A method for treating lymphoma with bispecific antibodies against CD3 and CD20.

Abstract

The present invention provides a regimen of bispecific antibodies targeting both CD3 and CD20 for use in the treatment of lymphoma, such as B-cell non-Hodgkin lymphoma (B-NHL).

Description

[Technical Field]

[0001] The present invention relates to bispecific antibodies targeting both CD3 and CD20, and to the use of such antibodies for the treatment of lymphomas, such as B-cell non-Hodgkin lymphoma (B-NHL), such as aggressive or indolent lymphoma. Advantageous treatment regimens are also provided. [Background technology]

[0002] Monoclonal antibodies (mAbs) have shown great success in treating cancer. A further promising approach to improving antibody therapy is to specifically recruit T cells to antigen-expressing cancer cells. This can be achieved by utilizing bsAbs that target both T cells and antigen-expressing cells. However, early clinical studies have been rather disappointing, mainly due to the low potency, serious adverse effects (cytokine storm), and immunogenicity of bispecific antibodies. Advances in the design and application of bispecific antibodies have partially overcome the initial barrier of cytokine release syndrome without dose-limiting toxicity and improved clinical efficacy (Garber, 2014, Nat. Rev. Drug Discov. 13:799-801).

[0003] The CD20 molecule, also known as human B lymphocyte-restrictive differentiation antigen or Bp35, is found on the surface of over 90% of B cells derived from peripheral blood or lymphoid organs. It is expressed during the early stages of B cell development and persists until plasma cell differentiation. CD20 is present in both normal and malignant B cells. In particular, CD20 is expressed in over 90% of B-cell non-Hodgkin lymphomas but is not found in hematopoietic stem cells, pro-B cells, normal plasma cells, or other normal tissues. Methods of treating cancer, as well as autoimmune and immunological diseases, by targeting CD20 are known in the art.

[0004] For example, the chimeric CD20 antibody rituximab is used or proposed for use in the treatment of cancers, such as aggressive or indolent lymphoma and chronic lymphocytic leukemia (CLL). The human monoclonal anti-CD20 antibody, ofatumumab, is used or proposed for use in the treatment of various CLL indications, follicular lymphoma (FL), neuromyelitis optica (NMO), and diffuse and relapsing-remitting multiple sclerosis (RRMS), among others.

[0005] Currently, bispecific antibodies targeting both CD20 and CD3 are under development. For example, International Publication No. 2011028952 describes, among other things, the generation of CD3xCD20 bispecific molecules using Xencor's XmAb bispecific Fc domain technology; International Publication No. 2014047231 describes REGN1979 and other CD3xCD20 bispecific antibodies generated using Regeneron Pharmaceuticals' FcΔAdp technology; and Sun et al. (2015, Science Translational Medicine 7, 287ra70) describe an anti-CD20 / CD3 T cell-dependent bispecific antibody targeting B cells, constructed using "knobs-into-holes" technology. Such bispecific antibodies are currently being tested in clinical trials for specific indications in humans.

[0006] A particularly interesting bispecific antibody under development is epcolitamab (Duobody CD3xCD20;GEN3013) (Engelberts et al., 2020, EBioMedicine, Vol. 52, 102625, International Publication No. 2016110576 and International Publication No. 2019155008, which are incorporated herein by reference). Epcolitamab has been shown to cause potent T cell-mediated killing of CD20-expressing cells. Compounds and drugs that engage T cells as a mechanism of action have also been shown to attract attention as a common adverse event (AE) known as cytokine release syndrome (CRS). CRS may or may not include fever, hypotension, hypoxia, and other symptoms. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] International Publication No. 2011028952 [Patent Document 2] International Publication No. 2014047231 [Patent Document 3] International Publication No. 2016110576 [Patent Document 4] International Publication No. 2019155008 [Non-patent literature]

[0008] [Non-Patent Document 1] Garber, 2014, Nat.Rev.Drug Discov.13:799-801 [Non-Patent Document 2] Engelberts et al., 2020, EBioMedicine, Vol.52, 102625 [Overview of the Initiative] [Problems that the invention aims to solve]

[0009] Therefore, there is a need for novel and effective therapies to reduce CRS and other adverse events associated with the administration of CD3xCD20 bispecific antibodies. [Means for solving the problem]

[0010] A method of treating a human subject (or "subject") having a lymphoma such as B-cell non-Hodgkin lymphoma (B-NHL), for example an aggressive or indolent lymphoma, by administering a bispecific antibody that binds to CD3 and CD20, such as epcoritamab, by utilizing an advantageous clinical treatment regimen is provided herein.

[0011] In one aspect, a method of treating B-NHL, including aggressive or indolent lymphoma, in a human subject is provided herein, the method comprising administering a bispecific antibody to the subject, the bispecific antibody comprising: (i) a first binding arm that binds to human CD3ε (epsilon) and comprises a first antigen-binding region comprising a variable heavy chain (VH) region and a variable light chain (VL) region, wherein the VH region comprises the CDR1, CDR2, and CDR3 sequences present in the VH region sequence of SEQ ID NO: 6, and the VL region comprises the CDR1, CDR2, and CDR3 sequences present 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 present in the VH region sequence of SEQ ID NO: 13, and the VL region comprises the CDR1, CDR2, and CDR3 sequences present in the VL region sequence of SEQ ID NO: 14, and the bispecific antibody is administered in a 28-day cycle comprising administering a first priming dose of about 0.05 mg to about 0.35 mg on day 1, a first intermediate dose of about 0.6 mg to 5 mg around day 8, a second intermediate dose of about 1 mg to about 10 mg around day 15, followed by a single weekly dose of about 20 - 100 mg.

[0012] Alternatively, the bispecific antibody is administered in a 35-day cycle comprising administering a first priming dose of about 0.05 mg to about 0.35 mg on day 1, a first intermediate dose of about 0.6 mg to 5 mg around day 8, a second intermediate dose of about 1 mg to about 10 mg around day 15, followed by at least two weekly doses of about 20 - 100 mg.

[0013] In some embodiments, the priming dose is about 0.16 mg. In some embodiments, the first intermediate dose is about 0.8 mg. In some embodiments, the second intermediate dose is about 3 mg. In some embodiments, the second intermediate dose is about 6 mg.

[0014] In one embodiment, the method further comprises administering the bispecific antibody once a week at a dose of about 24 to about 48 mg for at least 3 weeks. In some embodiments, the method comprises administering the bispecific antibody once a week at a dose of about 24 to about 48 mg for at least 4 weeks. In some embodiments, the method comprises administering the bispecific antibody once a week at a dose of about 24 to about 48 mg for at least 5 weeks, at least 6 weeks, at least 7 weeks or at least 8 weeks.

[0015] In some embodiments, the method further comprises administering the bispecific antibody once every two weeks (every other week) after each weekly administration, for example, over a 28-day cycle of 6 times. In some embodiments, the method further comprises administering the bispecific antibody once every four weeks after each every-other-week administration, for example, over at least 28-day cycles of 2 times.

[0016] In some embodiments, the bispecific antibody is administered as follows: (i) In cycle 1, a first priming dose of 0.16 mg is administered on day 1, a first intermediate dose of 0.8 mg is administered on day 8, a second intermediate dose of 3 mg is administered on day 15, and a dose of 24 mg is administered on days 22 and optionally day 28, (ii) In cycles 2 and 3, a dose of 24 mg is administered on days 1, 8, 15 and 22, (iii) In cycles 4-9, a dose of 24 mg is administered on days 1 and 15, (iv) In cycles 10 and subsequent cycles, a dose of 24 mg is administered on day 1.

[0017] In some embodiments, the bispecific antibody is administered as follows: (i) In cycle 1, a first priming dose of 0.16 mg is administered on day 1, a first intermediate dose of 0.8 mg is administered on day 8, a second intermediate dose of 3 mg is administered on day 15, and a dose of 48 mg is administered on day 22 and, if applicable, on day 28. (ii) In cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15 and 22. (iii) In cycles 4-9, a dose of 48 mg is administered on day 1 and day 15. (iv) In cycle 10 and subsequent cycles, a dose of 48 mg is administered on day 1.

[0018] In some embodiments, the bispecific antibody is administered as follows: (i) In cycle 1, a first priming dose of 0.16 mg is administered on day 1, a first intermediate dose of 0.8 mg is administered on day 8, a second intermediate dose of 6 mg is administered on day 15, and a dose of 24 mg is administered on day 22 and, if applicable, on day 28. (ii) In cycles 2 and 3, a dose of 24 mg is administered on days 1, 8, 15 and 22. (iii) In cycles 4-9, a dose of 24 mg is administered on day 1 and day 15. (iv) In cycle 10 and subsequent cycles, a dose of 24 mg is administered on day 1.

[0019] In some embodiments, the bispecific antibody is administered as follows: (i) In cycle 1, a first priming dose of 0.16 mg is administered on day 1, a first intermediate dose of 0.8 mg is administered on day 8, a second intermediate dose of 6 mg is administered on day 15, and a dose of 48 mg is administered on day 22 and, if applicable, on day 28. (ii) In cycles 2 and 3, a dose of 48 mg is administered on days 1, 8, 15 and 22. (iii) In cycles 4-9, a dose of 48 mg is administered on day 1 and day 15. (iv) In cycle 10 and subsequent cycles, a dose of 48 mg is administered on day 1.

[0020] In some embodiments, lymphoma is follicular lymphoma (FL), cutaneous T-cell lymphoma (CTCL), marginal zone lymphoma (MZL), chronic lymphocytic leukemia (CLL), or It is small cell lymphocytic lymphoma (SLL).

[0021] In one embodiment, the lymphoma is follicular lymphoma (FL).

[0022] In some embodiments, subjects are treated with a prophylactic measure for cytokine release syndrome (CRS). In some embodiments, the prophylactic measure includes, for example, administering a corticosteroid (including an oral dose, e.g., 15 mg of dexamethasone or its equivalent) on the same day as the bispecific antibody. In some embodiments, the corticosteroid is further administered 2, 3, and 4 days after the administration of the bispecific antibody.

[0023] In some embodiments, subjects are given premedication, such as an antihistamine (e.g., diphenhydramine, intravenously or orally, e.g., a dose of 50 mg or equivalent) and / or an antipyretic (e.g., acetaminophen, e.g., a dose of 560-1000 mg), to reduce their response to the injection. In some embodiments, the premedication is administered on the same day as the bispecific antibody.

[0024] In some embodiments, prophylaxis and premedication are administered during cycle 1. In some embodiments, if the subject experiences a Grade 1 or higher CRS after the last dose of bispecific antibodies in cycle 1, prophylaxis is administered during cycle 2. In some embodiments, prophylaxis is continued in subsequent cycles if the subject experiences a Grade 1 or higher CRS after the last dose of bispecific antibodies in the previous cycle. In further embodiments, premedication is administered during cycle 2. In even further embodiments, premedication is administered during subsequent cycles.

[0025] In some embodiments, if a subject develops Grade 1 CRS, the subject is administered antibiotics. In some embodiments, if a subject develops Grade 2 or Grade 3 CRS, the subject is administered vasopressors. In some embodiments, if a subject develops Grade 4 CRS, the subject is administered at least two vasopressors.

[0026] In some embodiments, if a subject develops grade 2, grade 3, or grade 4 CRS, the subject is administered tocilizumab. In some embodiments, the subject is further administered a steroid (e.g., dexamethasone or methylprednisolone). In some embodiments, if the subject is resistant to tocilizumab, tocilizumab is switched to an anti-IL-6 antibody (e.g., siltuximab) or an IL-1R antagonist (e.g., anakinra).

[0027] In some embodiments, subjects are administered a prophylactic treatment for tumor lysis syndrome (TLS). In some embodiments, the prophylactic treatment for TLS includes administering one or more uric acid-lowering agents before administering a bispecific antibody. In some embodiments, rasburicase and / or allopurinol are administered as uric acid-lowering agents. In some embodiments, if a subject shows signs of TLS, supportive therapies such as rasburicase may be used.

[0028] In some embodiments, subjects treated in the manner described herein achieve, for example, complete response, partial response, or stability as defined by the Lugano classification criteria or LYRIC.

[0029] In some embodiments, the first antigen-binding region of the bispecific antibody includes VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences shown in SEQ ID NOs: 1, 2, and 3, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences shown in SEQ ID NOs: 4, sequence GTN, and SEQ ID NOs: 5, respectively; and the second antigen-binding region includes VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences shown in SEQ ID NOs: 8, 9, and 10, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences shown in SEQ ID NOs: 11, sequence DAS, and SEQ ID NOs: 12, respectively.

[0030] In some embodiments, the first antigen-binding region of the bispecific antibody includes a VH region containing the amino acid sequence of SEQ ID NO: 6 and a VL region containing the amino acid sequence of SEQ ID NO: 7, and the second antigen-binding region includes a VH region containing the amino acid sequence of SEQ ID NO: 13 and a VL region containing the amino acid sequence of SEQ ID NO: 14.

[0031] 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 having a human IgG1 constant region.

[0032] In some embodiments, the bispecific antibody includes an inactive Fc region, for example, an Fc region where the amino acids at positions L234, L235, and D265 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 are F, E, and A, respectively. In some embodiments, the bispecific antibody includes substitutions that promote the formation of the bispecific antibody, for example, in the first heavy chain, the amino acid at position 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 K409 of the human IgG1 heavy chain constant region of SEQ ID NO: 15 is R, or vice versa. The amino acid positions in SEQ ID NO: 15 correspond to the amino acid positions of the human IgG1 heavy chain according to the Eu numbering scheme (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).

[0033] In some embodiments, the 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 further embodiments, the bispecific antibody includes a heavy chain constant region containing the amino acid sequences of SEQ ID NOs. 19 and 20.

[0034] In some embodiments, the bispecific antibody comprises a first heavy chain and a first light chain containing (or consisting of) the amino acid sequences shown in SEQ ID NOs. 24 and 25, respectively, and a second heavy chain and a second light chain containing (or consisting of) the amino acid sequences shown in SEQ ID NOs. 26 and 27, respectively. In some embodiments, the bispecific antibody is epcolitamab or a biosimilar thereof. [Brief explanation of the drawing]

[0035] [Figure 1]This is a schematic diagram of the Repeated Time-to-Event (RTTE) model. The hazard of repeated CRS AEs was explained using a combination of nonlinear transformations of epcolitamab plasma concentration (i.e., cytokine production) and an indirect effects compartment fluctuating between O and 1, thereby explaining the development of resistance. Production in the indirect effects compartment was inhibited by epcolitamab plasma concentration. [Figure 2] This is a schematic diagram of the model-predicted grade 2+ CRS risk within a certain range of SUD regimens. The probability of a grade 2+ CRS event during the first two cycles was simulated using a grid of 20 priming doses and 20 intermediate doses. The current SUD regimen (0.16 / 0.8 / 48 mg) is shown as a circle. [Figure 3] This graph shows the model-predicted risk of 2+ CRS under a 3-step SUD regimen. The probability of at least one grade 2+ event is shown. Full doses were repeated to complete two full cycles. The model-predicted probability of patients experiencing at least one grade 2+ CRS event during the first two cycles is represented by a filled circle (mean) and error bars (90% model prediction interval). The current SUD regimen (0.16 / 0.8 / 48 mg) is highlighted. [Modes for carrying out the invention]

[0036] As used herein, the term “immunoglobulin” 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, for example, Fundamental Immunology Ch.7 (Paul, W., 2nd ed. Raven Press, NY (1989))). Briefly, the heavy chain typically has a heavy chain variable region (VH or V in this specification). H (abbreviated as CH or C) and heavy chain constant region (CH or C in this specification) HIt consists of (abbreviated as ). The heavy chain constant region typically consists 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 highly flexible. The disulfide bond within the hinge region is part of the interaction between the two heavy chains in the IgG molecule. Each light chain typically consists of a light chain variable region (VL or VL in this specification). L (abbreviated as CL or C) and the light chain constant region (CL or C in this specification) L The light chain consists of the following (abbreviated as ). The constant light chain region typically consists of one domain, CL. The VH and VL regions can be further subdivided into hypervariable regions (or hypervariable regions that can be hypervariable in the form of sequencely and / or structurally defined loops), also called complementarity-determining regions (CDRs), and are interspersed with more conserved regions called framework regions (FRs). Each VH and VL typically consists of three CDRs and four FRs arranged in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from the amino terminus to the carboxy terminus (see also Chothia and Lesk J Mol Biol 1987;196:90117). Unless otherwise noted or unless otherwise noted, CDR sequences herein are identified according to 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 noted or unless otherwise noted, references to amino acid positions in the constant region follow 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, Sequence ID No. 15 shows amino acid positions 118-447 of the IgG1 heavy chain constant region according to EU numbering.

[0037] As used herein, the term “amino acid corresponding to position…” refers to the amino acid position number of the human IgG1 heavy chain. The 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 aligned with the other amino acid or segment using ALIGN, ClustalW, or a similar standard sequence alignment program, typically with default settings, and has at least 50%, at least 80%, at least 90%, or at least 95% identity with the human IgG1 heavy chain. Aligning sequences or segments within sequences and thereby determining the positions in the sequence corresponding to the amino acid positions according to the present invention is within the capabilities of those skilled in the art.

