Methods for treating cancer with bispecific anti-CD22 x anti-CD28 molecules

Abstract

The present disclosure provides methods for treating cancer, reducing the severity of cancer, or inhibiting the growth of cancer. In some embodiments, the method comprises administering to a subject in need thereof a therapeutically effective amount of a bispecific antibody or antigen-binding fragment thereof that specifically binds to CD22 and CD28, in combination with a bispecific antibody or antigen-binding fragment thereof that binds to CD20 and CD3. In some embodiments, the B-cell proliferative disorder is B-cell lymphoma. In some embodiments, the lymphoma is B-cell non-Hodgkin's lymphoma (B-NHL).

Description

[Technical Field]

[0001] Related Applications This application is related to and claims priority to U.S. Provisional Application No. 63 / 489,808, filed March 13, 2023. The entire contents of the aforementioned application, including all figures and sequence listings, are expressly incorporated herein by reference.

[0002] The present disclosure provides methods for treating, reducing the severity of, or inhibiting the growth of cancer in a subject (e.g., a human) in need thereof, comprising administering to the subject a therapeutically effective amount of a bispecific antibody, or antigen-binding fragment thereof, that specifically binds CD22 and CD28, in combination with a bispecific antibody, or antigen-binding fragment thereof, that binds CD20 and CD3.

[0003] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in XML format and is incorporated herein by reference in its entirety. The XML copy, created on March 5, 2024, is named 118003-59220.XML and is 39,470 bytes in size. [Background technology]

[0004] CD28 is a type I transmembrane protein expressed on the surface of T cells. It has a single extracellular IgV-like domain assembled as a homodimer. CD28 is a receptor for the CD80 (B7.1) and CD86 (B7.2) proteins and is activated by CD80 or CD86 expressed on antigen-presenting cells (APCs). Binding of CD28 to CD80 or CD86 provides a costimulatory signal important for T cell activation and survival. T cell stimulation via CD28 in addition to the T cell receptor (TCR) provides a potent signal for the production of various interleukins. CD28 also enhances cellular signals, such as those regulated by the NFκB transcription factor, after TCR activation. CD28 costimulation is important for effective T cell activation, including T cell differentiation, proliferation, cytokine release, and cell death.

[0005] Anti-CD28 antibodies have been proposed for therapeutic purposes, including T cell activation. One particular anti-CD28 antibody, TGN1412 (an anti-CD28 superagonist), was used in clinical trials in 2006. Six healthy volunteers were intravenously administered TGN1412 (an anti-CD28 superagonist) at a dose of 0.1 mg / kg. Within two hours, all six subjects developed a significant inflammatory response (cytokine storm), and all subjects developed multiple organ failure within 16 hours. When the subjects were treated with corticosteroids, cytokine levels returned to normal within two to three days. The starting dose of 0.1 mg / kg in the Phase 1 study (related to CRS) was based on 500 times the cynomolgus monkey "NOAEL" of 50 mg / kg (Suntharalingam, et al., Cytokine Storm in a Phase 1 Trial of the Anti-CD28 Monoclonal Antibody TGN1412, NEJM 355:1018-1028 (2006)). Unfortunately, TGN1412 induced a cytokine storm, which was not predicted in toxicology studies in cynomolgus monkeys or ex vivo human PBMC studies.

[0006] CD22 (also known as Siglec-2), a member of the Siglec family, is a transmembrane protein that specifically recognizes α2,6 sialic acid and is preferentially expressed on B lymphocytes (B cells). CD22 has many specific functions, including B cell homeostasis, B cell survival and migration, attenuation of TLR and CD40 signaling, inhibition of B cell receptor (BCR) signaling by recruiting SH2 domain-containing phosphatases via phosphorylation of immunoreceptor tyrosine-based inhibitory motifs (ITIMs) in the cytoplasmic region, and promotion of adhesion between B cells and other cell types. CD22 is not found on the surface of B cells early in development, nor is it expressed on stem cells. However, 60–70% of all B cell lymphomas and leukemias express CD22.

[0007] Anti-CD22 antibodies are being investigated for the treatment of B-cell lymphoma and leukemia. However, the monoclonal antibody, epratuzumab, has had limited success. (Grant, et al. (2013) Cancer 119(21):10.1002 / cncr.28299) Bispecific antigen binding molecules that bind to both CD28 and a target antigen (such as CD22) are useful in therapeutic settings where specific targeting and T cell-mediated killing of cells expressing the target antigen is desired. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Suntharalingam, et al., Cytokine Storm in a Phase 1 Trial of the Anti-CD28 Monoclonal Antibody TGN1412, NEJM 355:1018-1028(2006) [Non-patent document 2] Grant,et al.(2013)Cancer 119(21):10.1002 / cncr.28299 Summary of the Invention [Means for solving the problem]

[0009] The present disclosure provides a method for treating a B-cell proliferative disorder or malignancy, e.g., a CD20-expressing cell malignancy, in a subject, the method comprising administering to the subject a therapeutically effective amount of a bispecific CD22xCD28 antibody, or antigen-binding fragment thereof, in combination with a bispecific CD3xCD20 antibody, or antigen-binding fragment thereof, wherein the bispecific CD22xCD28 antibody, or antigen-binding fragment thereof, comprises a first antigen-binding domain that binds to cluster of differentiation factor 28 (CD28) and a second antigen-binding domain that binds to cluster of differentiation factor 22 (CD22), and the bispecific CD3xCD20 antibody, or antigen-binding fragment thereof, comprises a first antigen-binding domain that binds to cluster of differentiation factor 3 (CD3) and a second antigen-binding domain that binds to cluster of differentiation factor 20 (CD20), thereby treating the B-cell proliferative disorder or malignancy, e.g., a CD20-expressing cell malignancy, in the subject.

[0010] In some embodiments, the B-cell proliferative disorder is a B-cell lymphoma. In some embodiments, the lymphoma is a B-cell non-Hodgkin's lymphoma (B-NHL). In some embodiments, the non-Hodgkin's lymphoma is selected from the group consisting of diffuse large B-cell lymphoma (DLBCL), marginal zone lymphoma (MZL), high-grade B-cell lymphoma, Burkitt's lymphoma, primary mediastinal large B-cell lymphoma, and follicular lymphoma.

[0011] In some embodiments, the method further comprises selecting a subject, wherein the subject has aggressive B-NHL.

[0012] In some embodiments, the subject has at least one of the following criteria, or is selected based on at least one of the above criteria: has CD20+ aggressive B-NHL, has progressed after at least two lines of systemic therapy including a CD20 inhibitor and an alkylating agent, has measurable disease on cross-sectional imaging, has adequate bone marrow and liver function, and / or has any of the following cancer types: DLBCL, primary mediastinal (thymic) large B-cell lymphoma, T-cell / histiocyte-rich large B-cell lymphoma, grade 3b follicular lymphoma, and high-grade B-cell lymphoma (HGBL) with or without MYC, BCL2, or BCL6 translocations.

[0013] In some embodiments, the subject has been treated with a previous therapy and has relapsed or the disorder has progressed during or after the previous therapy, hi some embodiments, the subject is receiving CAR-T therapy.

[0014] In some embodiments, the subject has measurable CD20+ aggressive B-NHL that has progressed after two or more lines of systemic therapy that includes at least a CD20 inhibitor and an alkylating agent.

[0015] In some embodiments, the CD20 inhibitor is an anti-CD20 antibody.

[0016] In some embodiments, the subject is being treated with CAR-T cell therapy.

[0017] In some embodiments, the subject has not been previously treated with a CD3xCD20 bispecific antibody.

[0018] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered at a dose of about 0.01 mg to about 400 mg.

[0019] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered at a dose of about 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 240 mg, 280 mg, 300 mg, 320 mg, 350 mg, or 400 mg.

[0020] In some embodiments, the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered at a dose of about 0.1 mg to about 400 mg.

[0021] In some embodiments, the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered at a dose of about 0.1 mg, 0.2 mg, 0.3 mg, 0.5 mg, 0.7 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 240 mg, 280 mg, 300 mg, 320 mg, 350 mg, or 400 mg.

[0022] In some embodiments, the method comprises administering one or more doses of a bispecific CD22xCD28 antibody or antigen-binding fragment thereof in combination with one or more doses of a bispecific CD3xCD20 antibody or antigen-binding fragment thereof, in some embodiments, each of the one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is from about 0.01 mg to about 400 mg. In some embodiments, each of the one or more doses is about 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 240 mg, 280 mg, 300 mg, 320 mg, 350 mg, or 400 mg.

[0023] In some embodiments, each of the one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is from about 0.1 mg to about 400 mg.

[0024] In some embodiments, each of the one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is about 0.1 mg, 0.2 mg, 0.3 mg, 0.5 mg, 0.7 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 240 mg, 280 mg, 300 mg, 320 mg, 350 mg, or 400 mg.

[0025] In some embodiments, one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered 1 day to 8 weeks after the immediately preceding dose.

[0026] In some embodiments, each of the one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered once per week, once per two weeks, once per three weeks, once per four weeks, once per five weeks, or once per six weeks.

[0027] In some embodiments, one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof are administered once per week, hi some embodiments, one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof are administered once every two weeks.

[0028] In some embodiments, one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered once per week, hi some embodiments, one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered once every two weeks.

[0029] In some embodiments, the dose of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered in a single dose or divided and administered on two days no more than three days apart.

[0030] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered intravenously. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered subcutaneously.

[0031] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered on the same day.

[0032] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered on different days. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered before or after the bispecific CD3xCD20 antibody or antigen-binding fragment thereof. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered one day before the bispecific CD3xCD20 antibody or antigen-binding fragment thereof.

[0033] In some embodiments, the method comprises the steps of: (i) administering to the subject a bispecific CD3xCD20 antibody or antigen-binding fragment thereof subcutaneously or intravenously at a dose of 0.1 mg to 160 mg weekly for a duration of monotherapy, wherein the duration of monotherapy is at least 2 weeks; and (ii) administering to the subject a bispecific CD22xCD28 antibody or antigen-binding fragment thereof subcutaneously or intravenously at a dose of 0.01 mg to 400 mg weekly, and administering to the subject a bispecific CD3xCD20 or antigen-binding fragment thereof intravenously or subcutaneously at a dose of 80 mg to 160 mg weekly for a duration of induction combination therapy.

[0034] In some embodiments, the duration of monotherapy is at least 2 weeks, at least 3 weeks, at least 4 weeks, or at least 5 weeks.

[0035] In some embodiments, the dose of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof during the monotherapy period is split and administered on two different days no more than three days apart, or is administered in a single dose.

[0036] In some embodiments, the monotherapy period in step (i) comprises administering an initial dose of the bispecific CD3xCD20 antibody and escalating the dose to the full dose by the end of the monotherapy period. In some embodiments, the full dose of bispecific CD3xCD20 in step (i) is 80 mg or 160 mg.

[0037] In some embodiments, the induction combination therapy period in step (ii) comprises: (a) administering an initial dose of bispecific CD22×CD28 antibody, wherein the initial dose comprises between 0.03 mg and 2 mg; (b) administering an intermediate dose comprising between 0.1 mg and 20 mg of bispecific CD22×CD28 antibody; and (c) administering a full dose of bispecific CD22×CD28, wherein the full dose comprises between 0.3 mg and 160 mg.

[0038] In some embodiments, during step (ii), the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered on a different day than the bispecific CD22xCD28 antibody or antigen-binding fragment thereof.

[0039] In some embodiments, the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered one day after the bispecific CD22xCD28 antibody or antigen-binding fragment thereof.

[0040] In some embodiments, during step (ii), the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered on the same day as the bispecific CD22xCD28 antibody or antigen-binding fragment thereof.

[0041] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered in step (ii) in combination with the bispecific CD3xCD20 or antigen-binding fragment thereof for at least 9 weeks. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered in step (ii) in combination with the bispecific CD3xCD20 antibody or antigen-binding fragment thereof for at least 9, 10, 11, 12, 13, 14 or 15 weeks.

[0042] In some embodiments, the method further comprises (iii) administering after step (ii) the bispecific CD22xCD28 antibody or antigen-binding fragment thereof in combination with 160 mg or 320 mg of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof every two or more weeks for the duration of the maintenance combination therapy.

[0043] In some embodiments, in step (iii), the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered on the same day.

[0044] In some embodiments, in step (iii), the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and the bispecific CD3xCD20 or antigen-binding fragment thereof are administered once every two weeks or once every four weeks.

[0045] In some embodiments, the method further comprises administering to the subject one or more additional agents to treat or prevent one or more symptoms of the adverse event.

[0046] In some embodiments, the bispecific antibody is administered to the subject in combination with a second agent, wherein the second agent is selected from the group consisting of dexamethasone, diphenhydramine, acetaminophen, a steroid, an antihistamine, a nonsteroidal anti-inflammatory drug (NSAID), an IL-6 antagonist, and an IL-6R antagonist.

[0047] In some embodiments, a subject has stable disease, a partial response, or a complete response when administered a bispecific CD22xCD28 antibody, or antigen-binding fragment thereof, at a dose of about 0.01 mg to about 400 mg in combination with a bispecific CD3xCD20 antibody, or antigen-binding fragment thereof, for at least one week.

[0048] In some embodiments, the bispecific CD3xCD20 antibody or antigen-binding fragment thereof comprises i) a CD3-binding arm comprising heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 5, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 6, and ii) a CD20-binding arm comprising heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 4, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 6.

[0049] In some embodiments, the bispecific CD3xCD20 antibody or antigen-binding fragment thereof comprises i) a CD3-binding arm comprising three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 10, HCDR2 comprises the amino acid sequence of SEQ ID NO: 11, HCDR3 comprises the amino acid sequence of SEQ ID NO: 12, LCDR1 comprises the amino acid sequence of SEQ ID NO: 13, LCDR2 comprises the amino acid sequence of SEQ ID NO: 14, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 15. and ii) a CD20-binding arm comprising three HCDRs (HCDR1, HCDR2 and HCDR3) and three LCDRs (LCDR1, LCDR2 and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 7, HCDR2 comprises the amino acid sequence of SEQ ID NO: 8, HCDR3 comprises the amino acid sequence of SEQ ID NO: 9, LCDR1 comprises the amino acid sequence of SEQ ID NO: 13, LCDR2 comprises the amino acid sequence of SEQ ID NO: 14, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 15.

[0050] In some embodiments, the HCVR of the CD3-binding arm comprises the amino acid sequence of SEQ ID NO:5, the HCVR of the CD20-binding arm comprises the amino acid sequence of SEQ ID NO:4, and the consensus LCVR comprises the amino acid sequence of SEQ ID NO:6.

[0051] In some embodiments, the bispecific CD3xCD20 antibody or antigen-binding fragment thereof comprises a heavy chain of a CD3 binding arm comprising the amino acid sequence of SEQ ID NO:2, a heavy chain of a CD20 binding arm comprising the amino acid sequence of SEQ ID NO:1, and a common light chain comprising the amino acid sequence of SEQ ID NO:3.

[0052] In some embodiments, the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is odronextamab.

[0053] In some embodiments, the first antigen-binding domain that binds to CD28 comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 20, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 21. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO: 25, HCDR2 comprises the amino acid sequence of SEQ ID NO: 26, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 27. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 28, LCDR2 comprises the amino acid sequence of SEQ ID NO: 29, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 30.

[0054] In some embodiments, the first antigen-binding domain that binds to CD28 comprises an HCVR comprising the amino acid sequence of SEQ ID NO:20, and an LCVR comprising the amino acid sequence of SEQ ID NO:21.

[0055] In some embodiments, the second antigen-binding domain that binds to CD22 comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 19, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 21. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO: 22, HCDR2 comprises the amino acid sequence of SEQ ID NO: 23, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 24. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 28, LCDR2 comprises the amino acid sequence of SEQ ID NO: 29, and CDR-L3 comprises the amino acid sequence of SEQ ID NO: 30.

[0056] In some embodiments, the second antigen-binding domain that binds to CD22 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 19 and an LCVR comprising the amino acid sequence of SEQ ID NO: 21.

[0057] In some embodiments, (a) the first antigen-binding domain that binds to human CD28 comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 20 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21, and (b) the second antigen-binding domain that binds to CD22 comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21.

[0058] In some embodiments, the bispecific CD22xCD28 antibody comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO:17.

[0059] In some embodiments, the bispecific CD22xCD28 antibody comprises a second heavy chain comprising the amino acid sequence of SEQ ID NO:16.

[0060] In some embodiments, the bispecific CD22xCD28 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:18.

[0061] In some embodiments, the bispecific CD22xCD28 antibody comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 17, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 16, and a common light chain comprising the amino acid sequence of SEQ ID NO: 18.

[0062] In some embodiments, the first antigen-binding domain comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:17 and a light chain comprising the amino acid sequence of SEQ ID NO:18.

[0063] In some embodiments, the second antigen-binding domain comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:16 and a light chain comprising the amino acid sequence of SEQ ID NO:18.

[0064] In some embodiments, the bispecific CD22xCD28 antibody is REGN5837 or an antigen-binding fragment thereof. [Brief explanation of the drawings]

[0065] [Figure 1] A schematic diagram of the combination study is shown, with a 28-day screening period and induction period for odronextamab monotherapy, followed by combination with REGN5837. Each cycle in the induction period is 21 days, and each cycle in the maintenance period is 28 days. QW: weekly. Q2W: every 2 weeks. Q4W: every 4 weeks. [Figure 2] 1 shows a schematic diagram of an example of a combination dosing regimen for subjects at dose level 1 (DL1). [Figure 3] A schematic diagram of the pre-dose schedule for cycles 1 to 3 from initial odronextamab monotherapy to combination treatment with odronextamab and REGN5837 is shown. DETAILED DESCRIPTION OF THE INVENTION

[0066] Before describing the present disclosure, it is to be understood that the present disclosure is not limited to the particular methods and experimental conditions described, since such methods and conditions may vary. It is also to be understood that the terminology used herein is used only for the purpose of describing particular embodiments, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.

[0067] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this disclosure belongs.As used herein, the term "about" when used in relation to a specific listed numerical value means that the value can vary by 1% or less from the listed value.For example, as used herein, the expression "about 100" includes 99 and 101, and all values ​​therebetween (for example, 99.1, 99.2, 99.3, 99.4, etc.).

[0068] Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. All patents, applications, and non-patent publications mentioned herein are incorporated by reference in their entirety.

[0069] definition The term "CD28" as used herein refers to an antigen expressed on T cells as a costimulatory receptor. Human CD28 comprises the amino acid sequence set forth in NCBI accession number NP_006130.1. All references to proteins, polypeptides, and protein fragments herein are intended to refer to the human form of the respective protein, polypeptide, or protein fragment unless specifically identified as being derived from a non-human species. Thus, the term "CD28" refers to human CD28 unless specifically identified as being derived from a non-human species, such as "mouse CD28," "monkey CD28," etc.

[0070] As used herein, "antibodies that bind to CD28" or "anti-CD28 antibodies" include antibodies and antigen-binding fragments thereof that specifically recognize monomeric CD28, as well as antibodies and antigen-binding fragments thereof that specifically recognize dimeric CD28. The antibodies and antigen-binding fragments of the present disclosure can bind to soluble CD28 and / or cell surface-expressed CD28. Soluble CD28 includes native CD28 protein as well as recombinant CD28 protein variants that lack the transmembrane domain or are otherwise not associated with the cell membrane, such as monomeric and dimeric CD28 constructs.

[0071] As used herein, the phrase "cell surface-expressed CD28" refers to one or more CD28 proteins expressed on the surface of a cell in vitro or in vivo, where at least a portion of the CD28 protein is exposed on the extracellular surface of the cell membrane and is accessible to the antigen-binding portion of an antibody. "Cell surface-expressed CD28" includes CD28 protein in the context of a functional T cell costimulatory receptor within the cell membrane. "Cell surface-expressed CD28" includes CD28 protein expressed as part of a homodimer on the surface of a cell. "Cell surface-expressed CD28" can include or consist of CD28 protein expressed on the surface of a cell that normally expresses CD28 protein. Alternatively, "cell surface-expressed CD28" can include or consist of CD28 protein expressed on the surface of a cell that does not normally express human CD28 on its surface but has been artificially engineered to express CD28 on its surface.

[0072] As used herein, the phrase "anti-CD28 antibody" includes both monovalent antibodies having a single specificity, and bispecific antibodies comprising a first arm that binds to CD28 and a second arm that binds to a second (target) antigen, wherein the anti-CD28 arm comprises any of the HCVR / LCVR or CDR sequences set forth in Table 1 herein. Examples of anti-CD28 bispecific antibodies are described elsewhere herein. The term "antigen-binding molecule" includes antibodies and antigen-binding fragments of antibodies, including, for example, bispecific antibodies.

[0073] The term "CD22," as used herein, refers to the human CD22 protein unless specified as being derived from a non-human species (e.g., "mouse CD22," "monkey CD22," etc.). Human CD22 protein has the amino acid sequence set forth in accession number CAA42006. The sequence of recombinant human CD22 ecto(D20-R687) with a myc hexahistidine tag is set forth in accession number NP_001762.2. hCD22 ectodomain(D20-R687).hFc can also be purchased from R&D Systems, catalog #1968-SL-050.

[0074] As used herein, "antibodies that bind to CD22" or "anti-CD22 antibodies" include antibodies and antigen-binding fragments thereof that can bind to soluble CD22 and / or cell surface-expressed CD22. Soluble CD22 includes native CD22 protein as well as recombinant CD22 protein variants, e.g., CD22 constructs, that lack the transmembrane domain or are not associated with the cell membrane.

[0075] As used herein, the phrase "anti-CD22 antibody" includes both monovalent antibodies having a single specificity, and bispecific antibodies comprising a first arm that binds to CD22 and a second arm that binds to a second (target) antigen, wherein the anti-CD22 arm comprises any of the HCVR / LCVR or CDR sequences set forth in Table 1 herein. Examples of anti-CD22 bispecific antibodies are described elsewhere herein. The term "antigen-binding molecule" includes antibodies and antigen-binding fragments of antibodies, including, for example, bispecific antibodies.

[0076] The term "antigen-binding molecule" includes antibodies and antigen-binding fragments of antibodies, including, for example, bispecific antibodies.

