Dosage in combination treatment with bispecific Anti-CD20 / CD3 antibody and CD79b antibody-drug conjugate

Abstract

FIELD: oncology.SUBSTANCE: treatment of subjects with CD20-positive cell proliferative disorders. The use of polatuzumab and glofitamab is proposed for the treatment of a subject or subject population with a CD20-positive cell proliferative disorder. Polatuzumab and glofitamab are administered in a dosing regimen that includes a first dosing cycle and a second dosing cycle. The first dosing cycle includes the first dose (C1D1) and second dose (C1D2) of glofitamab, and a single dose of polatuzumab, to be administered on specific days of the dosing cycle. The second dosing cycle consists of a single dose (C2D1) of glofitamab and a single dose of polatuzumab, to be administered on specific days of the dosing cycle. Dosing cycles are 21-day dosing cycles.EFFECT: efficient method of dosing polatuzumab and glofitamab for the treatment of CD20-positive B-cell proliferative disorders, providing a more favorable benefit-risk profile.18 cl, 7 dwg, 21 tbl, 6 ex

Description

[0001] SEQUENCE LISTING

[0002] This application contains a sequence listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy, created on April 26, 2021, is designated 51177-033001 Sequence Listing 4.26.21 ST25 and is 32,706 bytes in size.

[0003] FIELD OF TECHNOLOGY

[0004] The present invention relates to methods for treating a disease, in particular a B-cell proliferative disease, by administering a bispecific antibody to CD20 / CD3 and a conjugate of an antibody to CD79b with a drug, as well as to methods for reducing adverse events in response to administering a bispecific antibody to CD20 / CD3 and a conjugate of an antibody to CD79b with a drug.

[0005] STATE OF THE ART

[0006] B-cell proliferative disorders are a heterogeneous group of malignancies that include both leukemias and lymphomas. Lymphomas develop from lymphoid cells and comprise two main categories: Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL). In the United States, lymphomas of B-cell origin account for approximately 80-85% of all non-Hodgkin lymphoma cases, with significant heterogeneity within the B-cell subgroup based on genotypic and phenotypic expression patterns in cells of B cell origin. For example, subgroups of B-cell lymphomas include slow-growing, indolent and incurable diseases such as follicular lymphoma (FL) or chronic lymphocytic leukemia (CLL), as well as more aggressive subtypes such as mantle cell lymphoma (MCL) and diffuse large B-cell lymphoma (DLBCL).Diffuse large B-cell lymphoma (DLBCL) is the most common type of NHL, accounting for approximately 30%–40% of all diagnosed NHL, followed by follicular lymphoma (FL; 20%–25% of all diagnosed NHL) and mantle cell lymphoma (MCL; 6%–10% of all diagnosed NHL). B-cell chronic lymphocytic leukemia (CLL) is the most common leukemia in adults, with approximately 15,000 new cases per year in the United States (American Cancer Society 2015).

[0007] Bispecific antibodies are capable of simultaneously binding cell surface antigens of cytotoxic cells (e.g., T cells, via binding to cluster of differentiation 3 (CD3)) and cancer cells (e.g., B cells, via binding to CD20) so that the bound cytotoxic cell kills the bound cancer cell. Glofitamab is a T-cell-capturing bispecific (TCB) antibody that targets CD20 expressed on B cells and the CD3 epsilon chain (CD3e) present on T cells.

[0008] However, immunotherapy using CD20 / CD3 bispecific antibodies such as glofitamab may be limited by adverse effects, including cytokine-mediated toxicities (e.g., cytokine release syndrome (CRS)), infusion-related reactions (IRR), severe tumor lysis syndrome (SLS), and central nervous system (CNS) toxicities.

[0009] There is thus an unmet need to develop effective dosing strategies for CD20 / CD3 bispecific antibody (e.g., glofitamab) for the treatment of CD20-positive B-cell proliferative disorders (e.g., non-Hodgkin lymphoma, NHL) that provide a more favorable benefit-risk profile.