[0038] As used herein in connection with the present invention, the term “antibody” (Ab) refers to an immunoglobulin molecule that has the ability to specifically bind to an antigen under typical physiological conditions and has a half-life of sufficient duration to induce, promote, enhance and / or modulate the physiological response associated with antibody binding to the antigen, and / or to mobilize effector activity. The variable regions of the heavy and light chains of the immunoglobulin molecule contain binding domains that interact with the antigen. Unless otherwise specified, the term antibody also includes polyclonal antibodies, monoclonal antibodies (mAbs), antibody-like polypeptides, chimeric antibodies, and humanized antibodies.

[0039] As used herein, the terms “antibody fragment” or “antigen-binding fragment” refer to fragments of immunoglobulin molecules that retain 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) monovalent fragments consisting of Fab' or Fab fragments, VL, VH, CL, and CH1 domains, or monovalent antibodies described in International Publication No. 2007059782 (Genmab); (ii) F(ab')2 fragments, bivalent fragments containing two Fab fragments linked by disulfide crosslinks in the hinge region; (iii) Fd fragments essentially consisting of the VH domain and CH1 domain; (iv) Fv fragments essentially consisting of the VL and VH domains of a single arm of the antibody; (v) dAb fragments essentially consisting of the VH domain, also called domain antibodies (Holt et al; Trends Biotechnol 2003;21:484-90) (Ward et al., Nature 1989;341:54446); (vi) camelid or nanobody (Revets et al; Expert Opin Biol Ther (vii) includes an isolated complementarity-determining region (CDR). Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they may be linked by a synthetic linker that allows the VL and VH regions to pair up and be produced as a single protein chain forming a monovalent molecule (known as a single-chain antibody or single-chain Fv (scFv), see, e.g., Bird et al., Science 1988;242:42326 and Huston et al., PNAS 1988;85:587983) using recombination. Such single-chain antibodies are included in the term antibody fragment unless otherwise specified or clearly indicated by the context.

[0040] As used herein, the terms “antibody-binding region” or “antigen-binding region” refer to a region that interacts with an antigen and includes both the VH and VL regions. The term “antibody,” as used herein, refers not only to monospecific antibodies but also to multispecific antibodies containing multiple, e.g., two or more, e.g., three or more different antigen-binding regions. Unless otherwise specified or clearly inconsistent with the context, the term “antigen-binding region” includes antigen-binding fragments, i.e., fragments of an antibody that retain the ability to specifically bind to an antigen.

[0041] As used herein, the term “isotype” refers to an immunoglobulin class (e.g., IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM) encoded by a heavy chain constant region gene. When a specific isotype, e.g., IgG1, is mentioned, the term is used to indicate that an antibody is sequence-closer to that isotype, e.g., IgG1, than to other isotypes, although it is not limited to a specific isotype sequence, e.g., a specific IgG1 sequence. Thus, an IgG1 antibody, for example, may be a sequence variant of a naturally occurring IgG1 antibody, which may contain mutations in the constant region.

[0042] As used herein, the terms “bispecific antibody,” “bs,” or “bsAb” refer to an antibody having two distinct antigen-binding regions defined by different antibody sequences. Bispecific antibodies may be in any form.

[0043] As used herein, the terms “half-pair,” “Fab arm,” and “arm” refer to a single heavy-chain-light-chain pair.

[0044] Where a bispecific antibody is described as comprising a humbnail antibody "derived from" a first parent antibody and a humbnail antibody "derived from" a second parent antibody, the term "derived from" indicates that the bispecific antibody was produced by recombining the humbnails from each of the first and second parent antibodies into the resulting bispecific antibody by any known method. In this context, "recombination" is not intended to be limited to any particular method of recombination, and therefore includes all methods for producing bispecific antibodies described herein, including, for example, recombination by humbnail exchange (also known as "controlled Fab-arm exchange"), as well as recombination at the nucleic acid level, and / or recombination by co-expression of two humbnails in the same cell.

[0045] When used herein in relation to antibodies, the term "full-length" indicates that the antibody is not a fragment but contains all of the specific isotype domains typically found in its native isotype, such as the VH, CH1, CH2, CH3, hinge, VL, and CL domains of an IgG1 antibody. Full-length antibodies can be designed. An example of a "full-length" antibody is epcolitamab.

[0046] As used herein, the term “Fc region” refers to an antibody region consisting of Fc sequences of two heavy chains of immunoglobulin, wherein the Fc sequences include at least a hinge region, a CH2 domain, and a CH3 domain.

[0047] As used herein, the term “heterodimer interaction between the first CH3 region and the second CH3 region” refers to the interaction between the first CH3 region and the second CH3 region in a first CH3 / second CH3 heterodimer protein.

[0048] As used herein, the term "homo-dimer interaction of the first CH3 region and the second CH3 region" refers to the interaction between a first CH3 region and another first CH3 region in a first CH3 / first CH3 homodimer protein, and the interaction between a second CH3 region and another second CH3 region in a second CH3 / second CH3 homodimer protein.

[0049] As used herein, the term "bind" in the context of the binding of an antibody to a given antigen typically refers to a K corresponding to about 10 -6 M or less, such as 10 -7 M or less, such as about 10 -8 M or less, such as about 10 -9 M or less, about 10 -10 M or less, or about 10 -11 M or less, and binds with an affinity corresponding to a K D where the antibody binds to a given antigen with a K that is at least 10-fold lower, such as at least 100-fold lower, such as at least 1,000-fold lower, such as at least 10,000-fold lower, such as at least 100,000-fold lower, than the K of binding to a non-specific antigen (e.g., BSA, casein) other than the given antigen or an antigen closely related thereto. Since the amount of K D of binding is lower and depends on the K D of the antibody, when the K D of the antibody is very low, the amount of K D of binding to the antigen that is lower than the K D of binding to a non-specific antigen can be at least 10,000-fold (i.e., the antibody is highly specific). D D D D

[0050] As used herein, the term "K D " (M) refers to the dissociation equilibrium constant of a specific antibody-antigen interaction. As used herein, affinity and K D are inversely related, i.e., a higher affinity corresponds to a lower K DThis is intended to refer to a lower affinity with a higher K D This is intended to refer to [something].

[0051] As used herein, the term “isolated antibody” refers to an antibody that substantially does not contain other antibodies having different antigen specificities. In a preferred embodiment, an isolated bispecific antibody that specifically binds to CD20 and CD3 further substantially does not contain a monospecific antibody that specifically binds to CD20 or CD3.

[0052] As used herein, the term “CD3” refers to the human differentiation antigen group 3 protein, which is part of the T cell coreceptor protein complex and consists of four distinct chains. Since CD3 is also found in other species, the term “CD3” is not limited to human CD3 unless otherwise specified in the context. In mammals, the complex consists of a CD3γ (gamma) chain (human CD3γ chain UniProtKB / Swiss-Prot No. P09693, or cynomolgus monkey CD3γ UniProtKB / Swiss-Prot No. Q95LI7), a CD3δ (delta) chain (human CD3δ UniProtKB / Swiss-Prot No. P04234, or cynomolgus monkey CD3δ UniProtKB / Swiss-Prot No. Q95LI8), two CD3ε (epsilon) chains (human CD3ε UniProtKB / Swiss-Prot No. P07766, SEQ ID NO: 28); cynomolgus monkey CD3ε UniProtKB / Swiss-Prot No. Q95LI5; or rhesus monkey CD3ε UniProtKB / Swiss-Prot No. G7NCB9), and a CD3ζ-chain (zeta) chain (human CD3ζ It contains UniProtKB / Swiss-Prot No. P20963 and cynomolgus monkey CD3ζ (UniProtKB / Swiss-Prot No. Q09TK0). These chains associate with molecules known as T cell receptors (TCRs) and generate activation signals in T lymphocytes. The TCR and CD3 molecules together constitute the TCR complex.

[0053] As used herein, the terms “CD3 antibody” or “anti-CD3 antibody” refer to antibodies that specifically bind to the antigen CD3, particularly human CD3ε (epsilon).

[0054] The terms "human CD20" or "CD20" refer to human CD20 (UniProtKB / Swiss-Prot No. P11836, SEQ ID NO: 29) and include all variants, isoforms, and species homologs of CD20 that are spontaneously expressed by cells, including tumor cells, or expressed on cells transfected with the CD20 gene or cDNA. Species homologs include rhesus macaque CD20 (macaca mulatta; UniProtKB / Swiss-Prot No. H9YXP1) and cynomolgus macaque CD20 (macaca fascicularis; UniProtKB No. G7PQ03).

[0055] As used herein, the terms “CD20 antibody” or “anti-CD20 antibody” refer to antibodies that specifically bind to the antigen CD20, particularly human CD20.

[0056] As used herein, the terms “CD3xCD20 antibody,” “anti-CD3xCD20 antibody,” “CD20xCD3 antibody,” or “anti-CD20xCD3 antibody” refer to a bispecific antibody comprising two distinct antigen-binding regions, one of which specifically binds to the antigen CD20 and the other specifically binds to the antigen CD3.

[0057] As used herein, the term “DuoBody-CD3xCD20” refers to an IgG1 bispecific CD3xCD20 antibody comprising a first heavy and light chain pair as defined in SEQ ID NOs. 24 and 25, respectively, and a second heavy and light chain pair as defined in SEQ ID NOs. 26 and 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 VH and VL sequences as defined by SEQ ID NOs. 6 and 7, and the second binding region comprises VH and VL sequences as defined by SEQ ID NOs. 13 and 14. This bispecific antibody can be prepared as described in International Publication No. 2016 / 110576.

[0058] The antibody comprises a functional variant of the heavy chain, light chain, VL region, VH region, or one or more CDRs of the antibody of the examples also provided herein. The functional variants of the heavy chain, light chain, VL, VH, or CDR used in the context of the antibody allow the antibody to still retain at least a substantial percentage (at least about 90%, 95% or more) of the functional features of the “reference” antibody and / or “parent” antibody, including affinity and / or specificity / selectivity for specific epitopes of CD20 and / or CD3, Fc inactivity, and PK parameters such as half-life, Tmax, and Cmax. Such functional variants typically have heavy and light chains of substantially similar length and retain significant sequence identity with the parent antibody. The percentage of 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 × 100), taking into account the number of gaps and the length of each gap (which must be introduced for optimal alignment of the two sequences). The percentage of 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), which is incorporated into the ALIGN program (version 2.0), using a PAM120 weighted residue table, gap length penalty 12, and gap penalty 4. Furthermore, the percentage of identity between two amino acid sequences can be determined using the algorithm of Needleman and Wunsch, J Mol Biol 1970; 48: 444-453. Exemplary variants include those that differ from the parent antibody sequence's heavy and / or light chain, VH and / or VL, and / or CDR regions primarily by conserved substitutions, such as 10 of the substitutions in the variant, e.g., 9, 8, 7, 6, 5, 4, 3, 2, or 1, being conserved amino acid residue substitutions.

[0059] Conservative substitutions can be defined by substitutions within a class of amino acids, as reflected in the table below. [Table 1]

[0060] Unless otherwise specified, the following nomenclature is used to describe mutations: i) an amino acid substitution at a given position is denoted as K409R, for example, meaning a substitution of lysine with arginine at position 409; and ii) for specific variants, a specific three-letter or one-letter code (including the codes Xaa and X to indicate any amino acid residue) is used. Thus, a substitution of lysine with arginine at position 409 is referred to as K409R, and a substitution of lysine with any amino acid residue at position 409 is referred to as K409X. In the case of a deletion of lysine at position 409, it is indicated as K409*.

[0061] As used herein, the term “humanized antibody” refers to a genetically engineered non-human antibody that comprises 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 six non-human antibody CDRs, which together form an antigen-binding site, onto a homologous human receptor framework region (FR) (see International Publication 92 / 22653 and European Patent No. 0629240). To completely reconstitute the binding affinity and specificity of the parental antibody, substitution (reversion mutation) of framework residues from the parental antibody (i.e., non-human antibody) into the human framework region may be required. Structural homology modeling can help identify amino acid residues within the framework region that are important for the antibody’s binding properties. Thus, a humanized antibody may comprise a non-human CDR sequence, a primarily human framework region which may contain one or more amino acid reverse mutations to the non-human amino acid sequence, and a fully human constant region. In DuoBody-CD3xCD20, VH and VL of the CD3 arm as used herein represent the humanized antigen-binding region. In some cases, further amino acid modifications (not necessarily reverse mutations) can be applied to obtain humanized antibodies with desirable characteristics, such as affinity and biochemical properties.

[0062] As used herein, the term “human antibody” refers to an antibody having a variable region and a constant region derived from a human germline immunoglobulin sequence. Human antibodies may contain amino acid residues not encoded by the human germline immunoglobulin sequence (e.g., mutations introduced by random or site-directed mutagenesis in vitro, or by somatic mutation in vivo). However, as used herein, the term “human antibody” is not intended to include antibodies in which a CDR sequence derived from the germline of another mammalian species, such as mouse, is grafted onto a human framework sequence. In DuoBody-CD3xCD20, VH and VL of the CD20 arm as used herein represent the human antigen-binding region. The human monoclonal antibodies of the present invention can be prepared by a variety of techniques, including conventional monoclonal antibody methodologies, e.g., the standard somatic hybridization technique described in Kohler and Milstein, Nature 256:495 (1975). While somatic cell hybridization procedures are preferred in principle, other techniques for producing monoclonal antibodies, such as viral or oncogenic transformation of B lymphocytes or phage display techniques using libraries of human antibody genes, can be used. The mouse is a suitable animal strain for preparing hybridomas that secrete human monoclonal antibodies. 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 produced using, for example, transgenic or transchromosomal mice or rats possessing a portion of the human immune system, rather than using mouse or rat strains. Therefore, 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 a human germline immunoglobulin sequence introduced into an animal, but the final antibody sequence is the result of further modification of the human germline immunoglobulin sequence by somatic hypermutation and affinity maturation via an endogenous animal antibody mechanism (see, e.g., Mendez et al. Nat Genet 1997;15:146-56). In DuoBody-CD3xCD20, the VH and VL regions of the CD20 arm as used herein represent human antigen-binding regions.

[0063] As used herein, the term “biosimilar” (of an approved reference product / biological drug) means a biological product that is similar to a reference product based on data from one or more clinical trials (including immunogenicity and pharmacokinetic or pharmacodynamic evaluations) sufficient to demonstrate the safety, purity and efficacy of the biological product in one or more appropriate use conditions for which the reference product has been approved, intended for use, and for which approval is sought, and is equivalent to the reference product (e.g., there is no clinically significant difference in terms of safety, purity and efficacy between the biological product and the reference product). In some embodiments, the biosimilar biological product and the reference product utilize the same one or more mechanisms of action for one or more states of use prescribed, recommended, or proposed in the proposed labeling, but one or more of these mechanisms of action are known only to the extent that they are known for the reference product. In some embodiments, one or more states of use prescribed, recommended, or proposed for the biological product in the proposed labeling are previously approved for the reference product. In some embodiments, the route of administration, dosage form, and / or potency of the biological product are the same as those of the reference product. The biosimilar may be, for example, a currently known antibody having the same primary amino acid sequence as a commercially available antibody, but may be produced for a different cell type or produced by different production, purification, or formulation methods.

[0064] As used herein, the terms “reducing conditions” or “reducing environment” refer to conditions or environments in which cysteine ​​residues in the hinge region of a substrate, in this case an antibody, are more likely to be reduced than oxidized.

[0065] As used herein, the term “recombinant host cell” (or simply “host cell”) is intended to refer to a cell into which an expression vector, such as an expression vector encoding an antibody as described herein, has been introduced. Examples of recombinant host cells include transfectomas, such as CHO, CHO-S, HEK, HEK293, HEK-293F, Expi293F, PER.C6, or NS0 cells, and lymphocytic cells.

[0066] As used herein, the term “indolent lymphoma” refers to a group of slow-growing non-Hodgkin lymphomas (NHLs), including follicular lymphoma grades 1–3A, cutaneous T-cell lymphoma, marginal zone lymphoma, chronic lymphocytic leukemia, and Waldenström macroglobulinemia. Indolent lymphoma accounts for approximately 41% of all non-Hodgkin lymphoma cases in North America and Northern Europe. In summary, indolent lymphoma responds to treatment, is maintained under control (remission), and has a long survival period, but is not curable.

[0067] As used herein, the term “aggressive lymphoma” refers to a group of non-Hodgkin lymphomas (NHLs) that typically require intensive treatment. Aggressive lymphomas include large B-cell lymphomas (LBCLs), such as diffuse large B-cell lymphoma (DLBCL), high-grade B-cell lymphoma (HGBCL), primary mediastinal large B-cell lymphoma (PMBCL), follicular lymphoma (FL) grade 3B, and mantle cell lymphoma (MCL). Diffuse large B-cell lymphoma (DLBCL) is the most common type of NHL, accounting for approximately 30%–40% of all NHL diagnoses.