[0077] As used herein, the term "antibody" refers to any antigen-binding molecule or molecular complex containing at least one complementarity-determining region (CDR) that specifically binds to or interacts with a particular antigen (e.g., CD28). The term "antibody" includes immunoglobulin molecules containing four polypeptide chains, two heavy (H) chains and two light (L) chains, interconnected by disulfide bonds, as well as multimers thereof (e.g., IgM). Each heavy chain contains a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region contains three domains: CH1, CH2, and CH3. Each light chain contains a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region contains one domain (CL1). The VH and VL regions can be further subdivided into regions of hypervariability called complementarity-determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. In different embodiments of the present disclosure, the FRs of an anti-CD28 antibody and / or an anti-CD22 antibody (or antigen-binding portion thereof) can be identical to human germline sequences or can be naturally or artificially modified. An amino acid consensus sequence can be defined based on a parallel analysis of two or more CDRs.

[0078] As used herein, the term "antibody" also includes antigen-binding fragments of intact antibody molecules. As used herein, the terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, and the like include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Antigen-binding fragments of antibodies can be derived from intact antibody molecules using any suitable standard technique, such as, for example, proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding antibody variable domains and, optionally, constant domains. Such DNA is known and / or readily available, for example, from commercial sources, DNA libraries (including, for example, phage antibody libraries), or can be synthesized. DNA can be sequenced and manipulated using chemical or molecular biology techniques, for example, to place one or more variable and / or constant domains in a suitable configuration, introduce codons, create cysteine ​​residues, or modify, add, or delete amino acids.

[0079] Non-limiting examples of antigen-binding fragments include (i) Fab fragments, (ii) F(ab')2 fragments, (iii) Fd fragments, (iv) Fv fragments, (v) single-chain Fv (scFv) molecules, (vi) dAb fragments, and (vii) minimal recognition units consisting of amino acid residues mimicking the hypervariable regions (e.g., isolated complementarity-determining regions (CDRs) such as CDR3 peptides) of an antibody, or constrained FR3-CDR3-FR4 peptides. Domain-specific antibodies, single-domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and other engineered molecules such as shark variable IgNAR domains are also encompassed by the term "antigen-binding fragment" as used herein.

[0080] Antigen-binding fragments of antibodies typically contain at least one variable domain. The variable domain can be of any size or amino acid composition and generally contains at least one CDR adjacent to or in-frame with one or more framework sequences. In antigen-binding fragments having a VH domain associated with a VL domain, the VH domain and the VL domain can be positioned relative to each other in any suitable configuration. For example, the variable region can be a dimer and can contain a VH-VH, VH-VL, or VL-VL dimer. Alternatively, the antigen-binding fragment of an antibody can contain a monomeric VH or VL domain.

[0081] In certain embodiments, an antigen-binding fragment of an antibody may comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that may be found in an antigen-binding fragment of an antibody herein include: (i) VH-CH1, (ii) VH-CH2, (iii) VH-CH3, (iv) VH-CH1-CH2, (v) VH-CH1-CH2-CH3, (vi) VH-CH2-CH3, (vii) VH-CL, (viii) VL-CH1, (ix) VL-CH2, (x) VL-CH3, (xi) VL-CH1-CH2, (xii) VL-CH1-CH2-CH3, (xiii) VL-CH2-CH3, and (xiv) VL-CL. In any arrangement of variable and constant domains, including any of the exemplary arrangements listed above, the variable and constant domains can be either directly linked to each other or linked by a complete or partial hinge or linker region. The hinge region can consist of at least two (e.g., 5, 10, 15, 20, 40, 60, or more) amino acids that provide a flexible or semi-flexible link between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, antigen-binding fragments can comprise homodimers or heterodimers (or other multimers) of any of the variable and constant domain configurations listed above in non-covalent association with each other and / or with one or more monomeric VH or VL domains (e.g., via disulfide bond(s)).

[0082] As with intact antibody molecules, antigen-binding fragments can be monospecific or multispecific (e.g., bispecific). Multispecific antigen-binding fragments of antibodies typically contain at least two different variable domains, each capable of specifically binding to a separate antigen or a different epitope on the same antigen. Any multispecific antibody format, including the exemplary bispecific antibody formats disclosed herein, can be adapted for use in the context of the antigen-binding fragments of antibodies of the present disclosure using routine techniques available in the art.

[0083] The antibodies of the present disclosure may function through complement-dependent cytotoxicity (CDC) or antibody-dependent cell-mediated cytotoxicity (ADCC). "Complement-dependent cytotoxicity" (CDC) refers to the lysis of antigen-expressing cells by the antibodies of the present disclosure in the presence of complement. "Antibody-dependent cell-mediated cytotoxicity" (ADCC) refers to a cell-mediated reaction in which nonspecific cytotoxic cells expressing Fc receptors (FcRs) (e.g., natural killer (NK) cells, neutrophils, and macrophages) recognize bound antibodies on target cells, thereby resulting in lysis of the target cells. CDC and ADCC can be measured using assays that are well known and available in the art. (See, e.g., U.S. Patent Nos. 5,500,362 and 5,821,337, and Clynes et al. (1998) Proc. Natl. Acad. Sci. (USA) 95:652-656.) The constant region of an antibody is important for the ability of the antibody to fix complement and mediate cell-dependent cytotoxicity. Thus, the antibody isotype can be selected based on whether it is desirable for the antibody to mediate cytotoxicity.

[0084] In certain embodiments of the present disclosure, the anti-CD28 and / or anti-CD22 antibodies (monospecific or bispecific) of the present disclosure are human antibodies. As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies of the present disclosure may include, for example, amino acid residues in the CDRs, particularly CDR3, that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-specific mutagenesis or in vivo somatic mutation). However, the term "human antibody," as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0085] The antibodies of the present disclosure may, in some embodiments, be recombinant human antibodies. As used herein, the term "recombinant human antibody" is intended to include all human antibodies prepared, expressed, generated, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described below), antibodies isolated from a recombinant combinatorial human antibody library (described below), antibodies isolated from an animal (e.g., a mouse) transgenic for human immunoglobulin genes (see, e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295), or antibodies prepared, expressed, generated, or isolated by any other means, including splicing of human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis), and therefore the amino acid sequences of the VH and VL regions of the recombinant antibodies, while derived from and related to human germline VH and VL sequences, are sequences that may not naturally occur in the human antibody germline repertoire in vivo.

[0086] Human antibodies can exist in two forms related to hinge heterogeneity. In one form, the immunoglobulin molecule contains a stable four-chain construct of approximately 150-160 kDa in which dimers are held together by interchain heavy chain disulfide bonds. In the second form, the dimers are not linked by interchain disulfide bonds, forming approximately 75-80 kDa molecules consisting of covalently linked light and heavy chains (half antibodies). These forms have been extremely difficult to separate, even after affinity purification.

[0087] The frequency of occurrence of the second form in various intact IgG isotypes depends on, but is not limited to, structural differences associated with the antibody hinge region isotype. A single amino acid substitution in the hinge region of a human IgG4 hinge can significantly reduce the occurrence of the second form to the level typically observed using a human IgG1 hinge (Angal et al. (1993) Molecular Immunology 30:105). The present disclosure encompasses antibodies with one or more mutations in the hinge, CH2 region, or CH3 region, which may be desirable, for example, to improve the yield of the desired antibody form during production.

[0088] The antibody of the present disclosure may be an isolated antibody. As used herein, an "isolated antibody" refers to an antibody that has been separated and / or recovered from the identified antibody and at least one component of its natural environment. For example, an antibody that has been separated or removed from at least one component of an organism, or from a tissue or cell in which it naturally occurs or is naturally produced, is an "isolated antibody" for purposes of this disclosure. An isolated antibody also includes an antibody in situ within a recombinant cell. An isolated antibody is an antibody that has undergone at least one purification or isolation step. According to certain embodiments, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0089] The anti-CD28 and / or anti-CD22 antibodies, or antigen-binding domains thereof, of the present disclosure may contain one or more amino acid substitutions, insertions, and / or deletions in the framework and / or CDR regions of the heavy and light chain variable domains compared to the corresponding germline sequences from which the antigen-binding protein or antigen-binding domain was derived. Such mutations can be readily ascertained by comparing the amino acid sequences disclosed herein to germline sequences available, for example, from public antibody sequence databases. The present disclosure includes antibodies, and antigen-binding domains thereof, derived from any of the amino acid sequences disclosed herein, in which one or more amino acids in one or more framework and / or CDR regions are mutated to the corresponding residue(s) in the germline sequence from which the antibody was derived, or to the corresponding residue(s) in another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue(s) (such sequence changes are collectively referred to herein as "germline mutations"). Starting from the heavy and light chain variable region sequences disclosed herein, one skilled in the art can readily produce numerous antibodies and antigen-binding fragments containing one or more individual germline mutations or combinations thereof. In certain embodiments, all framework and / or CDR residues in the VH and / or VL domains are mutated back to the residue found in the original germline sequence from which the antibody was derived. In other embodiments, only certain residues are mutated back to the original germline sequence, e.g., a mutated residue is found within the first 8 amino acids of FR1, or a mutated residue is found within the last 8 amino acids of FR4, or a mutated residue is found only in CDR1, CDR2, or CDR3. In other embodiments, one or more of the framework and / or CDR residue(s) is mutated to the corresponding residue(s) in a different germline sequence (i.e., a germline sequence different from the germline sequence from which the antibody was originally derived).Furthermore, antibodies or antigen-binding domains thereof of the present disclosure may contain any combination of two or more germline mutations within the framework and / or CDR regions, e.g., certain individual residues are mutated to the corresponding residue in a particular germline sequence, while certain other residues that differ from the original germline sequence are maintained or mutated to the corresponding residue in a different germline sequence. Once obtained, antibodies or antigen-binding fragments thereof containing one or more germline mutations can be readily tested for one or more desired properties, such as improved binding specificity, increased binding affinity, improved or enhanced antagonist or agonist biological properties (as the case may be), reduced immunogenicity, etc. Antibodies or antigen-binding fragments thereof obtained in this general manner are encompassed by the present disclosure.

[0090] The present disclosure also includes anti-CD28 and / or anti-CD22 antibodies and antigen-binding molecules comprising variants of any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein. Exemplary variants included in this aspect of the disclosure include variants of any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein with one or more conservative substitutions. For example, the present disclosure includes anti-CD28 antibodies and antigen-binding molecules having HCVR, LCVR, and / or CDR amino acid sequences with, for example, 10 or fewer, 8 or fewer, 6 or fewer, or 4 or fewer conservative amino acid substitutions relative to any of the HCVR, LCVR, and / or CDR amino acid sequences listed in Table 1 herein.

[0091] The term "epitope" refers to an antigenic determinant that interacts with a specific antigen-binding site in the variable region of an antibody molecule, known as the paratope. A single antigen can have multiple epitopes. Thus, different antibodies may bind to different regions on the antigen and have different biological effects. Epitopes can be either conformational or linear. Conformational epitopes are produced by spatially juxtaposed amino acids from different segments of a linear polypeptide chain. Linear epitopes are produced by adjacent amino acid residues in a polypeptide chain. In certain circumstances, epitopes can include carbohydrate, phosphoryl, or sulfonyl moieties on an antigen.

[0092] The terms "substantial identity" or "substantially identical," when referring to a nucleic acid or fragment thereof, indicate that when optimally aligned with another nucleic acid (or its complementary strand) using appropriate nucleotide insertions or deletions, there is at least about 95%, more preferably about 96%, 97%, about 98%, or 99% nucleotide sequence identity of the nucleotide bases as measured by any well-known algorithm for sequence identity, such as FASTA, BLAST, or Gap, as discussed below. A nucleic acid molecule having substantial identity to a reference nucleic acid molecule can, in certain cases, encode a polypeptide having the same or substantially similar amino acid sequence as the polypeptide encoded by the reference nucleic acid molecule.

[0093] When applied to polypeptides, the term "substantial similarity" or "substantially similar" means that two peptide sequences share at least 95% sequence identity, and even more preferably at least 98% or 99% sequence identity, when optimally aligned, such as by the programs GAP or BESTFIT, using a predetermined gap weight. Preferably, non-identical residue positions differ by conservative amino acid substitutions. A "conservative amino acid substitution" is one in which an amino acid residue is replaced by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). Generally, conservative amino acid substitutions do not substantially alter the functional properties of a protein. When two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity or degree of similarity may be adjusted upward to correct for the conservative nature of the substitution. Means for making this adjustment are well known to those of skill in the art. See, e.g., Pearson (1994) Methods Mol. Biol. 24:307-331. Examples of groups of amino acids having side chains with similar chemical properties include: (1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine, (2) aliphatic-hydroxyl side chains: serine and threonine, (3) amide-containing side chains: asparagine and glutamine, (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan, (5) basic side chains: lysine, arginine, and histidine, (6) acidic side chains: aspartic acid and glutamic acid, and (7) sulfur-containing side chains: cysteine ​​and methionine. Preferred conservative amino acid substitution groups are valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine. Alternatively, a conservative replacement is any change that has a positive value in the PAM250 log-likelihood matrix as disclosed in Gonnet et al (1992) Science 256:1443-1445. A "reasonably conservative" replacement is any change that has a non-negative value in the PAM250 log-likelihood matrix.

[0094] Sequence similarity for polypeptides, also referred to as sequence identity, is typically measured using sequence analysis software. Protein analysis software matches similar sequences using similarity measures assigned to various substitutions, deletions, and other modifications, including conservative amino acid substitutions. For example, GCG software includes programs such as Gap and Bestfit, which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms, or between a wild-type protein and its mutant protein. See, for example, GCG version 6.1. Polypeptide sequences can also be compared using FASTA, a program in GCG version 6.1, using default or recommended parameters. FASTA (e.g., FASTA2 and FASTA3) provides alignments and percent sequence identity of the regions of best overlap between the query and search sequences (Pearson (2000) supra). Another preferred algorithm for comparing the sequences of the present disclosure to a database containing a large number of sequences from different organisms is the computer program BLAST, particularly BLASTP or TBLASTN, using default parameters. See, e.g., Altschul et al. (1990) J. Mol. Biol. 215:403-410 and Altschul et al. (1997) Nucleic Acids Res. 25:3389-402.

[0095] The terms "cell proliferative disorder" and "proliferative disorder" refer to disorders associated with some degree of abnormal cell proliferation that would benefit from treatment with an anti-CD28 / anti-CD22 bispecific antigen-binding molecule or a method of the present disclosure. This includes chronic and acute disorders, including those pathological conditions that predispose a mammal to the disorder in question. In one embodiment, the cell proliferative disorder is cancer, a physiological condition in a mammal that is typically characterized by unregulated cell growth / proliferation.

[0096] "Tumor," as used herein, refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor" are not mutually exclusive when referred to herein.

[0097] "B-cell proliferative disorder" refers to any B-cell disorder, including Hodgkin's lymphoma, non-Hodgkin's lymphoma (NHL), e.g., aggressive NHL, relapsed aggressive NHL, low-grade / follicular NHL, small lymphocytic (SL) NHL, intermediate-grade / follicular NHL, intermediate-grade diffuse NHL, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small non-cleaved cell NHL, bulky disease NHL, relapsed asymptomatic NHL, and asymptomatic NHL, including rituximab-refractory asymptomatic NHL. This includes NHL, refractory NHL, refractory asymptomatic NHL, mantle cell lymphoma, AIDS-related lymphoma, and leukemias including Waldenstrom's macroglobulinemia, lymphocyte-predominant Hodgkin's disease (LPHD), small lymphocytic lymphoma (SLL), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), hairy cell leukemia, chronic myeloid leukemia, and other hematologic disorders.

[0098] As used herein, the term "non-Hodgkin's lymphoma" or "NHL" refers to a cancer of the lymphatic system other than Hodgkin's lymphoma. Hodgkin's lymphoma can generally be distinguished from non-Hodgkin's lymphoma by the presence of Reed-Sternberg cells in Hodgkin's lymphoma and the absence of such cells in non-Hodgkin's lymphoma. Examples of non-Hodgkin's lymphomas encompassed by the term as used herein include any identified as such by a skilled artisan (e.g., an oncologist or pathologist) according to classification schemes known in the art, such as the Revised European-American Lymphoma (REAL) scheme as described in Color Atlas of Clinical Hematology (3rd edition), A. Victor Hoffbrand and John E. Pettit (eds.) (Harcourt Publishers Ltd., 2000). See in particular the lists in Figures 11.57, 11.58, and 11.59.More specific examples are relapsed or refractory NHL, frontline low-grade NHL, stage III / IV NHL, chemotherapy-resistant NHL, precursor B-lymphoblastic leukemia and / or lymphoma, small lymphocytic lymphoma, B-cell chronic lymphocytic leukemia and / or prolymphocytic leukemia and / or small lymphocytic lymphoma, B-cell prolymphocytic lymphoma, immunocytoma and / or lymphoplasmacytic lymphoma, lymphoplasmocytic lymphoma, marginal zone B-cell lymphoma, splenic marginal zone lymphoma, extranodal marginal zone-MALT lymphoma, nodal marginal zone lymphoma, hairy cell leukemia, plasmacytoma and / or plasma cell myeloma, low-grade / follicular lymphoma, intermediate-grade / follicular NHL, mantle cell lymphoma, follicular center lymphoma, including, but not limited to, lymphoma (follicular), intermediate-grade diffuse NHL, diffuse large B-cell lymphoma, aggressive NHL (including aggressive frontline NHL and aggressive relapsed NHL), relapsed or refractory NHL after autologous stem cell transplant, primary mediastinal large B-cell lymphoma, primary effusion lymphoma, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small non-cleaved cell NHL, large-disease NHL, Burkitt lymphoma, precursor (peripheral) large granular lymphocytic leukemia, mycotic syndrome and / or Sézary syndrome, epidermal (skin) lymphoma, anaplastic large cell lymphoma, angiocentric lymphoma.

[0099] As used herein, the phrase "in combination with" means that a first therapeutic agent, such as a bispecific CD22xCD28 antibody or antigen-binding fragment thereof, is administered before, after, or simultaneously with a second therapeutic agent, such as a bispecific CD3xCD20 antibody or antigen-binding fragment thereof. The term "in combination with" also includes sequential or simultaneous administration of a first therapeutic agent, such as a bispecific CD22xCD28 antibody or antigen-binding fragment thereof, and a second therapeutic agent, such as a bispecific CD3xCD20 antibody or antigen-binding fragment thereof.

[0100] Combination Therapies and Formulations The present disclosure includes a method for treating, ameliorating, or reducing the severity of at least one symptom or sign of cancer in a subject, or inhibiting the growth of cancer, comprising administering to a subject in need thereof a therapeutic composition comprising a multispecific (e.g., bispecific) antigen-binding molecule that specifically binds to CD28 and CD22 in combination with a bispecific antibody that binds to CD3 and CD20, e.g., odronextamab. The therapeutic composition may comprise any of the multispecific antibodies or bispecific antigen-binding molecules disclosed herein and a pharmaceutically acceptable carrier or diluent. As used herein, the phrase "subject in need thereof" refers to a human or non-human animal that exhibits one or more symptoms or signs of cancer (e.g., a subject that develops a tumor or is afflicted with any of the cancers described herein below), or that would otherwise benefit from inhibition or reduction of CD22 activity or depletion of CD22+ cells.

[0101] The antibodies and bispecific antigen-binding molecules of the present disclosure (and therapeutic compositions comprising them) are useful, inter alia, for treating any disease or disorder in which stimulating, activating, and / or targeting the immune response may be beneficial. In particular, the anti-CD28 / anti-CD22 bispecific antigen-binding molecules of the present disclosure can be used to treat, prevent, and / or ameliorate any disease or disorder associated with or mediated by CD22 expression or activity or the proliferation of CD22+ cells. The mechanism of action achieved by the therapeutic methods of the present disclosure involves the killing of cells expressing CD22 in the presence of effector cells, e.g., T cells. Cells expressing CD22 that can be inhibited or killed using the bispecific antigen-binding molecules of the present disclosure include, for example, cancerous B cells.

[0102] The antigen-binding molecules of the present disclosure can be used to treat primary and / or metastatic tumors occurring in, for example, the blood, bone marrow, lymph nodes (e.g., thymus, spleen), colon, liver, lung, breast, kidney cancer, central nervous system, and bladder cancer. According to certain exemplary embodiments, the bispecific antigen-binding molecules of the present disclosure are used to treat B-cell proliferative disorders.

[0103] In some embodiments, the B-cell proliferative disorder is a B-cell lymphoma, e.g., B-cell non-Hodgkin's lymphoma (B-NHL). In some embodiments, the B-cell lymphoma is diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), high-grade B-cell lymphoma, Burkitt's lymphoma, primary mediastinal large B-cell lymphoma, or follicular lymphoma.

[0104] The present disclosure also includes methods for treating residual cancer in a subject. As used herein, the term "residual cancer" refers to the presence or persistence of one or more cancerous cells in a subject after treatment with an anti-cancer therapy.

[0105] According to certain aspects, the present disclosure provides methods for treating a disease or disorder associated with CD22 expression (e.g., a B-cell proliferative disorder) comprising administering to a subject one or more of the bispecific antigen-binding molecules described elsewhere herein after the subject has been shown to be non-responsive to other types of anti-cancer therapy.

[0106] For example, the disclosure includes a method for treating a B cell proliferative disorder, comprising administering to a subject an anti-CD28 / anti-CD22 bispecific antigen binding molecule in combination with an anti-CD3 / anti-CD20 bispecific antigen binding molecule 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 4 weeks, 2 months, 4 months, 6 months, 8 months, 1 year or more after the subject has received a standard of care treatment for a subject suffering from a B cell proliferative disorder, such as cancer, e.g., DLBCL.

[0107] In certain embodiments, administration of a bispecific CD22xCD28 antibody or antigen-binding fragment thereof in combination with a bispecific CD3xCD20 antibody or antigen-binding fragment thereof results in increased cancer regression, tumor shrinkage and / or elimination. In certain embodiments, administration of a bispecific CD22xCD28 or antigen-binding fragment thereof and a bispecific CD3xCD20 antibody or antigen-binding fragment thereof results in a delay in tumor growth and development, for example, tumor growth may be delayed by about 3 days, more than 3 days, about 7 days, more than 7 days, more than 15 days, more than 1 month, more than 3 months, more than 6 months, more than 1 year, more than 2 years, or more than 3 years compared to untreated subjects or subjects treated with either antibody as monotherapy. In certain embodiments, administration of the bispecific CD22xCD28 or antigen-binding fragment thereof and the bispecific CD3xCD20 antibody or antigen-binding fragment thereof prevents recurrence of cancer and / or increases survival of the subject, for example, by more than 15 days, more than 1 month, more than 3 months, more than 6 months, more than 12 months, more than 18 months, more than 24 months, more than 36 months, or more than 48 months compared to an untreated subject or a subject receiving either antibody as monotherapy.

