, wherein the second binding arm of the bispecific antibody is derived from a human antibody, preferably a full-length IgG1, κ (kappa) antibody.

[0222] 56. The method of embodiment 55, wherein the second binding arm comprises a kappa light chain constant region comprising the amino acid sequence set forth in SEQ ID NO:23.

[0223] 57. The method of any one of embodiments 1 to 56, wherein the bispecific antibody is a full-length antibody having a human IgG1 constant region.

[0224] 58. The method of any one of embodiments 1 to 57, wherein the bispecific antibody comprises an inactive Fc region.

[0225] 59. The method of any one of embodiments 1 to 58, 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.

[0226] 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, 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.

[0227] 61. 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 of any one of embodiments 1 to 60, 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.

[0228] 62. The method of embodiment 61, wherein the bispecific antibody comprises a heavy chain constant region comprising the amino acid sequences of SEQ ID NOs: 19 and 20.

[0229] 63. The method according to any one of embodiments 1 to 62, 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.

[0230] 64. The method according to any one of embodiments 1 to 63, 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.

[0231] 65. The method of any one of embodiments 1 to 64, wherein the bispecific antibody is epcolitamab, or a biosimilar thereof.

[0232] [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.

[0233] 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.

[0234] 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.

[0235] [Example 1] Antitumor activity of epcolitamab in samples from NHL patients in the presence of anti-CD20 antibodies in vivo and after anti-CD20 treatment The effect of the presence of anti-CD20 antibodies on the anti-tumor activity of epcolitamab in a humanized mouse xenograft model is described in Engelberts et al., EBioMedicine 2020;52:10265, as summarized below.

[0236] Epcolitamab was found to effectively reduce tumor growth in a xenograft model (NOD-SCID mice injected with CD20-expressing Raji-luc tumor cells and PBMCs) even in the presence of excess rituximab variants with inactive Fc domains (IgG1-RTX-FEAR containing L234F, L235E, D265A, and K409R mutations). Rituximab and IgG1-CD20, from which the CD20 arm of epcolitamab is derived, compete for CD20 binding despite binding to different epitopes, indicating that epcolitamab can induce effective antitumor activity in the presence of circulating anti-CD20 antibodies that can compete for target binding.

[0237] Moreover, epcolitamab induced T cell-mediated cytotoxicity in biopsies from primary DLBCL and follicular lymphoma patients over a period of time following administration of anti-CD20 antibodies (Van der Horst et al., Blood (2019) 134 (Supplement_1): 4066). Even in biopsies taken 2 weeks after administration of anti-CD20 antibodies, epcolitamab was able to induce up to 40% tumor cell death.

[0238] [Example 2] Effect of CHP on epcolitamab-induced in vitro T cell-mediated cytotoxicity This experiment was performed to test the impact of the CHP components separately to evaluate the effect of each component on epcolitamab-induced T cell-mediated cytotoxicity.

[0239] Briefly, T cells were preincubated with cyclophosphamide, doxorubicin or prednisone for 16 hours, after which epcolitamab and CD20-expressing Daudi cells were used as target cells (E:T ratio 2:1) in the cytotoxicity assay, and cyclophosphamide, doxorubicin or prednisone were added at the same concentrations as during preincubation, respectively. Data are presented as percent viable target cells (CD4-CD8-CD22+) normalized to medium control (no Ab, no CHP components). Doxorubicin pretreatment affected T cell viability, so all concentrations of epcolitamab could not be tested.

[0240] Figure 1A-1C show the dose-response curves of DuoBody®-CD3xCD20 for a representative donor in the left panel, and the right panel shows the response to a dose of 333ng / ml of DuoBody®-CD3xCD20 for four different donors, with or without various concentrations of CHP components. Pretreatment of T cells with cyclophosphamide or prednisone, respectively, did not affect T cell viability (data not shown). As mentioned above, pretreatment with doxorubicin resulted in a decrease in T cell viability (not shown), but the extent observed in vitro seemed exaggerated compared to that observed in patients treated with R-CHOP (Oncology 2016;91:302-10 and Hematol Oncol 2011;29:5-9). As shown in Figure 1A, C, T cells pretreated with cyclophosphamide (A) or prednisone (C) were able to mediate epcolitamab-induced cytotoxic responses against CD20-expressing target cells, as shown by the dose-dependent cytotoxicity (left panel) and the very low percentage of viable B cells remaining after incubation (right panel). As shown in Figure 1B, the remaining T cells pretreated with doxorubicin were also able to mediate epcolitamab-induced cytotoxicity against target cells, indicating that the remaining T cells were functional.