[0068] When used herein, “follicular lymphoma” (FL) refers to a lymphoproliferative disorder generally associated with slow progression (Freedman, et al., American Journal of Hematology 95(3):316-317, 2020). FL originates from follicular central B cells, and approximately 85% of cases exhibit the t(14;18)(q32;q21) chromosomal translocation, which causes overexpression of the anti-apoptotic protein Bcl-2. FL is characterized by diffuse lymphadenopathy, bone marrow infiltration, and splenomegaly. Infiltration into non-lymphoid areas is less common. FL grades 1–3A are the most common forms of indolent lymphoma, accounting for 70% of indolent cases and 20–30% of all non-Hodgkin lymphoma cases, with an annual incidence of 1.6–3.1 per 100,000 people. This condition is most frequently diagnosed among people in their 50s and 60s, and is more common in Caucasians than in Black or Asian individuals.

[0069] As used herein, “cutaneous T-cell lymphoma” (CTCL) is a form of lymphoma derived from T cells. Mycosis fungoides, which attacks the skin, is the most common form of CTCL (Bagherini et al. F1000 Research 5:1882, 2016). When cancer cells infiltrate and accumulate in the bloodstream, it is known as Sézary syndrome.

[0070] Marginal zone lymphoma (MZL) refers to a heterogeneous group of indolent B-cell lymphomas that originate in the marginal zone of lymphoid tissue. It accounts for 5–10% of all NHL cases, with an annual incidence of 0.4–1.0 per 100,000 people in Western countries. The World Health Organization classifies MZL into three subtypes: nodal, extranodal, and splenic. Nodal MZL occurs within lymph nodes. Extranodal MZL occurs in the area outside the lymph nodes, with the stomach being the most common site. Splenic MZL occurs in the spleen and can spread into the bloodstream.

[0071] Chronic lymphocytic leukemia (CLL) and small cell lymphocytic lymphoma (SLL) refer to different manifestations of the same disease. When abnormal lymphocytes are primarily located in the lymph nodes, it is called SLL; when abnormal lymphocytes are primarily located in the blood and bone marrow, it is called CLL. CLL is the most common type of leukemia in Western countries. The median age at diagnosis is 72 years.

[0072] The terms “DLBCL” and “HGBCL” as defined herein refer to B-NHL classified as either diffuse large B-cell lymphoma (DLBCL) or high-grade B-cell lymphoma (HGBCL), according to the WHO classification as defined in Swerdlow SH, Campo E, Harris NL, et al. WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues (ed. 4th). Lyon, France: IARC Press (2008) and 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), which are incorporated herein by reference.

[0073] "MCL," or mantle cell lymphoma, includes B-cell lymphomas with a chromosomal translocation t(11;14) that leads to the expression of cyclin D1, which also contains CD5+. MCL as defined herein includes B-NHL classified as MCL according to the WHO classification as defined in Swerdlow SH, Campo E, Harris NL, et al. WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues (ed. 4th). Lyon, France: IARC Press (2008) and 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), which are incorporated herein by reference.

[0074] Using the Lugano modification of the Ann Arbor system, lymphomas are categorized as follows:

[0075] Stage I: Lymphoma is located in one lymph node or group of lymph nodes; or, in rare cases, in one organ of the lymphatic system, such as Waldeyer's ring, thymus, or spleen; or in one extra-surface site of the lymphatic system (IE).

[0076] Stage II: Lymphoma is located in a region near one of the outer lymphatic systems, with or without infiltration of two or more lymph node groups or other lymph nodes (IIE). In either case, the lymphoma site is on the same side of the diaphragm. In Stage II, "bulky disease" refers to a tumor mass larger than a certain size, the threshold of which depends on the type of lymphoma.

[0077] Stage III: Lymphoma is present on both sides of the diaphragm, in either the lymph nodes above and below the diaphragm, or in the lymph nodes above the diaphragm and the spleen.

[0078] Stage IV: Lymphoma is located in one or more organs beyond the lymphatic system, such as the liver, lungs, bone marrow, or cerebrospinal fluid.

[0079] The patient's stage can be determined by blood tests, bone marrow biopsy, chest X-ray, computed tomography (CT) scan, positron emission tomography (PET) scan, and magnetic resonance imaging (MRI).

[0080] As used herein, the term “relapsed lymphoma” refers to lymphoma that has progressed after achieving a partial response (PR) or complete response (CR) to prior antineoplastic therapy.

[0081] As used herein, the term “refractory lymphoma” refers to lymphoma that has been treated with at least one prior antineoplasmic therapy but has not achieved at least a partial response to that therapy.

[0082] As used herein, the terms “autologous stem cell transplantation” or “ASCT” refer to stem cells that are collected from an individual and returned to that individual.

[0083] The term "treatment" refers to administering an effective dose of the therapeutically active antibody described herein for the purpose of alleviating, improving, inhibiting, or eradicating (curing) symptoms or conditions 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 classification criteria and / or LYRIC. Treatment may be continued, for example, until progression or unacceptable toxicity occurs.

[0084] As used herein, the terms “administer” or “dosage” refer to the physical delivery of a composition (or formulation) containing a therapeutic agent to a subject using any of the various methods and delivery systems known to those skilled in the art. Preferred routes of administration for the antibodies described herein include, for example, intravenous, intraperitoneal, intramuscular, subcutaneous, spinal, or other parenteral administration routes by injection or infusion. As used herein, the term “parenteral administration” means, but is not limited to, a mode of administration other than enteral and topical administration, usually by injection, including intravenous, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intrafocal, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions, as well as in vivo electroporation. Alternatively, the therapeutic agents described herein may be administered via routes other than parenteral administration, for example, topical, epidermal, or mucosal administration routes, such as intranasal, oral, vaginal, rectal, sublingual, or topically. The administration may be, for example, a single dose, multiple doses, and / or one or more long-term doses. In the methods described herein, a bispecific antibody (e.g., epcolitamab) is administered subcutaneously. Other agents used in combination with the bispecific antibody, such as GemOx, cytokine release syndrome prophylaxis, and / or oncolytic syndrome (TLS) prophylaxis, may be administered via other routes, such as intravenous or oral.

[0085] The term “effective dose” or “therapeutic effective dose” refers to the amount that is effective in the required dosage and duration to achieve the desired therapeutic outcome. For example, the doses defined herein for a subcutaneously administered bispecific antibody (e.g., epcolitamab), namely 24 mg or 48 mg, can be defined as such an “effective dose” or “therapeutic effective dose.” The therapeutic effective dose of an antibody may vary depending on factors such as the individual’s disease state, age, sex, and weight, as well as the antibody’s ability to induce the desired response in the individual. The therapeutic effective dose is also the amount in which the therapeutically beneficial effects outweigh any toxic or adverse effects of the antibody or antibody portion. In some embodiments, patients treated with the methods described herein show improvement in ECOG performance status. The therapeutic effective dose or dosage of a drug includes a “preventive effective dose” or “preventive effective dosage,” which is any amount of a drug that, when administered alone or in combination with another therapeutic agent, inhibits the onset or recurrence of a disease in subjects at risk of developing a disease or disorder (e.g., cytokine release syndrome) or at risk of disease recurrence.

[0086] As used herein, the term “inhibits tumor growth” includes any measurable reduction of tumor growth, e.g., inhibition of tumor growth by at least about 10%, 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%.

[0087] As used herein, the term “subject” (or “subject”) refers to a human patient, for example, a human patient with B-NHL such as FL or LBCL, including DLBCL. The terms “subject” (or “subject”) and “patient” are used interchangeably herein.

[0088] As used herein, the term "buffer" refers to a pharmaceutically acceptable buffer. The term "buffer" encompasses agents that maintain the pH value of a solution within an acceptable range, and includes, but is not limited to, acetates, histidines, TRIS® (tris(hydroxymethyl)aminomethane), citrates, succinates, glycolates, etc. Generally, as used herein, "buffers" have a pKa and buffering capacity suitable for a pH range of about 5 to about 6, preferably about 5.5.

[0089] As used herein, “progression” or “PD” refers to a situation in which one or more indicators of lymphoma indicate that the disease is progressing despite treatment. In one embodiment, progression is defined based on the Lugano Response Criteria ("Lugano Classification Criteria") and / or Response to Immunomodulatory Therapy Criteria (LYRIC) for malignant lymphoma. Details of the Lugano Classification Criteria / Classification System, including the definitions of complete response (CR), partial response (PR), no response / stable disease (NR / SD), and progression (PD), are provided in Cheson et al. J Clin Oncol 2014;32:3059-68, which are incorporated herein by reference (see, in particular, Table 3 of Cheson et al., 2014). Details of LYRIC are shown in Table 7.

[0090] As used herein, “surfactants” are compounds typically used in pharmaceutical formulations to prevent drug adsorption and / or aggregation to surfaces. Furthermore, 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, meaning they are typically composed of both hydrophilic and hydrophobic or lipophilic groups, and can therefore form micelles or similar self-assembling structures in aqueous solutions. Known surfactants for pharmaceutical use include glycerol monooleate, benzethonium chloride, sodium doxate, phospholipids, polyethylene alkyl ethers, sodium lauryl sulfate, and tricaprylin (anionic surfactants); benzalkonium chloride, citrimide, cetylpyridinium chloride, and phospholipids (cationic surfactants); α-tocopherol, glycerol monooleate, myristyl alcohol, phospholipids, poloxamer, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stellarate, polyoxyl hydroxystearate, polyoxylglycerides, polysorbates, e.g., polysorbate 20 or polysorbate 80, propylene glycol dilaurate, propylene glycol monolaurate, sorbitan ester sucrose palmitate, sucrose stearate, tricaprylin, and TPGS (nonionic and amphoteric surfactants).

[0091] As used herein, “diluents” are pharmaceutically acceptable (safe and non-toxic for administration to humans) and useful for preparing dilutions of pharmaceutical compositions or pharmaceutical formulations (the terms “composition” and “formulation” are used interchangeably herein). Preferably, such dilutions of a composition dilute only the antibody concentration and not the buffer and stabilizer. Thus, in one embodiment, the diluent contains the buffer and stabilizer at the same concentrations as those present in the pharmaceutical composition of the present invention. Further exemplary diluents include sterile water, bacteriostatic water for injection (BWFI), pH buffered solution (preferably acetate buffer), physiological saline (e.g., physiological saline for injection), Ringer's solution, or dextrose solution. In one embodiment, the diluent contains or is essentially composed of acetate buffer and sorbitol.

[0092] As used herein, the term “approximately” refers to values ​​that are 10% above and 10% below a given value.

[0093] In one embodiment, the present invention provides a method for treating indolent lymphoma in a human subject, the method comprising administering a bispecific antibody to the subject, the bispecific antibody being (i) A first binding arm comprising a first antigen-binding region that binds to human CD3ε (epsilon) and includes a variable heavy chain (VH) region and a variable light chain (VL) region, wherein the VH region includes the CDR1, CDR2, and CDR3 sequences in the VH region sequence of SEQ ID NO: 6, and the VL region includes the CDR1, CDR2, and CDR3 sequences in the VL region sequence of SEQ ID NO: 7, and (ii) A second binding arm that binds to human CD20 and includes a second antigen-binding region comprising a VH region and a VL region, wherein the VH region comprises the CDR1, CDR2, and CDR3 sequences in the VH region sequence of SEQ ID NO: 13, and the VL region comprises the CDR1, CDR2, and CDR3 sequences in the VL region sequence of SEQ ID NO: 14, The bispecific antibody is administered in a 28-day cycle, including a priming dose of approximately 0.05 mg to 0.35 mg on day 1, a first intermediate dose of approximately 0.6 mg to 5 mg around day 8, a second intermediate dose of approximately 1 mg to 10 mg around day 15, and a subsequent single weekly dose of approximately 20 to 100 mg.

[0094] Alternatively, the bispecific antibody may be administered in a 35-day cycle, including a priming dose of approximately 0.05 mg to 0.35 mg on day 1, a first intermediate dose of approximately 0.6 mg to 5 mg around day 8, a second intermediate dose of approximately 1 mg to 10 mg around day 15, followed by two weekly doses of approximately 20 to 100 mg.

[0095] In some embodiments, the priming dose is approximately 0.16 mg.

[0096] In some embodiments, the first intermediate dose is approximately 0.6 to 1.2 mg, for example, approximately 0.8 mg.

[0097] In some embodiments, the second intermediate dose is approximately 3 to 6 mg, for example, about 3 mg or about 6 mg.

[0098] In some embodiments, the weekly dose is approximately 20–60 mg, for example, 24 mg or 48 mg.

[0099] In some embodiments, weekly administration is performed at least four times.

[0100] Further embodiments provide a dosing regimen in which a bispecific antibody is administered every two weeks after a weekly dose. In particular, the bi-weekly administration of the bispecific antibody may be performed for at least six times.

[0101] Furthermore, this specification provides a dosing regimen in which a bispecific antibody is administered every four weeks after administration every other week.

[0102] Preferably, a bispecific antibody is administered subcutaneously.

[0103] In some embodiments, the method includes: (a) The first cycle of 28 days (i) Administer a priming dose of the bispecific antibody on day 1; (ii) On day 8, administer the first intermediate dose of the bispecific antibody; (iii) On day 15, administer a second intermediate dose of the bispecific antibody; (iv) Administer a dose of 24 mg of bispecific antibody on day 22; (b) Cycles 2-3, each consisting of 28 days, in which a 24 mg dose of bispecific antibody is administered on days 1, 8, 15, and 22; (c) Each cycle consists of 4-9 days, with each receiving a 24 mg dose of bispecific antibody on days 1 and 15; and (d) A further 28-day cycle is followed by administration of a 24 mg dose of bispecific antibody on day 1.

[0104] In some embodiments, the method includes: (a) The first cycle of 35 days (i) Administer a priming dose of the bispecific antibody on day 1; (ii) On day 8, administer the first intermediate dose of the bispecific antibody; (iii) On day 15, administer a second intermediate dose of the bispecific antibody; (iv) Administer a dose of 24 mg of bispecific antibody on days 22 and 28; (b) Cycles 2-3, each consisting of 28 days, in which a 24 mg dose of bispecific antibody is administered on days 1, 8, 15, and 22; (c) Each cycle consists of 4-9 days, with each receiving a 24 mg dose of bispecific antibody on days 1 and 15; and (d) A further 28-day cycle is followed by administration of a 24 mg dose of bispecific antibody on day 1.

[0105] In some embodiments, the method includes: (a) The first cycle of 28 days (i) Administer a priming dose of the bispecific antibody on day 1; (ii) On day 8, administer the first intermediate dose of the bispecific antibody; (iii) On day 15, administer a second intermediate dose of the bispecific antibody; (iv) Administer a dose of 48 mg of bispecific antibody on day 22; (b) Each of the 28-day cycles consists of 2-3, each administering a 48 mg dose of bispecific antibody on days 1, 8, 15, and 22; (c) Each cycle consists of 4-9 days, with each receiving a 48 mg dose of bispecific antibody on days 1 and 15; and (d) A further 28-day cycle is followed by administration of a 48 mg dose of bispecific antibody on day 1.

[0106] In some embodiments, the method includes: (a) The first cycle of 35 days (i) Administer a priming dose of the bispecific antibody on day 1; (ii) On day 8, administer the first intermediate dose of the bispecific antibody; (iii) On day 15, administer a second intermediate dose of the bispecific antibody; (iv) Administer a dose of 48 mg of bispecific antibody on days 22 and 28; (b) Each of the 28-day cycles consists of 2-3, each administering a 48 mg dose of bispecific antibody on days 1, 8, 15, and 22; (c) Each cycle consists of 4-9 days, with each receiving a 48 mg dose of bispecific antibody on days 1 and 15; and (d) A further 28-day cycle is followed by administration of a 48 mg dose of bispecific antibody on day 1.

[0107] In some embodiments, the method includes: (a) The first cycle of 28 days (i) Administer a priming dose of 0.16 mg of bispecific antibody on day 1; (ii) On day 8, administer the first intermediate dose of 0.8 mg of the bispecific antibody; (iii) On day 15, administer a second intermediate dose of 3 mg of bispecific antibody; (iv) Administer a dose of 48 mg of bispecific antibody on day 22; (b) Each of the 28-day cycles consists of 2-3, each administering a 48 mg dose of bispecific antibody on days 1, 8, 15, and 22; (c) Each cycle consists of 4-9 days, with each receiving a 48 mg dose of bispecific antibody on days 1 and 15; and (d) A further 28-day cycle is followed by administration of a 48 mg dose of bispecific antibody on day 1.

[0108] In some embodiments, the method includes: (a) The first cycle of 35 days (i) Administer a priming dose of 0.16 mg of bispecific antibody on day 1; (ii) On day 8, administer the first intermediate dose of 0.8 mg of the bispecific antibody; (iii) On day 15, administer a second intermediate dose of 3 mg of bispecific antibody; (iv) Administer a dose of 48 mg of bispecific antibody on days 22 and 28; (b) Each of the 28-day cycles consists of 2-3, each administering a 48 mg dose of bispecific antibody on days 1, 8, 15, and 22; (c) Each cycle consists of 4-9 days, with each receiving a 48 mg dose of bispecific antibody on days 1 and 15; and (d) A further 28-day cycle is followed by administration of a 48 mg dose of bispecific antibody on day 1.