[0108] In certain embodiments, administration of the bispecific CD22xCD28 or antigen-binding fragment thereof and the bispecific CD3xCD20 antibody or antigen-binding fragment thereof increases the subject's response and duration of response by, for example, more than 2%, more than 3%, more than 4%, more than 5%, more than 6%, more than 7%, more than 8%, more than 9%, more than 10%, more than 20%, more than 30%, more than 40%, or more than 50% compared to untreated subjects or subjects administered either antibody as monotherapy.

[0109] In certain embodiments, administration of the bispecific CD22xCD28 or antigen-binding fragment thereof and the bispecific CD3xCD20 antibody or antigen-binding fragment thereof to a subject with cancer results in at least a 30% or greater reduction in tumor cells or tumor size (a "partial response"). In certain embodiments, administration of the bispecific CD22xCD28 or antigen-binding fragment thereof and the bispecific CD3xCD20 antibody or antigen-binding fragment thereof to a subject with cancer results in the complete disappearance of all evidence of tumor cells (a "complete response"). In certain embodiments, administration of the bispecific CD22xCD28 or antigen-binding fragment thereof and the bispecific CD3xCD20 antibody or antigen-binding fragment thereof to a subject with cancer results in the complete or partial disappearance of tumor cells / lesions, including new measurable lesions. Tumor reduction can be measured by any method known in the art, for example, by X-ray, positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), cytology, histology, or molecular genetic analysis.

[0110] In certain embodiments, administration of the bispecific CD22xCD28 or antigen-binding fragment thereof and the bispecific CD3xCD20 antibody or antigen-binding fragment thereof to a subject with cancer results in a reduction in the size or number of lymphoma lesions.

[0111] In certain embodiments, administration of the bispecific CD22xCD28 or antigen-binding fragment thereof and the bispecific CD3xCD20 antibody or antigen-binding fragment thereof increases progression-free survival or overall survival.

[0112] In certain embodiments, the methods of the disclosure comprise administering to a subject in need thereof a therapeutically effective amount of a bispecific CD22xCD28 or antigen-binding fragment thereof in combination with a bispecific CD3xCD20 antibody or antigen-binding fragment, wherein administration of the combination results in an increase in overall survival (OS) or progression-free survival (PFS) in the patient compared to patients administered standard of care (SOC) therapies (e.g., chemotherapy, surgery, or radiation). In certain embodiments, PFS is increased by at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to patients administered any one or more SOC therapies. In certain embodiments, OS is increased by at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to patients administered any one or more SOC therapies.

[0113] (i) Dosage and timing An "effective" or "therapeutically effective" dose of a bispecific CD22xCD28 antibody or antigen-binding fragment thereof, e.g., REGN5837, or a bispecific CD3xCD20 antibody or antigen-binding fragment thereof, e.g., odronextamab, for treating or preventing cancer, such as a CD22-expressing cancer, is that amount of the antibody or antigen-binding fragment sufficient to alleviate one or more signs and / or symptoms of the disease in the treated subject, whether by inducing regression or elimination of such signs and / or symptoms, or by inhibiting the progression of such signs and / or symptoms.

[0114] The dose of the antigen-binding molecule administered to a subject may vary depending on the subject's age and physique, target disease, condition, administration route, etc. The preferred dose is typically calculated based on body weight or body surface area. The frequency and duration of treatment can be adjusted depending on the severity of the condition.

[0115] In one embodiment, the antigen-binding molecule (e.g., a bispecific antigen-binding molecule that specifically binds to CD22 and CD28) is administered to a subject as a weight-based dose. A "weight-based dose" (e.g., a dose in mg / kg) is a dose of an antibody or antigen-binding fragment thereof or bispecific antigen-binding molecule that varies depending on the subject's weight.

[0116] In another embodiment, the antibody or antigen-binding fragment thereof, or bispecific antigen-binding molecule is administered to the subject as a fixed dose. A "fixed dose" (e.g., a dose in mg) means that one dose of the antibody or antigen-binding fragment thereof, or bispecific antigen-binding molecule is used for all subjects, regardless of any specific subject-related factors, such as body weight. In one specific embodiment, the fixed dose of the antibody or antigen-binding fragment thereof, or bispecific antigen-binding molecule of the present disclosure is based on a predetermined weight or age.

[0117] Generally, suitable doses of antigen-binding molecules of the present disclosure can be within the range of about 0.001 to about 200.0 milligrams per kilogram of recipient body weight, generally within the range of about 1 to 50 mg per kilogram of body weight. For example, antibodies or antigen-binding fragments thereof, or bispecific antigen-binding molecules can be administered at about 0.1 mg / kg, about 0.2 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 1.5 mg / kg, about 2 mg / kg, about 3 mg / kg, about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 40 mg / kg, or about 50 mg / kg per single dose. Values ​​and ranges intermediate to the recited values ​​are also intended to be part of the present disclosure.

[0118] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered at a dose of about 0.01 mg to about 400 mg. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered at a dose of about 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 240 mg, 280 mg, 300 mg, 320 mg, 350 mg, or 400 mg.

[0119] In some embodiments, the bispecific CD22xCD28 antibody is administered intravenously (IV). In some embodiments, the IV infusion is administered over about 1 hour, 2 hours, 3 hours, or 4 hours. In some embodiments, the bispecific CD22xCD28 antibody is administered subcutaneously.

[0120] In some embodiments of the present disclosure, a therapeutically effective dose of a bispecific CD22xCD28 antibody, e.g., REGN5837, is 0.01-400 mg IV or SC once per week, once every two weeks, or once every four weeks.

[0121] In some embodiments, the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered at a dose of about 0.1 mg to about 400 mg. In some embodiments, the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered at a dose of about 0.1 mg, 0.2 mg, 0.3 mg, 0.5 mg, 0.7 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 240 mg, 280 mg, 300 mg, 320 mg, 350 mg, or 400 mg.

[0122] In some embodiments, the bispecific CD3xCD20 antibody is administered intravenously (IV). In some embodiments, the IV infusion is administered over about 1 hour, 2 hours, 3 hours, or 4 hours. In some embodiments, the bispecific CD3xCD20 antibody is administered subcutaneously.

[0123] In some embodiments of the present disclosure, a therapeutically effective dose of a bispecific CD3xCD20 antibody, e.g., odronextamab, is 0.1 to 320 mg IV or SC once per week, once every two weeks, or once every four weeks.

[0124] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding molecule is administered simultaneously with the bispecific CD3xCD20 antibody or antigen-binding portion thereof. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding molecule is administered on the same day as the bispecific CD3xCD20 antibody or antigen-binding portion thereof. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding molecule is administered 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, or 21 days before administration of the bispecific CD3xCD20 antibody or antigen-binding portion thereof. In some embodiments, the bispecific CD3xCD20 antibody or antigen-binding molecule is administered before, for example 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, or 21 days before, the administration of the bispecific CD22xCD28 antibody or antigen-binding portion thereof.

[0125] According to certain embodiments of the present disclosure, multiple doses of an antigen-binding molecule (e.g., a bispecific antigen-binding molecule that specifically binds to CD22 and CD28) may be administered to a subject over a predetermined period of time. A method according to this aspect of the present disclosure comprises sequentially administering multiple doses of an antigen-binding molecule of the present disclosure to a subject. As used herein, "sequentially administering" means that each dose of the antigen-binding molecule is administered to a subject at different time points, for example, on different days separated by a predetermined interval (e.g., several hours, several days, weeks, or months). The present disclosure includes methods comprising sequentially administering to a subject a single initial dose of an antigen-binding molecule, followed by one or more secondary doses of the antigen-binding molecule, and then optionally one or more tertiary doses of the antigen-binding molecule.

[0126] The terms "initial dose," "secondary dose," and "tertiary dose" refer to the time sequence of administration of the antigen-binding molecules of the present disclosure. Thus, the "initial dose" is the dose administered at the beginning of a treatment regimen (also referred to as a "baseline dose"), the "secondary dose" (also referred to as an "intermediate dose") is the dose administered after the initial dose, and the "tertiary dose" is the dose administered after the secondary dose. The initial, secondary, and tertiary doses may all contain the same amount of antigen-binding molecule, but generally may differ from each other in terms of administration frequency. However, in certain embodiments, the amount of antigen-binding molecule contained in the initial, secondary, and / or tertiary doses differs from each other (e.g., adjusted up or down as appropriate) during the course of treatment. In certain embodiments, two or more doses (e.g., 2, 3, 4, or 5) are administered as "loading doses" at the beginning of a treatment regimen, followed by subsequent doses (e.g., "maintenance doses") administered less frequently.

[0127] In one exemplary embodiment of the present disclosure, each secondary dose and / or tertiary dose is within 1 to 26 (e.g., 1, 1 1 / 2, 2, 2 1 / 2, 3, 3 1 / 2, 4, 4 1 / 2, 5, 5 1 / 2, 6, 6 1 / 2, 7, 7 1 / 2, 8, 8 1 / 2, 9, 9 1 / 2, 10, 10 1 / 2, 11, 11 1 / 2, 12, 12 1 / 2, 13, 13 1 / 2, 14, 14 1 / 2, 15, 15 1 / 2, 16, 16 1 / 2, 17, 17 1 / 2, 18, 18 1 / 2, 19, 19 1 / 2, 20, 20 1 / 2, 21, 21 1 / 2, 22, 22 1 / 2, 23, 23 The immediately preceding dose is administered 24, 24 1 / 2, 25, 25 1 / 2, 26, 26 1 / 2, or more weeks later. The phrase "immediately preceding dose" as used herein refers to a dose of an antigen-binding molecule administered to a subject prior to administration of a subsequent dose in a multiple administration series, with no intervening doses.

[0128] The method according to this aspect of the disclosure may include administering any number of secondary and / or tertiary doses of an antigen-binding molecule (e.g., an anti-CD28 antibody or a bispecific antigen-binding molecule that specifically binds to CD22 and CD28) to a subject. For example, in certain embodiments, only a single secondary dose is administered to a subject. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 or more) secondary doses are administered to a subject. Similarly, in certain embodiments, only a single tertiary dose is administered to a subject. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 or more) tertiary doses are administered to a subject.

[0129] In embodiments involving multiple secondary doses, each secondary dose may be administered at the same frequency as the other secondary doses. For example, each secondary dose may be administered to a subject 1 to 2 weeks after the immediately preceding dose. Similarly, in embodiments involving multiple tertiary doses, each tertiary dose may be administered at the same frequency as the other tertiary doses. For example, each tertiary dose may be administered to a subject 2 to 4 weeks after the immediately preceding dose. Alternatively, the frequency with which the secondary and / or tertiary doses are administered to a subject may vary over the course of the treatment regimen. The administration frequency may also be adjusted by a physician during the course of treatment depending on the individual subject's needs after clinical examination.

[0130] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered in a small initial dose to observe the subject's tolerance to the antibody, and the amount of antibody in one or more subsequent doses is increased as the subject tolerates the administration up to a therapeutically effective dose.

[0131] In some embodiments, a dose of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered in a single administration. In some other embodiments, a dose of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is split and administered over two or more days. In some embodiments, the split doses are administered on two different days, no more than three days apart.

[0132] In some embodiments, the method of treatment comprises the steps of: (i) administering to the subject a bispecific CD3xCD20 antibody or antigen-binding fragment thereof at a dose of 0.1 mg to 160 mg subcutaneously or intravenously weekly for a period of monotherapy, wherein the period of monotherapy is at least 2 weeks; and (ii) administering to the subject a bispecific CD22xCD28 antibody or antigen-binding fragment thereof at a dose of 0.01 mg to 400 mg subcutaneously or intravenously weekly for a period of induction combination therapy, wherein the bispecific CD3xCD20 or antigen-binding fragment thereof at a dose of 80 mg to 160 mg is administered intravenously or subcutaneously weekly for a period of induction combination therapy.

[0133] In some embodiments, the duration of monotherapy is at least 2 weeks, at least 3 weeks, at least 4 weeks, or at least 5 weeks.

[0134] In some embodiments, the monotherapy period in step (i) comprises escalating the dose of bispecific CD3xCD20 from a low initial dose to a therapeutically effective dose by the end of the monotherapy period.

[0135] In some embodiments, the therapeutically effective dose of bispecific CD3xCD20 in step (i) is 80 mg or 160 mg.

[0136] In some embodiments, the induction combination therapy period of step (ii) comprises increasing the dose of bispecific CD22xCD28 from a small initial dose to a therapeutically effective dose.

[0137] In some embodiments, during step (ii), the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered on a different day (e.g., one day later) than the bispecific CD22xCD28 antibody or antigen-binding fragment thereof.

[0138] In some embodiments, during step (ii), the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered on the same day as the bispecific CD22xCD28 antibody or antigen-binding fragment thereof.

[0139] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered in step (ii) in combination with the bispecific CD3xCD20 or antigen-binding fragment thereof for at least 9 weeks (e.g., at least 9, 10, 11, 12, 13, 14 or 15 weeks).

[0140] In some embodiments, the method further comprises (iii) administering the bispecific CD22xCD28 antibody or antigen-binding fragment thereof in combination with 180 mg or 320 mg of the bispecific CD3xCD20 or antigen-binding fragment thereof every two or more weeks following step (ii) for the duration of the maintenance combination therapy.

[0141] In some embodiments, in step (iii), the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and the bispecific CD3xCD20 or antigen-binding fragment thereof are administered on the same day. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and the bispecific CD3xCD20 or antigen-binding fragment thereof are administered once every two weeks or once every four weeks.

[0142] (ii) Route of administration The present disclosure provides methods for administering a bispecific CD22xCD28 antibody, e.g., REGN5837, or an antigen-binding fragment thereof, or an anti-CD22 HCVR paired with an HCVR from any of the CD28 antibodies described herein, or any combination thereof, alone or in combination with a bispecific CD3xCD20 antibody, e.g., odronextamab, or an antigen-binding fragment thereof, to a subject (e.g., a human suffering from cancer), comprising introducing the antigen-binding protein or pharmaceutical composition into the subject's (e.g., human) body, e.g., intravenously or subcutaneously. For example, the method comprises puncturing the subject's body with a syringe needle and injecting the antigen-binding protein or pharmaceutical composition into the subject's body, e.g., into a vein, artery, skin, tumor, muscle tissue, or subcutaneous tissue of the subject.

[0143] The mode of administration of an antibody or pharmaceutical composition thereof can vary, including parenteral, non-parenteral, oral, rectal, transmucosal, enteral, parenteral, intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intraocular, inhalation, insufflation, topical, dermal, intraocular, intravitreal, transdermal, or intra-arterial.

[0144] In some embodiments, the bispecific CD22xCD28 antibody, or antigen-binding portion thereof, is administered intravenously and the bispecific CD3xCD20 antibody, or antigen-binding portion thereof, is administered subcutaneously. In some embodiments, the bispecific CD22xCD28 antibody, or antigen-binding portion thereof, is administered subcutaneously and the CD3xCD20 antibody, or antigen-binding portion thereof, is administered intravenously. In some embodiments, the bispecific CD22xCD28 antibody, or antigen-binding portion thereof, is administered intravenously and the bispecific CD3xCD20 antibody, or antigen-binding portion thereof, is administered intravenously. In some embodiments, the bispecific CD22xCD28 antibody, or antigen-binding portion thereof, is administered subcutaneously and the bispecific CD3xCD20, or antigen-binding portion thereof, is administered subcutaneously.

[0145] In some further embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered to the subject over about 10 to 300, 20 to 240, 30 to 180, 45 to 150, or 60 to 120 minutes. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered to the subject over about 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 120, 150, 180, 210, 240, 270, or 300 minutes.

[0146] In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered to the subject for about 1 hour. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered to the subject for about 2 hours. In some embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered to the subject for about 4 hours.

[0147] The present disclosure also provides a container (e.g., a plastic or glass vial or ampoule with a cap or chromatography column, a hollow bore needle or a syringe cylinder, e.g., a cap or chromatography column, a hollow bore needle or a syringe cylinder) comprising a bispecific CD22xCD28 antigen binding protein of the disclosure or a pharmaceutical composition thereof.

[0148] The present disclosure also provides an injection device comprising one or more antigen-binding proteins (e.g., antibodies or antigen-binding fragments) that specifically bind to CD22 and CD28 (CD22×CD28), or pharmaceutical formulations thereof. The injection device can be packaged in a kit. An injection device is a device that introduces a substance into a subject's body via a parenteral route, for example, intramuscularly, subcutaneously, or intravenously. For example, the injection device can be a syringe or auto-injector (e.g., pre-filled with a pharmaceutical formulation) that includes, for example, a cylinder or barrel for holding the fluid to be injected (e.g., comprising an antibody or fragment thereof, or a pharmaceutical formulation thereof), a needle for penetrating the skin, blood vessel, or other tissue for injection of the fluid, and a plunger for forcing the liquid from the cylinder through the bore of the needle and into the subject's body.

[0149] A pre-filled syringe is a syringe that is filled with a composition (e.g., a pharmaceutical composition comprising a multispecific antigen-binding protein and a pharmaceutically acceptable carrier) prior to sale or transfer to an end user, e.g., a physician or caregiver, who will administer the composition (e.g., a pharmaceutical composition comprising a multispecific antigen-binding protein and a pharmaceutically acceptable carrier) to a subject.

[0150] Pharmaceutical compositions are described in more detail below.

[0151] (iii) Selection of subjects In some embodiments, the methods described herein further comprise one or more steps of selecting subjects. Patients may be selected, for example, based on inclusion criteria, or excluded, for example, based on exclusion criteria. The inclusion and exclusion criteria are described in more detail in Example 2 below.

[0152] In some embodiments, the methods include selecting a subject having a disease or disorder associated with CD22 expression.

[0153] In some embodiments, the methods include selecting a subject with a B cell proliferative disorder.

[0154] In some embodiments, the method includes selecting a subject with lymphoma. In some embodiments, the method includes selecting a subject with B-cell non-Hodgkin's lymphoma (B-NHL).

[0155] In some embodiments, the subject has aggressive B-NHL, which is selected from the group consisting of DLBCL, primary mediastinal (thymic) large B-cell lymphoma, T-cell / histiocytic-rich large B-cell lymphoma, grade 3b follicular lymphoma, and high-grade B-cell lymphoma (HGBL) with or without MYC and BCL2 or BCL6 translocations.

[0156] In some embodiments, the subject has at least one of the following criteria, or is selected based on at least one of the above criteria: (i) has CD20+ aggressive B-NHL, (ii) has progressed after at least two lines of systemic therapy including an anti-CD20 antibody and an alkylating agent, (iii) has measurable disease on cross-sectional imaging, (iv) has adequate bone marrow and liver function, and / or (v) has any of the following cancer types: DLBCL, primary mediastinal (thymic) large B-cell lymphoma, T-cell / histiocyte-rich large B-cell lymphoma, grade 3b follicular lymphoma, and high-grade B-cell lymphoma (HGBL).

[0157] In some embodiments, the subject has been treated with a previous therapy and has relapsed or the disorder has progressed during or after the previous treatment.

[0158] In some embodiments, the subject is undergoing CAR-T therapy.

[0159] (iv) Adverse events As used herein, an "adverse event" is an unfavorable and unintended sign (including abnormal laboratory findings), symptom, or disease temporally associated with the use of an investigational drug, whether or not considered related to the investigational drug.

[0160] In some embodiments, the subject develops one or more mild symptoms of an adverse event after administration of the bispecific CD22xCD28 antibody alone or in combination with the bispecific CD3xCD20 antibody, hi some embodiments, the one or more symptoms of the adverse event are symptoms of an infusion reaction (IR) or cytokine release syndrome (CRS), or a combination thereof.

[0161] In some embodiments, symptoms of an infusion reaction are selected from the group consisting of prolonged / severe cough, persistent chills / chills, rash, pruritus (itching), urticaria (hives, aches, rubella), diaphoresis (sweating), hypotension, dyspnea (shortness of breath), vomiting, and flushing.

[0162] In some embodiments, the symptoms of cytokine release syndrome are selected from the group consisting of fever, tachypnea, headache, tachycardia, hypotension, rash, and / or hypoxia.

[0163] In some embodiments, the subject receives one or more additional therapies to treat one or more symptoms of the adverse event.

[0164] In some embodiments, treatment with the bispecific CD22xCD28 antibody alone or in combination with the bispecific CD3xCD20 antibody is paused if the subject develops one or more mild symptoms of an adverse event and is resumed when the one or more symptoms have resolved.

[0165] Multispecific CD22×CD28 antigen binding molecule The present disclosure provides methods of using antigen binding proteins that are multispecific (e.g., bispecific) and bind to at least CD28 and CD22 in combination with a bispecific anti-CD3 and anti-CD20 antibody or antigen-binding fragment thereof, e.g., odronextamab, to treat cancer.

[0166] Multispecific binding refers to binding to two or more different epitopes (CD22 and CD28 or more), which may be on the same or different antigens. Multispecificity includes bispecificity, trispecificity, and tetraspecificity. An antibody or fragment thereof can be operatively linked (e.g., by chemical bonding, genetic fusion, noncovalent bonding, or other methods) to one or more other molecular entities, such as another antibody or antibody fragment, to produce a bispecific or multispecific antibody having a second binding specificity.

[0167] In certain embodiments, the multispecific antigen-binding protein comprises a bispecific antigen-binding protein. As used herein, the term "bispecific antigen-binding protein" refers to a protein, polypeptide, or molecular complex (e.g., an antibody or antigen-binding fragment thereof) comprising at least a first antigen-binding domain and a second antigen-binding domain. Each antigen-binding domain in a bispecific antigen-binding molecule comprises at least one CDR that specifically binds to a particular antigen, either alone or in combination with one or more additional CDRs and / or FRs. In the context of the present disclosure, the first antigen-binding domain specifically binds to CD28, and the second antigen-binding domain specifically binds to CD22.

[0168] According to certain exemplary embodiments, the present disclosure includes bispecific antigen-binding molecules that specifically bind to CD28 and CD22. Such molecules may be referred to herein as, for example, "anti-CD28 / anti-CD22," or "anti-CD28xCD22," or "CD28xCD22," or "anti-CD22 / anti-CD28," or "anti-CD22xCD28," or "CD22xCD28" bispecific molecules, or "αCD22xαCD28," or "αCD28xαCD22," or other similar terms.