[0241] Taken together, the above data and observations indicate that epcolitamab can be combined with CHP and R-CHP to induce highly effective antitumor activity against CD20-expressing target cells, as rituximab does not interfere with epcolitamab activity.

[0242] [Example 3] A Phase 1b / 2 Open-Label Study to Evaluate the Safety and Tolerability of Epcolitamab in Combination with Anti-Neoplastic Agents in Subjects with Diffuse Large B-Cell Lymphoma (DLBCL) A Phase 1b / 2, open-label, multicenter, interventional study evaluating the safety, tolerability, and preliminary efficacy of epcolitamab in combination with polatuzumab vedotin with rituximab, cyclophosphamide, doxorubicin, and prednisone (pola-R-CHP) in subjects diagnosed with DLBCL. The study will include a dose escalation phase followed by an expansion phase.

[0243] 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).

[0244] 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.

[0245] The phase 2 study included subjects with R / R NHL, including DLBCL, and treatment-naïve DLBCL evaluated at doses of 24 mg and 48 mg in the expansion 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).

[0246] An ongoing phase 1 / 2 trial (NCT04663347) in newly diagnosed high-risk patients with DLBCL showed that epcolitamab + R-CHOP had promising efficacy and a manageable safety profile in patients with International Prognostic Index (IPI) scores 3 to 5. Among efficacy-evaluable patients (n=31), ORR was 100% with complete metabolic response in 77%; cytokine release syndrome (CRS) events (n / N=17 / 33; 52%) were mostly low-grade and did not lead to treatment discontinuation (Falchi et al, ASCO 2022, abstract 7523).

[0247] Objectives and Endpoints Main purpose To characterize the safety and toxicity profile of epcolitamab when co-administered with polatuzumab vedotin and rituximab, cyclophosphamide, doxorubicin, and prednisone (pola-R-CHP) in subjects with DLBCL. To determine the recommended dose for further investigation of epcolitamab when coadministered with pola-R-CHP in subjects with DLBCL.

[0248] Secondary Objectives To evaluate the anti-NHL activity of epcolitamab when administered in combination with pola-R-CHP in subjects with DLBCL. To characterize the pharmacokinetics of epcolitamab when administered in combination with pola-R-CHP in subjects with DLBCL.

[0249] 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).

[0250] Primary Endpoint The primary endpoint was dose-limiting toxicity (DLT) of epcolitamab in combination with pola-R-CHP.

[0251] Secondary Endpoints - Overall response rate (ORR) by Lugano 2014 criteria as assessed by the investigator for epcolitamab in combination with pola-R-CHP. Anti-lymphoma activity of epcolitamab in combination with pola-R-CHP: 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)

[0252] Safety Endpoints Safety and tolerability evaluations during the study may include, but are not limited to, the following:

[0253] Monitoring the severity and incidence of adverse events (AEs), including adverse events of special interest (AESIs) CRS, ICANS, and 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

[0254] Pharmacokinetic endpoints Pharmacokinetic (PK) parameter values, including maximum observed plasma concentration (Cmax), time to Cmax (Tmax), and area under the plasma concentration versus time curve (AUC), will be determined using non-compartmental methods for epcolitamab in combination with pola-R-CHP.

[0255] · Epcolitamab anti-drug antibodies (ADA) and neutralizing ADA in combination with pola-R-CHP.

[0256] Overview of study design A schematic of the overall study design is shown in Figure 2.