[0010] SUMMARY OF THE INVENTION

[0011] In one aspect of the present invention, a method of treating a subject having a CD20-positive cell proliferative disorder is described, comprising administering to the subject a drug conjugate of an anti-CD79b antibody and a bispecific antibody that binds to CD20 and CD3, in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein C1D1 of the bispecific antibody is approximately 2.5 mg, and C1D2 of the bispecific antibody is approximately 10 mg; and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody, wherein C2D1 of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0012] In another aspect, the present invention provides a drug conjugate of an anti-CD79b antibody and a bispecific antibody that binds to CD20 and CD3 for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein the drug conjugate of the anti-CD79b antibody and the bispecific antibody that binds to CD20 and CD3 are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein C1D1 of the bispecific antibody is about 2.5 mg and C1D2 of the bispecific antibody is about 10 mg; and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0013] In another aspect, the present invention provides a use of a drug conjugate of an anti-CD79b antibody and a bispecific antibody that binds to CD20 and CD3 in the treatment of a subject having a CD20-positive cell proliferative disorder, wherein the drug conjugate of the anti-CD79b antibody and the bispecific antibody that binds to CD20 and CD3 are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein C1D1 of the bispecific antibody is approximately 2.5 mg and C1D2 of the bispecific antibody is approximately 10 mg; and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0014] In another aspect, the present invention provides a use of a drug conjugate of an anti-CD79b antibody and a bispecific antibody that binds to CD20 and CD3 in the manufacture of a medicament for treating a subject having a CD20-positive cell proliferative disorder, wherein the drug conjugate of the anti-CD79b antibody and the bispecific antibody that binds to CD20 and CD3 are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein C1D1 of the bispecific antibody is approximately 2.5 mg and C1D2 of the bispecific antibody is approximately 10 mg;and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0015] In some embodiments, the C2D1 of the bispecific antibody is approximately 10 mg. In some embodiments, the C2D1 of the bispecific antibody is approximately 16 mg. In some embodiments, the C2D1 of the bispecific antibody is approximately 30 mg.

[0016] In some embodiments, the first dosing cycle comprises a single dose of the C1D1 anti-CD79b antibody drug conjugate.In some embodiments, the single dose of the C1D1 anti-CD79b antibody drug conjugate is from about 0.1 mg / kg to about 2.4 mg / kg (e.g., from about 0.1 mg / kg to about 2.2 mg / kg, from about 0.1 mg / kg to about 2.0 mg / kg, from about 0.5 mg / kg to about 2.2 mg / kg, from about 0.8 mg / kg to about 2.2 mg / kg, from about 1 mg / kg to about 2.2 mg / kg, from about 1.2 mg / kg to about 2.2 mg / kg, from about 1.4 mg / kg to about 2.2 mg / kg, from about 1.6 mg / kg to about 2.2 mg / kg, from about 1.8 mg / kg to about 2.0 mg / kg, from about 0.1 mg / kg to about 1.6 mg / kg, from about 0.5 mg / kg to about 1.6 mg / kg or from about 1 mg / kg to about 1.8 mg / kg; for example, about 1 mg / kg, about 1.2 mg / kg, about 1.6 mg / kg, or about 1.8 mg / kg).In some embodiments, a single dose of the C1D1 anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg. In some embodiments, the C1D1 anti-CD79b antibody drug conjugate is administered or intended to be administered on approximately day 2 (±1 day) of the dosing cycle. In some embodiments, the second dosing cycle includes a single dose of the C2D1 anti-CD79b antibody drug conjugate.In some embodiments, a single dose of the C2D1 anti-CD79b antibody drug conjugate is from about 0.1 mg / kg to about 2.4 mg / kg (e.g., from about 0.1 mg / kg to about 2.2 mg / kg, from about 0.1 mg / kg to about 2.0 mg / kg, from about 0.5 mg / kg to about 2.2 mg / kg, from about 0.8 mg / kg to about 2.2 mg / kg, from about 1 mg / kg to about 2.2 mg / kg, from about 1.2 mg / kg to about 2.2 mg / kg, from about 1.4 mg / kg to about 2.2 mg / kg, from about 1.6 mg / kg to about 2.2 mg / kg, from about 1.8 mg / kg to about 2.0 mg / kg, from about 0.1 mg / kg to about 1.6 mg / kg, from about 0.5 mg / kg to about 1.6 mg / kg or from about 1 mg / kg to about 1.8 mg / kg; for example, about 1 mg / kg, about 1.2 mg / kg, about 1.6 mg / kg, or about 1.8 mg / kg).In some embodiments, a single dose of the C2D1 anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg.