[0169] According to certain exemplary embodiments, a bispecific antigen-binding molecule (e.g., a bispecific antibody) may have an effector arm and a targeting arm. The effector arm may be a first antigen-binding domain (e.g., an anti-CD28 antibody) that binds to an antigen on an effector cell (e.g., a T cell). The targeting arm may be a second antigen-binding domain (e.g., an anti-CD22 antibody) that binds to an antigen on a target cell (e.g., a tumor cell). According to certain exemplary embodiments, the effector arm binds to CD28, and the targeting arm binds to CD22. The bispecific anti-CD28 / CD22 may provide a costimulatory signal to an effector cell (e.g., a T cell). The effector arm is ineffective at stimulating T cells without clustering. The effector arm alone has little effect on stimulating T cells unless combined with the targeting arm. The tumor-targeting arm may have incomplete tumor specificity. The antigen (e.g., CD22) targeted by the targeting arm may be expressed on a subset of tumor cells. The specificity of the tumor-targeting arm can be increased by overlapping it in combination with an anti-CD3 bispecific antigen-binding molecule (e.g., an anti-CD3 / CD20 bispecific antibody).

[0170] As used herein, the term "antigen-binding molecule" refers to a protein, polypeptide, or molecular complex comprising or consisting of at least one complementarity-determining region (CDR), which, alone or in combination with one or more additional CDRs and / or framework regions (FRs), specifically binds to a particular antigen. In certain embodiments, the antigen-binding molecule is an antibody or an antibody fragment, as those terms are defined elsewhere herein.

[0171] As used herein, the term "bispecific antigen-binding molecule" refers to a protein, polypeptide, or molecular complex comprising at least a first antigen-binding domain and a second antigen-binding domain. Each antigen-binding domain in a bispecific antigen-binding molecule comprises at least one CDR that specifically binds to a particular antigen, either alone or in combination with one or more additional CDRs and / or FRs. In the context of the present disclosure, the first antigen-binding domain specifically binds to a first antigen (e.g., CD28), and the second antigen-binding domain specifically binds to a second, distinct antigen (e.g., CD22).

[0172] In certain exemplary embodiments of the present disclosure, the bispecific antigen-binding molecule is a bispecific antibody. Each antigen-binding domain of a bispecific antibody comprises a heavy chain variable domain (HCVR) and a light chain variable domain (LCVR). In the context of a bispecific antigen-binding molecule (e.g., a bispecific antibody) comprising a first and a second antigen-binding domain, the CDRs of the first antigen-binding domain may be designated with the prefix "D1", and the CDRs of the second antigen-binding domain may be designated with the prefix "D2". Thus, the CDRs of the first antigen-binding domain may be referred to herein as D1-HCDR1, D1-HCDR2, and D1-HCDR3, and the CDRs of the second antigen-binding domain may be referred to herein as D2-HCDR1, D2-HCDR2, and D2-HCDR3.

[0173] The first and second antigen-binding domains can be directly or indirectly connected to each other to form the bispecific antigen-binding molecule of the present disclosure. Alternatively, the first and second antigen-binding domains can each be connected to a separate multimerization domain. The association of one multimerization domain with another multimerization domain promotes the association between the two antigen-binding domains, thereby forming the bispecific antigen-binding molecule. As used herein, a "multimerization domain" refers to any polymer, protein, polypeptide, peptide, or amino acid capable of associating with a second multimerization domain of the same or similar structure or configuration. For example, the multimerization domain may be a polypeptide containing an immunoglobulin CH3 domain. Non-limiting examples of multimerization domains include the Fc portion of an immunoglobulin (including the CH2-CH3 domain), such as the Fc domain of an IgG selected from the isotypes IgG1, IgG2, IgG3, and IgG4, as well as any allotype within each isotype group.

[0174] The bispecific antigen-binding molecules of the present disclosure typically comprise two multimerization domains, e.g., two Fc domains, each of which is an individual portion of a separate antibody heavy chain. The first and second multimerization domains may be of the same IgG isotype, e.g., IgG1 / IgG1, IgG2 / IgG2, or IgG4 / IgG4. Alternatively, the first and second multimerization domains may be of different IgG isotypes, e.g., IgG1 / IgG2, IgG1 / IgG4, or IgG2 / IgG4.

[0175] In certain embodiments, the multimerization domain is an Fc fragment or an amino acid sequence of 1 to about 200 amino acids in length containing at least one cysteine ​​residue. In other embodiments, the multimerization domain is a cysteine ​​residue or a short cysteine-containing peptide. Other multimerization domains include peptides or polypeptides comprising or consisting of a leucine zipper, a helix-loop motif, or a coiled-coil motif.

[0176] Any bispecific antibody format or technology can be used to generate the bispecific antigen-binding molecules of the present disclosure. For example, an antibody or fragment thereof having a first antigen-binding specificity can be operatively linked (e.g., by chemical bonding, genetic fusion, non-covalent bonding, or other methods) to one or more other molecular entities, such as another antibody or antibody fragment having a second antigen-binding specificity, to produce the bispecific antigen-binding molecule. Certain exemplary bispecific formats that may be used in the context of the present disclosure include, but are not limited to, for example, scFv-based or diabody bispecific formats, IgG-scFv fusions, dual variable domain (OVO)-Ig, quadroma, knobs-into-holes, common light chain (such as a common light chain with knobs-into-holes), CrossMab, CrossFab, (SEEO)body, leucine zipper, Duobody, IgG1 / IgG2, dual acting Fab (OAF)-IgG, and Mab2 bispecific formats (for a review of the aforementioned formats, see, e.g., Klein et al. 2012, mAbs 4:6, 1-11, and references cited therein).

[0177] In the context of the bispecific antigen-binding molecules of the present disclosure, the multimerization domain, e.g., the Fc domain, may contain one or more amino acid changes (e.g., insertions, deletions, or substitutions) compared to a wild-type, naturally occurring Fc domain. For example, the present disclosure includes bispecific antigen-binding molecules containing one or more modifications in the Fc domain that result in a modified Fc domain with a modified binding interaction (e.g., enhanced or decreased) between the Fc and FcRn. In one embodiment, the bispecific antigen-binding molecule contains a modification in the CH2 or CH3 region that increases the affinity of the Fc domain for FcRn in an acidic environment (e.g., within an endosome at a pH range of about 5.5 to about 6.0). Non-limiting examples of such Fc modifications include, for example, modifications at positions 250 (e.g., E or Q), 250 and 428 (e.g., L or F), 252 (e.g., LN / FIW or T), 254 (e.g., S or T), and 256 (e.g., S / R / Q / EID or T), or at positions 428 and / or 433 (e.g., UR / S / P / Q or K) and / or 434 (e.g., H / F or V), or modifications at positions 250 and / or 428, or at positions 307 or 308 (e.g., 308F, V308F), and 434. In one embodiment, the modifications include 428L (e.g., M428L) and 434S (e.g., N434S) modifications, 428L, 2591 (e.g., V2591), and 308F (e.g., V308F) modifications, 433K (e.g., H433K) and 434 (e.g., 434Y) modifications, 252, 254, and 256 (e.g., 252Y, 254T, and 256E) modifications, 250Q and 428L modifications (e.g., T250Q and M428L), 307 and / or 308 modifications (e.g., 308F or 308P).

[0178] The present disclosure also includes bispecific antigen-binding molecules comprising a first CH3 domain and a second Ig CH3 domain, wherein the first Ig CH3 domain and the second Ig CH3 domain differ from each other by at least one amino acid, and wherein the at least one amino acid difference reduces binding of the bispecific antibody to Protein A compared to a bispecific antibody lacking the amino acid difference. In one embodiment, the first Ig CH3 domain binds to Protein A and the second Ig CH3 domain comprises a mutation that reduces or eliminates Protein A binding, e.g., an H95R modification (according to IMGT exon numbering, H435R according to EU numbering). The second CH3 may further comprise a Y96F modification (according to IMGT, Y436F according to EU). Further modifications that may be found within the second CH3 include, for IgG1 antibodies, D16E, L18M, N44S, K52N, V57M, and V821 (by IMGT; D356E, L358M, N384S, K392N, V397M, and V4221 (by EU)); for IgG2 antibodies, N44S, K52N, and V821 (by IMGT; N384S, K392N, and V4221 (by EU)); and for IgG4 antibodies, Q15R, N44S, K52N, V57M, R69K, E79Q, and V821 (by IMGT; Q355R, N384S, K392N, V397M, R409K, E419Q, and V4221 (by EU)).

[0179] In certain embodiments, the Fc domain may be a chimera combining Fc sequences from two or more immunoglobulin isotypes. For example, the chimeric Fc domain may comprise part or all of the CH2 sequence from the CH2 region of human IgG1, human IgG2, or human IgG4, and part or all of the CH3 sequence from human IgG1, human IgG2, or human IgG4. The chimeric Fc domain may also comprise a chimeric hinge region. For example, the chimeric hinge may comprise an "upper hinge" sequence from a human IgG1 hinge region, a human IgG2 hinge region, or a human IgG4 hinge region, combined with a "lower hinge" sequence from a human IgG1 hinge region, a human IgG2 hinge region, or a human IgG4 hinge region. A specific example of a chimeric Fc domain that can be included in any of the antigen-binding molecules described herein comprises, from N-terminus to C-terminus, [IgG4 CH1]-[IgG4 upper hinge]-[IgG2 lower hinge]-[IgG4 CH2]-[IgG4 CH3]. Another example of a chimeric Fc domain that can be included in any of the antigen-binding molecules described herein comprises, from N-terminus to C-terminus, [IgG1 CH1]-[IgG1 upper hinge]-[IgG2 lower hinge]-[IgG4 CH2]-[IgG1 CH3]. These and other examples of chimeric Fc domains that can be included in any of the antigen-binding molecules of the present disclosure are described in WO2014 / 022540A1, the entire contents of which are incorporated herein by reference. Chimeric Fc domains with these general structural arrangements, and variants thereof, can alter Fc receptor binding, which in turn affects Fc effector function.

[0180] According to certain exemplary embodiments of the present disclosure, a bispecific CD22xCD28 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region, a light chain variable region, and / or complementarity determining regions (CDRs) comprising any of the amino acid sequences of the bispecific CD22xCD28 antibodies described in U.S. Patent Publication No. 11,396,544. In certain exemplary embodiments, a bispecific CD22xCD28 antibody, or antigen-binding fragment thereof, that can be used in the context of the methods of the present disclosure comprises: (a) a heavy chain complementarity determining region (CD28-HCDR1, CD28-HCDR2, and CD28-HCDR3) of a heavy chain variable region (CD28-HCVR) comprising the amino acid sequence of SEQ ID NO: 20, and a light chain complementarity determining region (CD28-LCDR1, CD28-LCDR2, and CD28-LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 21. and CD28-LCDR3), and (b) a second antigen-binding arm that binds to CD22, comprising heavy chain CDRs (CD22-HCDR1, CD22-HCDR2, and CD22-HCDR3) of an HCVR (CD22-HCVR) comprising the amino acid sequence of SEQ ID NO: 19, and light chain CDRs (CD22-LCDR1, CD22-LCDR2, and CD22-LCDR3) of an LCVR (CD22-LCVR) comprising the amino acid sequence of SEQ ID NO: 21.

[0181] According to certain embodiments, CD28-HCDR1 comprises the amino acid sequence of SEQ ID NO: 25, CD28-HCDR2 comprises the amino acid sequence of SEQ ID NO: 26, CD28-HCDR3 comprises the amino acid sequence of SEQ ID NO: 27, CD28-LCDR1 comprises the amino acid sequence of SEQ ID NO: 28, CD28-LCDR2 comprises the amino acid sequence of SEQ ID NO: 29, CD28-LCDR3 comprises the amino acid sequence of SEQ ID NO: 30, CD22-HCDR1 comprises the amino acid sequence of SEQ ID NO: 22, CD28-HCDR2 comprises the amino acid sequence of SEQ ID NO: 23, CD28-HCDR3 comprises the amino acid sequence of SEQ ID NO: 24, CD22-LCDR1 comprises the amino acid sequence of SEQ ID NO: 28, CD22-LCDR2 comprises the amino acid sequence of SEQ ID NO: 29, and CD22-LCDR3 comprises the amino acid sequence of SEQ ID NO: 30.

[0182] In yet other embodiments, the bispecific CD22xCD28 antibody or antigen-binding fragment thereof comprises (a) a first antigen-binding arm comprising a HCVR comprising SEQ ID NO: 20 (CD28-HCVR) and a LCVR comprising SEQ ID NO: 21 (CD28-LCVR), and (b) a second antigen-binding arm comprising a HCVR comprising SEQ ID NO: 19 (CD22-HCVR) and a LCVR comprising SEQ ID NO: 21 (CD22-LCVR).

[0183] In certain exemplary embodiments, the bispecific CD22xCD28 antibody comprises a CD28-binding arm comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 17 and a light chain comprising the amino acid sequence of SEQ ID NO: 18, and a CD22-binding arm comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 16 and a light chain comprising the amino acid sequence of SEQ ID NO: 18.

[0184] An exemplary CD22xCD28 bispecific antibody for use in the methods of the disclosure comprises a HCVR in the CD28 binding arm comprising the amino acid sequence of SEQ ID NO: 20, a HCVR in the CD22 binding arm comprising the amino acid sequence of SEQ ID NO: 19, and a consensus LCVR comprising the amino acid sequence of SEQ ID NO: 21. In one embodiment, the CD22xCD28 bispecific antibody is REGN5837, or an antigen-binding fragment thereof.

[0185] (i) Sequence variants The antibodies and bispecific antigen-binding molecules of the present disclosure may contain one or more amino acid substitutions, insertions, and / or deletions in the framework and / or CDR regions of the heavy and light chain variable domains compared to the corresponding germline sequences from which the individual antigen-binding domains were derived. Such mutations can be readily identified by comparing the amino acid sequences disclosed herein to germline sequences available, for example, from public antibody sequence databases. The antigen-binding molecules of the present disclosure may comprise antigen-binding fragments derived from any of the exemplary amino acid sequences disclosed herein, in which one or more amino acids in one or more framework and / or CDR regions are mutated to the corresponding residue(s) in the germline sequence from which the antibody was derived, to the corresponding residue(s) in another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue(s) (such sequence changes are collectively referred to herein as "germline mutations"). Starting from the heavy and light chain variable region sequences disclosed herein, one skilled in the art can easily produce numerous antibodies and antigen-binding fragments containing one or more individual germline mutations or combinations thereof. In certain embodiments, all framework and / or CDR residues in the VH and / or VL domains are mutated back to the residue found in the original germline sequence from which the antigen-binding domain was originally derived. In other embodiments, only certain residues are mutated back to the original germline sequence, e.g., the mutated residue is found within the first 8 amino acids of FR1, or the mutated residue is found within the last 8 amino acids of FR4, or the mutated residue is found only in CDR1, CDR2, or CDR3. In other embodiments, one or more of the framework and / or CDR residue(s) is mutated to the corresponding residue(s) in a different germline sequence (i.e., a germline sequence that differs from the germline sequence from which the antigen-binding domain was originally derived). Furthermore, the antigen-binding domain may contain any combination of two or more germline mutations in the framework and / or CDR regions, e.g., certain individual residues are mutated to the corresponding residue in a particular germline sequence, while certain other residues that differ from the original germline sequence are maintained or mutated to the corresponding residue in a different germline sequence.Once obtained, antigen-binding domains containing one or more germline mutations can be readily tested for one or more desired properties, such as improved binding specificity, increased binding affinity, improved or enhanced antagonist or agonist biological properties (as the case may be), reduced immunogenicity, etc. Bispecific antigen-binding molecules comprising one or more antigen-binding domains obtained in this general manner are encompassed by the present disclosure.

[0186] The present disclosure also includes antigen-binding molecules in which one or both antigen-binding domains comprise variants of any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein with one or more conservative substitutions. For example, the present disclosure includes antigen-binding molecules comprising an antigen-binding domain having an HCVR, LCVR, and / or CDR amino acid sequence with, for example, 10 or less, 8 or less, 6 or less, 4 or less, etc., conservative amino acid substitutions relative to any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein. A "conservative amino acid substitution" is one in which an amino acid residue is replaced with another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). Generally, conservative amino acid substitutions do not substantially alter the functional properties of a protein. Examples of groups of amino acids having side chains with similar chemical properties include: (1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine, (2) aliphatic-hydroxyl side chains: serine and threonine, (3) amide-containing side chains: asparagine and glutamine, (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan, (5) basic side chains: lysine, arginine, and histidine, (6) acidic side chains: aspartic acid and glutamic acid, and (7) sulfur-containing side chains: cysteine ​​and methionine. Preferred conservative amino acid substitution groups are valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine. Alternatively, a conservative replacement is any change that has a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256:1443-1445, incorporated herein by reference. A "moderately conservative" replacement is any change that has a non-negative value in the PAM250 log-likelihood matrix.

[0187] The present disclosure also includes antigen-binding molecules comprising an antigen-binding domain having an HCVR, LCVR, and / or CDR amino acid sequence substantially identical to any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein. When referring to amino acid sequences, the term "substantial identity" or "substantially identical" means that two amino acid sequences, when optimally aligned using a program such as GAP or BESTFIT with a predetermined gap weight, share at least 95% sequence identity, and even more preferably at least 98% or 99% sequence identity. Preferably, residue positions that are not identical differ by conservative amino acid substitutions. When two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity or degree of similarity may be adjusted upward to correct for the conservative nature of the substitutions. Means for making this adjustment are well known to those skilled in the art. See, for example, Pearson (1994) Methods Mol. Biol. 24:307-331.

[0188] Sequence similarity for polypeptides, also referred to as sequence identity, is typically measured using sequence analysis software. Protein analysis software matches similar sequences using similarity measures assigned to various substitutions, deletions, and other modifications, including conservative amino acid substitutions. For example, GCG software includes programs such as Gap and Bestfit, which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms, or between a wild-type protein and its mutant protein. See, for example, GCG version 6.1. Polypeptide sequences can also be compared using FASTA, a program in GCG version 6.1, using default or recommended parameters. FASTA (e.g., FASTA2 and FASTA3) provides alignments and percent sequence identity of the regions of best overlap between the query and search sequences (Pearson (2000) supra). Another preferred algorithm for comparing the sequences of the present disclosure to a database containing a large number of sequences from different organisms is the computer program BLAST, particularly BLASTP or TBLASTN, using default parameters. See, e.g., Altschul et al. (1990) J. Mol. Biol. 215:403-410 and Altschul et al. (1997) Nucleic Acids Res. 25:3389-402.

[0189] (ii) pH-dependent binding The present disclosure includes anti-CD28 / anti-CD22 bispecific antigen-binding molecules with pH-dependent binding properties. For example, an anti-CD28 antibody of the present disclosure may exhibit reduced binding to CD28 at acidic pH compared to neutral pH. Alternatively, an anti-CD22 antibody of the present disclosure may exhibit enhanced binding to CD22 at acidic pH compared to neutral pH. The term "acidic pH" includes pH values ​​below about 6.2, e.g., about 6.0, 5.95, 5.9, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0, or less. As used herein, the term "neutral pH" refers to a pH of about 7.0 to about 7.4. The expression "neutral pH" includes pH values ​​of about 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35, and 7.4.

[0190] In certain instances, "reduced binding...at acidic pH compared to neutral pH" is expressed in terms of the ratio of the K value of an antibody that binds to its antigen at acidic pH to the K value of the antibody that binds to its antigen at neutral pH (or vice versa). For example, for purposes of this disclosure, an antibody or antigen-binding fragment thereof can be considered to exhibit "reduced binding to CD28 at acidic pH compared to neutral pH" if the antibody or antigen-binding fragment thereof exhibits an acidic / neutral K ratio of about 3.0 or greater. In certain exemplary embodiments, the acidic / neutral KD ratio for an antibody or antigen-binding fragment of the disclosure can be about 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 20.0, 25.0, 30.0, 40.0, 50.0, 60.0, 70.0, 100.0, or more.

[0191] Antibodies with pH-dependent binding properties may be obtained, for example, by screening a population of antibodies for reduced (or enhanced) binding to a particular antigen at acidic pH compared to neutral pH. Furthermore, modification of the antigen-binding domain at the amino acid level may generate antibodies with pH-dependent properties. For example, substituting one or more amino acids in the antigen-binding domain (e.g., within the CDRs) with histidine residues may result in antibodies with reduced antigen binding at acidic pH relative to neutral pH.

[0192] (iii) an antibody containing an Fc variant According to certain embodiments of the present disclosure, anti-CD28 / anti-CD22 bispecific antigen-binding molecules are provided, comprising an Fc domain containing one or more mutations that enhance or decrease antibody binding to the FcRn receptor, e.g., at acidic pH compared to neutral pH. For example, the present disclosure includes antibodies and antigen-binding molecules containing mutations in the CH2 or CH3 region of the Fc domain, where the mutation(s) increase the affinity of the Fc domain for FcRn in acidic environments (e.g., in endosomes at a pH ranging from about 5.5 to about 6.0). Such mutations can result in an extended serum half-life of the antibody when administered to an animal. Non-limiting examples of such Fc modifications include, for example, modifications at positions 250 (e.g., E or Q), 250 and 428 (e.g., L or F), 252 (e.g., L / Y / F / W or T), 254 (e.g., S or T), and 256 (e.g., S / R / Q / E / D or T), or modifications at positions 428 and / or 433 (e.g., H / L / R / S / P / Q or K) and / or 434 (e.g., H / F or Y), or modifications at positions 250 and / or 428, or at positions 307 or 308 (e.g., 308F, V308F), and 434. In one embodiment, the modifications include 428L (e.g., M428L) and 434S (e.g., N434S) modifications, 428L, 259I (e.g., V259I), and 308F (e.g., V308F) modifications, 433K (e.g., H433K) and 434 (e.g., 434Y) modifications, 252, 254, and 256 (e.g., 252Y, 254T, and 256E) modifications, 250Q and 428L modifications (e.g., T250Q and M428L), 307 and / or 308 modifications (e.g., 308F or 308P).

[0193] For example, the present disclosure includes anti-CD28 / anti-CD22 bispecific antigen-binding molecules comprising an Fc domain containing one or more pairs or groups of mutations selected from the group consisting of: 250Q and 248L (e.g., T250Q and M248L); 252Y, 254T and 256E (e.g., M252Y, S254T and T256E); 428L and 434S (e.g., M428L and N434S); and 433K and 434F (e.g., H433K and N434F). All possible combinations of the foregoing Fc domain mutations, and other mutations in antibody variable domains disclosed herein, are contemplated as being within the scope of the present disclosure.

[0194] Bispecific CD3×CD20 antibodies and antigen-binding fragments thereof According to certain exemplary embodiments of the present disclosure, the method comprises administering a therapeutically effective amount of a bispecific antigen-binding molecule or antigen-binding fragment thereof that binds to CD3 and CD20, in combination with a bispecific CD22xCD28 antigen-binding molecule or antigen-binding fragment thereof.