[0257] The following regimens will be initially evaluated in the corresponding population: epcolitamab in combination with pola-R-CHP in subjects with DLBCL

[0258] Study treatment Epcolitamab is administered in combination with pola-R-CHP using a step-up dosing regimen: a priming dose of 0.16 mg (Cycle 1, Day 1), followed by an intermediate dose of 0.8 mg (Cycle 1, Day 8), and a total dose of 24 or 48 mg at the assigned dose level (Cycle 1, Day 15 onwards). Epcolitamab is administered as a SC injection once weekly (QW) in Cycles 2-4, then once every 3 weeks (Q3W) in Cycles 5-8. On days when subjects receive epcolitamab in combination with other study agent(s), the other study agent should be administered before epcolitamab. Additionally, because infusion reactions due to rituximab are more common than with polatuzumab, polatuzumab should be administered after prednisone and rituximab.

[0259] Polatuzumab 1.8 mg / kg will be administered on days 1 of cycles 1-6 Rituximab 375mg / m 2 will be administered on Day 1 of Cycles 1-6 Cyclophosphamide 750mg / m 2 will be administered on Day 1 of Cycles 1-6 Doxorubicin 50mg / m 2 will be administered on Day 1 of Cycles 1-6 Prednisone 100 mg administered on days 1-5 of cycles 1-6 Epcolitamab will be administered for a total of 8 cycles as above

[0260] 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.

[0261] Dose Escalation Phase The dose escalation phase is designed to evaluate the initial safety and tolerability of epcolitamab in combination with other antineoplastic agents.

[0262] Dose escalation will be guided by a Bayesian optimal interval (BOIN) design. Initial enrollment into the dose escalation cohort will consist of at least three DLT-evaluable subjects. Epcolitamab will be administered initially in combination with the corresponding antineoplastic agent. If acceptable safety and tolerability are observed during the DLT period, the dose of epcolitamab will be escalated to the next dose level, 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 experiencing dose-limiting toxicity (DLT).

[0263] 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).

[0264] [Table 2]

[0265] Dose-limiting toxicities (DLTs) will be evaluated during dose escalation 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.

[0266] 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.

[0267] 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.

[0268] Expansion Stage The objective of the expansion phase is to evaluate the safety, tolerability and preliminary clinical activity of epcolitamab at the recommended dose in combination with pola-R-CHP.

[0269] The expansion phase of the study will enroll a total of approximately 20 subjects. Epcolitamab will be administered at the determined recommended phase 2 dose (RP2D) in combination with pola-R-CHP in the same manner as was done in dose escalation.

[0270] The toxicity monitoring rule will be implemented after six subjects are enrolled. The rule will monitor the occurrence of DLT and will pause enrollment if the posterior probability of a DLT rate exceeding 0.25 is greater than 80%. The prior distribution of DLT rates is assumed to follow a beta (1.5, 4.5) distribution, reflecting a prior average 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).

[0271] If the number of subjects experiencing DLTs exceeds the toxicity boundary at any time after six subjects have been enrolled, further enrollment will be paused and a comprehensive safety review of all available data will be conducted. Based on the toxicology monitoring rules, enrollment will be paused if the number of subjects experiencing DLTs meets any of the following boundaries:

[0272] 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

[0273] 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.

[0274] Disease / Condition Activity Diagnosis of DLBCL (de novo or histologically transformed from follicular lymphoma or nodal marginal zone lymphoma) with histologically confirmed CD20+ disease (recorded on the pathology report, including the following according to the WHO 2016 classification): 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:

[0275] 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 allergy or intolerance to any component or excipient of epcolitamab or any component of the study drug combination (e.g., lenalidomide, rituximab, 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%.

[0276] 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.

[0277] 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.

[0278] 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.

[0279] 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. 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.

[0280] 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:

[0281] 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). Subjects must not have undergone major surgery within 4 weeks of the first dose of study drug.

[0282] Additional eligibility criteria: Subjects must have newly diagnosed, untreated (no prior treatment for transformed indolent lymphoma) DLBCL. Subjects must be suitable, in the opinion of the investigator, for treatment with polatuzumab, rituximab, cyclophosphamide, doxorubicin and prednisone.

[0283] Dose-limiting toxicity 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.

[0284] 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.

[0285] 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:

[0286] 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. 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.

[0287] 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.

[0288] 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)

[0289] 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.

[0290] 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).

[0291] 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.

[0292] 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).

[0293] 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.

[0294] [Table 3] TIFF2025503177000005.tif205154TIFF2025503177000006.tif88154

[0295] 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).

[0296] · 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).

[0297] 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.

[0298] 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.