[0109] In some embodiments, the method includes: (a) The first cycle of 35 days (i) Administer a priming dose of 0.16 mg of bispecific antibody on day 1; (ii) On day 8, administer the first intermediate dose of 0.8 mg of the bispecific antibody; (iii) On day 15, administer a second intermediate dose of 6 mg of bispecific antibody; (iv) Administer a dose of 48 mg of bispecific antibody on days 22 and 28; (b) Each of the 28-day cycles consists of 2-3, each administering a 48 mg dose of bispecific antibody on days 1, 8, 15, and 22; (c) Each cycle consists of 4-9 days, with each receiving a 48 mg dose of bispecific antibody on days 1 and 15; and (d) A further 28-day cycle is followed by administration of a 48 mg dose of bispecific antibody on day 1.

[0110] In short, B-NHL is typically divided into indolent (slow-growing) and aggressive subtypes. Aggressive B-NHL has high Ki67 expression, while indolent B-NHL has relatively low Ki67 expression. In summary, indolent lymphoma responds to treatment, is maintained under control (remission), and has a long survival period, but is not cured. Aggressive lymphoma usually requires intensive treatment and some have a good prospect of permanent cure. Aggressive B-NHL includes diffuse large B-cell lymphoma (DLBCL), high-grade B-cell lymphoma (HGBCL), primary mediastinal large B-cell lymphoma (PMBCL), follicular lymphoma (FL) grade 3B, and mantle cell lymphoma (MCL). Indolent B-NHL includes FL grades 1–3A, marginal zone lymphoma (MZL), and small lymphocytic lymphoma (SLL). Diffuse large B-cell lymphoma (DLBCL) is the most common type of NHL, accounting for approximately 30–40% of all NHL diagnoses, followed by FL (20–25% of all NHL diagnoses). Many B-cell lymphomas express B-cell markers, such as CD19, CD20, CD22, and CD79b. The biological heterogeneity of B-cell malignancies is reflected in the clinical course and outcomes of individual diseases. Indolent diseases such as FL G1–3A, MZL, and SLL progress slowly, with a median survival of 8–10 years. In contrast, more aggressive diseases such as DLBCL / HGBCL have a median survival of 6 months if left untreated. The median age at diagnosis for most patients with lymphoma is approximately 60–65 years (WHO, 2008).

[0111] Aggressive lymphoma treatment regimens This specification provides further methods for treating aggressive lymphomas, such as large B-cell lymphoma (LBCL), in human subjects using a bispecific antibody that binds to CD3 and CD20 ("anti-CD3xCD20 antibody"), such as an isolated anti-CD3xCD20 antibody such as epcolitamab that binds to human CD3 and human CD20, in a treatment regimen comprising at least one administration of a first priming dose of 0.5–0.35 mg, a first intermediate dose of 0.6–5 mg, a second intermediate dose of 1–10 mg of the anti-CD3xCD20 antibody, followed by a bispecific antibody at a dose of 20–100 mg.

[0112] In some embodiments, the priming dose is approximately 0.16 mg.

[0113] In some embodiments, the first intermediate dose is approximately 0.6 to 1.2 mg, for example, approximately 0.8 mg.

[0114] In some embodiments, the second intermediate dose is approximately 3 to 6 mg. Specifically, the second intermediate dose may be approximately 3 mg, or it may be approximately 6 mg.

[0115] In certain embodiments, the bispecific antibody is administered in an initial cycle, for example, a 21-day cycle, which includes administering a first priming dose of about 0.05 mg to about 0.35 mg on day 1, a first intermediate dose of about 0.6 mg to about 5 mg around day 8, and a second intermediate dose of about 1 mg to about 10 mg around day 15.

[0116] After the first cycle, the bispecific antibody, (a) Cycles 2-4, each consisting of weekly administration of approximately 24 or 48 mg of bispecific antibody, for example, around day 1, day 8, and day 15, for a total of 21 days; (b) Cycles 5-6, each consisting of two doses of bispecific antibody at approximately 24 or 48 mg, administered around day 1 and around day 15, respectively, over a 21-day period; and (c) A bispecific antibody may be administered in a single dose of approximately 24 or 48 mg, for example, on day 1, in subsequent 28-day cycles.

[0117] The method according to this embodiment may further include the administration of therapeutically effective doses of rituximab, cyclophosphamide, doxorubicin, and vincristine. In particular, this method may be for the treatment of DLBCL, for example, previously untreated DLBCL.

[0118] An alternative embodiment provides a method in which, after the first cycle, the bispecific antibody, (a) Cycles 2-4, each consisting of weekly administration of approximately 24 or 48 mg of bispecific antibody, for example, around day 1, day 8, and day 15, for a total of 21 days; (b) Cycles 5-9, each consisting of two doses of bispecific antibody at approximately 24 or 48 mg, for example, around day 1 and day 15, over a 21-day period; and (c) A bispecific antibody is administered in a single dose of approximately 24 or 48 mg over a 28-day period, for example, on day 1.

[0119] The method according to this embodiment may further include the administration of therapeutically effective doses of rituximab, dexamethasone, cytarabine, and oxaliplatin / carboplatin. In particular, the method may be for the treatment of DLBCL, e.g., relapsed / refractory DLBCL, especially DLBCL eligible for autologous stem cell transplantation (ASCT).

[0120] In a further alternative embodiment, the method is provided, and after the first cycle, the bispecific antibody, (a) Cycle 2, consisting of 21 days, in which a bispecific antibody dose of approximately 24 or 48 mg is administered weekly, for example, around day 1, day 8, and day 15; (b) Cycles 3-6, each consisting of two doses of bispecific antibody at approximately 24 or 48 mg, administered around day 1, for example, over 21 days; and ()The bispecific antibody is administered in a single dose of approximately 24 or 48 mg over a 28-day period, for example, on day 1.

[0121] The method according to this embodiment may further include the administration of therapeutically effective doses of rituximab, cyclophosphamide, doxorubicin, and vincristine. In particular, the method may be for treating DLBCL in subjects who have previously untreated DLBCL and are ineligible to receive the full therapeutic dose of anthracyclines.

[0122] Indolent lymphoma treatment regimen This specification provides a method for treating FL in human subjects using a bispecific antibody that binds to CD3 and CD20 ("anti-CD3xCD20 antibody"), such as an isolated anti-CD3xCD20 antibody such as epcolitamab that binds to human CD3 and human CD20, in a treatment regimen comprising the administration of a first priming dose of 0.5–0.35 mg, a first intermediate dose of 0.6–5 mg, a second intermediate dose of 1–10 mg of the anti-CD3xCD20 antibody, followed by a bispecific antibody at least twice weekly in doses of 20–100 mg.

[0123] Therefore, in one embodiment, a method is provided for treating indolent lymphoma (e.g., FL) in human subjects, the method comprising administering a bispecific antibody, the bispecific antibody is (i) A first binding arm comprising a first antigen-binding region that binds to human CD3ε (epsilon) and includes a variable heavy chain (VH) region and a variable light chain (VL) region, wherein the VH region includes the CDR1, CDR2, and CDR3 sequences in the VH region sequence of SEQ ID NO: 6, and the VL region includes the CDR1, CDR2, and CDR3 sequences in the VL region sequence of SEQ ID NO: 7, and (ii) A second binding arm that binds to human CD20 and includes a second antigen-binding region comprising a VH region and a VL region, wherein the VH region comprises the CDR1, CDR2, and CDR3 sequences in the VH region sequence of SEQ ID NO: 13, and the VL region comprises the CDR1, CDR2, and CDR3 sequences in the VL region sequence of SEQ ID NO: 14, The bispecific antibody is administered in a priming dose of approximately 0.05 mg to 0.35 mg, a first intermediate dose of approximately 0.6 mg to 5 mg, a second intermediate dose of approximately 1 mg to 10 mg, followed by a dose of 24 mg or 48 mg.

[0124] In some embodiments, the priming dose is approximately 0.16 mg.

[0125] In some embodiments, the first intermediate dose is approximately 0.6 to 1.2 mg, for example, approximately 0.8 mg.

[0126] In some embodiments, the second intermediate dose is approximately 3 to 6 mg. Specifically, the second intermediate dose may be approximately 3 mg, or it may be approximately 6 mg.

[0127] In some embodiments, the bispecific antibody is administered in a dose of 24 mg (or approximately 24 mg). In some embodiments, the bispecific antibody is administered in a dose of 48 mg (or approximately 48 mg).

[0128] In certain embodiments, the bispecific antibody is administered in an initial cycle, for example a 28-day cycle, which includes administering a first priming dose of about 0.05 mg to about 0.35 mg on day 1, a first intermediate dose of about 0.6 mg to about 5 mg around day 8, a second intermediate dose of about 1 mg to about 10 mg around day 15, and a dose of about 24 or about 48 mg around day 22.

[0129] After the first cycle, the bispecific antibody, (b) Cycle 2, consisting of 28 days, in which a bispecific antibody dose of approximately 24 or 48 mg is administered weekly, for example, around day 1, day 8, day 15, and day 22; (c) The bispecific antibody may be administered in cycles 3 to 26, each cycle consisting of a single dose of approximately 24 or 48 mg, for example, on day 1, over a period of 28 days.

[0130] The method according to this embodiment may further include the administration of therapeutically effective doses of rituximab and lenalidomide. This method may be particularly for the treatment of relapsed / refractory FL. Alternatively, this method may be for the treatment of previously untreated, advanced FL.

[0131] In an alternative embodiment, a method is provided in which a bispecific antibody is administered once every 8 weeks at a dose of approximately 24 or approximately 48 mg after the initial cycle.

[0132] This method is primarily intended as maintenance therapy for patients with FL who have achieved a complete or partial response after first-line or second-line treatment.

[0133] In some embodiments, the bispecific antibody is a full-length antibody. In other embodiments, the bispecific antibody is an antibody having an inactive Fc region. In yet another embodiment, the bispecific antibody is a full-length antibody having an inactive Fc region.

[0134] With respect to a dose of 24 mg or 48 mg (or approximately 24 mg or approximately 48 mg) of the bispecific antibody to be administered, or any other specific dose, this amount is understood to refer to the amount of bispecific antibody representative of the full-length antibody, such as epcolitamab as defined in the Examples section. Therefore, administering a 24 mg dose of bispecific antibody can be referred to as administering the dose of bispecific antibody described herein, which corresponds to a 24 mg dose of epcolitamab. Those skilled in the art can easily determine the amount of antibody to be administered, for example, if the molecular weight of the antibody used is substantially different from that of a full-length antibody such as epcolitamab. For example, the amount of 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. Insofar as a bispecific antibody (e.g., a functional variant of DuoBody CD3xCD20) has characteristics highly similar to DuoBody CD3xCD20 with respect to its plasma half-life, Fc inactivity, and / or binding properties to CD3 and CD20, i.e., CDR and epitope binding characteristics, such an antibody is suitable for use in the methods provided herein, at doses described for full-length antibodies such as epcolitamab.

[0135] In some embodiments, the dose of the bispecific antibody is administered once a week in a 28-day cycle (weekly administration). In one embodiment, a weekly dose of 24 mg or 48 mg is administered in 2.5 28-day cycles (i.e., 10 doses; on days 15 and 22 of cycle 1, and on days 1, 8, 15 and 22 of cycles 2 and 3). In other embodiments, after weekly administration, the administration interval can be reduced to once every two weeks (bi-weekly administration). In one embodiment, bi-weekly administration is performed in 6 28-day cycles (i.e., 12 doses). In some embodiments, after bi-weekly administration, the administration interval can be reduced to once every four weeks. In one embodiment, administration once every four weeks can be carried out over a long period, for example, over at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen, at least seventeen cycles of a 28-day cycle, for example, cycles 1-20, 1-19, 1-18, 1-17, 1-16, 1-15, 1-14, 1-13, 1-12, 1-10, 1-5, 5-20, 5-15, or 5-10. In some embodiments, epcolitamab is administered as monotherapy (i.e., without GemOx) starting from cycle 10 of a 28-day cycle. In some embodiments, epcolitamab is administered as monotherapy from cycle 10 to cycle 26 of a 28-day cycle. In some embodiments, epcolitamab is administered as monotherapy from cycle 7 of a 28-day cycle until progression or unacceptable toxicity occurs (for example, as defined by the Lugano classification criteria or LYRIC).

[0136] In one embodiment, weekly doses of the bispecific antibody are administered in 28-day cycles, cycles 1-3 (which may include priming and intermediate doses, as described below); bi-weekly doses of the bispecific antibody are administered in cycles 4-9; and once every four weeks doses are administered from cycle 10 onward, for example, in cycles 10-15, 10-20, 10-25, 10-30, or more, until, for example, progressive or unacceptable toxicity is observed in the subject. In some embodiments, once every four weeks doses are administered in cycles 10-26.

[0137] It is understood that the doses referred to herein may also be called full doses or flat doses in the above scenario where, for example, weekly doses, bi-weekly doses, and / or quarter-weekly doses are administered at the same level. Therefore, when selecting a dose of 48 mg, preferably the same dose of 48 mg is administered weekly, bi-weekly, and quarter-weekly, respectively. A priming dose or a priming dose followed by first and second intermediate doses is administered before administering the full dose. This may be advantageous as it may help mitigate the risk and severity of cytokine release syndrome (CRS), a possible side effect that can occur during treatment with the bispecific anti-CD3xCD20 antibody described herein. Such a priming dose, or priming dose and intermediate doses, are lower doses compared to the flat or full dose.

[0138] Therefore, in some embodiments, a priming dose of the bispecific antibody can be administered before administering the weekly dose of 24 mg or 48 mg. In one embodiment, the priming dose in cycle 1 is administered two weeks before administering the first weekly dose of 24 mg or 48 mg. In one embodiment, the priming dose is 0.16 mg (or about 0.16 mg) of the full-length bispecific antibody.

[0139] In some embodiments, a first intermediate dose of the bispecific antibody is administered after a priming dose and before a weekly dose of 24 mg or 48 mg. In one embodiment, the priming dose is administered one week before the first intermediate dose (i.e., on day 1 of cycle 1), the first intermediate dose is administered one week before the second intermediate dose (i.e., on day 8 of cycle 1), and the second intermediate dose is administered before the first weekly dose of 24 mg or 48 mg (i.e., on day 15 of cycle 1). In one embodiment, the priming dose is 0.16 mg of the full-length bispecific antibody. In one embodiment, the first intermediate dose is 800 μg (0.8 mg) or about 800 μg (0.8 mg) of the full-length bispecific antibody. In one embodiment, the second intermediate dose is 3 mg of the full-length bispecific antibody. In one embodiment, the second intermediate dose is 6 mg of the full-length bispecific antibody.

[0140] In some embodiments, subjects are given premedication and / or prophylactic treatment for CRS prior to administration of the bispecific antibody.

[0141] In one embodiment, a subject receiving treatment according to the method herein has recorded indolent lymphoma 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), the contents of which are incorporated herein by reference). In some embodiments, the subject has FL. In some embodiments, the subject has follicular lymphoma grade 3B. In further embodiments, the subject has relapsed after at least one prior therapy or is resistant to that therapy. In further embodiments, the subject has failed a previous self-administered HSCT. In further embodiments, the subject is ineligible to undergo a self-administered HSCT for reasons such as age, performance status, comorbidities, and / or inadequate response to prior treatment.

[0142] In some embodiments, participants have an East Coast Cancer Clinical Trials Group (ECOG) Performance Status (ECOG PS) of 0, 1, or 2. Information regarding ECOG PS scores can be found, for example, in Oken et al, Am J Clin Oncol 1982 Dec;5(6):649-55.

[0143] In some embodiments, the subject has a measurable disease defined as (a) one or more measurable nodular lesions (long axis > 1.5 cm and short axis > 1.0 cm) or one or more measurable extranodal lesions (long axis > 1 cm) as detected by CT or MRI.

[0144] In some embodiments, the subject is (a) ANC ≥ 1.0 × 10 9 / L, (b) Platelet count>75×10 9 / L, or ≥50 × 10 9 The patient has acceptable organ function defined as (c) ALT level ≤ 2.5 times ULN, (d) total bilirubin level ≤ 2 times ULN, (e) eGFR > 50 mL / min (according to the Cockcroft-Gault formula), and (f) PT, INR, and aPTT ≤ 1.5 times ULN (without anticoagulant administration).

[0145] In some embodiments, the subjects do not have a severe allergic reaction or anaphylactic reaction to anti-CD20 antibody therapy or bispecific antibodies, nor do they have a known allergy or intolerance to any component or excipient of the bispecific antibody preparation.

[0146] In some embodiments, subjects do not have clinically significant cardiac disease, such as (a) myocardial infarction within one year of the first administration of a bispecific antibody, or unstable or uncontrolled disease / condition related to or affecting cardiac function (e.g., unstable angina, congestive heart failure, NYHA class III-IV), cardiac arrhythmia (CTCAE version 4 grade 2 or higher), or clinically significant ECG abnormalities, and / or (b) a 12-lead ECG showing baseline QTcF > 470 milliseconds.

[0147] Human subjects receiving the treatments described herein may be patients who meet one or more of the inclusion criteria described in the examples.

[0148] The methods described herein are advantageous for the treatment of indolent lymphomas such as FL. Treatment is maintained continuously, for example, using the treatment regimens described herein, until progression occurs or unacceptable toxicity occurs.