[0195] CD3 is a homodimeric or heterodimeric antigen expressed on T cells in conjunction with the T cell receptor complex (TCR) and is required for T cell activation. Functional CD3 is formed from the dimeric association of two of four different chains: epsilon, zeta, delta, and gamma. Dimeric configurations of CD3 include gamma / epsilon, delta / epsilon, and zeta / zeta. As used herein, molecules that "bind to CD3" include antibodies and antigen-binding fragments thereof that specifically recognize a single CD3 subunit (e.g., epsilon, delta, gamma, or zeta), as well as antibodies and antigen-binding fragments thereof that specifically recognize a dimeric complex of two CD3 subunits (e.g., gamma / epsilon, delta / epsilon, and zeta / zeta CD3 dimers). The bispecific CD3xCD20 antibodies and antigen-binding fragments used in the methods of the present disclosure may bind to soluble CD3 and / or cell-surface-expressed CD3. Soluble CD3 includes native CD3 protein as well as recombinant CD3 protein variants that lack the transmembrane domain or are not otherwise associated with the cell membrane, such as monomeric and dimeric CD3 constructs.

[0196] CD20 is a nonglycosylated phosphoprotein expressed on the plasma membrane of mature B cells. It is expressed by more than 95% of B-cell non-Hodgkin's lymphomas (NHLs) and other B-cell malignancies, but is absent from precursor B cells, dendritic cells, and plasma cells, and is therefore considered a B-cell tumor-associated antigen.

[0197] As used in the context of this disclosure, an antibody that "specifically binds" to CD3 or CD20 includes an antibody or antigen-binding fragment that binds to CD3 or CD20, or a portion thereof, with a KD of less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, or less than about 0.5 nM, as measured in a surface plasmon resonance assay. However, an isolated antibody or antigen-binding fragment that specifically binds to human CD3 or CD20 may have cross-reactivity to other antigens, such as CD3 or CD20 molecules from other (non-human) species.

[0198] According to certain exemplary embodiments of the present disclosure, the bispecific CD3xCD20 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR), and / or a complementarity determining region (CDR) comprising any of the amino acid sequences of the anti-CD3 antibodies and anti-CD20 antibodies described in U.S. Pat. No. 9,657,102.

[0199] In certain exemplary embodiments, the CD3-binding arm of a bispecific CD3xCD20 antibody or antigen-binding fragment thereof that can be used in the context of the methods of the present disclosure comprises a heavy chain complementarity-determining region (HCDR) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:5, and a light chain complementarity-determining region (LCDR) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:6.

[0200] According to certain embodiments, the CD3-binding arm of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2 and HCDR3) and three LCDRs (LCDR1, LCDR2 and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 10, HCDR2 comprises the amino acid sequence of SEQ ID NO: 11, HCDR3 comprises the amino acid sequence of SEQ ID NO: 12, LCDR1 comprises the amino acid sequence of SEQ ID NO: 13, LCDR2 comprises the amino acid sequence of SEQ ID NO: 14 and LCDR3 comprises the amino acid sequence of SEQ ID NO: 15.

[0201] In still other embodiments, the CD3 binding arm of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof comprises an HCVR comprising SEQ ID NO: 5 and an LCVR comprising SEQ ID NO: 6. In certain embodiments, the methods of the disclosure comprise the use of a bispecific CD3xCD20 antibody, wherein the CD3 binding arm comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the CD3 binding arm comprises a light chain comprising the amino acid sequence of SEQ ID NO: 3.

[0202] In certain exemplary embodiments, the CD20-binding arm of a bispecific CD3xCD20 antibody or antigen-binding fragment thereof that can be used in the context of the methods of the present disclosure comprises a heavy chain complementarity-determining region (HCDR) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:4, and a light chain complementarity-determining region (LCDR) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:6.

[0203] According to certain embodiments, the CD20-binding arm of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2 and HCDR3) and three LCDRs (LCDR1, LCDR2 and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 7, HCDR2 comprises the amino acid sequence of SEQ ID NO: 8, HCDR3 comprises the amino acid sequence of SEQ ID NO: 9, LCDR1 comprises the amino acid sequence of SEQ ID NO: 13, LCDR2 comprises the amino acid sequence of SEQ ID NO: 14 and LCDR3 comprises the amino acid sequence of SEQ ID NO: 15.

[0204] In still other embodiments, the CD20-binding arm of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof comprises an HCVR comprising SEQ ID NO: 4 and an LCVR comprising SEQ ID NO: 6. In certain embodiments, the methods of the disclosure comprise the use of a bispecific CD3xCD20 antibody, wherein the CD20-binding arm comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the CD20-binding arm comprises a light chain comprising the amino acid sequence of SEQ ID NO: 3.

[0205] An exemplary antibody comprising an HCVR in the CD3 binding arm comprising the amino acid sequence of SEQ ID NO: 5, an HCVR in the CD20 binding arm comprising the amino acid sequence of SEQ ID NO: 4, and a common LCVR comprising the amino acid sequence of SEQ ID NO: 6 is the antibody known as odronextamab (also known as REGN1979).

[0206] According to certain exemplary embodiments, the methods of the present disclosure include the use of odronextamab or a biological equivalent thereof. As used herein, the term "biological equivalent" refers to an antigen-binding protein (e.g., an antibody or antigen-binding fragment thereof) that binds to CD3 and CD20 and is a pharmaceutical equivalent or pharmaceutical substitute that does not exhibit a significantly different absorption rate and / or extent from odronextamab when administered in the same molar amount under similar experimental conditions, either in a single dose or multiple doses. In the context of the present disclosure, this term refers to an antigen-binding protein that binds to CD3 and CD20 and that does not have clinically meaningful differences from odronextamab in its safety, purity, and / or efficacy.

[0207] biological equivalent The present disclosure encompasses antigen-binding molecules having amino acid sequences that differ from those of the described antibodies but retain the ability to bind to CD28 and CD22 or CD3 and CD20. Such variant molecules contain one or more additions, deletions, or substitutions of amino acids compared to the parent sequence, but exhibit essentially equivalent biological activity to that of the described antigen-binding molecules. Similarly, the DNA sequences encoding the antigen-binding molecules of the present disclosure encompass sequences that contain one or more additions, deletions, or substitutions of nucleotides compared to the disclosed sequences, but encode antigen-binding molecules that are essentially biologically equivalent to the described antigen-binding molecules of the present disclosure. Examples of such variant amino acid sequences and DNA sequences are discussed above.

[0208] The present disclosure includes antigen-binding molecules that are biologically equivalent to any of the exemplary antigen-binding molecules described herein.Two antigen-binding proteins or antibodies are considered to be pharmaceutical equivalents or pharmaceutical substitutes if, for example, they are administered at the same molar dose, either in a single dose or multiple doses, under similar experimental conditions, and the absorption rate and extent of absorption do not show significant differences.Some antibodies are considered to be equivalents or pharmaceutical substitutes if their absorption rate is equivalent but their absorption rate is not, and such differences in absorption rate are intentional and reflected in the labeling, and therefore can be considered biologically equivalent, for example, they are not essential for achieving effective body drug concentrations in long-term use and are not considered medically significant for the particular formulation studied.

[0209] In one embodiment, two antigen binding proteins are bioequivalent if there are no clinically significant differences in their safety, purity, and efficacy.

[0210] In one embodiment, two antigen binding proteins are bioequivalent if a subject can be switched one or more times compared to therapy sustained without switching between the reference product and the biological product without an expected increase in the risk of adverse effects, including clinically significant changes in immunogenicity or reduced efficacy.

[0211] In one embodiment, two antigen binding proteins are biologically equivalent if they both operate by a common mechanism or mode of action for the condition(s) of use, to the extent that such mechanism is known.

[0212] Bioequivalence may be demonstrated by in vivo and / or in vitro methods. Bioequivalence measurements include, for example, (a) in vivo studies in humans or other mammals in which the concentration of an antibody or its metabolites is measured as a function of time in blood, plasma, serum, or other biological fluids, (b) in vitro studies that correlate with and reasonably predict human bioavailability data, (c) in vivo studies in humans or other mammals in which the relevant acute pharmacological effect of the antibody (or its target) is measured as a function of time, and (d) well-controlled clinical trials that establish the safety, efficacy, or bioavailability or bioequivalence of the antibody.

[0213] Biologically equivalent variants of the exemplary bispecific antigen-binding molecules described herein can be constructed, for example, by making various substitutions of residues or sequences or by deleting terminal or internal residues or sequences that are not required for biological activity. For example, cysteine ​​residues that are not essential for biological activity can be deleted or replaced with other amino acids to prevent unnecessary or incorrect intramolecular disulfide bridge formation during renaturation. In other contexts, biologically equivalent antibodies can include exemplary bispecific antigen-binding molecules described herein that contain amino acid changes that modify the glycosylation characteristic of antibodies, for example, mutations that eliminate or remove glycosylation.

[0214] Therapeutic Formulations and Administration The present disclosure provides pharmaceutical compositions comprising the antigen-binding molecules of the present disclosure. The pharmaceutical compositions of the present disclosure are formulated with suitable carriers, excipients, and other agents that improve transport, delivery, and tolerance. Many suitable formulations can be found in formularies known to all pharmacists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic)-containing vesicles (such as LIPOFECTIN™, Life Technologies, Carlsbad, CA), DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, emulsion carbowax (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. See also Powell et al. "Compendium of excipients for parenteral formulations" PDA (1998) J Pharm Sci Technol 52:238-311.

[0215] The dose of an antigen-binding molecule administered to a subject may vary depending on the subject's age and physique, the target disease, condition, route of administration, etc. A preferred dose is typically calculated based on body weight or body surface area. When a bispecific antigen-binding molecule of the present disclosure is used for therapeutic purposes in an adult subject, it may be advantageous to administer the bispecific antigen-binding molecule of the present disclosure intravenously, typically in a single dose of about 0.01 to 400 mg. The frequency and duration of treatment can be adjusted depending on the severity of the condition.

[0216] Various delivery systems, such as liposomes, microparticles, encapsulation in microcapsules, recombinant cells capable of expressing mutant viruses, and receptor-mediated endocytosis, are known and can be used to administer the pharmaceutical compositions of the present disclosure (see, for example, Wu et al., 1987, J. Biol. Chem. 262:4429-4432). Introduction methods include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered by any convenient route, for example, by infusion or bolus injection, or by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal, and intestinal mucosa), and can be administered together with other biologically active agents. Administration can be systemic or local.

[0217] The pharmaceutical composition of the present disclosure can be delivered subcutaneously or intravenously using a standard needle and syringe. In addition, for subcutaneous delivery, a pen delivery device is easily used to deliver the pharmaceutical composition of the present disclosure. Such a pen delivery device can be reusable or disposable. Reusable pen delivery devices generally utilize a replaceable cartridge containing the pharmaceutical composition. Once the pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen delivery device can then be reused. In disposable pen delivery devices, there is no replaceable cartridge. Rather, disposable pen delivery devices are pre-filled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is empty of the pharmaceutical composition, the entire device is discarded.

[0218] Numerous reusable pen delivery devices and autoinjector-type delivery devices have application in the subcutaneous delivery of the pharmaceutical compositions of the present disclosure. Examples include the AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), the DISETRONIC™ pen (Disetronic Medical Systems, Bergdorf, Switzerland), the HUMALOG MIX 75 / 25™ pen, the HUMALOG™ pen, the HUMALIN 70 / 30™ pen (Eli Lilly and Co., Indianapolis, IN), the NOVOPEN™ I, II, and III (Novo Nordisk, Copenhagen, Denmark), the NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), the BD™ pen (Becton Dickinson, Franklin Lakes, NJ), the OPTIPEN™, the OPTIPEN PRO™, the OPTIPEN™, to name a few. Examples of disposable pen delivery devices that have applications in the subcutaneous delivery of pharmaceutical compositions of the present disclosure include, but are not limited to, the SOLOSTAR pen (Sanofi-Aventis), FLEXPEN (Novo Nordisk), and KWIKPEN (Eli Lilly), the SURECLICK auto-injector (Amgen, Thousand Oaks, CA), the PENLET pen (Haselmeier, Stuttgart, Germany), the EPIPEN pen (Dey, LP), and the HUMIRA pen (Abbott Labs, Abbott Park, IL), to name a few.

[0219] In certain circumstances, pharmaceutical compositions can be delivered in a sustained-release system. In one embodiment, a pump can be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, a polymeric material can be used; see Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Press, Boca Raton, Florida. In yet another embodiment, a sustained-release system can be placed near the target of the composition, thereby requiring only a fraction of the systemic dose (see, e.g., Goodson, 1984, Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138). Other sustained-release systems are discussed in the review by Langer, 1990, Science 249:1527-1533.

[0220] The injectable preparations may include dosage forms for intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection, drip injection, etc. These injectable preparations may be prepared by publicly known methods. For example, the injectable preparations may be prepared by dissolving, suspending, or emulsifying the above-mentioned antibody or its salt in a sterile aqueous medium or oily medium conventionally used for injections. The injections prepared in this manner are preferably filled into appropriate ampoules.

[0221] Advantageously, the pharmaceutical compositions for oral or parenteral use described above are prepared into a suitable unit dosage form to accommodate the dose of the active ingredient. Examples of such unit dosage forms include tablets, pills, capsules, injections (ampoules), suppositories, etc. The amount of the antibody contained is generally about 5 to about 500 mg per dosage form in a unit dose. In particular, for injections, the antibody is preferably contained in an amount of about 5 to about 100 mg, and for other dosage forms, it is preferably contained in an amount of about 10 to about 250 mg.

[0222] Additional combination therapy The present disclosure includes methods for treating, ameliorating, or reducing the severity of at least one symptom or sign, or inhibiting the growth of cancer in a subject, comprising administering to a subject in need thereof a therapeutic composition comprising a multispecific (e.g., bispecific) antigen binding molecule that specifically binds CD28 and CD22, alone as monotherapy, or with a bispecific antibody that binds CD3 and CD20 (e.g., odronextamab) as combination therapy, and optionally with one or more therapeutic agents, e.g., at least a third therapeutic agent or therapy.

[0223] Exemplary third therapeutic agents or therapies that may be combined or administered in combination with the antigen binding molecules of the present disclosure include, for example, surgery, chemotherapy, radiation therapy, checkpoint inhibitors that target PD-1 (e.g., anti-PD-1 antibodies such as pembrolizumab, nivolumab, or cemiplimab), costimulatory agonist bivalent antibodies that target molecules such as CTLA-4, LAG3, TIM3, GITR, OX40, 4-1BB, and the like, CD3x bispecific antibodies (e.g., see US9,657,102, WO2017 / 053856A1, WO2014 / 047231A1, WO2018 / 0673 31A1 and WO2018 / 058001A1), other antibodies targeting CD22×CD3, CD22×CD28, or targeting CD20×CD3, other costimulatory CD28× bispecific antibodies, oncolytic viruses, cancer vaccines, tamoxifen, aromatase inhibitors, cytokine inhibitors including small molecule cytokine inhibitors, and antibodies that bind to cytokines such as IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-9, IL-11, IL-12, IL-13, IL-17, IL-18, or their respective receptors.Anti-CD28 / anti-CD22 antigen binding molecules of the present disclosure (e.g., pharmaceutical compositions comprising the anti-CD28 / anti-CD22 bispecific antigen binding molecules disclosed herein) can also be used in combination with other anti-CD28 / anti-CD22 drugs, including "ICE": ifosfamide (e.g., Ifex®), carboplatin (e.g., Paraplatin®), etoposide (e.g., Etopophos®, Toposar®, VePesid®, VP-16), "DHAP": dexamethasone (e.g., Decadron®), cytarabine (e.g., Cyto It may be administered as part of a treatment regimen that includes a combination of one or more therapeutic agents selected from "ESHAP": etoposide (e.g., Etopophos®, Toposar®, VePesid®, VP-16), methylprednisolone (e.g., Medrol®), high-dose cytarabine, cisplatin (e.g., Platinol®-AQ), sar-U®, cytosine arabinoside, ara-C), cisplatin (e.g., Platinol®-AQ), and "ESHAP": etoposide (e.g., Etopophos®, Toposar®, VePesid®, VP-16), methylprednisolone (e.g., Medrol®), high-dose cytarabine, cisplatin (e.g., Platinol®-AQ). The anti-CD28 / anti-CD22 antigen binding molecules of the present disclosure may also be administered in combination with any of the antigen binding molecules mentioned herein and one or more inhibitors of VEGF, Ang2, DLL4, EGFR, ErbB2, ErbB3, ErbB4, EGFRvIII1, cMet, IGF1 R, B-raf, PDGFR-o, PDGFR-I3, FOLH1, PRLR, STEAP1, STEAP2, TMPRSS2, MSLN, CA9, uroplakin, or any of the aforementioned cytokines, wherein the inhibitor is an aptamer, antisense molecule, ribozyme, siRNA, peptibody, nanobody, or antibody fragment (e.g., Fab fragment, F(ab')2 fragment, Fd fragment, Fv fragment, scFv, dAb fragment, or other engineered molecules such as diabodies, triabodies, tetrabodies, minibodies, and minimal recognition units). The anti-CD28 / anti-CD22 antigen binding molecules of the present disclosure may also be administered in combination with and / or co-formulated with antivirals, antibiotics, analgesics, corticosteroids and / or NSAIDs.The antigen binding molecules of the present disclosure may also be administered as part of a treatment regimen that also includes radiation therapy and / or conventional chemotherapy, or treatment with biologics, including checkpoint inhibitors or other bispecific antibodies.

[0224] The present disclosure includes compositions and therapeutic formulations comprising any of the antigen-binding molecules described herein in combination with one or more chemotherapeutic agents. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (Cytoxan™); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine (trimethylolpropane). ethylenimines and methylamelamine, including methylolomelamine; nitrogen mustards, e.g., chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoure Antibiotics such as aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norloquinone; isin, doxorubicin, epirubicin, esorubicin, idarubicin, marcelomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, chelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU);Folic acid analogues, for example, denopterin, methotrexate, pteropterin, trimetrexate; purine analogues, for example, fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues, for example, ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens, for example, calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; antiadrenal drugs, for example, aminoglutethimide, mitotane, trilostane; folic acid supplements, for example, florinic acid acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestravsil; bisantrene; edatrexate; defofamine; demecolcine; diaziconazole; elfornithine; elliptinium acetate; etoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; fenamet; pirarubicin; podophyllinic acid acid); 2-ethylhydrazide; procarbazine; PSK™; razoxane; sizofiran; spirogermanium; tenuazonic acid; triazicon; 2,2',2''-trichlorotriethylamine; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxanes, such as paclitaxel (Taxol™, Bristol-Myers Squibb Oncology, Princeton, NJ) and docetaxel (Taxotere™; Aventis Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide;Included within this definition are daunomycin, aminopterin, xeloda, ibandronate, CPT-11, the topoisomerase inhibitor RFS 2000, difluoromethylornithine (DMFO), retinoic acid, esperamicin, capecitabine, and pharmaceutically acceptable salts, acids, or derivatives of any of the above. Also included within this definition are antihormonal agents that act to regulate or inhibit hormone action on tumors, such as antiestrogens, including tamoxifen, raloxifene, aromatase-inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene (Fareston), and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin, and pharmaceutically acceptable salts, acids, or derivatives of any of the above.

[0225] The additional therapeutically active ingredient(s) may be administered immediately before, simultaneously with, or immediately after administration of the antigen-binding molecule of the present disclosure (for purposes of the present disclosure, such an administration regimen is considered administration of the antigen-binding molecule "in combination" with the additional therapeutically active ingredient).

[0226] The present disclosure includes pharmaceutical compositions in which the antigen-binding molecules of the present disclosure are co-formulated with one or more of the additional therapeutically active ingredient(s) described elsewhere herein. [Example]

[0227] The following examples are put forward so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the disclosed methods and compositions, and are not intended to limit the scope of what the inventors regard as the present disclosure.

[0228] Example 1: Bispecific CD22xCD28 and CD3xCD20 antibodies Bispecific CD22xCD28 antibodies are described in WO2020 / 132066, the entire contents of which are expressly incorporated herein by reference. An exemplary bispecific CD22xCD28 antibody used in the examples below is REGN5837.

[0229] Table 1 lists the amino acid sequence identifiers for the heavy and light chain variable regions and CDRs of REGN5837. [Table 1]

[0230] Sequence of the heavy chain of the CD22 binding arm of REGN5837: HCVR:EVQLVQSGAEVKKPGESLKISCKGSGYNFATYWIAWVRQMPGKGLELMGIIYPGDSETTYNPSFQGQVTISADKSISNAYLQWSSLKASDTAMYYCARVGGYCSGTSCHNWFDPWGLGTLVTVSS (SEQ ID NO: 19) HCDR1: GYNFATYW (SEQ ID NO: 22) HCDR2: IYPGDSET (SEQ ID NO: 23) HCDR3: ARVGGYCSGTSCHNWFDP (SEQ ID NO: 24) HC:EVQLVQSGAEVKKPGESLKISCKGSGYNFATYWIAWVRQMPGKGLELMGIIYPGDSETTYNPSFQGQVTISADKSISNAYLQWSSLKASDTAMYYCARVGGYCSGTSCHNWF DPWGLGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 16)

[0231] Sequence of the heavy chain of the CD28 binding arm of REGN5837: HCVR:QVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGLEWIGYIYYSGITHYNPSLKSRVTISVDTSKIQFSLKLSSVTAADTAVYYCARWGVRRDYYYYGMDVWGQGTTVTVSS (SEQ ID NO: 20) HCDR1: GGSISSYY (SEQ ID NO: 25) HCDR2: IYYSGIT (SEQ ID NO: 26) HCDR3: ARWGVRRDYYYYGMDV (SEQ ID NO: 27) HCVR:QVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGLEWIGYIYYSGITHYNPSLKSRVTISVDTSKIQFSLKLSSVTAADTAVYYCARWGVRRDYYYYGMD VWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNRFTQKSLSLSPGK (SEQ ID NO: 17)

[0232] The sequence of the common light chain for both the CD22 and CD28 binding arms of REGN5837: LCVR: EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK (SEQ ID NO: 21) LCDR1: QSVSSSY (SEQ ID NO: 28) LCDR2:GAS (SEQ ID NO: 29) LCDR3: QQYGSSPWT (SEQ ID NO: 30) LC:EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 18)

[0233] Bispecific CD3xCD20 antibodies are described in US 9,657,102, the entire contents of which are expressly incorporated herein by reference. An exemplary bispecific CD3xCD20 antibody used in the examples below is REGN1979.