[0299] 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."

[0300] The association of epcolitamab (GEN3013; DuoBody®-CD3xCD20) with pseudoprogression is currently unknown, but its mechanism of action means that pseudoprogression would be expected.

[0301] 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.

[0302] LYRIC is a modification of the Lugano response evaluation criteria, adapted for immune-based therapy, and implements a new palliative response category, the "indeterminate response" (IR) designation. This IR designation was introduced to identify potentially "atypical responding" 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 trial.

[0303] Indeterminate Response (IR) Category Lugano classification 7 A subject exhibiting PD due to IR is considered to have IR in one or more of the following three situations:

[0304] 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.

[0305] 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.

[0306] IR(3): Increased FDG uptake in one or more lesions without a concomitant increase in lesion size or number.

[0307] Screening evaluation: At screening, FDG-PET / CT and diagnostic CT or MRI scans should be read according to the Lugano classification, as described above.

[0308] 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).

[0309] 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.

[0310] 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).

[0311] 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.

[0312] 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.

[0313] 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.

[0314] 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.

[0315] 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).

[0316] 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").

[0317] The number of responders along with a narrative summary of TTR will be provided for each arm.

[0318] 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.

[0319] Kaplan-Meier estimates of the distribution of deaths and OS are provided.

[0320] 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.

[0321] 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.

[0322] 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.

[0323] 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.

[0324] Preliminary results Dose escalation (epcolitamab 24mg + Pola-R-CHP) Number of subjects registered: 8 Number of subjects expected to have at least one post-baseline efficacy assessment: 8 Number of subjects with available post-baseline efficacy assessments: 4 ORR=100%(4 / 4) CRR=75%(3 / 4) Dose escalation (epcolitamab 48mg + Pola-R-CHP) Number of subjects registered: 4 Number of subjects expected to have at least one post-baseline efficacy assessment: 0 Number of subjects with available post-baseline efficacy assessments: 0

[0325] [Table 4] TIFF2025503177000008.tif235154TIFF2025503177000009.tif224154

[0326] 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; for the treatment of diffuse large B-cell lymphoma (DLBCL) in a human subject; the treatment comprises administering to the subject the bispecific antibody and effective amounts of (a) polatuzumab vedotin, (b) rituximab, (c) cyclophosphamide, (d) doxorubicin, and (e) prednisone or an equivalent thereof; wherein the bispecific antibody is administered at a dose of 24 mg or 48 mg, and wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, prednisone or an equivalent thereof, and the bispecific antibody are administered in a 21 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 occurs over three and one-third 21-day cycles.

6. 6. The pharmaceutical composition of claim 4 or 5, wherein after said weekly administration, the bispecific antibody is administered once every three weeks, for example in a 21 day cycle, on day 1 of each 21 day cycle.

7. 7. The pharmaceutical composition of claim 6, wherein said administration once every three weeks occurs for at least four 21-day cycles.

8. 8. The pharmaceutical composition of claim 7, wherein said administration once every three weeks occurs over four 21-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 21 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 a first weekly dose of 24 mg or 48 mg on day 15 of cycle 1.

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

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

16. 10. The pharmaceutical composition of claim 1, wherein polatuzumab vedotin is administered once every three weeks.

17. 10. The pharmaceutical composition of claim 1, wherein polatuzumab vedotin is administered once every three weeks for six 21-day cycles.

18. 2. The pharmaceutical composition of claim 1, wherein polatuzumab vedotin is administered at a dose of 1.8 mg / kg.

19. 2. The pharmaceutical composition of claim 1, wherein polatuzumab vedotin is administered on day 1 of each 21-day cycle.

20. 10. The pharmaceutical composition of claim 1, wherein rituximab is administered once every three weeks.

21. 21. The pharmaceutical composition of claim 20, wherein rituximab is administered once every three weeks for six 21-day cycles.

22. Rituximab 375 mg / m ２ The pharmaceutical composition of claim 1 , wherein the composition is administered at a dose of

23. 10. The pharmaceutical composition of claim 1, wherein rituximab is administered on day 1 of each 21-day cycle.

24. 10. The pharmaceutical composition of claim 1, wherein cyclophosphamide is administered once every three weeks.