[0149] The response of subjects with indolent lymphoma to treatment using the methods described herein may be evaluated according to the Lugano Response Criteria for Malignant Lymphoma (also referred to herein as the “Lugano Classification Criteria”) and / or Lymphoma Response to Immunomodulatory Therapy Criteria (also referred to herein as “LYRIC”), as described in the Examples. In one embodiment, the Lugano Classification Criteria is used to evaluate complete response (CR), partial response (PR), and stable (SD). In some embodiments, patients showing disease progression, also referred to as progression (PD) according to the Lugano Classification Criteria, are further evaluated according to LYRIC. Details regarding the Lugano Classification Criteria / Classification System, including the definitions of complete response, partial response, no response / stable, and progression, are provided in Cheson et al. J Clin Oncol 2014;32:3059-68 (see, in particular, Table 3 of Cheson et al., 2014). Details regarding LYRIC are shown in Table 7.

[0150] In some embodiments, subjects are treated in the manner described herein until they exhibit disease progression (PD) as defined, for example, by the Lugano classification criteria and / or LYRIC. In one embodiment, subjects are treated in the manner described herein until they exhibit disease progression (PD) as defined by both the Lugano classification criteria and LYRIC.

[0151] Subjects treated according to the methods described herein preferably experience improvement in at least one sign of indolent lymphoma (e.g., FL). In one embodiment, improvement is measured by a reduction in the amount and / or size of a measurable tumor lesion. In some embodiments, the lesion can be measured on CT, PET-CT, or MRI films. In some embodiments, cytology or histology can be used to assess the response to therapy. In some embodiments, bone marrow aspiration and bone marrow biopsy can be used to assess the response to therapy.

[0152] In one embodiment, the treated subject exhibits a complete response (CR), partial response (PR), or stable disease (SD) as defined by the Lugano classification criteria or LYRIC (see, for example, Table 7). In some embodiments, the methods described herein result in at least one therapeutic effect, selected from an extension of survival, e.g., progression-free survival or overall survival, compared to, optionally, another therapy or placebo.

[0153] In some embodiments, T cell activity (e.g., CD4+ and / or CD8+ T cell activity) is increased in subjects treated with a combination of a bispecific antibody, gemcitabine, and oxaliplatin. In some embodiments, CD69, CD25, PD-1, and / or LAMP-1 expression is increased in CD4+ and / or CD8+ T cells from subjects treated with a combination of a bispecific antibody, gemcitabine, and oxaliplatin.

[0154] In some embodiments, antitumor activity (e.g., B cell cytotoxicity) is increased in subjects treated with a combination of a bispecific antibody, gemcitabine, and oxaliplatin, for example, compared to subjects treated with the bispecific antibody alone or with a combination of the bispecific antibody and gemcitabine or oxaliplatin.

[0155] Cytokine release syndrome (CRS) can occur in human subjects when methods utilizing approaches based on immune cells and bispecific antibodies that function by engaging, for example, CD3, through the activation of immune effector cells (Lee et al., Biol Blood Marrow Transplant 2019;25:625-38, incorporated herein by reference). Therefore, in some embodiments, CRS relief is performed in conjunction with the methods described herein. As part of CRS relief, the selection of a priming dose and first and second intermediate doses is made before administering the full dose (e.g., 24 or 48 mg), as described herein. CRS can be classified according to standard practice (e.g., as outlined in Lee et al., Biol Blood Marrow Transplant 2019;25:625-38, incorporated herein by reference). CRS may include the overrelease of cytokines, such as pro-inflammatory cytokines, e.g., IL-6, TNF-α, or IL-8, and may result in adverse effects such as fever, nausea, vomiting, and chills. Therefore, despite the unique antitumor activity of bispecific antibodies such as epcolitamab, their immunological mode of action may cause undesirable side effects, namely the induction of undesirable inflammatory responses. Thus, patients may be further subjected to concurrent treatment, prophylaxis, and / or premedication with, for example, analgesics, antipyretics, and / or anti-inflammatory drugs to alleviate potential CRS symptoms.

[0156] Accordingly, in one embodiment, human subjects in the method described herein are treated with a prophylactic treatment for CRS. In some embodiments, the prophylactic treatment includes the administration of a corticosteroid. In one embodiment, the prophylactic treatment is administered on the same day as the bispecific antibody. The prophylactic treatment may also be administered on the following day, more preferably on the second, third, and fourth days thereafter. It is understood that the second, third, and fourth days, when related to further dosing, e.g., prophylactic treatment, are related to the administration of the bispecific antibody administered on day 1. For example, if the antibody is administered on day 15 in a cycle and the prophylactic treatment is also administered, the prophylactic treatment corresponding to days 2, 3, and 4 would be on days 16, 17, and 18 of that cycle. In some embodiments, the prophylactic treatment is administered on the day the bispecific antibody is administered and on the second to fourth days thereafter. When the prophylactic treatment is administered on the same day as the bispecific antibody, the prophylactic treatment is preferably administered 30 to 120 minutes before the administration of the bispecific antibody. A suitable exemplary corticosteroid for use in the methods and uses described herein is dexamethasone. In some embodiments, prednisolone is administered in an intravenous dose of 15 mg or in an equivalent dose including an oral dose. Exemplary corticosteroid equivalents of dexamethasone that can be used for the prophylaxis of CRS are shown in Table 4, along with their administered equivalents.

[0157] Furthermore, in some embodiments, human subjects in the methods described herein are treated with premedication to reduce their response to injection. In one embodiment, premedication includes the administration of an antihistamine. In some embodiments, premedication includes the administration of an antipyretic. In further embodiments, premedication includes systemic administration of an antihistamine and an antipyretic.

[0158] An exemplary antihistamine suitable for use as premedication is diphenhydramine. In one embodiment, diphenhydramine is administered intravenously or orally in doses of 50 mg or equivalent. An exemplary antipyretic suitable for use as premedication is acetaminophen. In one embodiment, acetaminophen is administered orally in doses of 650 to 1000 mg or equivalent. In some embodiments, premedication is administered on the same day as the bispecific antibody, for example, before the injection of the bispecific antibody, for example, 30 to 120 minutes before the administration of the bispecific antibody.

[0159] Premedication and / or prophylaxis for CRS may be administered at least in the early stages of treatment. In some embodiments, premedication and / or prophylaxis are administered during the first four doses of the bispecific antibody. For example, prophylaxis may be administered during the first 28-day cycle of bispecific antibody administration, as described herein. In some embodiments, premedication is administered during the first cycle.

[0160] Typically, the risk of reaction during initial treatment is reduced after several doses, for example, after the first four doses (first cycle). Therefore, if a human subject does not experience CRS by the fourth dose, CRS prophylaxis may be discontinued. However, CRS prophylaxis may be continued, especially if the human subject experiences CRS of grade 1 or higher. Similarly, premedication may also be continued. CRS grading can be performed as described in Tables 5 and 6.

[0161] In further embodiments, in the methods described herein, the prophylaxis for CRS is administered during the second 28-day cycle if a human subject experiences CRS of grade 1 or higher after the fourth dose of the bispecific antibody in cycle 1. Furthermore, the prophylaxis may be continued in subsequent cycles if a human subject experiences CRS of grade 1 or higher after the last dose of the bispecific antibody in the previous cycle. Optional premedication may be administered during the second cycle. Further premedication may also be administered during subsequent cycles.

[0162] In one embodiment, premedication and prophylaxis for CRS is administered, comprising an antihistamine, such as diphenhydramine (e.g., 50 mg intravenously or orally, or its equivalent), an antipyretic, such as acetaminophen (e.g., 650-1000 mg or its equivalent or orally), and a corticosteroid, such as dexamethasone (e.g., a 15 mg dose or its equivalent). In some embodiments, premedication and prophylaxis are administered 30-120 minutes before administration of the bispecific antibody. Further prophylaxis, including systemic administration of a corticosteroid such as dexamethasone (e.g., a 15 mg dose or its equivalent), is administered on the following days: days 2, 3, and optionally day 4. In some embodiments, the schedule of premedication and prophylaxis is preferably administered during the first four administrations of the bispecific antibody, for example, during the first 28-day cycle of bispecific antibody administration as described herein. Furthermore, subsequent cycles may include the same dosing schedule, for example, if a Grade 1 or higher CRS occurs during the last dose of the previous cycle, premedication as part of the dosing schedule may be optional.

[0163] During treatment of human subjects with indolent lymphoma (e.g., FL) using the doses and treatment regimens described herein, indolent lymphoma can be effectively controlled and / or treated while adequately managing CRS. Subjects treated in the manner described herein may experience manageable CRS, as described in the examples. In some cases, subjects receiving the treatment described herein may develop Grade 1 CRS as defined according to standard practice. In other cases, subjects may develop Grade 2 manageable CRS as defined according to standard practice. Thus, subjects receiving the treatment described herein may have Grade 1 or Grade 2 manageable CRS as defined according to standard practice. According to the standard classification of CRS, Grade 1 CRS includes fever up to at least 38°C, without hypotension or hypoxia, and Grade 2 CRS includes fever up to at least 38°C, plus hypotension requiring oxygen via low-flow nasal cannula or blow-by, without requiring vasopressors and / or hypoxia. Such manageable CRS may occur during Cycle 1. Human subjects receiving the treatments described herein may also experience CRS of grade 2 or higher, as defined according to standard practice, during treatment. Therefore, human subjects receiving the treatments described herein may also experience CRS of grade 3, as defined according to standard practice, during said treatment. Such manageable CRS may further occur during cycle 1 and subsequent cycles.

[0164] Human subjects treated according to the methods described herein may also experience fever, fatigue, and injection site reactions. Subjects may also experience neurotoxicity, partial seizures, agraphia associated with CRS, or confusional states associated with CRS.

[0165] As described above, subjects may develop CRS during treatment according to the methods described herein, even if they have received CRS prophylaxis. The CRS grading criteria are shown in Tables 5 and 6.

[0166] In one embodiment, subjects who develop Grade 1 CRS are treated with antibiotics if they present with an infection. In some embodiments, antibiotics are continued until neutropenia (if present) is resolved. In some embodiments, subjects with Grade 1 CRS exhibiting systemic symptoms are treated with NSAIDs.

[0167] In one embodiment, subjects who develop grade 2 CRS are treated with intravenous fluid bolus and / or supplemental oxygen. In some embodiments, subjects who develop grade 2 CRS are treated with vasopressors. In some embodiments, subjects with grade 2 CRS along with comorbidities are treated with tocilizumab (a humanized antibody against the IL-6 receptor, e.g., marketed as ACTEMRA®) and / or steroids (e.g., dexamethasone or its equivalent of methylprednisolone). In further embodiments, subjects presenting with concurrent ICANS are administered dexamethasone. In further embodiments, if a subject does not show improvement in CRS symptoms, for example within 6 hours, or if a subject begins to worsen after initial improvement, a second dose of tocilizumab is administered together with a dose of corticosteroids. In some embodiments, if a subject is resistant to tocilizumab after three doses, additional cytokine therapy, such as an anti-IL-6 antibody (e.g., siltuximab) or an IL-1R antagonist (e.g., anakinra), is administered to the subject.

[0168] In one embodiment, subjects who develop grade 3 CRS are treated with vasopressor (e.g., norepinephrine) support and / or supplemental oxygen. In some embodiments, subjects with grade 3 CRS are treated with tocilizumab, or tocilizumab in combination with a steroid (e.g., dexamethasone or its equivalent of methylprednisolone). In some embodiments, subjects presenting with concurrent ICANS are administered dexamethasone. In further embodiments, if a subject is resistant to tocilizumab after three doses, additional cytokine therapy, such as an anti-IL-6 antibody (e.g., siltuximab) or an IL-1R antagonist (e.g., anakinra), is administered to the subject.

[0169] In one embodiment, subjects who develop Grade 4 CRS are treated with vasopressor support and / or supplemental oxygen (e.g., by positive pressure ventilation, e.g., CPAP, BiPAP, intubation, or mechanical ventilation). In some embodiments, subjects are administered at least two vasopressors. In some embodiments, subjects are administered tocilizumab and steroids. In further embodiments, subjects presenting with concurrent ICANS are administered dexamethasone. In further embodiments, if a subject is resistant to tocilizumab after three doses, additional cytokine therapy, e.g., an anti-IL-6 antibody (e.g., siltuximab) or an IL-1R antagonist (e.g., anakinra) is administered to the subject.

[0170] In some embodiments, human subjects receive prophylactic treatment for tumor lysis syndrome (TLS). Classification and grading of tumor lysis syndrome can be performed using methods known to those skilled in the art, such as those described, for example, in Howard et al. N Engl J Med 2011;364:1844-54 and Coiffier et al., J Clin Oncol 2008;26:2767-78. In some embodiments, prophylactic treatment for TLS includes administering a uric acid-lowering agent before administering a bispecific antibody. Exemplary uric acid-lowering agents include rasburicase and allopurinol. Thus, in one embodiment, prophylactic treatment for TLS includes administering rasburicase before administering a bispecific antibody. In some embodiments, if a subject shows signs of TLS, supportive therapies such as rasburicase may be used.

[0171] In one embodiment, the bispecific antibody is administered subcutaneously and is therefore formulated into a pharmaceutical composition having a formulation and / or concentration suitable for subcutaneous (sc) administration, i.e., enabling pharmaceutically acceptable sc administration at the doses described herein. In some embodiments, subcutaneous administration is carried out by injection. For example, formulations for DuoBody CD3xCD20 suitable for subcutaneous formulation and usable in the methods described herein have been previously described (see, for example, International Publication No. 2019155008, incorporated herein by reference). In some embodiments, the bispecific antibody may be formulated using 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 antibody is provided as a concentrate of 5 mg / mL or 60 mg / mL. In other embodiments, the desired dose of the bispecific antibody is reconstituted into a volume of about 1 mL for subcutaneous injection.

[0172] In one embodiment, a pharmaceutical composition suitable for a bispecific antibody may comprise a bispecific antibody, 20-40 mM acetate, 140-160 mM sorbitol, and a surfactant, such as polysorbate 80, with a pH of 5.3-5.6. In some embodiments, the pharmaceutical formulation may comprise a bispecific antibody with an antibody concentration ranging from 5-100 mg / mL, for example, 48 or 60 mg / mL, 30 mM acetate, 150 mM sorbitol, 0.04% w / v polysorbate 80, with a pH of 5.5. Such formulations may be diluted, for example, with a formulation buffer to enable appropriate administration and subcutaneous delivery.

[0173] 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. Therefore, 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 1.7 mL or about 1.7 mL. In some embodiments, the administered volume is 2 mL or about 2 mL. In some embodiments, the administered volume is 2.5 mL or about 2.5 mL.

[0174] It is understood that the methods according to the present invention may be the first treatment offered to such a patient, or part of the first treatment. However, the patient may have received prior treatment for indolent lymphoma. Prior treatment may include, but is not limited to, one or more of the following: chemotherapy, radiotherapy, immunotherapy, and targeted therapy, or combinations thereof. Most commonly, standard treatment includes treatment using CD20 monoclonal antibodies, alkylating agents, and anthracyclines, either alone or in combination. It is also understood that the methods and uses according to the present invention may be used in combination with other appropriate treatments.

[0175] Therefore, in a further embodiment, in the method according to the present invention, a human subject having indolent lymphoma has received at least one line of treatment prior to treatment according to the present invention. In another embodiment, a human subject having indolent lymphoma has received one line of treatment prior to treatment according to the present invention. In yet another further embodiment, a human subject having indolent lymphoma has received two lines of treatment prior to treatment according to the present invention. In yet another further embodiment, a human subject having indolent lymphoma has received three lines of treatment prior to treatment according to the present invention. In yet another further embodiment, a human subject having indolent lymphoma has received four or more lines of treatment prior to treatment according to the present invention. In yet another further embodiment, a human subject having indolent lymphoma has received one, two, three or more lines of treatment prior to treatment according to the present invention.

[0176] In one embodiment, the bispecific antibody used in the method described herein includes: (i) A first binding arm comprising a first antigen-binding region comprising a variable heavy chain (VH) region and a variable light chain (VL) region, wherein the VH region comprises the CDR1, CDR2, and CDR3 sequences in the amino acid sequence of SEQ ID NO: 6, and the VL region comprises the CDR1, CDR2, and CDR3 sequences in the amino acid sequence of SEQ ID NO: 7; and (ii) A second binding arm that binds to human CD20 and comprises a second antigen-binding region including a VH region and a VL region, wherein the VH region comprises the CDR1, CDR2 and CDR3 sequences in the amino acid sequence of SEQ ID NO: 13, and the VL region comprises the CDR1, CDR2 and CDR3 sequences in the amino acid sequence of SEQ ID NO: 14.

[0177] The CDR1, CDR2, and CDR3 regions can be identified from the variable heavy chain region and the variable light chain region using methods known in the art. The CDR regions from the variable heavy chain and light chain regions can be annotated according to IMGT (see Lefranc et al., Nucleic Acids Research 1999;27:209-12, 1999) and Brochet. Nucleic Acids Res 2008;36:W503-8).

[0178] In one embodiment, the bispecific antibody includes: (i) A first binding arm comprising a first antigen-binding region comprising VHCDR1, VHCDR2 and VHCDR3, which bind to human CD3ε (epsilon) and include amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively, and VLCDR1, VLCDR2 and VLCDR3, which include amino acid sequences shown in SEQ ID NOs: 4, sequence GTN, and 5, respectively; and (ii) A second binding arm comprising a second antigen-binding region comprising VHCDR1, VHCDR2, and VHCDR3, which bind to human CD20 and include amino acid sequences shown in SEQ ID NOs: 8, 9, and 10, respectively, and VLCDR1, VLCDR2, and VLCDR3, which include amino acid sequences shown in SEQ ID NOs: 11, sequence DAS, and SEQ ID NOs: 12, respectively.