[0234] Table 2 lists the amino acid sequence identifiers for the heavy and light chain variable regions and CDRs of REGN1979. [Table 2]

[0235] Sequence of the heavy chain of the CD3 binding arm of REGN1979: HCVR:EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYTMHWVRQAPGKGLEWVSGISWNSGSIGYADSVKGRFTISRDNAKKSLYLQMNSLRAEDTALYYCAKDNSGYGHYYYGMDVWGQGTTVTVAS (SEQ ID NO: 5) HCDR1: GFTFDDYT (SEQ ID NO: 10) HCDR2: ISWNSGSI (SEQ ID NO: 11) HCDR3: AKDNSGYGHYYYGMDV (SEQ ID NO: 12) HC:EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYTMHWVRQAPGKGLEWVSGISWNSGSIGYADSVKGRFTISRDNAKKSLYLQMNSLRAEDTALYYCAKDNSGYGHYYYGMD VWGQGTTVTVASASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNRFTQKSLSLSLGK (SEQ ID NO: 2)

[0236] Sequence of the heavy chain of the CD20 binding arm of REGN1979: HCVR:EVQLVESGGGLVQPGRSLRLSCVASGFTFNDYAMHWVRQAPGKGLEWVSVISWNSDSIGYADSVKGRFTISRDNAKNSLYLQMHSLRAEDTALYYCAKDNHYGSGSYYYYQYGMDVWGQGTTVTVSS (SEQ ID NO: 4) HCDR1: GFTFNDYA (SEQ ID NO: 7) HCDR2: ISWNSDSI (SEQ ID NO: 8) HCDR3: AKDNHYGSGSYYYYQYGMDV (SEQ ID NO: 9) HC:EVQLVESGGGLVQPGRSLRLSCVASGFTFNDYAMHWVRQAPGKGLEWVSVISWNSDSIGYADSVKGRFTISRDNAKNSLYLQMHSLRAEDTALYYCAKDNHYGSGSYYYYQY GMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESK YGPPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 1)

[0237] The sequence of the common light chain of both the CD3 and CD20 binding arms of REGN1979: LCVR: EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQHYINWPLTFGGGTKVEIKR (SEQ ID NO: 6) LCDR1:QSVSSN (SEQ ID NO: 13) LCDR2:GAS (SEQ ID NO: 14) LCDR3: QHYINWPLT (SEQ ID NO: 15) LC:EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQHYINWPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 3)

[0238] Example 2: Clinical Evaluation of Treatment with REGN5837 and REGN1979 in Combination This example describes a Phase 1 clinical trial of REGN5837 (a CD22×CD28 costimulatory bispecific antibody) in combination with REGN1979 (odronextamab) in subjects with relapsed or refractory aggressive B-cell non-Hodgkin's lymphoma.

[0239] The primary objectives of the study are to evaluate safety, tolerability, and dose-limiting toxicities (DLTs) and to determine the recommended phase 2 dose (RP2D) regimen(s) (maximum tolerated dose (MTD) regimen or lower dose regimen) of REGN5837 in combination with odronextamab in subjects with relapsed or refractory aggressive B-NHL.

[0240] Secondary objectives of this study are to (i) characterize the pharmacokinetics (PK) of REGN5837 when given in combination with REGN1979 (odronextamab), (ii) evaluate the PK of REGN1979 (odronextamab) when given in combination with REGN5837, (iii) evaluate the immunogenicity of REGN5837 and odronextamab, and (iv) evaluate the preliminary antitumor activity of REGN5837 in combination with REGN1979 (odronextamab) in subjects with relapsed or refractory aggressive B-NHL.

[0241] The exploratory objectives of this study are to (i) evaluate the association between biomarkers of systemic immune activation (serum cytokine levels, T cell counts and activation markers) and clinical efficacy and safety; (ii) evaluate the association between disease response and / or recurrence and change from baseline in counts and phenotypes of tumor-infiltrating T cells and tumor B cell target antigen (CD20, CD22) expression; (iii) evaluate the association between molecular minimal residual disease (MRD) status and progression-free survival (PFS) and overall survival (OS) in subjects with a clinical complete response (CR); (iv) evaluate other biomarkers (pharmacodynamic, predictive, and prognostic), antitumor activity, and safety potentially relevant to REGN5837 in combination with REGN1979 (odronextamab) exposure; and (v) evaluate the relationship between pharmacodynamics, drug concentrations, and clinical safety and efficacy measures.

[0242] Test Design The objective of this Phase 1, open-label, FIH study is to evaluate the safety, PK and pharmacodynamic properties, and preliminary clinical activity of the anti-CD22 × anti-CD28 bispecific antibody REGN5837 in combination with the anti-CD20 × anti-CD3 bispecific antibody odronextamab (REGN1979) in subjects with relapsed or refractory aggressive B-NHL (excluding mantle cell lymphoma (MCL) and hereafter simply referred to as "aggressive B-NHL"). The study has two parts: dose escalation and dose expansion.

[0243] To reduce the likelihood and severity of cytokine release and mitigate the risk of TLS, REGN1979 (odronextamab) is introduced as monotherapy from day 1 of cycle 1 to day 8 of cycle 2. REGN5837 is initiated on day 15 of cycle 2. REGN5837 will not be initiated until a QW dose of odronextamab has been administered as a single infusion the previous week without signs or symptoms of cytokine release syndrome (CRS), infusion-related reaction (IRR), or tumor lysis syndrome (TLS). Sequential introduction of REGN1979 (odronextamab) and REGN5837 will allow subjects who developed CRS, IRR, or TLS with odronextamab monotherapy to recover before receiving REGN5837. The dose of odronextamab is escalated through day 1 of cycle 2, with an initial dose administered on days 1 and 2 of cycle 1, followed by an intermediate dose on days 8 and 9 of cycle 1, a second intermediate dose on days 15 and 16 of cycle 1, and full weekly doses on day 1 and thereafter of cycle 2. In ongoing trials with odronextamab, no severe CRS has been observed in subjects with diffuse large B-cell lymphoma (DLBCL) once subjects tolerate two full nominal doses of odronextamab.

[0244] To further reduce the risk of severe CRS, IRR, or TLS when REGN5837 is administered in the presence of odronextamab, following the same approach as escalating dosing with odronextamab, REGN5837 will also be administered using an escalating dosing regimen, with a lower initial dose on day 15 of cycle 2, an intermediate dose on day 1 of cycle 3, and a full weekly dose on day 8 of cycle 3.

[0245] Weekly dosing of REGN5837 and odronextamab continues until Day 8 of Cycle 6, after which dosing changes to a Q2W schedule. The weekly (QW) dosing period constitutes the induction period (Cycles 1-6), followed by Q2W administration as the maintenance period. Subjects who achieve and maintain a complete response (CR) for 9 months will transition to Q4W dosing. Subjects must have received at least three prior doses of their assigned dose on the Q2W dosing schedule before switching from Q2W to Q4W dosing.

[0246] The dose-escalation portion of the study will evaluate the safety of REGN5837 in combination with odronextamab and select the recommended Phase 2 dose (RP2D) regimen(s) for REGN5837 in combination with odronextamab according to a Bayesian optimal interval design (BOIN). The DLT observation period will be 35 days from the start of REGN5837 administration (nominally from Day 15 of Cycle 2 to Day 7 of Cycle 4) and will consist of at least three full doses of REGN5837 in combination with odronextamab.

[0247] The dose expansion portion will have two expansion cohorts: A and B. Cohort A will consist of subjects with aggressive B-NHL who have not previously received chimeric antigen receptor T-cell (CAR-T) therapy. Cohort B will consist of subjects with aggressive B-NHL who have progressed after failure of CAR-T therapy. Subjects enrolled in the expansion cohort will receive the RP2D of the combination treatment. Initial antitumor activity will be investigated, and safety and tolerability, PK properties, and biomarker responses will be further characterized.

[0248] Exam period After a 28-day screening period, study treatment will involve six 21-day cycles of induction dosing, including weekly therapy with odronextamab monotherapy from day 1 of cycle 1 to day 8 of cycle 2, followed by combination treatment with REGN5837 and odronextamab from day 15 of cycle 2 to day 8 of cycle 6.

[0249] During the maintenance period, which begins two weeks after the last induction, REGN5837 in combination with odronextamab will be administered every two weeks (Q2W) according to a 28-day cycle schedule. REGN5837 in combination with odronextamab will continue on the Q2W schedule until disease progression or other protocol-defined reasons for treatment discontinuation. Subjects who achieve and maintain a complete response (CR) for at least nine months will be switched to a Q4W schedule of REGN5837 and odronextamab until disease progression or other protocol-defined reasons for treatment discontinuation. Subjects must have received at least three prior doses of the assigned dose on the Q2W dosing schedule before switching from Q2W to Q4W dosing.

[0250] Safety Follow-Up: The safety follow-up period consists of three Q4W safety follow-up visits: 4 weeks after the last dose (Safety Follow-Up Visit 1), 8 weeks after the last dose (Safety Follow-Up Visit 2), and 12 weeks after the last dose (Safety Follow-Up Visit 3). Safety follow-up will continue until all three visits are completed, or until initiation of non-protocol antilymphoma therapy, or subject withdrawal of consent, whichever occurs first.

[0251] Extended Follow-Up: Extended follow-up will include subjects who discontinue study drug for reasons other than disease progression, initiation of non-protocol anti-lymphoma therapy, withdrawal of consent, or death. Disease response will be assessed until disease progression, death, initiation of non-protocol anti-lymphoma therapy, or withdrawal of subject consent for follow-up of disease status, whichever occurs first.

[0252] Survival Follow-Up: After the safety follow-up period and (if applicable) the extension follow-up period, all study subjects will be followed up at Q12W intervals for survival until death, loss to follow-up, withdrawal of subject consent for follow-up, or termination of the study by the sponsor, whichever occurs first. Survival follow-up status may be determined at a clinic visit or remotely (e.g., by telephone) by the study site.

[0253] The end of the study for an individual subject is when the subject discontinues the study, until the end of extended follow-up, and before survival follow-up. All subjects will continue survival follow-up until death, loss to follow-up, withdrawal of consent for follow-up, or the end of the study, whichever occurs first.

[0254] The study and all follow-up will end when all study subjects have been withdrawn from the study or at the end of the study, whichever occurs first.

[0255] Induction medication period Odronextamab Monotherapy (Cycle 1 Day 1–Cycle 2 Day 8): Participating sites should ensure that two doses of anti-IL6 therapy (e.g., tocilizumab) are available for each subject before any study drug is administered. From Cycle 1 Day 1 through Cycle 2 Day 8 of study treatment, odronextamab will be infused weekly as monotherapy. For the initial dose (0.7 mg divided as 0.2 mg / 0.5 mg), intermediate dose 1 (4 mg given as 2 mg / 2 mg divided infusions), and intermediate dose 2 (20 mg given as 10 mg / 10 mg divided infusions), odronextamab will be administered as split infusions, preferably on consecutive days but no more than three days apart, to improve tolerability and mitigate against CRS, IRR, and TLS. For example, the dosing days for odronextamab are: the initial dose on days 1 and 2 of cycle 1; the first intermediate dose on days 8 and 9 of cycle 1; and the second intermediate dose on days 15 and 16 of cycle 1. Each split dose is administered over 4 hours on each of the two days. The initial dose and both intermediate doses should always be split, even if a treatment delay causes administration beyond day 15 of cycle 1. In cycle 2, a full dose of 160 mg (note: 80 mg QW on DL1) is administered as a single infusion over 4 hours on day 1. On day 8 of cycle 2, another full dose of odronextamab is administered. If the administration of the first full QW dose of odronextamab is tolerated as a single infusion, subsequent doses (nominally from day 8 of cycle 2 onward) may be administered as single infusions over 1–4 hours, depending on prior tolerability. To be able to proceed to combination therapy, the most recent odronextamab dose must have been tolerated as a single infusion without grade CRS, IRR, or TLS.

[0256] Subjects should remain hospitalized during the odronextamab infusion and for at least 24 hours after the end of the infusion (including days of split dosing) until the full dose has been administered without CRS. Each infusion should be administered over 4 hours.

[0257] Weekly dosing of REGN5837 / odronextamab (Cycle 2 Day 15 - Cycle 6 Day 8): Initiation of REGN5837 is on Day 15 of Cycle 2. REGN5837 is also administered at escalating doses. All doses of REGN5837 are administered as a single infusion. For all dose levels, REGN5837 dosing will not begin until the subject has tolerated two full QW doses of odronextamab, as defined for each dose level (DL) (Table 4). Additionally, the most recent QW dose of odronextamab monotherapy must have been administered as a single infusion without CRS, IRR, or TLS grade before the subject can start REGN5837. Given that combination therapy with REGN5837 and odronextamab can worsen CRS, subjects who develop grade 3 CRS during the monotherapy run-in period are not eligible to receive REGN5837 and may continue on single-agent odronextamab for the duration of the study.

[0258] On Day 15 of Cycle 2 of study treatment, REGN5837 will be introduced at a lower initial dose, followed by QW dosing of odronextamab on Day 16. On Day 1 of Cycle 3, REGN5837 will be escalated to the intermediate dose. On Day 8 of Cycle 3, REGN5837 will be escalated to the full dose. When combination therapy is initiated, REGN5837 will be administered one day before odronextamab administration during Cycles 2 and 3. This staggered dosing period includes escalating dosing of REGN5837 on Day 15 of Cycle 2, Day 1 of Cycle 3, and Day 8 of Cycle 3, plus a second full combination dose on Day 15 of Cycle 3. If the staggered dosing is well tolerated without Grade 2 or higher CRS, same-day administration of REGN5837 and odronextamab will begin on Day 1 of Cycle 4. However, if Grade 2 or higher CRS occurs during staggered administration, same-day administration of the combination will be postponed until two full combination doses of REGN5837 and odronextamab are tolerated without such an event. Once initiated, same-day administration of REGN5837 and odronextamab will continue for the remainder of study treatment. During same-day administration of the combination, REGN5837 will always be administered first, followed by the initiation of odronextamab up to 60 minutes after the end of the REGN5837 infusion.

[0259] For doses less than 1 mg, the duration of the REGN5837 infusion is 1 hour, and REGN5837 is administered via a syringe pump. For doses equal to or greater than 1 mg, the duration of the REGN5837 infusion is 2 hours via an IV pump throughout the staggered dosing period and for the dose on Day 1 of Cycle 4. If the subject does not experience IRR, CRS, or TLS, the duration of the REGN5837 infusion may be reduced to 1 hour starting on Day 8 of Cycle 4. The duration of the infusion may be extended based on clinical judgment. The odronextamab infusion time and the second QW total dose during the REGN5837 escalation dosing period (Cycle 2 Day 15, Cycle 3 Day 1, Cycle 3 Day 8, and Cycle 3 Day 15) remain consistent with the infusion time on Day 8 of Cycle 2.

[0260] Once combination therapy is initiated, subjects should remain hospitalized for REGN5837 and odronextamab infusions until at least 48 hours after the completion of the REGN5837 infusion (24 hours after the completion of the odronextamab infusion). The observation period in the protocol varies based on the rationale that the highest risk of CRS development is within the first 24 hours after the end of the bispecific antibody infusion. During the hospitalization period, subjects should be monitored for IRR, CRS, and TLS during and after each infusion in accordance with institutional guidelines for observation. Monitoring includes, but is not limited to, vital signs (including temperature, blood pressure, and oxygen saturation), clinical and laboratory assessments (including at least serum chemistry).

[0261] Hospitalization for combination therapy administration will continue until both full doses of the combination therapy have been administered without the occurrence of CRS. Beginning on Day 1 of Cycle 4, hospitalization is not required as long as the subject receives and tolerates a full dose of REGN5837 in combination with a full dose of odronextamab. However, if the subject experiences Grade 2 or higher CRS on or after Day 8 of Cycle 3, the subject must be monitored in the hospital for at least 48 hours from the end of the REGN5837 infusion until they are tolerating both study drugs without the occurrence of a Grade 2 or higher CRS event.

[0262] After Day 1 of Cycle 4 or when hospitalization is no longer required (whichever occurs later), and during the first 4 weeks of outpatient infusion visits, subjects must be observed for at least 2 hours after the end of their last infusion. Clinical status assessments, including vital sign checks, must be performed at a minimum hourly frequency. Subjects must receive a safety assessment call the day after their first 4 outpatient infusion visits.

[0263] If the occurrence or persistence of a safety event prevents a subject from initiating combination treatment with REGN5837 by Day 15 of Cycle 3, odronextamab monotherapy may be continued at the assigned dose, provided the criteria for permanent discontinuation of odronextamab have not been met. In such cases, there will be no further attempts to introduce REGN5837, and the subject in the dose-escalation cohort may be replaced for DLT evaluation.

[0264] Subjects receive 12 weekly doses of REGN5837 and odronextamab combination therapy during the REGN5837 / odronextamab induction dosing period. Assuming REGN5837 is introduced on day 15 of cycle 2, this means that subjects will receive a total of 17 doses of odronextamab upon completion of the REGN5837 / odronextamab weekly dosing period. If there is a dosing interruption, the REGN5837 / odronextamab weekly dosing period will be extended until the subject has received 12 doses of REGN5837 in combination with odronextamab.

[0265] Prior Treatment: Prior treatment medication requirements to complete escalating doses of REGN5837 are described in Figure 3 and below.

[0266] The following premedication applies to initial odronextamab monotherapy from the initial dose through the first full weekly (QW) dose as a single infusion: If a subject has an IRR and / or CRS of any grade with the first full QW dose, continue premedication until a full QW dose is administered without experiencing an IRR and / or CRS. a) 12 to 24 hours prior to the planned start time of the first fractional infusion or the first QW full dose as a single infusion, and prior to the planned start of the second fractional infusion if each odronextamab dose is administered on non-consecutive days: i. Dexamethasone 10 mg PO or equivalent dose of steroid* b) Premedication on each day of odronextamab split infusions and on the day of the full QW dose as a single infusion: i. Dexamethasone 20 mg IV 1 to 3 hours before the start of the infusion ii. Diphenhydramine 25 mg IV or PO 30-60 minutes before infusion (may be substituted with another equivalent antihistamine) iii. Acetaminophen 650 mg PO 30-60 minutes prior to infusion (unless the subject received acetaminophen within 4 hours prior to the odronextamab infusion or is allergic to acetaminophen) c) 24 (±4) hours from the end of the second split infusion or the end of the first full QW dose as a single infusion: i. Dexamethasone 10 mg PO or equivalent dose of steroid*

[0267] The following premedication applies on the odronextamab treatment day of Cycle 2, Day 8 (or the second full dose of odronextamab), provided that the subject has not experienced any grade of IRR and / or CRS during the past 4 weeks of odronextamab monotherapy. One-day premedication of odronextamab as a single infusion: i. Dexamethasone 10 mg IV 1 to 3 hours before the start of the infusion on the day of treatment. ii. Diphenhydramine 25 mg IV or PO 30–60 minutes before (may be substituted with another equivalent antihistamine) iii. Acetaminophen 650 mg PO 30-60 minutes prior (unless the subject received odronextamab within 4 hours prior to the infusion or is allergic to acetaminophen)

[0268] The following premedication applies to REGN5837 in combination with odronextamab between Cycle 2 Day 15 and Cycle 3 Day 8 (escalating doses of REGN5837 in combination with odronextamab) (Figure 3). If a subject has an IRR and / or CRS of any grade with the first full REGN5837 QW dose, continue premedication until the full QW dose is tolerated without experiencing an IRR and / or CRS. a) 12 to 24 hours before the planned start of your REGN5837 infusion: i. Dexamethasone 10 mg PO or equivalent dose of steroid* b) Premedication on each day of REGN5837 / odronextamab combination therapy: ii. Dexamethasone 10 mg IV 1-3 hours before the start of the REGN5837 and odronextamab infusion on the treatment day iii. Diphenhydramine 25 mg IV or PO (may be substituted with another equivalent antihistamine) 30-60 minutes prior to REGN5837 and odronextamab infusion iv. Acetaminophen 650 mg PO 30-60 minutes prior to REGN5837 and odronextamab infusion (unless the subject received acetaminophen within 4 hours prior to REGN5837 or odronextamab infusion or is allergic to acetaminophen) c) 24 (± 4) hours from the end of odronextamab infusion: i. Dexamethasone 10 mg PO or equivalent dose of steroid* *Steroid doses equivalent to dexamethasone 10 mg include prednisone / prednisolone 60 mg or methylprednisolone 50 mg (PO dose only).

[0269] The following premedication applies to REGN5837 in combination with odronextamab on days 15 and 16 of Cycle 3 or the second full combination dose of REGN5837 and odronextamab without grade IRR and / or CRS, whichever occurs later. Premedication on each infusion day: i. Dexamethasone 10 mg IV 1 to 3 hours before the start of the REGN5837 and odronextamab infusion on the treatment day ii. Diphenhydramine 25 mg IV or PO (may be substituted with another equivalent antihistamine) 30-60 minutes prior to REGN5837 and odronextamab infusion iii. Acetaminophen 650 mg PO 30-60 minutes prior to REGN5837 and odronextamab infusion (unless the subject has received REGN5837 or odronextamab within 4 hours prior to the infusion or is allergic to acetaminophen)

[0270] For subsequent doses of REGN5837 in combination with odronextamab: a) If the subject continues to adequately tolerate REGN5837 and odronextamab combination treatment without grade IRR and / or CRS at dexamethasone doses as described above, discontinuation of subsequent REGN5837 and odronextamab combination administration of corticosteroids (along with diphenhydramine and acetaminophen) is encouraged. b) If continuous corticosteroid administration is warranted, another intermediate or long-acting corticosteroid (e.g., methylprednisolone) may be substituted for dexamethasone.

[0271] If at any point during treatment the subject experiences CRS or IRR of any grade, premedication with acetaminophen / antihistamine is also required for the next dose.

[0272] Additional premedication with antihistamines, acetaminophen, and / or corticosteroid equivalents may also be considered.

[0273] If two subjects experience a grade 3 or higher CRS event, the dose of oral dexamethasone (or equivalent) before and after dosing for that cohort will be increased to 20 mg.

[0274] Maintenance medication period Q2W and Q4W Dosing of REGN5837 / odronextamab: After completion of the weekly induction dosing period of REGN5837 / odronextamab, subjects will proceed to the Q2W treatment period and receive their designated QW REGN5837 dose and 320 mg odronextamab (or 160 mg odronextamab Q2W for DL1) Q2W. During the Q2W treatment period, REGN5837 should be administered on the same day as odronextamab (or on 2 consecutive days, if necessary for scheduling or other reasons). REGN5837 in combination with odronextamab will be continued on a Q2W schedule until disease progression or other protocol-defined reason for treatment discontinuation. However, if the subject achieves a CR sustained for at least 9 months, the subject will be transitioned to a Q4W dosing schedule for both the QW dose of REGN5837 and 320 mg of odronextamab (Note: 160 mg for DL1). Subjects must have received their assigned REGN5837 / odronextamab dose on a Q2W dosing schedule for at least three prior doses before switching from Q2W to Q4W dosing.