25. 25. The pharmaceutical composition of claim 24, wherein the administration of cyclophosphamide occurs once every three weeks for six 21-day cycles.

26. Cyclophosphamide 750 mg / m ２ The pharmaceutical composition of claim 1 , wherein the composition is administered at a dose of

27. 10. The pharmaceutical composition of claim 1, wherein cyclophosphamide is administered on day 1 of each 21-day cycle.

28. 10. The pharmaceutical composition of claim 1, wherein doxorubicin is administered once every three weeks.

29. 29. The pharmaceutical composition of claim 28, wherein doxorubicin is administered once every three weeks for six 21-day cycles.

30. Doxorubicin 50 mg / m ２ The pharmaceutical composition of claim 1 , wherein the composition is administered at a dose of

31. 10. The pharmaceutical composition of claim 1, wherein doxorubicin is administered on day 1 of each 21-day cycle.

32. 2. The pharmaceutical composition of claim 1, wherein the equivalent amount of prednisone is prednisolone.

33. 10. The pharmaceutical composition of claim 1, wherein prednisone or prednisolone is administered once daily on days 1 through 5 of a 21-day cycle.

34. 34. The pharmaceutical composition of claim 33, wherein the prednisone or prednisolone is administered in six 21-day cycles.

35. 10. The pharmaceutical composition of claim 1, wherein prednisone or prednisolone is administered at a dose of 100 mg / day.

36. 10. The pharmaceutical composition of claim 1, wherein prednisone or prednisolone is administered on days 1 to 5 of each 21-day cycle.

37. 2. The pharmaceutical composition of claim 1, wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, prednisone or equivalents thereof, and the bispecific antibody are administered on the same day (e.g., day 1 of cycles 1-6 or cycles 1-8 of a 21-day cycle).

38. Administration is given in 21-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 a dose of 24 mg is administered on day 15; (ii) In cycles 2-4, a dose of 24 mg is administered on days 1, 8, and 15; (iii) In Cycles 5-8, a dose of 24 mg is administered on Day 1; (b) polatuzumab vedotin, rituximab, cyclophosphamide, and doxorubicin are administered on Day 1 of Cycles 1-6; and (c) prednisone or its equivalent is administered on days 1-5 of cycles 1-6; The pharmaceutical composition of claim 1.

39. Administration is given in 21-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 a dose of 48 mg is administered on day 15; (ii) In cycles 2-4, a dose of 48 mg is administered on days 1, 8, and 15; (iii) In Cycles 5-8, a dose of 48 mg is administered on day 1; (b) polatuzumab vedotin, rituximab, cyclophosphamide, and doxorubicin are administered on Day 1 of Cycles 1-6; and (c) prednisone or its equivalent is administered on days 1-5 of cycles 1-6; The pharmaceutical composition of claim 1.

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

41. 10. The pharmaceutical composition of claim 1, wherein rituximab is administered intravenously.

42. 10. The pharmaceutical composition of claim 1, wherein the cyclophosphamide is administered intravenously.

43. 10. The pharmaceutical composition of claim 1, wherein doxorubicin is administered intravenously.

44. 10. The pharmaceutical composition of claim 1, wherein prednisone or prednisolone is administered intravenously or orally.

45. 2. The pharmaceutical composition of claim 1, wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, prednisone and the bispecific antibody are administered sequentially.

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

47. 2. The pharmaceutical composition of 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).

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

49. 2. The pharmaceutical composition of claim 1, wherein the subject has an International Prognostic Index (IPI) score of 2-5.

50. 10. The pharmaceutical composition of claim 1, wherein the subject has not received prior treatment for DLBCL or follicular lymphoma grade 3B.

51. (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.

52. (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.

53. 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.

54. 54. The pharmaceutical composition of claim 53, 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.

55. 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.

56. 56. The pharmaceutical composition of claim 55, wherein the second binding arm comprises a kappa light chain constant region comprising the amino acid sequence set forth in SEQ ID NO:

23.

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

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

59. 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.

60. 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.

61. 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; 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 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. 62. The pharmaceutical composition of claim 61 , wherein the bispecific antibody comprises a heavy chain constant region comprising the amino acid sequences of SEQ ID NOs: 19 and 20.

63. 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.

64. 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.

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

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