[0179] In some embodiments, the bispecific antibody includes: (i) A first binding arm comprising a first antigen-binding region comprising 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) A second binding arm comprising a second antigen-binding region which binds to human CD20 and includes a VH region containing the amino acid sequence of SEQ ID NO: 13 and a VL region containing the amino acid sequence of SEQ ID NO: 14.

[0180] In one embodiment, the bispecific antibody is a full-length antibody and may have an inactive Fc region. In some embodiments, the first binding arm for CD3 is derived from a humanized antibody, e.g., a full-length IgG1, λ (lambda) antibody, e.g., H1L1 described in International Publication No. 2015001085 incorporated herein by reference, and / or the second binding arm for CD20 is derived from a human antibody, e.g., a full-length IgG1, κ (kappa) antibody, e.g., clone 7D8 described in International Publication No. 2004035607 incorporated herein by reference. The bispecific antibody may be prepared from two halves of an antibody. Each of the two halves of an antibody includes, for example, the first and second binding arms shown in SEQ ID NOs. 24 and 25, and SEQ ID NOs. 26 and 27. The half-antibodies may be prepared in CHO cells, and the bispecific antibody may be produced, for example, by Fab arm exchange. In one embodiment, the bispecific antibody is a functional variant of Duobody CD3xCD20.

[0181] Therefore, in some embodiments, the bispecific antibody comprises a first binding arm comprising (i) a first antigen-binding region comprising a VH region containing an amino acid sequence that binds to human CD3ε (epsilon) and is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 6, or a VH region containing the amino acid sequence of SEQ ID NO: 6 but having one, two, or three mutations (e.g., amino acid substitutions), and a VL region containing 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 containing the amino acid sequence of SEQ ID NO: 7 but having one, two, or three mutations (e.g., amino acid substitutions); and (ii) A second binding arm comprising a second antigen-binding region comprising a VH region that binds to human CD20 and includes an amino acid sequence that is at least 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO: 13, or a VH region that includes the amino acid sequence of SEQ ID NO: 13 but has one, two, or three mutations (e.g., amino acid substitutions), and a VL region that includes an amino acid sequence that is at least 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO: 14, or a VL region that includes the amino acid sequence of SEQ ID NO: 14 but has one, two, or three mutations (e.g., amino acid substitutions).

[0182] In one embodiment, the bispecific antibody includes: (i) A first binding arm comprising a first antigen-binding region comprising 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, which binds to human CD3ε (epsilon); and (ii) A second binding arm comprising a second antigen-binding region which binds to human CD20 and includes a VH region containing the amino acid sequence of SEQ ID NO: 26 and a VL region containing the amino acid sequence of SEQ ID NO: 27.

[0183] In some embodiments, the bispecific antibody includes: (i) A first binding arm comprising a first antigen-binding region comprising a heavy chain containing an amino acid sequence that binds to human CD3ε (epsilon) and is at least 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO: 24, or a heavy chain containing the amino acid sequence of SEQ ID NO: 24 but having one, two, or three mutations (e.g., amino acid substitutions), and a light chain region containing an amino acid sequence that is at least 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO: 25, or a light chain region containing the amino acid sequence of SEQ ID NO: 25 but having one, two, or three mutations (e.g., amino acid substitutions); and (ii) A second binding arm comprising a second antigen-binding region comprising a heavy chain containing an amino acid sequence that binds to human CD20 and is at least 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO: 26, or a heavy chain containing the amino acid sequence of SEQ ID NO: 26 but having one, two, or three mutations (e.g., amino acid substitutions), and a light chain containing an amino acid sequence that is at least 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO: 27, or a light chain region containing the amino acid sequence of SEQ ID NO: 27 but having one, two, or three mutations (e.g., amino acid substitutions).

[0184] Various constant regions or variants thereof may be used in bispecific antibodies. In one embodiment, the antibody comprises an IgG constant region, e.g., a human IgG1 constant region, e.g., the human IgG1 constant region defined in SEQ ID NO: 15, or any other suitable IgG1 allotype. In one embodiment, the first binding arm of the bispecific antibody is derived from a humanized antibody, e.g., a full-length IgG1, λ (lambda) antibody, and therefore comprises a λ light chain constant region. In some embodiments, the first binding arm comprises a λ light chain constant region defined in SEQ ID NO: 22. In some embodiments, the second binding arm of the bispecific antibody may be derived from a human antibody, preferably a full-length IgG1, κ (kappa) antibody, and therefore comprises a κ light chain constant region. In some embodiments, the second binding arm comprises a κ light chain constant region defined in SEQ ID NO: 23.

[0185] It is understood that the constant region portion of a bispecific antibody may include modifications that enable the efficient formation / production of the bispecific antibody and / or provide an inactive Fc region. Such modifications are known in the art.

[0186] Various formats of bispecific antibodies are known in the art (outlined by Kontermann, Drug Discov Today 2015;20:838-47;MAbs,2012;4:182-97). Therefore, the bispecific antibodies used in the methods and uses described herein are not limited to any particular bispecific format or method of preparation thereof. For example, bispecific antibodies may include, but are not limited to, bispecific antibodies having a complementary CH3 domain that forces heterodimerization, Knobs-into-Holes molecules (Genentech, International Publication No. 9850431), CrossMAb (Roche, International Publication No. 2011117329), or electrostatically matched molecules (Amgen, European Patent No. 1870459 and International Publication No. 2009089004; Chugai Pharmaceutical, U.S. Patent Application Publication No. 201000155133; Oncomed, International Publication No. 2010129304).

[0187] Preferably, the bispecific antibody comprises an Fc region comprising a first heavy chain having a first Fc sequence containing a first CH3 region and a second heavy chain having a second Fc sequence containing a second CH3 region, wherein the sequences of the first and second CH3 regions are different, thereby the heterodimer interaction between the first and second CH3 regions is stronger than the homodimer interaction between each of the first and second CH3 regions. Further details on these interactions and how they can be achieved are provided, for example, in International Publication No. 2011131746 and International Publication No. 2013060867 (Genmab), which are incorporated herein by reference. In one embodiment, the bispecific antibody comprises, in the first heavy chain, (i) amino acid L at the position corresponding to F405 of the human IgG1 heavy chain constant region of SEQ ID NO: 15, and in the second heavy chain, amino acid R at the position corresponding to K409 of the human IgG1 heavy chain constant region of SEQ ID NO: 15, and vice versa.

[0188] Bispecific antibodies may include modifications within the Fc region to inactivate or deactivate the Fc region. Therefore, in the bispecific antibodies disclosed herein, one or both heavy chains may be modified to reduce the degree of Fc-mediated effector function induced by the antibody compared to bispecific antibodies without modifications. Fc-mediated effector function can be measured by determining Fc-mediated CD69 expression on T cells (i.e., CD3 antibody-mediated, CD69 expression as a result of Fcγ receptor-dependent CD3 crosslinking), by binding to the Fcγ receptor, by binding to C1q, or by inducing Fc-mediated crosslinking of FcγR. In particular, the heavy chain constant region sequence can 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% compared to a wild-type (unmodified) antibody, and the Fc-mediated CD69 expression is determined by a PBMC-based functional assay, for example, as described in Example 3 of International Publication No. 2015001085. Modification of the heavy and light chain constant region sequences can also result in a reduction in C1q binding to the antibody. Compared to an 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 can be determined, for example, by ELISA. Furthermore, the Fc region may be modified such that antibody-mediated Fc-mediated T cell proliferation is reduced by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 99%, or 100% compared to an unmodified antibody, and the T cell proliferation is measured by a PBMC-based functional assay. For example, examples of amino acid positions that can be modified in an IgG1 isotype antibody include positions L234 and L235. Thus, in one embodiment, a bispecific antibody may comprise a first heavy chain and a second heavy chain, and in both the first and second heavy chains, the amino acid residues at positions L234 and L235 of the human IgG1 heavy chain by Eu numbering are F and E, respectively.In addition, the D265A amino acid substitution can reduce binding to all Fcγ receptors and prevent ADCC (Shields et al., JBC 2001;276:6591-604). Therefore, bispecific antibodies may contain a first heavy chain and a second heavy chain, and in both the first and second heavy chains, the amino acid residue at the position corresponding to position D265 of the human IgG1 heavy chain according to Eu numbering is A.

[0189] In one embodiment, in the first and second heavy chains of a bispecific antibody, the amino acids at positions L234, L235, and D265 in the human IgG1 heavy chain are F, E, and A, respectively. Antibodies having these amino acids at these positions are an example of an antibody having an inactive or non-active Fc region.

[0190] With respect to the bispecific antibodies described herein, those having the K409R or F405L mutation in addition to the three amino acid substitutions L234F, L235E, and D265A, as described above, may be referred to with the suffix "FEAR" or "FEAL," respectively.

[0191] 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, a bispecific antibody may comprise an IgG1 heavy chain constant region having the F405L substitution, the amino acid sequence shown in SEQ ID NO: 17 and / or the K409R substitution, the amino acid sequence shown in SEQ ID NO: 18, and further substitutions that inactivate or deactivate the Fc region. Thus, in one embodiment, a bispecific antibody comprises a combination of the amino acid sequence of one IgG1 heavy chain constant region having the L234F, L235E, D265A and F405L substitutions (e.g., shown in SEQ ID NO: 19) and the amino acid sequence of the other IgG1 heavy chain constant region having the L234F, L235E, D265A and K409R substitutions (e.g., shown in SEQ ID NO: 20).

[0192] In some embodiments, the bispecific antibody used in the methods and uses described herein comprises a first binding arm containing a heavy chain and a light chain as defined by SEQ ID NOs: 24 and 25, respectively, and a second binding arm containing a heavy chain and a light chain as defined by SEQ ID NOs: 26 and 27, respectively. Such an antibody is referred to herein as DuoBody CD3xCD20. Variants of such an antibody are also intended for use in the methods and uses described herein. In some embodiments, the bispecific antibody is epcolitamab (CAS 2134641-34-0) or a biosimilar thereof.

[0193] kit This specification also provides kits, which include pharmaceutical compositions containing a bispecific antibody conjugating to CD3 and CD20 according to the present invention, such as DuoBody CD3xCD20 or epcolitamab, and a pharmaceutically acceptable carrier in a therapeutically effective amount suitable for use in the method described herein. The kits may also include instructions, possibly including, for example, a dosing schedule, to enable practitioners (e.g., physicians, nurses, or patients) to administer one or more of the compositions contained therein to patients with DLBCL. The kits may also include one or more syringes.

[0194] The kit may comprise multiple packages of single-dose pharmaceutical compositions, each containing an effective amount of a bispecific antibody for single-dose administration by the method described herein. They may also comprise multiple packages of single-dose pharmaceuticals. Instruments or devices necessary for administering (one or more) pharmaceutical compositions may also be included in the kit.

[0195] This disclosure is further illustrated by the following embodiments, which should not be construed as further limitations. All figures and all references, Genbank sequences, journal publications, patents and published patent applications cited throughout this application are expressly incorporated herein by reference.

[0196] [Examples] [Example 1] Step-up dosing (SUD) is a strategy used to reduce the incidence and severity of CRS events. A SUD regimen typically consists of a target full dose followed by one or more lower doses to desensitize the patient's immune system and "prepare" the patient for the target full dose.

[0197] A model-based approach was applied to identify the optimal step-up dosing regimen for the trial of epcolitamab in DLBCL and FL patients. A repeated time-to-event (RTTE) model was developed using PK and CRS event data collected across the EPCORE NHL-1 (NCT03625037; monotherapy) and EPCORE NHL-3 (NCT04542824; monotherapy in Japanese patients) studies (Hutchings M, et al. Lancet. 2021; 398:1157-69; Thieblemont C, et al. J Clin Oncol. 2022; DOI: 10.1200 / JCO.22.01725; Izutsu K, et al. JSMO 2023. Abstract O13-3).

[0198] Based on the data collected during these dose escalation studies, it became clear that at least two steps (priming and intermediate doses) are necessary to effectively mitigate the risk of CRS. A broad priming (0.004–0.16 mg) and intermediate (0.25–1.6 mg) dose range was investigated, and based on the observed incidence and severity of CRS events, a 0.16 / 0.8 / 48 mg SUD regimen was selected for escalation.

[0199] The model considered in the development of the RTTE analysis was one that could incorporate long-term exposure into the survival framework. Mechanistically, the risk of CRS is related to cytokine production and the tumor-killing activity of the compound. To capture these dynamics, the hazard function (risk of CRS event) was modeled as the product of two components, allowing for the flexibility to empirically capture the dynamics of epcolitamab exposure onset and resistance that affect the hazard of CRS events.

[0200] The first (STIM) regimen, addressing the development of AE risk, allowed for an increase in hazards in several (possibly time-delayed) functions of epcolitamab plasma concentration, while the second (EFF) regimen described the inhibition of hazards in several (again, possibly time-delayed) functions of epcolitamab plasma concentration, capturing resistance kinetics. Simulations using the developed models were performed to predict the probability of grade 2+ CRS over the first two cycles of epcolitamab treatment and to evaluate different SUD regimens. Furthermore, three-step SUD regimens were evaluated, and the optimal SUD regimen was identified based on the simulation results and selected for further investigation in the clinic (Figure 1).

[0201] Using this model, we predicted the risk of grade 2+ CRS in regimens beyond the priming and intermediate dose permutations tested with EPCORE NHL-1 dose escalation. Based on the model predictions, further increases in priming or intermediate doses may result in only a slight reduction in the risk of grade 2+ CRS in DLBCL (Figure 2). Based on the simulation results, two alternative SUD regimens were selected for investigation in practice.

[0202] Based on model predictions, a 3-step SUD regimen could potentially reduce the risk of grade 2+ CRS (Figure 3). 3-step SUD was predicted to improve the benefit-risk in low-grade aggressive NHL (e.g., FL). Based on the simulation results, two different 3-step SUD regimens were selected and investigated in FL patients.

[0203] [Example 2] GCT3013-01: Phase 1 / 2 open-label dose-escalation study of GEN3013 in patients with relapsed, advanced, or refractory B-cell lymphoma. GCT3013-01 is an open-label, multicenter, phase 1 / 2 trial of epcolitamab in subjects with relapsed, advanced, or refractory B-cell lymphoma. The trial includes dose expansion of epcolitamab in subjects with FL grade 1–3A (hereinafter referred to as the "pivotal cohort"). The trial further includes two additional step-up dosing (SUD) regimens of epcolitamab in subjects with FL grade 1–3A (arm A: 0.16 / 0.8 / 3 / 48 mg, arm B: 0.16 / 0.8 / 6 / 48 mg), and an optimization part to evaluate their effects on the rate of grade 2 or higher cytokine release syndrome (CRS) events.

[0204] Epcolitamab administration regimen [Table 2]

[0205] [Table 3]

[0206] Epcolitamab is administered as follows:

[0207] Cycles 1-3: Day 1, Day 8, Day 15, and Day 22 (once a week) Cycles 4-9: Day 1 and Day 15 (every two weeks) From cycle 10 until PD, unacceptable toxicity, or end of the test: Day 1 (every 4 weeks) A single preferred SUD regimen was identified using decision rules based on the percentage of Grade 2 or higher CRS, safety, PK, and PD data. Further modifications to the protocol included recommendations for adequate hydration and the use of dexamethasone (15 mg) instead of prednisolone or other corticosteroid equivalents as prophylactic corticosteroids administered to minimize CRS during treatment with epcolitamab in cycle 1.

[0208] For FL grade 1–3A optimization, subjects are enrolled in two arms in parallel, with a maximum of approximately 10 subjects in each arm. Dose optimization for FL grade 1–3A involves evaluating two alternative second intermediate doses in parallel. The decision rules based on CRS after Stage 1 are as follows:

[0209] If two or fewer out of ten subjects experience a Grade 2 or higher CRS event, the SUD regimen is considered acceptable for further evaluation.

[0210] If three or more out of ten subjects experience a Grade 2 or higher CRS event, the SUD escalation will be terminated.

[0211] CRS prevention A 4-day course of corticosteroids is administered to reduce / prevent the severity of symptoms from potential CRS for each dose of epcolitamab. For epcolitamab administration from cycle 2 onward, prophylaxis to CRS with corticosteroids is optional. Corticosteroid administration may be intravenous or oral at the recommended dose or equivalent. [Table 4]

[0212] Supportive therapy for cytokine release syndrome CRS will be graded according to the ASTCT grading system for CRS (Tables 6 and 7), and subjects will receive supportive care for the treatment of CRS. Supportive care includes, but is not limited to, • Infusion of physiological saline solution Systemic glucocorticosteroids, antihistamines, antipyretics • Blood pressure support (vasopressin, vasopressors) • Support for low-flow and high-flow oxygen and positive pressure ventilation • Intravenous administration of a monoclonal antibody against IL-6R, such as tocilizumab. If repeated tocilizumab treatment is unsuccessful, a monoclonal antibody against IL-6, such as siltuximab IV, may be used. It may include. [Table 5] TIFF2026513993000006.tif9154

[0213] [Table 6] TIFF2026513993000008.tif53169

[0214] Prevention and management of tumor lysis syndrome To prevent tumor lysis syndrome (TLS), subjects receive hydration and uric acid-lowering medication before administration of epcolitamab. If signs of tumor lysis syndrome (TLS) occur, supportive care including rasburicase is used.