[0275] The safety of Q4W dosing will be evaluated in the first 16 subjects after receiving two doses of the combination at this frequency. If 4 or fewer subjects experience Grade 2 or higher CRS events, continue Q4W dosing in subsequent subjects If 5 or more subjects experience Grade 2 or higher CRS events, discontinue Q4W dosing and continue Q2W dosing for all subjects.

[0276] This rule is based on the lower limit of the one-sided 80% confidence interval. If the lower limit of the one-sided 80% confidence interval of the rate of Grade ≥ 2 CRS is > 20%, the transition from Q2W to Q4W will be suspended. Subjects who experience Grade ≥ 2 CRS after the first dose of any investigational drug after transitioning from Q2W to Q4W dosing will be included in the analysis.

[0277] Dose Escalation and Study Cohorts The dose escalation rules are based on a Bayesian optimal interval design (BOIN) with a target dose-limiting toxicity (DLT) rate of 28%. This target DLT rate corresponds to the same taper boundary as the 3+3 design. Every dose level will enroll at least 3 subjects. The decision rules at dose level j are summarized as follows: If the observed DLT rate is less than or equal to 0.221, escalate to dose level j+1. The observed DLT rate is calculated by dividing the number of subjects with a DLT by the number of DLT-evaluable subjects at the dose level. If the observed DLT rate is ≥ 0.334, taper to dose level j-1 and enroll 3 additional subjects at dose level j-1. If the observed DLT rate is greater than 0.221 and less than 0.334, remain at dose level j and enroll 3 additional subjects for further evaluation. If the number of subjects with DLT among evaluable subjects reaches a boundary defined as a posterior probability of 0.95 or greater that the true DLT rate is higher than the target DLT rate, dose levels j and above are excluded from the trial. If the first dose level is excluded, the trial is terminated. The posterior probability can be estimated based on the beta-binomial mode, assuming a non-informative prior distribution of beta(1,1).

[0278] The decision rule is repeated until the pre-specified maximum sample size of 54 subjects is exhausted, at which point the MTD is selected as the dose with an isotonic estimate of DLT probability closest to the target DLT rate of 28%.

[0279] Please note that the actual number of evaluable subjects per cohort may vary.The decision rule during the study applies as long as at least one subject is evaluable at a dose level (except DL1 and DL2, which require at least three DLT-evaluable subjects), and varies depending on the number of evaluable subjects at each dose level and the number of observed subjects with DLT.Table 3 shows the detailed decision rule for various numbers of subjects with DLT and evaluable subjects. [Table 3]

[0280] For scenarios with true toxicity probabilities (0.05, 0.15, 0.28, 0.45, 0.6), the operating characteristics based on 1000 simulated trials are as follows: ●Selection percentages (%) at each dose level: 1.2, 27.3, 50.6, 18.4, 2.5 Number of subjects treated at each dose level: 3.954, 6.921, 6.933, 2.817, 0.375 Number of toxicities observed at each dose level: 0.177, 1.002, 1.910, 1.274, 0.226 Average toxicity number: 4.589 Average number of targets: 21 Percentage of early terminations due to toxicity: 0.0% Risk of overmedication (>60% of subjects treated beyond the MTD): 0.0% Risk of overmedication (>80% of subjects treated beyond the MTD): 0.0% [Table 4]

[0281] If the initial (Cycle 2, Day 15) dose of REGN5837 in a dose-escalation cohort is determined to be intolerable, the initial dose of subsequent cohorts will be reduced to the maximum tolerated initial dose of REGN5837. Similarly, if the continued escalation of the intermediate dose of REGN5837 (Cycle 3, Day 1) is not tolerated, the dose on Day 1 of Cycle 3 will be fixed at the maximum tolerated intermediate dose.

[0282] If the initial and / or intermediate doses of REGN5837 are fixed, the purpose of the DLT window is to evaluate the safety of all doses of REGN5837 in combination with odronextamab. If a subject experiences any toxicity requiring discontinuation before the first full dose of REGN5837 in combination with odronextamab, the subject is considered inevaluable for DLT. Adverse events occurring before the first full dose of REGN5837 in combination with odronextamab inform the overall safety profile of REGN5837 in combination with odronextamab and affect dose escalation according to the dose escalation limit rules (outlined below). Subjects considered inevaluable for DLT may be replaced.

[0283] According to the rule-based dose escalation guidelines, a dose of REGN5837 between the values ​​in Table 4 but not exceeding the predetermined maximum dose value may be explored based on observed safety events. Dose levels are defined by the dose on Day 8 of Cycle 3. Safety events occurring on Day 15 of Cycle 2 with a designated dose of REGN5837 and tolerated at the dose on Day 1 of Cycle 3 at the previous dose level may be discussed by the SSET, and a decision may be made to reduce the dose on Day 15 of Cycle 2. The dose on Day 15 of Cycle 2 may be fixed at a lower dose to allow dose escalation to continue through the dose on Day 1 of Cycle 3. Similarly, safety events occurring on Day 1 of Cycle 3 with a designated dose of REGN5837 and tolerated at the dose on Day 8 of Cycle 3 at the previous dose level may be evaluated, and a decision may be made to reduce the dose on Day 1 of Cycle 3. The dose on Day 1 of Cycle 3 may be fixed at a lower dose to allow dose escalation to continue through the dose on Day 8 of Cycle 3. In all cases, modifications will only be applied to implement more conservative dose escalation between cohorts (e.g., a less abrupt increase between two consecutive DLs, or splitting the initial or subsequent doses over two administrations).

[0284] Prior to escalation to dose level 3, safety will be assessed for a minimum of three DLT-evaluable subjects in each cohort. Escalation to DL3 will proceed only if the safety profile (including the frequency and severity of CRS / IRR events with the combination) is comparable between DL1 and DL2. However, if a higher proportion of subjects in DL2 experience Grade 2 or higher CRS / IRR events with the combination compared to DL1, dose escalation of REGN5837 in combination with odronextamab 80 mg QW from DL3 onward will proceed. Preliminary pharmacodynamic data from DL1 and DL2 may be evaluated once available but will not be considered in this decision.

[0285] The following dose increment restrictions for REGN5837 will also be implemented: a) If any subject within a given cohort experiences a Grade 2 or higher AE (excluding AEs clearly attributable to the underlying disease or external cause) during the DLT period, the maximum dose increment of REGN5837 will be limited to 100%, including the escalating dose and the full therapeutic dose of the next dosing cohort. b) If two or more subjects within a given cohort experience a Grade 2 or higher AE (excluding AEs clearly attributable to the underlying disease or external cause) during the DLT period, the maximum dose increment of REGN5837 will be limited to 50%, including the escalating dose and the full therapeutic dose of the next dosing cohort. c) If any subject within a given cohort experiences a DLT during the DLT Period, the maximum dose increment of REGN5837 will be limited to 50%, including the escalating dose and the full therapeutic dose of the next dosing cohort. d) If two or more subjects within a given cohort experience a DLT during the DLT Period, the maximum dose increment of REGN5837 will be limited to 30%, including the escalating dose and the full therapeutic dose of the next dosing cohort.

[0286] The following AEs are commonly observed with odronextamab monotherapy and may be present at low grade at the time of REGN5837 administration: asthenia, cytopenia, clinically significant electrolyte disturbances, fatigue, decreased appetite, myalgia, arthralgia, nausea, vomiting, diarrhea, rash, and headache. For these AEs, the dose increment restriction guidelines #(a) and (b) above require the occurrence of Grade 3 or higher events rather than Grade 2 or higher events. Furthermore, the rules (a) and (b) listed above do not apply to infections except in the case of Grade 4 or elevated liver enzymes and bilirubin that resolve within 72 hours.

[0287] No intrasubject dose escalation is permitted (Table 4).

[0288] Dose-limiting toxicity (DLT) A DLT is defined as any of the toxicities listed below, except when the event is clearly attributable to the underlying disease or an external cause (including concomitant medications).

[0289] The grades of these toxicities are defined according to the Common Terminology Criteria for Adverse Events (CTCAE) version 5.0, except for cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS), which are defined according to the American Society for Transplantation and Cellular Therapy (ASTCT) criteria.

[0290] DLT is defined as follows: Non-hematological toxicity: Any grade 5 toxicity Seizures of any grade Grade 4 alanine aminotransferase / aspartate aminotransferase (ALT / AST) levels persisting for more than 3 consecutive days Any other grade 3 or greater non-hematologic toxicity, except for the following: Alopecia Nausea, vomiting, fatigue, or diarrhea lasting less than 72 hours with supportive care prescribed by a treating physician Grade 3 tumor lysis syndrome (TLS) Grade 3 infusion-related reaction (IRR) or CRS that responds to medical management and acute effects and resolves to Grade 1 or baseline within 72 hours. Note: Associated laboratory abnormalities can remain the same grade for 7 days without suggestion of persistent organ damage. Isolated laboratory abnormalities in the absence of clinical symptoms: Grade 2 or less alkaline phosphatase, gamma-glutamyltransferase, amylase, lipase, INR, activated partial thromboplastin time, hypertriglyceridemia, electrolyte abnormalities not indicating hospitalization Hematological toxicity: Any grade 5 hematologic toxicity Grade 4 neutropenia lasting more than 7 days despite granulocyte colony-stimulating factor (G CSF) Grade 4 febrile neutropenia Grade 4 thrombocytopenia lasting more than 7 days Grade 3 or greater thrombocytopenia associated with Grade 2 or greater bleeding (excluding Grade 2 epistaxis) Grade 3 or greater neutropenia with documented infection

[0291] Evaluate treatment-emergent adverse events (TEAEs) that may meet the definition of a DLT. Discuss any inability to administer two full doses of the REGN5837 and odronextamab combination within 35 days due to study drug toxicity. A final determination of whether a TEAE meets the definition of a DLT will be made based on a careful review of all relevant data.

[0292] Continued treatment may occur in subjects who do not meet protocol-defined criteria for permanent discontinuation only if it is determined that it is in the subject's best interest to resume concomitant study treatment after the occurrence of a DLT.

[0293] Dose expansion A total of 37 subjects will be enrolled in the dose expansion phase of RP2D, which will be determined in the dose escalation portion of the study across two expansion cohorts, A and B. Cohorts A and B will enroll subjects with aggressive B-NHL without prior CAR-T therapy, as well as subjects who have progressed after failure of prior CAR-T therapy, evaluable for safety and efficacy, respectively. Analyses of these subjects will be combined with subjects treated with the recommended phase 2 dose regimen (RP2D) in the dose escalation portion, resulting in a total of 20 subjects in each cohort treated with RP2D.

[0294] Subjects enrolled in the dose expansion cohorts will undergo the RP2D to further evaluate the preliminary antitumor activity, safety, PK properties, and biomarker changes associated with the combination of REGN5837 and odronextamab therapy.

[0295] Subjects in the dose expansion cohort will not be replaced if they have received at least one dose of REGN5837.

[0296] Study population The study population consisted of subjects with aggressive B-NHL lymphoma according to WHO criteria that had progressed after at least two lines of therapy, including anti-CD20 antibodies and alkylating agents.

[0297] Selection Criteria Subjects must meet the following criteria to be eligible for inclusion in the study: 1. 18 years of age or older 2. Has disease that has progressed after at least two lines of systemic therapy, including anti-CD20 antibodies and alkylating agents, +Aggressive B-NHL was documented. Lymphoma subtypes were based on the World Health Organization (WHO) classification. Eligible subtypes included: DLBCL, primary mediastinal (thymic) large B-cell lymphoma, T-cell / histiocyte-rich large B-cell lymphoma, grade 3b follicular lymphoma, and high-grade B-cell lymphoma (HGBL) with and without MYC and BCL2 or BCL6 translocations. Subjects must require systemic therapy for their lymphoma at the time of study enrollment, as determined by the investigator. Note: • Subjects who relapse after their most recent prior line or who progress during or after prior therapy are eligible. Subjects who have received CAR-T therapy are eligible. Ongoing studies have demonstrated that odronextamab can be tolerated by these subjects and provides efficacy. CAR-T naive and post-CAR-T failure subjects will be evaluated separately in Cohorts A and B, respectively, during the dose expansion phase. DLBCL transformed from a low-grade neoplasm (e.g., FL or CLL) may be enrolled. Subjects with DLBCL transformed from a prior CLL may be enrolled only if there is no leukemic CLL component. For subjects with transformed DLBCL, prior systemic therapy administered for the low-grade neoplasm will not be considered among the prior lines of treatment for purposes of determining eligibility. 3. Disease measurable on cross-sectional images recorded by imaging (computed tomography [CT] or magnetic resonance imaging [MRI]) (defined as at least one bidimensionally measurable lymph node lesion ≥ 1.5 cm in greatest transverse diameter (GTD), regardless of short-axis diameter) 4. Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1 5. Adequate bone marrow function documented by:. a. Platelet count ≥ 50 x 109 / L. Subjects may not have received platelet transfusion therapy within 7 days prior to the first dose of odronextamab to meet the platelet eligibility criteria. b. Hemoglobin ≥ 9.0 g / dL; transfusions to meet this criterion are permitted per protocol. c. Absolute neutrophil count (ANC) ≥ 1.0 x 109 / L. Subjects may not have received G CSF within 2 days prior to the first dose of odronextamab to meet the ANC eligibility criteria. NOTE: Subjects with bone marrow involvement or splenic sequestration should meet the following hematological parameters: - Platelet count ≥ 25 x 109 / L. Subjects may not have received platelet transfusion therapy within 3 days prior to the first dose of odronextamab to meet the platelet eligibility criteria. - Hemoglobin greater than 7.0 g / dL - Absolute neutrophil count (ANC) ≥ 0.5 x 109 / L. Subjects may not have received G-CSF within 2 days prior to the first dose of odronextamab to meet the ANC eligibility criteria. 6. Proper Liver Function: a. Total bilirubin below 1.5 × upper limit of normal (ULN) (below 3 × ULN if caused by lymphoma infiltration of the liver) b. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels below 3 × ULN (or below 5 × ULN if caused by lymphomatous infiltration of the liver) c. Alkaline phosphatase (ALP) level of 2.5 × ULN or less (5 × ULN or less if caused by lymphoma infiltration of the liver) Note: *Subjects with total bilirubin greater than 1.5 x ULN and concomitant AST greater than 3 x ULN and / or ALT greater than 3 x ULN, regardless of the presence of lymphomatous infiltration of the liver, are excluded. *Subjects with known Gilbert's syndrome are excluded if their total bilirubin level is >4 x ULN in the local general population. Creatinine clearance calculated by the Cockcroft-Gault formula of 7.50 mL / min or greater: Note: Subjects with a calculated creatinine clearance less than 50 mL / min may be considered for enrollment if their measured creatinine clearance (based on a 24-hour urine collection or other reliable method) is 50 mL / min or greater. 8. Subjects should be willing to undergo mandatory tumor biopsy during the dose expansion phase of the study if, in the opinion of the investigator, the subject has an accessible lesion that can be biopsied without significant risk to the subject. In the absence of such a lesion at screening, archival tissue samples up to 6 months prior (and without interventional treatment) may be considered acceptable for the subject's study eligibility (after approval by the medical monitor). 9. Ability to understand the purpose and risks of the study and provide signed and dated informed consent and permission to use protected health information (in accordance with state and local subject privacy regulations). 10. Willingness and ability to comply with clinic visits and study-related procedures. 11. Provide informed consent signed by the study subject or legally acceptable representative

[0298] Exclusion criteria Subjects who meet any of the following criteria will be excluded from the study. 1. Pre-treatment: Previous allogeneic stem cell transplant or solid organ transplant - Subjects who have received prior treatment with anti-CD20 × anti-CD3 bispecific antibodies such as odronextamab 2. Diagnosis of Mantle Cell Lymphoma (MCL) 3. Known involvement by primary central nervous system (CNS) lymphoma or non-primary CNS lymphoma (even if treated to complete remission). Suspected CNS lymphoma should be evaluated with mandatory head CT or MRI plus lumbar puncture if indicated. Treatment with any systemic anti-lymphoma therapy within 4.5 half-lives or 14 days prior to the first dose of investigational drug, whichever is shorter 5. Standard radiotherapy within 14 days of the first dose of study drug. Note: Palliative radiotherapy to symptomatic lymph nodes / lesions is permitted if the irradiated lesion(s) or node(s) are not included as target lesions in the tumor evaluation 6. Ongoing systemic corticosteroid treatment with more than 10 mg prednisone or corticosteroid equivalent per day within 72 hours of initiating odronextamab 7. Comorbidities: History of neurodegenerative condition or CNS movement disorder. Subjects with a history of seizures within 12 months prior to study enrollment will be excluded. b. Another malignancy within the past 5 years, except for tumors that are localized (e.g., nonmelanoma skin cancer or cervical intraepithelial neoplasia) and have been effectively treated with definitive local control (with or without ongoing adjuvant hormonal therapy). c. Cardiac ejection fraction of less than 40% by echocardiogram (ECHO) or multiple acquisition (MUGA) scan d. Any other significant concurrent disease or medical condition that, in the opinion of the investigator, may interfere with the conduct of the study or pose a significant risk to the subject, including, but not limited to, significant cardiovascular (e.g., New York Heart Association Class III or IV heart disease, myocardial infarction within the past 6 months, unstable arrhythmia, or unstable angina), pulmonary (e.g., history of obstructive pulmonary disease and symptomatic bronchospasm), gastrointestinal, hepatic, renal, endocrine, hematological, autoimmune, psychiatric, or neurological disorders 8. Infection: a. Infection requiring hospitalization or treatment with IV anti-infectives within 2 weeks of the first dose of study drug b. Uncontrolled infection with human immunodeficiency virus (HIV), hepatitis B (HBV), or hepatitis C (HCV) infection; or other uncontrolled infection Subjects with HIV who have controlled infection (undetectable viral load spontaneously or on a stable antiviral regimen and CD4 count greater than 350 cells / microliter) are allowed Subjects with hepatitis B (HepBsAg+) who have controlled infection (serum hepatitis B virus DNA polymerase chain reaction [PCR] below the limit of detection and receiving hepatitis B antiviral therapy) are allowed Subjects who are hepatitis C virus antibody positive (HCV Ab+) and have controlled infection (undetectable HCV RNA by PCR, either spontaneously or in response to a previous successful course of anti-HCV therapy) are allowed 9. Cytomegalovirus infection as demonstrated by detectable levels by peripheral blood PCR assay. Subjects who demonstrate detectable levels of CMV at screening must be treated with appropriate antiviral therapy and demonstrate at least two undetectable levels of CMV by PCR assay (at least 7 days apart) before being reconsidered for eligibility. 10. Allergy / Hypersensitivity: Known hypersensitivity to both allopurinol and rasburicase 11. History of severe allergic reaction caused by compounds with similar chemical or biological composition as the investigational drug or excipients 12. Vaccination with a replication-competent vector within 28 days prior to the first study drug administration 13. Members of the clinical site trial team or their immediate families, unless prior approval by the sponsor. 14. Women of childbearing potential (WOCBP) with a positive serum beta hCG pregnancy test. 15. Pregnant or lactating women. 16. Women of childbearing potential who are unwilling to practice highly effective contraception before the first dose / initiation of first treatment, during the study, and for at least 6 months after the last dose. Sperm donation is prohibited during the study and for 6 months after the last dose of investigational drug. Highly effective contraception includes: a. Stable use of combined (estrogen and progesterone-containing) hormonal contraception (oral, intravaginal, transdermal) or progesterone-only hormonal contraception (oral, injectable, implantable) associated with ovulation inhibition for at least two menstrual cycles prior to screening b. Intrauterine device (IUD), intrauterine hormone-releasing system (IUS) C. Bilateral tubal ligation d. Vasectomized partner† (if the male vasectomized partner was the study participant's only sexual partner and the partner had a medical evaluation of the surgical success of the operation). e. and / or sexual abstinence‡,§. f. Male study participants with WOCBP partners must use condoms unless they have a vasectomy† or practice sexual abstinence. ‡, § *Women of childbearing potential are defined as women who are capable of bearing children after menarche and after menopause, unless they are permanently infertile. Permanent methods of contraception include hysterectomy, bilateral salpingectomy, and bilateral oophorectomy. Postmenopausal status is defined as the absence of menstruation for 12 months without an alternative medical cause. High follicle-stimulating hormone (FSH) levels in the postmenopausal range can be used to confirm postmenopausal status in women not using hormonal contraception or hormone replacement therapy. However, a single FSH measurement in the absence of 12 months of amenorrhea is insufficient to determine the occurrence of postmenopausal status. The above definition follows guidelines from the Clinical Trials Facilitation Group (CTFG). Pregnancy testing and contraception are not required for women with a documented hysterectomy. †Vastectomized partners or vasectomized study participants must have undergone a medical evaluation of the success of their surgery. ‡Sexual abstinence is considered highly effective only if it is defined as abstinence from heterosexual intercourse for the entire period of risk associated with the investigational drug. The reliability of sexual abstinence must be evaluated in relation to the duration of the clinical trial and the subject's preferred usual lifestyle. Periodic abstinence (calendar, symptom-temperature, or ovulation method), withdrawal (abortive intercourse), spermicide alone, and lactating amenorrhea (LAM) are not acceptable methods of contraception. Female condoms and male condoms should not be used together. 17. A subject who is institutionalized by order issued by either a judicial or administrative authority.

[0299] research treatment REGN5837 and odronextamab (REGN1979) drug products are supplied as liquids in sterile, single-use vials for administration by IV infusion.

[0300] A pharmacist or other qualified individual will be identified at each site and prepared to administer REGN5837 and odronextamab. The dose administered is a fixed dose and is not determined by the subject's weight or body surface area.

[0301] During the first 5 weeks of study treatment, subjects will receive weekly odronextamab as monotherapy. The escalating doses consist of an initial dose of 0.7 mg, followed by a first intermediate dose of 4 mg and a second intermediate dose of 20 mg. The initial dose and both intermediate doses are always split into two separate infusions: the initial dose is split at 0.2 mg / 0.5 mg, the first intermediate dose is split at 2 mg per infusion, and the second intermediate dose is split at 10 mg per infusion. Each split dose is preferably administered over 4 hours on each of two consecutive days, with an interval of no more than 3 days (e.g., for the initial dose, on days 1 and 2 of cycle 1; for the first intermediate dose, on days 8 and 9 of cycle 1; and for the second intermediate dose, on days 15 and 16 of cycle 1). These escalating doses should be split even if there is a treatment delay that causes administration beyond day 15 of cycle 1. A total dose of 160 mg (Note: 80 mg QW on DL1) will be administered on Day 1 of Cycle 2. Another total dose of odronextamab will be administered on Day 8 of Cycle 2. The first QW dose will be administered as a single infusion on Day 1 of Cycle 2 and continued that way thereafter.