[0215] Test evaluation Demographic assessment and baseline assessment Demographic details of subjects will be collected, as well as information such as the date of lymphoma diagnosis, Ann Arbor stage classification at diagnosis including systemic symptoms (B symptoms), and prior evidence of CD20 positivity. Information will also be collected regarding medical history, past and concomitant medications, concomitant treatments, and past cancer therapies and surgeries (including past anti-cancer therapies for NHL, e.g., surgery, radiotherapy, chemoradiotherapy, and systemic treatment regimens).

[0216] Effectiveness evaluation Eligible subjects have at least one measurable disease site (as indicated in the inclusion criteria) for disease assessment. Measurable sites for lymphoma are defined as lymph nodes, lymphadenopathy, or extranodal sites. Up to six measurable sites are tracked as target lesions for each subject, and measurements are determined by imaging assessment. Sites that are not measurable as defined above are considered evaluable by objective evidence of the disease (i.e., radiography, physical examination, or other procedures). Examples of evaluable diseases include, for example, bone marrow infiltration, bone lesions, exudation, or thickening of the intestinal wall.

[0217] Tumor and bone marrow biopsy For all subjects with accessible tumors, two fresh core tumor biopsies will be collected prior to epcolitamab treatment (during the screening period), and two fresh core tumor biopsies will be collected on day 15 (±1 week) of the start of cycle 2. Archived tumor biopsies are acceptable if fresh biopsies cannot be collected at the time of screening, provided they were collected within 3 months prior to enrollment. Biopsies may be whole lymph node or core biopsies. Tumor biopsies must be FFPE. Tumor biopsies will be tested for MRD assessment and exploratory biomarkers.

[0218] Radiation assessment During screening, an FDG PET-CT scan (or CT / MRI and FDG PET if a PET-CT scan is unavailable) will be performed. For subjects with FDG-avid tumors at screening, all subsequent disease assessments will include FDG PET using the 5-point scale described by Barrington et al. (J Clin Oncol 2014;32:3048-58; Score 1: No uptake; Score 2: Uptake not extending beyond the mediastinum; Score 3: Uptake extending beyond the mediastinum but not beyond the liver; Score 4: Moderate uptake beyond the liver; Score 5: Significant uptake beyond the liver and / or new lesion; Score X: New area of ​​uptake unlikely to be associated with lymphoma). For subjects with non-avid or variable FDG-avid tumors, a CT scan including IV contrast of the neck / chest / abdomen / pelvis / additional known lesions may be performed. The CT component of PET-CT may be used instead of a standalone CT / MRI if the CT component has similar diagnostic quality to a contrast-enhanced CT performed without PET. If contrast-enhanced PET-CT is unavailable, standalone diagnostic CT / MRI and standard FDGPET will be performed. Subjects intolerant to IV CT contrast agents will undergo a CT scan with an oral contrast agent.

[0219] MRI can be used to evaluate diseased areas that cannot be adequately imaged using CT, or for subjects who are intolerant to CT contrast agents. When MRI is the imaging modality of choice, MRI images are obtained at screening and at all subsequent response assessments.

[0220] Bone marrow evaluation Bone marrow biopsies (archived or fresh), whether or not performed by aspiration, are to be obtained for all patients at screening to record bone marrow infiltration for lymphoma. Bone marrow biopsies obtained as a standard status of care (SOC) may be used if taken up to 42 days prior to the first dose of epcolitamab. Once bone marrow aspirates are obtained, flow cytometry can be used to confirm the determination of bone marrow infiltration. Bone marrow biopsies are taken (1) at screening; (2) for subjects with bone marrow infiltration at screening who later achieve complete remission (CR) by imaging (bone marrow evaluation includes morphological examination to confirm the presence or absence of lymphoma (complete remission) and, if necessary, either flow cytometry or IHC); and (3) for subjects with bone marrow infiltration recorded at screening who later achieve CR by imaging (a portion of the aspirates collected to confirm CR is used for MRD evaluation).

[0221] Evaluation of minimal residual disease MRD is assessed by tracking the presence of DNA encoding a B-cell receptor (BCR) specifically expressed by cancer cells. The DNA sequence of this BCR is identified by a tumor biopsy submitted at screening. After the initiation of treatment, blood samples are taken at predetermined time points and at complete response (CR) to assess whether the amount of cancer DNA has decreased as a potential measure of (early) response and to assess MRD. As an exploratory analysis, if a subject reaches metabolic / radiological CR and has bone marrow infiltration recorded at screening, a portion of the fine-needle aspiration collected to confirm CR is used to assess MRD.

[0222] Assessment of disease response and progression The disease response is evaluated according to both the Lugano classification criteria (described in Cheson et al., J Clin Oncol 2014;32:3059-68 (see Table 3 in Cheson et al., 2014 in particular)) and LYRIC (Table 7) to inform the decision on whether to continue treatment.

[0223] The definitions of the evaluation items are as follows:

[0224] The overall response rate (ORR) is defined as the proportion of subjects who achieved a PR or CR response prior to the initiation of subsequent therapy.

[0225] The time to response (TTR) is defined in responders as the time between the first dose of epcoritamab (day 1, cycle 1) and the initial documentation of a PR or CR.

[0226] The duration of response (DOR) is defined in responders as the time from the initial documentation of a PR or CR to the earlier of progression or death.

[0227] The progression-free survival (PFS) is defined as the time from the first administration date of epcoritamab (day 1, cycle 1) to the earlier of progression or death.

[0228] The overall survival (OS) is defined as the time from the first administration date of epcoritamab (day 1, cycle 1) to the date of death.

[0229] The time to next antilymphoma therapy (TTNT) is defined as the number of days from day 1 of cycle 1 to the first documented administration of subsequent antilymphoma therapy.

[0230] The MRD negativity rate is defined as the proportion of subjects who had at least one non-detectable MRD result according to a specific threshold prior to the initiation of subsequent therapy.

[0231] For the Lugano classification criteria (definitions of complete response, partial response, non-response / stable, and progression, see, for example, Cheson et al., J Clin Oncol 2014;32:3059-68).

[0232] (a) Target and non-target lesions The target lesions for the Lugano classification criteria include up to six of the largest dominant nodules, nodular masses, or other lymphomatous lesions, which are measurable in two diameters and preferably originate from various body regions representing the subject's overall disease burden, including mediastinal and retroperitoneal lesions, where applicable. At baseline, the maximum diameter (LDi) of a measurable nodule is greater than 15 mm. Measurable extranodal lesions may be included in the six representative target lesions. At baseline, measurable extranodal lesions are defined as having an LDi greater than 10 mm.

[0233] All other lesions (including nodular, extranodal, and evaluable diseases) may be tracked as non-target lesions (e.g., skin, GI, bone, spleen, liver, kidney, pleural or pericardial effusion, ascites, bone, bone marrow).

[0234] (b) Split lesions and confluent lesions The lesion may be split or become confluent over time. In the case of a split lesion, the product of the vertical diameters (PPDs) of each segment is summed to represent the PPD of the split lesion, and this PPD is added to the sum of the PPDs of the remaining lesion to measure the response. If any or all of these individual segments subsequently grow, the minimum value of each individual segment is used to determine progression. In the case of a confluent lesion, the PPD of the confluent mass is compared to the sum of the PPDs of the individual segments, and progression (PD) is indicated if the increase in the PPD of the confluent mass compared to the sum of the individual segments exceeds 50%. LDi and minimum diameter (SDi) are no longer necessary for determining progression.

[0235] LYRIC Clinical trials have shown that cancer immunotherapy can cause early apparent radiographic progression (including the appearance of new lesions) and subsequent delayed responses. This initial increase in tumor size may be caused by immune cell infiltration in the setting of the T cell response, and therefore this progression may not represent true progression and is therefore called "pseudo-progression" (Wolchok et al., Clin Cancer Res 2009;15:7412-20).

[0236] The current Lugano response evaluation criteria (Cheson et al., J Clin Oncol 2014;32:3059-68) do not consider pseudo-progression, and there is a significant risk of premature discontinuation of potentially effective immunomodulatory drugs after observing an atypical response. An atypical response is characterized by either early progression and subsequent response of an existing lesion, or the development of a new lesion, regardless of whether tumor shrinkage occurs elsewhere.

[0237] LYRIC is a modified Lugano response assessment criterion adapted for immunotherapy and introduces a new mitigating response category: “indeterminate response” (IR) designation (Cheson et al., Blood 2016;128:2489-96). This IR designation was introduced to identify potentially “atypical response” cases until confirmed as flare / pseudoprogression or true PD by either biopsy or subsequent imaging.

[0238] Subjects exhibiting PD according to the Lugano classification criteria / classification are likely to have one or more of the following three conditions:

[0239] IR(1): Total tumor volume (assessed by the sum of the product of diameters [SPD]) of up to six target lesions increases by 50% or more during the first 12 weeks of therapy, with no clinical deterioration.

[0240] IR(2): A new lesion appears at any point during treatment, or one or more existing lesions grow by 50% or more; this occurs when the total tumor volume, as measured by SPD of up to six lesions at any point during treatment, has not progressed overall (SPD increase of less than 50%).

[0241] IR(3): Increased FDG uptake in one or more lesions, without co-occurring increases in lesion size or number.

[0242] It is possible that a subject may meet the criteria for either IR(1) or both IR(2) and IR(3) at a single point in time, for example, a new FDG-avid lesion (IR[2]) in the absence of overall progression and increased FDG uptake in a separate lesion (IR[3]) may be present simultaneously. In such cases, the designation of IR(1) or IR(2) shall take precedence (e.g., IR[2] in the example above).

[0243] Subjects categorized as having any of the IR types will undergo repeated imaging 12 weeks later (or earlier if clinically indicated). At that time, the response will be re-evaluated, and the subject will be considered to have true PD, taking the following considerations into account.

[0244] Follow-up IR(1): In the case of IR(1), the initial IR(1) should be compared with the current SPD. IR(1) is consistent with the Lugano classification criteria and is classified as PD if (a) SPD has increased by 10% or more from the initial IR1, and (b) one or more lesions smaller than 2 cm have increased by 5 mm or more (in any dimension), and lesions larger than 2 cm have increased by 10 mm or more.

[0245] Follow-up IR(2): In the case of IR(2), new or growing lesions (one or more) are added to the target lesions (one or more), up to a total of six lesions. IR(2) becomes PD if (a) the SPD (newly defined set of target lesions) increases by 50% or more from the lowest value.

[0246] Follow-up IR (3): IR (3) becomes PD when the lesion with increased FDG uptake also shows an increase in size. [Table 7]

[0247] Clinical safety evaluation Safety is evaluated by measuring adverse events, clinical test results, ECG, vital sign measurements, physical examination findings, and ECOG performance status. Immune effector cell-related neurotoxicity syndrome (e.g., as described in Lee et al., Biol Blood Marrow Transplant 2019;25:625-638), systemic symptoms (B symptoms), tumor flare reaction, and survival period are also evaluated.

[0248] Summary of safety data By April 2023, 16 subjects with R / R FL in the optimization arm had received at least one dose (i.e., priming) of epcoritamab, and 11 subjects with FL had received at least one full dose of epcoritamab. Of the 11 subjects who received the first full dose of epcoritamab, 6 subjects were enrolled in Arm A and 5 subjects were enrolled in Arm B. Only grade 1 CRS events were recorded. Four (36.4%) subjects had the reported grade 1 CRS event (1 in Arm A and 3 in Arm B). One subject in Arm B had a grade 1 CRS event after the second intermediate (6 mg) dose, and two subjects had grade 1 CRS events after the first full dose (1 in Arm A and 1 in Arm B).

[0249] These data showed that the addition of the second intermediate dose, combined with the preventive measures incorporated, reduced the CRS risk of epcoritamab.

[0250] After six subjects were enrolled in each arm, a preliminary analysis revealed that the CRS rate in arm A was numerically lower than that in arm B, and priority was given to further enrollment in arm A.

[0251] As of the data cutoff date of January 8, 2024, 86 subjects in Arm A and 6 subjects in Arm B had received at least one dose of epcolitamab. Overall, 64 subjects (74.4%) in Arm A were continuing epcolitamab treatment as of the DCO date, while 22 subjects (25.6%) had discontinued treatment (17 subjects discontinued treatment due to progression, and 3 subjects discontinued treatment due to adverse events). A summary of TEAEs for all subjects is presented in Table 8. [Table 8] TIFF2026513993000011.tif44165

[0252] In Arm A, 85 subjects (98.8%) experienced at least one TEAE, and 78 subjects (90.7%) experienced at least one TEAE that the researchers considered to be related to epcolitamab. The most common (≥20%) TEAEs reported by the PT were CRS (48.8%), injection site reactions (26.7%), and constipation (20.9%).

[0253] In Arm A, a total of 46 subjects (53.5%) experienced at least one Grade 3 or higher TEAE, of which 29 subjects experienced at least one Grade 3 or higher TEAE that the researchers considered to be associated with epcolitamab (Table 9). In Arm A, Grade 3 or 4 TEAE PTs that occurred in two or more subjects each included neutropenia (14 subjects; 16.3%), decreased lymphocyte count (6 subjects; 7.0%), lymphocyte count and COVID-19 (4 subjects each; 4.7%), decreased neutropenia (3 subjects; 3.5%), and anemia, thrombocytopenia, elevated ALT, hyperglycemia, and hypokalemia (2 subjects each; 2.3%). The most common (≥2 subjects) grade 3 or 4 TEAEs thought to be associated with epcolitamab were neutropenia (11 subjects; 12.8%), decreased lymphocyte count (4 subjects; 4.7%), lymphopenia (3 subjects; 3.5%), decreased neutropenia (3 subjects; 3.5%), and elevated ALT (2 subjects; 2.3%).

[0254] Serious TEAEs were reported in 38 subjects (44.2%) in Arm A, and 31 subjects were considered epcolitamab-related by the researchers. The most common (≥20%) serious TEAE was CRS (24 subjects; 27.9%). Three subjects (3.5%) in Arm A had at least one TEAE that led to discontinuation of epcolitamab. One subject (1.2%) experienced grade 2 bronchopulmonary aspergillosis, and two subjects (2.3%) had grade 2 interstitial pneumonia. No subjects had fatal (grade 5) TEAEs. One subject in Arm A died from disease progression 19 days after the last dose of epcolitamab.

[0255] The AESI included CRS, ICANS, and CTLS. 42 subjects (48.8%) in Arm A experienced at least one CRS event (Table 9). 34 subjects (39.5%) had CRS with a maximum severity grade of 1, 8 subjects (9.3%) had CRS with a maximum severity grade of 2, and no subjects experienced CRS events of grade 3 or higher.

[0256] Overall, the 42 subjects in Arm A had a total of 69 CRS events (Table 9). For these 69 CRS events, the median time to CRS onset was 4.0 days from the most recent epcolitamab dose across all doses (range: 1, 8). All 69 CRS events resolved within a median time to resolution of 2.0 days (range: 1, 14). No subjects had ICANS or CTLS (Table 8). [Table 9] TIFF2026513993000013.tif215165

[0257] [Table 10]

[0258] Summary of effectiveness data pivotal cohort The majority of subjects with FL (N=128) were male (79 subjects; 61.7%). The median age was 65.0 years (range: 39, 84), with 67 subjects (52.3%) aged 65 years or older and 17 subjects (13.3%) aged 75 years or older. The majority of subjects (77 subjects; 60.2%) were Caucasian. Most subjects with FL had advanced-stage lymphoma (Ann Arbor stage III-IV in 85.2% of subjects), a FLIPI score of 3 or higher (60.9% of subjects), and were bi-resistant to both anti-CD20 and alkylating agents (70.3% of subjects). Furthermore, subjects were heavily pre-treated, with a median of 3.0 (range: 2, 9) previously received systemic anti-lymphoma treatment lines, and 52.3% of subjects with FL had experienced disease progression (POD24) within 24 months of any initial treatment line. Therefore, the FL subjects treated in the trial represent a population for whom treatment is clinically very difficult and who have poor response and survival outcomes based on their medical history.

[0259] In subjects with FL, the ORR was 82.0% (95% CI: 74.3, 88.3) and the CR rate was 62.5% (95% CI: 53.5, 70.9). Efficacy was consistent across pre-specified subgroups, including older subjects, heavily pre-treated subjects, and subjects with dual resistance or POD24 disease.

[0260] After a median DOR follow-up of 14.8 months (range: 0.0+, 27.2+), subjects with FL did not reach the median DOR or complete duration of response (DOCR). The estimated proportion of subjects maintaining a response at 12 months was 68.7% (overall responders) and 82.2% (complete responders).

[0261] The median time to response (TTR) was 1.4 months (range: 1.0, 3.0) for all responders with FL, and the median time to complete response (TTCR) was 1.5 months (range: 1.2, 11.1) for complete responders. The median progression-free survival (PFS) (primary definition) was 15.4 months in subjects with FL (95% CI: 10.9, NR). Subjects with complete response (CR) did not reach the median PFS (95% CI: 22.8, NR).

[0262] During a median follow-up period of 17.4 months, subjects with FL did not reach the median OS, and it is estimated that 81.1% of subjects were alive at 12 months.

[0263] Optimization part A total of 92 subjects received at least one dose of epcolitamab in the FL-optimized part, of which 86 subjects were assigned to receive the proposed dosing regimen of epcolitamab 0.16 / 0.8 / 3 / 48 mg (Arm A) and 6 subjects were assigned to receive epcolitamab 0.16 / 0.8 / 6 / 48 mg (Arm B). Below, we present data from both Arm A and Arm B, but primarily focus on the results from the 86 subjects in Arm A who received the 3-step SUD dosing regimen.

[0264] Of the 86 subjects treated in Arm A, a total of 49 (57.0%) were male. The median age was 63.5 years (range: 33, 90), 39 subjects (45.3%) were 65 years or older, and 10 subjects (11.6%) were 75 years or older. 64 subjects (74.4%) were white, 2 subjects (2.3%) were Asian (both Korean), 1 subject (1.2%) was Black or African American, and the race of 19 subjects (22.1%) was not reported due to country-specific data protection laws.

[0265] Participants in Arm A had highly refractory and high-risk FL disease. Advanced-stage disease (Ann Arbor stages III and IV) was present in 79 participants (91.9%) at study enrollment, and 44 participants (51.2%) had a FLIPI score of 3 or higher. 54 participants (62.8%) were bi-resistant to anti-CD20 and alkylating agents, and 42 participants (48.8%) experienced POD24. The median number of prior lines of systemic anti-lymphoma therapy was 2.0 (range: 2, 9), and 17 participants (19.8%) received four or more prior lines of anti-lymphoma therapy.

[0266] Although efficacy was not the primary endpoint of the optimization part of GCT3013-01, ORR(CR+PR) and CR rate were assessed by researchers as secondary endpoints using the Lugano classification criteria. After a median follow-up of 5.7 months (range: 0.4, 11.8), the ORR in subjects in Arm A was 86.0% (95% CI: 76.9, 92.6), and the CR rate was 64.0% (95% CI: 52.9, 74.0). In addition, of the 78 subjects who underwent tumor evaluation at week 6, 35 (44.9%) showed CR and 26 (33.3%) had PR. At week 12, of the 60 subjects who underwent tumor evaluation, 39 (65.0%) had CR and 10 (16.7%) had PR. These data are consistent with efficacy results observed in subjects with FL in a pivotal cohort who received a two-step SUD regimen of epcolitamab 0.16 / 0.8 / 48 mg.

[0267] In subjects in Arm A, the median TTR was 1.4 months (range: 1.2, 4.4), and the median TTCR was 1.5 months (range: 1.2, 4.7). [Table 11] TIFF2026513993000016.tif253166 TIFF2026513993000017.tif252165 TIFF2026513993000018.tif105166

[0268] Bold and underlined text is FE;A;L and R, corresponding to positions 234 and 235;265;405 and 409, respectively, where the positions are based on EU numbering. In the variable region, the CDR region is underlined and annotated according to the IMGT definition.

Claims

1. A method for treating B-cell non-Hodgkin lymphoma, such as aggressive or indolent lymphoma, in human subjects, comprising administering a bispecific antibody to the subject, wherein the bispecific antibody is (i) A first binding arm comprising a first antigen-binding region that binds to human CD3ε (epsilon) and includes a variable heavy chain (VH) region and a variable light chain (VL) region, wherein the VH region includes the CDR1, CDR2, and CDR3 sequences in the VH region sequence of SEQ ID NO: 6, and the VL region includes the CDR1, CDR2, and CDR3 sequences in the VL region sequence of SEQ ID NO: 7, and (ii) A second binding arm that binds to human CD20 and includes a second antigen-binding region comprising a VH region and a VL region, wherein the VH region comprises CDR1, CDR2, and CDR3 sequences in the VH region sequence of SEQ ID NO: 13, and the VL region comprises CDR1, CDR2, and CDR3 sequences in the VL region sequence of SEQ ID NO: 14, The bispecific antibody is administered in a 28-day cycle, including a priming dose of approximately 0.05 mg to 0.35 mg on day 1, a first intermediate dose of approximately 0.6 mg to 5 mg around day 8, a second intermediate dose of approximately 1 mg to 10 mg around day 15, and a subsequent single weekly dose of approximately 20 to 100 mg, or A method comprising administering the bispecific antibody in a 35-day cycle, including a priming dose of approximately 0.05 mg to 0.35 mg on day 1, a first intermediate dose of approximately 0.6 mg to 5 mg around day 8, a second intermediate dose of approximately 1 mg to 10 mg around day 15, and at least two subsequent weekly doses of approximately 20 to 100 mg.

2. The method according to claim 1, wherein the priming dose is approximately 0.16 mg.

3. The method according to claim 1 or 2, wherein the first intermediate dose is approximately 0.6 to 1.2 mg.

4. The method according to claim 3, wherein the first intermediate dose is approximately 0.8 mg.

5. The method according to any one of claims 1 to 4, wherein the second intermediate dose is approximately 3 to 6 mg.

6. The method according to claim 5, wherein the second intermediate dose is 3 mg.

7. The method according to claim 5, wherein the second intermediate dose is 6 mg.

8. The method according to any one of claims 1 to 7, wherein the weekly dose is approximately 20 to 60 mg.

9. The method according to claim 8, wherein the weekly dose is approximately 24 mg.

10. The method according to claim 8, wherein the weekly dose is approximately 48 mg.

11. The method according to any one of claims 1 to 10, wherein the indolent lymphoma is follicular lymphoma (FL), cutaneous T-cell lymphoma (CTCL), marginal zone lymphoma (MZL), chronic lymphocytic leukemia (CLL), or small cell lymphocytic lymphoma (SLL), and / or the aggressive lymphoma is large B-cell lymphoma (LBCL), such as diffuse large B-cell lymphoma (DLBCL), high-grade B-cell lymphoma (HGBCL), primary mediastinal large B-cell lymphoma (PMBCL), follicular lymphoma (FL) grade 3B, or mantle cell lymphoma (MCL).

12. The method according to claim 11, wherein the indolent lymphoma is FL.

13. The method according to any one of claims 1 to 12, wherein the subject has previously received antineoplastic therapy.

14. The method according to claim 13, wherein the subject has received prior treatment with a CD20 monospecific antibody.

15. The method according to any one of claims 1 to 14, wherein the indolent lymphoma is recurrent or refractory.

16. The method according to any one of claims 1 to 15, wherein the weekly administration is performed at least four times.

17. The method according to any one of claims 1 to 16, wherein the bispecific antibody is administered once every two weeks after the weekly administration.

18. The method according to claim 17, wherein the bispecific antibody is administered every other week for at least six times.

19. The method according to claim 17 or 18, wherein the bispecific antibody is administered once every four weeks after the bi-weekly administration.

20. The method according to any one of claims 1 to 19, wherein the bispecific antibody is administered subcutaneously.

21. (a) The first cycle of 28 days, (i) On day 1, administer the priming dose of the bispecific antibody; (ii) On the 8th day, administer the first intermediate dose of the bispecific antibody; (iii) On the 15th day, administer the second intermediate dose of the bispecific antibody; (iv) A cycle in which a dose of 24 mg of the bispecific antibody is administered on day 22; (b) Cycles 2-3, each comprising 28 days in which a dose of 24 mg of the bispecific antibody is administered on days 1, 8, 15, and 22; (c) Cycles 4 to 9, each comprising 28 days in which a dose of 24 mg of the bispecific antibody is administered on day 1 and day 15; and (d) Administer a dose of 24 mg of the bispecific antibody on day 1, followed by a further subsequent 28-day cycle. The method according to any one of claims 1 to 20, including the method described in any one of claims 1 to 20.

22. (a) The first cycle of 28 days, (i) On day 1, administer the priming dose of the bispecific antibody; (ii) On the 8th day, administer the first intermediate dose of the bispecific antibody; (iii) On the 15th day, administer the second intermediate dose of the bispecific antibody; (iv) A cycle in which a dose of 48 mg of the bispecific antibody is administered on day 22; (b) Cycles 2-3, each comprising 28 days in which a dose of 48 mg of the bispecific antibody is administered on days 1, 8, 15, and 22; (c) Cycles 4 to 9, each comprising 28 days in which a dose of 48 mg of the bispecific antibody is administered on day 1 and day 15; and (d) A further subsequent 28-day cycle in which 48 mg of the bispecific antibody is administered on day 1. The method according to any one of claims 1 to 20, including the method described in any one of claims 1 to 20.

23. (a) The first cycle of 28 days, (i) On day 1, administer a priming dose of 0.16 mg of the bispecific antibody; (ii) On day 8, administer a first intermediate dose of 0.8 mg of the bispecific antibody; (iii) On day 15, administer a second intermediate dose of 3 mg of the bispecific antibody; (iv) A cycle in which a dose of 48 mg of the bispecific antibody is administered on day 22; (b) Cycles 2-3, each comprising 28 days in which a dose of 48 mg of the bispecific antibody is administered on days 1, 8, 15, and 22; (c) Cycles 4 to 9, each comprising 28 days in which a dose of 48 mg of the bispecific antibody is administered on day 1 and day 15; and (d) A further subsequent 28-day cycle in which 48 mg of the bispecific antibody is administered on day 1. The method according to any one of claims 1 to 20, including the method described in any one of claims 1 to 20.

24. (a) The first cycle of 28 days, (i) On day 1, administer a priming dose of 0.16 mg of the bispecific antibody; (ii) On day 8, administer a first intermediate dose of 0.8 mg of the bispecific antibody; (iii) On day 15, administer a second intermediate dose of 6 mg of the bispecific antibody; (iv) A cycle in which a dose of 48 mg of the bispecific antibody is administered on day 22; (b) Cycles 2-3, each comprising 28 days in which a dose of 48 mg of the bispecific antibody is administered on days 1, 8, 15, and 22; (c) Cycles 4 to 9, each comprising 28 days in which a dose of 48 mg of the bispecific antibody is administered on day 1 and day 15; and (d) A further subsequent 28-day cycle in which 48 mg of the bispecific antibody is administered on day 1. The method according to any one of claims 1 to 20, including the method described in any one of claims 1 to 20.

25. The method according to any one of claims 21 to 24, wherein the indolent lymphoma is FL.

26. The method according to any one of claims 1 to 25, wherein the subject has a manageable grade 1 or grade 2 cytokine release syndrome (CRS) during administration of the bispecific antibody.

27. The method according to any one of claims 1 to 26, wherein the subject has not experienced tumor lysis syndrome.

28. The method according to any one of claims 1 to 27, wherein the subject is treated with a method for preventing CRS.

29. The method according to claim 28, wherein the preventive method includes the administration of a corticosteroid.

30. The method according to claim 29, wherein the corticosteroid is dexamethasone.

31. The method according to claim 30, wherein the dexamethasone is administered in a dose of approximately 2 to 20 mg.

32. The method according to claim 31, wherein the dexamethasone is administered in a dose of approximately 15 mg.

33. The method according to any one of claims 28 to 32, wherein the preventive method is administered on the same day as the bispecific antibody.

34. The method according to any one of claims 28 to 33, wherein the preventive method is administered on the second to third day thereafter, optionally on the fourth day, or on the second to fourth day thereafter.

35. The method according to any one of claims 28 to 34, wherein, if the prophylactic method is administered on the same day as the bispecific antibody, the prophylactic method is administered 30 to 120 minutes before the administration of the bispecific antibody.

36. The method according to any one of claims 1 to 35, wherein the aforementioned human subject is treated with premedication to reduce the response to injection.

37. The method according to claim 36, wherein the premedication includes the administration of an antihistamine.

38. The method according to claim 36 or 37, wherein the premedication includes the administration of an antipyretic.

39. The method according to any one of claims 36 to 37, wherein the antihistamine is, for example, 50 mg of diphenhydramine administered intravenously or orally, or an equivalent thereof.

40. The method according to claim 38, wherein the antipyretic is, for example, acetaminophen or an equivalent thereof in an oral dose of 650 to 1000 mg.

41. The method according to any one of claims 36 to 40, wherein the premedication is administered on the same day as the bispecific antibody.

42. The method according to claim 41, wherein the premedication is administered 30 to 120 minutes before the administration of the bispecific antibody.

43. The method according to one of claims 28 to 42, wherein the preventive method is administered during the first cycle.

44. The method according to any one of claims 28 to 43, wherein the premedication is administered during the first cycle.

45. The method according to claim 43 or 44, wherein the preventive measure is administered during the second cycle if the human subject experiences CRS of grade 1 or higher after the fourth administration of the bispecific antibody in cycle 1.

46. The method according to any one of claims 43 to 43, wherein the preventive measure is continued during a subsequent cycle if the human subject experiences a CRS of grade 1 or higher in the last dose of the bispecific antibody in the previous cycle.

47. The method according to any one of claims 44 to 46, wherein the premedication may be administered during the second cycle.

48. The method according to claim 47, wherein the premedication may be administered during a subsequent cycle.

49. The aforementioned bispecific antibody a. A first binding arm comprising a first antigen-binding region that binds to human CD3ε (epsilon), wherein the first antigen-binding region comprises heavy chain variable (VH) regions CDR1, CDR2, and CDR3 having sequences shown in SEQ ID NOs: 1, 2, and 3, respectively, and light chain variable (VL) regions CDR1, CDR2, and CDR3 having sequences shown in SEQ ID NOs: 4, GTN, and 5, respectively, and b. A second binding arm comprising a second antigen-binding region for binding to human CD20, wherein the second antigen-binding region comprises heavy chain variable (VH) regions CDR1, CDR2, and CDR3 having sequences shown in SEQ ID NOs: 8, 9, and 10, respectively, and light chain variable (VL) regions CDR1, CDR2, and CDR3 having sequences shown in SEQ ID NOs: 11, DAS, and 12, respectively. The method according to any one of claims 1 to 48, including the method described in any one of claims 1 to 48.

50. The aforementioned bispecific antibody a. A first binding arm comprising a first antigen-binding region for binding to human CD3ε (epsilon), comprising a variable heavy chain region and a variable light chain region, wherein the variable heavy chain region comprises SEQ ID NO: 6 and the variable light chain region comprises SEQ ID NO: 7; and b. A second binding arm comprising a second antigen-binding region for binding to human CD20, comprising a variable heavy chain region and a variable light chain region, wherein the variable heavy chain region comprises SEQ ID NO: 13 and the variable light chain region comprises SEQ ID NO:

14. The method according to any one of claims 1 to 49, including the method described in any one of claims 1 to 49.

51. The method according to any one of claims 1 to 50, wherein the first binding arm of the bispecific antibody is derived from a humanized antibody, preferably a full-length IgG1, λ (lambda) antibody.

52. The method according to claim 51, wherein the bispecific antibody includes a λ light chain constant region as defined in SEQ ID NO:

22.

53. The method according to claim 51, wherein the second binding arm of the bispecific antibody is derived from a human antibody, preferably a full-length IgG1, κ (kappa) antibody.

54. The method according to claim 53, wherein the bispecific antibody includes a κ light chain constant region as defined in SEQ ID NO:

23.

55. The method according to any one of claims 1 to 54, wherein the bispecific antibody is a full-length antibody having a human IgG1 constant region.

56. The method according to any one of claims 1 to 55, wherein the bispecific antibody includes an inactive Fc region.

57. The method according to claim 56, wherein the bispecific antibody contains amino acids F, E, and A in the first and second heavy chains at positions corresponding to L234, L235, and D265 of the human IgG1 heavy chain of SEQ ID NO: 15 in both the first and second heavy chains.

58. The method according to any one of claims 1 to 57, wherein the bispecific antibody contains amino acid L in the first heavy chain at a position corresponding to F405 of the human IgG1 heavy chain of SEQ ID NO: 15, and the second heavy chain contains amino acid R at a position corresponding to K409 of the human IgG1 heavy chain of SEQ ID NO: 15, or vice versa.

59. The method according to any one of claims 1 to 58, wherein the bispecific antibody contains amino acids F, E, and A in the first and second heavy chains at positions corresponding to L234, L235, and D265 in the human IgG1 heavy chain of SEQ ID NO: 15, respectively, in the first heavy chain, amino acid L at a position corresponding to F405 in the human IgG1 heavy chain of SEQ ID NO: 15, and the second heavy chain contains amino acid R at a position corresponding to K409 in the human IgG1 heavy chain of SEQ ID NO: 15, or vice versa.

60. The method according to claim 59, wherein the bispecific antibody includes constant regions defined in SEQ ID NOs: 19 and 20.

61. The method according to any one of claims 1 to 60, wherein the bispecific antibody comprises a heavy chain and a light chain defined in SEQ ID NOs. 24 and 25, and a heavy chain and a light chain defined in SEQ ID NOs. 26 and 27, respectively.

62. The method according to any one of claims 1 to 61, wherein the bispecific antibody comprises a heavy chain and a light chain defined by SEQ ID NOs. 24 and 25, and a heavy chain and a light chain defined by SEQ ID NOs. 26 and 27, respectively.

63. The method according to any one of claims 1 to 62, wherein the bispecific antibody is epcolitamab or a biosimilar thereof.

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