[0302] REGN5837 will begin on Day 15 of Cycle 2 to allow subjects to tolerate odronextamab monotherapy before initiating the combination therapy. REGN5837 will also be administered initially at an escalating dose. All doses of REGN5837 will be administered as a single infusion. On Day 15 of Cycle 2 of study treatment, REGN5837 will be introduced at a lower initial dose in combination with odronextamab at 160 mg (Note: 80 mg on DL1). On Day 1 of Cycle 3, REGN5837 will be administered at an intermediate dose. On Day 8 of Cycle 3, REGN5837 will be escalated to the full QW dose.

[0303] Prior to escalation to DL3, safety data from DL1 and DL2 will be evaluated. Escalation to DL3 will occur only if the safety profile is comparable between DL1 and DL2. However, if the safety profile of DL2 is deemed worse, escalation of REGN5837 from DL3 will proceed in combination with odronextamab 80 mg QW.

[0304] Subjects will receive 12 doses of the combination of REGN5837 and odronextamab during the induction period of REGN5837 / odronextamab. Subjects will receive a total of 17 doses of odronextamab upon completion of the induction period of REGN5837 / odronextamab.

[0305] After completion of the REGN5837 / odronextamab induction period, subjects will proceed to the maintenance period where dosing is Q2W and subjects will receive their assigned QW REGN5837 dose and 320 mg of odronextamab (160 mg for DL1).

[0306] Study endpoints The primary endpoints are: Incidence of DLTs from the first dose of REGN5837 in combination with odronextamab to the end of the DLT observation period The incidence and severity of treatment-emergent adverse events (TEAEs) and adverse events of special interest (AESIs) during treatment with REGN5837 in combination with odronextamab

[0307] Secondary endpoints are: Serum concentrations of REGN5837 and odronextamab Immunogenicity as measured by anti-drug antibodies (ADA) to REGN5837 and odronextamab Overall response rate (ORR), which is the proportion of patients who achieved the best overall response (CR) or PR during or after study treatment according to the Lugano Classification of Responses in Malignant Lymphoma as assessed by the investigator. Complete response (CR) rate, which is the proportion of patients who achieved the best overall response (CR) during or after study treatment according to the Lugano classification as assessed by the investigator. • Progression-free survival (PFS) according to the Lugano classification assessed by the investigator, determined as the time from the start of study treatment to the first date of progressive disease or death from any cause, whichever occurs first. Overall survival (OS), measured from the start of study treatment to death from any cause Duration of response (DOR) based on the investigator-assessed Lugano classification, determined from the date of the first documented CR or PR to the first date of progressive disease or death from any cause, whichever occurs first.

[0308] The exploratory endpoints are as follows: Changes in absolute numbers of peripheral T cells, including activation and proliferation phenotypes of T cell subsets measured by multiparameter flow cytometry, along with changes in serum cytokine levels Comparison of changes in lymph node T-cell density, both at baseline levels and during treatment / relapse, as measured by multiplex IHC and / or RNAscope, and response (according to Lugano classification) by expression of immune markers such as CD28, 41BB, programmed death receptor-1 (PD-1), Lag3, GzmB, IFN-γ, Ki67, and B-cell markers (CD20, CD22). Correlation between the proportion of subjects with negative molecular MRD at clinical complete response by next-generation sequencing and PFS and OS Assessment of the relationship between clinical drug concentrations and pharmacodynamic, safety, and efficacy measures

[0309] Procedures and Evaluation Data will be summarized by dose level, disease subtype, and prior treatment arm(s). Demographic and baseline characteristics will be summarized descriptively. Safety observations and measurements, including drug exposure, adverse events, laboratory data, vital signs, and European Collaborating Group oncology performance status, will be summarized.

[0310] Evaluate treatment using the following procedures and assessments Procedures performed only at the screening / baseline visit ●Safety procedures Immunosafety evaluation ● Laboratory testing ●Validity Procedures Drug concentration and measurement Immunogenicity measurements and samples Pharmacodynamic and exploratory biomarkers Future Biomedical Research (Optional) Pharmacogenomic analysis (optional)

[0311] Statistical Planning and Analysis This section provides the basis for the study's statistical analysis plan (SAP), which is revised prior to study completion to accommodate amendments to the clinical trial protocol and to accommodate unforeseen issues during study execution and data that may affect the planned analyses.

[0312] Sample size justification A total of 91 subjects are planned to be enrolled.

[0313] Dose Escalation Portion: A maximum of 54 subjects will be enrolled in the dose escalation portion, assuming an average of 6 subjects per dose level can be enrolled across 9 dose levels. The actual sample size of these dose escalation cohorts will vary depending on the number of observed subjects with DLT and the number of dose levels administered.

[0314] Dose Expansion Portion: There will be two dose expansion cohorts of subjects with aggressive B-NHL: one cohort with no prior CAR-T therapy and one cohort of subjects who have progressed after failure of prior CAR-T therapy. Each cohort will require 20 subjects, including three subjects treated at RP2D in the dose escalation portion. Therefore, a maximum of 37 subjects will be enrolled in the dose expansion portion.

[0315] A sample size of 20 subjects in each cohort will be determined to further investigate the safety of the combination of REGN5837 and odronextamab in subjects treated at RP2D and to better understand the preliminary antitumor activity.

[0316] Preliminary Assessment of Antitumor Activity: Twenty subjects treated with RP2D in the dose escalation and expansion portions will provide a preliminary assessment of antitumor activity.

[0317] Analysis Set Efficacy Analysis Set: The Full Analysis Set (FAS) includes all subjects who received any study drug. Efficacy endpoints will be analyzed using the FAS.

[0318] Safety Analysis Set: The Safety Analysis Set (SAF) includes all subjects who received any study drug and is based on treatment received. Treatment compliance / administration and all clinical safety variables will be analyzed using the SAF.

[0319] Pharmacokinetic Analysis Set: The PK analysis set includes all subjects who received at least one dose of study drug and had at least one non-missing drug concentration result after the first dose of study drug.

[0320] Immunogenicity Analysis Set: The anti-drug antibody analysis set (AAS) was defined separately for each investigational drug and included all treated subjects who received any amount of investigational drug (safety analysis set) and had at least one non-missing ADA result after the first dose of the respective investigational drug.

[0321] Dose-Limiting Toxicity Analysis Set: The dose-limiting toxicity (DLT) analysis set includes all subjects treated with REGN5837 and odronextamab who enrolled in the DLT-evaluable dose escalation, defined as subjects who completed the DLT observation period, as well as subjects who discontinued early due to the onset of a DLT during the DLT observation period. This analysis set will be used to evaluate DLTs for dose escalation decisions.

[0322] Efficacy analysis Primary Efficacy Analysis: All efficacy endpoints are secondary endpoints in this study.

[0323] Secondary efficacy analyses: ORR and CR rates based on the Lugano classification based on local investigator review will be summarized with two-sided 95% confidence intervals. Subjects who cannot be assessed for best overall response will be considered non-responders. Other secondary efficacy endpoints, including DOR, PFS, and OS, will be summarized by median and its 95% confidence interval, when applicable, using the Kaplan-Meier method.

[0324] Example 3: Results Administration of REGN5837 in combination with REGN1979 is expected to lead to enhanced tumor regression and improved disease control. Furthermore, administration of REGN5837 in combination with REGN1979 is expected to be safe, without an increased incidence of adverse events and / or increased toxicity over monotherapy.

[0325] The trial is currently enrolling, and preliminary results indicate that REGN5837 and REGN1979 are safe to administer.

[0326] The present disclosure is not limited in scope by the specific embodiments described herein. Indeed, various modifications of the present disclosure in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

Claims

1. 1. A method for treating a B-cell proliferative disorder or a CD20-expressing cell malignancy in a subject, the method comprising administering to the subject a therapeutically effective amount of a bispecific CD22xCD28 antibody or antigen-binding fragment thereof in combination with a bispecific CD3xCD20 antibody or antigen-binding fragment thereof; the bispecific CD22xCD28 antibody or antigen-binding fragment thereof comprises a first antigen-binding domain that binds to cluster of differentiation factor 28 (CD28) and a second antigen-binding domain that binds to cluster of differentiation factor 22 (CD22); the bispecific CD3xCD20 antibody or antigen-binding fragment thereof comprises a first antigen-binding domain that binds to cluster of differentiation 3 (CD3) and a second antigen-binding domain that binds to cluster of differentiation 20 (CD20); thereby treating said B-cell proliferative disorder or CD20-expressing cell malignancy in said subject.

2. 10. The method of claim 1, wherein the B-cell proliferative disorder is a B-cell lymphoma.

3. 3. The method of claim 2, wherein the lymphoma is B-cell non-Hodgkin's lymphoma (B-NHL).

4. 4. The method of claim 3, wherein the non-Hodgkin's lymphoma is selected from the group consisting of diffuse large B-cell lymphoma (DLBCL), marginal zone lymphoma (MZL), high-grade B-cell lymphoma, Burkitt's lymphoma, primary mediastinal large B-cell lymphoma, and follicular lymphoma.

5. The method of any one of claims 1 to 4, further comprising selecting a subject, wherein the subject has aggressive B-NHL.

6. The subject meets the following criteria: a. have CD20+ aggressive B-NHL; b. Progressed after at least two lines of systemic therapy, including a CD20 inhibitor and an alkylating agent; c. Have disease that can be measured on cross-sectional imaging d. have adequate bone marrow and liver function; and / or e. Have any of the following cancer types: DLBCL, primary mediastinal (thymic) large B-cell lymphoma, T-cell / histiocyte-rich large B-cell lymphoma, grade 3b follicular lymphoma, and high-grade B-cell lymphoma (HGBL) with or without MYC, BCL2, or BCL6 translocations; 6. The method of claim 1, wherein the first and second electrodes have at least one of the following characteristics:

7. 7. The method of any one of claims 1 to 6, wherein the subject has been treated with a previous therapy and has relapsed or the disorder has progressed during or after the previous treatment.

8. The method of any one of claims 1 to 7, wherein the subject is undergoing CAR-T therapy.

9. 9. The method of any one of claims 1-8, wherein the subject has measurable CD20+ aggressive B-NHL that has progressed after two or more lines of systemic therapy including at least a CD20 inhibitor and an alkylating agent.

10. The method of claim 9 , wherein the CD20 inhibitor is an anti-CD20 antibody.

11. The method of claim 9 or 10, wherein the subject is being treated with CAR-T cell therapy.

12. The method of any one of claims 9 to 11, wherein the subject has not been previously treated with a CD3 antibody, a CD3 bispecific antibody, or a CD3xCD20 bispecific antibody.

13. The method of any one of claims 1 to 12, wherein the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered at a dose of about 0.01 mg to about 400 mg.

14. 14. The method of any one of claims 1 to 13, wherein the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered at a dose of about 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 240 mg, 280 mg, 300 mg, 320 mg, 350 mg, or 400 mg.

15. The method of any one of claims 1 to 14, wherein the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered at a dose of about 0.1 mg to about 400 mg.

16. 16. The method of any one of claims 1 to 15, wherein the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered at a dose of about 0.1 mg, 0.2 mg, 0.3 mg, 0.5 mg, 0.7 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 240 mg, 280 mg, 300 mg, 320 mg, 350 mg, or 400 mg.

17. 17. The method of any one of claims 1 to 16, wherein the method comprises administering one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof in combination with one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof.

18. 18. The method of claim 17, wherein each of the one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is from about 0.01 mg to about 400 mg.

19. 19. The method of claim 18, wherein each of the one or more doses is about 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 240 mg, 280 mg, 300 mg, 320 mg, 350 mg, or 400 mg.

20. 20. The method of any one of claims 17 to 19, wherein each of the one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is from about 0.1 mg to about 400 mg.

21. 21. The method of any one of claims 17 to 20, wherein each of the one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is about 0.1 mg, 0.2 mg, 0.3 mg, 0.5 mg, 0.7 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 240 mg, 280 mg, 300 mg, 320 mg, 350 mg, or 400 mg.

22. 22. The method of any one of claims 17 to 21, wherein the one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered 1 day to 8 weeks after the immediately preceding dose.

23. 23. The method of any one of claims 17 to 22, wherein each of the one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered once per week, once per two weeks, once per three weeks, once per four weeks, once per five weeks, or once per six weeks.

24. The method of any one of claims 17 to 23, wherein one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof are administered once a week.

25. The method of any one of claims 17 to 24, wherein one or more doses of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof are administered once every two weeks.

26. The method of any one of claims 17 to 25, wherein one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered once a week.

27. The method of any one of claims 17 to 26, wherein one or more doses of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered once every two weeks.

28. 28. The method of any one of claims 17 to 27, wherein the dose of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered in a single dose or divided and administered on two days no more than three days apart.

29. 29. The method of any one of claims 1 to 28, wherein the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered intravenously.

30. 30. The method of any one of claims 1 to 29, wherein the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered subcutaneously.

31. 31. The method of any one of claims 1 to 30, wherein the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered on the same day.

32. 32. The method of any one of claims 1 to 31, wherein the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and / or the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered on different days.

33. 33. The method of claim 32, wherein the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered before or after the bispecific CD3xCD20 antibody or antigen-binding fragment thereof.

34. 34. The method of claim 33, wherein the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered one day before the bispecific CD3xCD20 antibody or antigen-binding fragment thereof.

35. (i) administering to the subject the bispecific CD3xCD20 antibody or antigen-binding fragment thereof subcutaneously or intravenously weekly at a dose of 0.1 mg to 160 mg for a duration of monotherapy, wherein the duration of monotherapy is at least two weeks; (ii) administering to the subject the bispecific CD22xCD28 antibody or antigen-binding fragment thereof subcutaneously or intravenously every week at a dose of 0.01 mg to 400 mg, and administering to the subject the bispecific CD3xCD20 or antigen-binding fragment thereof intravenously or subcutaneously every week for the duration of induction combination therapy at a dose of 80 mg to 160 mg.

36. 36. The method of claim 35, wherein the duration of the monotherapy is at least 2 weeks, at least 3 weeks, at least 4 weeks, or at least 5 weeks.

37. 38. The method of claim 36 or 37, wherein the dose of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof during the monotherapy period is split and administered on two different days no more than three days apart, or is administered in a single dose.

38. 38. The method of any one of claims 35 to 37, wherein the monotherapy period in step (i) comprises administering an initial dose of the bispecific CD3xCD20 antibody and escalating the dose to a full dose by the end of the monotherapy period.

39. 39. The method of claim 38, wherein the total dose of the bispecific CD3xCD20 in step (i) is 80 mg or 160 mg.

40. The induction combination therapy period in step (ii) comprises: (a) administering an initial dose of the bispecific CD22xCD28 antibody, wherein the initial dose comprises 0.03 mg to 2 mg; (b) administering an intermediate dose comprising between 0.1 mg and 20 mg of a bispecific CD22xCD28 antibody; (c) administering a total dose of bispecific CD22xCD28, wherein said total dose comprises between 0.3 mg and 160 mg.

41. 41. The method of any one of claims 35 to 40, wherein during step (ii) the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered on a different day than the bispecific CD22xCD28 antibody or antigen-binding fragment thereof.

42. 42. The method of claim 41 , wherein the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered one day after the bispecific CD22xCD28 antibody or antigen-binding fragment thereof.

43. 41. The method of any one of claims 35 to 40, wherein during step (ii), the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is administered on the same day as the bispecific CD22xCD28 antibody or antigen-binding fragment thereof.

44. 44. The method of any one of claims 35 to 43, wherein the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered in step (ii) in combination with the bispecific CD3xCD20 or antigen-binding fragment thereof for at least 9 weeks.

45. 45. The method of claim 44, wherein the bispecific CD22xCD28 antibody or antigen-binding fragment thereof is administered in step (ii) in combination with the bispecific CD3xCD20 antibody or antigen-binding fragment thereof for at least 9, 10, 11, 12, 13, 14 or 15 weeks.

46. (iii) After step (ii), the method of any one of claims 35 to 44 further comprises administering the bispecific CD22xCD28 antibody or antigen-binding fragment thereof in combination with 160 mg or 320 mg of the bispecific CD3xCD20 antibody or antigen-binding fragment thereof every two or more weeks for the duration of the maintenance combination therapy.

47. 47. The method of claim 46, wherein in step (iii) the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and the bispecific CD3xCD20 antibody or antigen-binding fragment thereof are administered on the same day.

48. 48. The method of claim 46 or 47, wherein in step (iii) the bispecific CD22xCD28 antibody or antigen-binding fragment thereof and the bispecific CD3xCD20 or antigen-binding fragment thereof are administered once every two weeks or once every four weeks.

49. 49. The method of any one of claims 1-48, further comprising administering to the subject one or more additional agents for treating or preventing one or more symptoms of the adverse event.

50. 50. The method of any one of claims 1 to 49, wherein the bispecific antibody is administered to the subject in combination with a second agent, wherein the second agent is selected from the group consisting of dexamethasone, diphenhydramine, acetaminophen, a steroid, an antihistamine, a nonsteroidal anti-inflammatory drug (NSAID), an IL-6 antagonist, and an IL-6R antagonist.

51. 51. The method of any one of claims 1 to 50, wherein the subject has stable disease, a partial response, or a complete response upon at least one week of administration of the bispecific CD22xCD28 antibody or antigen-binding fragment thereof at a dose of about 0.01 mg to about 400 mg in combination with the bispecific CD3xCD20 antibody or antigen-binding fragment thereof.

52. the bispecific CD3xCD20 antibody or antigen-binding fragment thereof i) a CD3-binding arm comprising a heavy chain complementarity determining region (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 5, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 6; ii) a CD20-binding arm comprising heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 4, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

6.

53. the bispecific CD3xCD20 antibody or antigen-binding fragment thereof i) a CD3 binding arm comprising three HCDRs (HCDR1, HCDR2 and HCDR3) and three LCDRs (LCDR1, LCDR2 and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 10, HCDR2 comprises the amino acid sequence of SEQ ID NO: 11, HCDR3 comprises the amino acid sequence of SEQ ID NO: 12, LCDR1 comprises the amino acid sequence of SEQ ID NO: 13, LCDR2 comprises the amino acid sequence of SEQ ID NO: 14, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 15; and ii) a CD20-binding arm comprising three HCDRs (HCDR1, HCDR2 and HCDR3) and three LCDRs (LCDR1, LCDR2 and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 7, HCDR2 comprises the amino acid sequence of SEQ ID NO: 8, HCDR3 comprises the amino acid sequence of SEQ ID NO: 9, LCDR1 comprises the amino acid sequence of SEQ ID NO: 13, LCDR2 comprises the amino acid sequence of SEQ ID NO: 14, and LCDR3 comprises the amino acid sequence of SEQ ID NO:

15.

54. 54. The method of claim 53, wherein the HCVR of the CD3-binding arm comprises the amino acid sequence of SEQ ID NO: 5, the HCVR of the CD20-binding arm comprises the amino acid sequence of SEQ ID NO: 4, and the common LCVR comprises the amino acid sequence of SEQ ID NO:

6.

55. 55. The method of any one of claims 52 to 54, wherein the bispecific CD3xCD20 antibody or antigen-binding fragment thereof comprises a heavy chain of the CD3 binding arm comprising the amino acid sequence of SEQ ID NO:2, a heavy chain of the CD20 binding arm comprising the amino acid sequence of SEQ ID NO:1, and a common light chain comprising the amino acid sequence of SEQ ID NO:

3.

56. 56. The method of any one of claims 1 to 55, wherein the bispecific CD3xCD20 antibody or antigen-binding fragment thereof is odronextamab.

57. The method of any one of claims 1 to 56, wherein the first antigen-binding domain that binds to CD28 comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 20, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

21.

58. 58. The method of claim 57, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 25, HCDR2 comprises the amino acid sequence of SEQ ID NO: 26, and HCDR3 comprises the amino acid sequence of SEQ ID NO:

27.

59. 59. The method of claim 58, wherein LCDR1 comprises the amino acid sequence of SEQ ID NO: 28, LCDR2 comprises the amino acid sequence of SEQ ID NO: 29, and LCDR3 comprises the amino acid sequence of SEQ ID NO:

30.

60. The method of any one of claims 57 to 59, wherein the first antigen-binding domain that binds to CD28 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 20 and an LCVR comprising the amino acid sequence of SEQ ID NO:

21.

61. The method of any one of claims 1 to 60, wherein the second antigen-binding domain that binds to CD22 comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 19, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

21.

62. 62. The method of claim 61, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 22, HCDR2 comprises the amino acid sequence of SEQ ID NO: 23, and HCDR3 comprises the amino acid sequence of SEQ ID NO:

24.

63. 63. The method of claim 62, wherein LCDR1 comprises the amino acid sequence of SEQ ID NO: 28, LCDR2 comprises the amino acid sequence of SEQ ID NO: 29, and CDR-L3 comprises the amino acid sequence of SEQ ID NO:

30.

64. The method of any one of claims 61 to 63, wherein the second antigen-binding domain that binds to CD22 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 19 and an LCVR comprising the amino acid sequence of SEQ ID NO:

21.

65. (a) the first antigen-binding domain that binds to human CD28 comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 20 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21; (b) the second antigen-binding domain that binds to CD22 comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 19, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:

21.

66. 66. The method of any one of claims 57 to 65, wherein the bispecific CD22xCD28 antibody comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO:

17.

67. 67. The method of any one of claims 57 to 66, wherein the bispecific CD22xCD28 antibody comprises a second heavy chain comprising the amino acid sequence of SEQ ID NO:

16.

68. 68. The method of any one of claims 57 to 67, wherein the bispecific CD22xCD28 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:

18.

69. 69. The method of any one of claims 57 to 68, wherein the bispecific CD22xCD28 antibody comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 17, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 16, and a common light chain comprising the amino acid sequence of SEQ ID NO:

18.

70. 70. The method of any one of claims 57 to 69, wherein the first antigen-binding domain comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 17 and a light chain comprising the amino acid sequence of SEQ ID NO:

18.

71. 71. The method of any one of claims 57 to 70, wherein the second antigen-binding domain comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 16 and a light chain comprising the amino acid sequence of SEQ ID NO:

18.

72. 72. The method of any one of claims 1 to 71, wherein the bispecific CD22xCD28 antibody is REGN5837 or an antigen-binding fragment thereof.

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