[0017] In some embodiments, the C1D1 bispecific antibody and the C1D2 bispecific antibody are administered or intended to be administered to the subject on approximately days 8 (±1 day) and 15 (±1 day), respectively, of the first dosing cycle. In some embodiments, the C2D1 bispecific antibody is administered or intended to be administered to the subject on approximately day 1 (±1 day) of the second dosing cycle. In some embodiments, the C2D1 bispecific antibody is administered or intended to be administered after completion of administration of the C2D1 anti-CD79b antibody drug conjugate.In some embodiments, the C2D1 bispecific antibody is administered or is intended to be administered about 60-120 minutes (e.g., about 60-100 minutes, about 60-90 minutes, about 60-80 minutes, about 90-120 minutes, about 80-100 minutes, about 80-120 minutes, about 75-105 minutes, or about 85-95 minutes; e.g., about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes, about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes) after completion of administration of the C2D1 anti-CD79b antibody drug conjugate.In some embodiments, the C2D1 bispecific antibody is administered or intended to be administered approximately 90 minutes after completion of administration of the C2D1 anti-CD79b antibody drug conjugate. In some embodiments, the C1D1 anti-CD79b antibody drug conjugate is administered or intended to be administered to the subject on or about day 2 (±1 day) of the first dosing cycle, and the C2D1 anti-CD79b antibody drug conjugate is administered or intended to be administered to the subject on or about day 1 (±1 day) of the second dosing cycle. In some embodiments, the first and second dosing cycles are 14-day (e.g., 14±3 days) dosing cycles. In some embodiments, the first and second dosing cycles are 21-day (e.g., 21±3 days) dosing cycles.

[0018] In some embodiments, the dosing regimen includes one or more additional dosing cycles. In some embodiments, the dosing regimen includes six to ten additional dosing cycles (e.g., six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles). In some embodiments, the dosing regimen includes ten additional dosing cycles. In some embodiments, the additional dosing cycles are 14-day (e.g., 14 ± 3 days) dosing cycles. In some embodiments, the additional dosing cycles are 21-day (e.g., 21 ± 3 days) dosing cycles.

[0019] In some embodiments, one or more additional dosing cycles include an additional single dose of the bispecific antibody and an additional single dose of the anti-CD79b antibody drug conjugate. In some embodiments, the additional single dose of the anti-CD79b antibody drug conjugate is approximately equivalent in amount to the C2D1 anti-CD79b antibody drug conjugate. In some embodiments, the additional single dose of the anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg. In some embodiments, the additional single dose of the anti-CD79b antibody drug conjugate is administered or intended to be administered to the subject on approximately day 1 (±1 day) of each additional dosing cycle including the additional dose of the anti-CD79b antibody drug conjugate.

[0020] In some embodiments, the additional single dose of the bispecific antibody of each additional dosing cycle including the additional dose of the CD79b antibody drug conjugate is administered or is intended to be administered after completion of the administration of the additional single dose of the CD79b antibody drug conjugate.In some embodiments, the additional single dose of the bispecific antibody of each additional dosing cycle comprising an additional dose of the anti-CD79b antibody drug conjugate is administered or is intended to be administered about 60-120 minutes (e.g., about 60-100 minutes, about 60-90 minutes, about 60-80 minutes, about 90-120 minutes, about 80-100 minutes, about 80-120 minutes, about 75-105 minutes, or about 85-95 minutes; e.g., about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes, about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes) after completion of the additional single dose of the anti-CD79b antibody drug conjugate. medicinal product.In some embodiments, the additional single dose of the bispecific antibody of each additional dosing cycle including the additional dose of the anti-CD79b antibody drug conjugate is administered or is intended to be administered approximately 90 minutes after completion of the administration of the additional single dose of the anti-CD79b antibody drug conjugate.

[0021] In some embodiments, the dosing regimen includes at least four additional dosing cycles comprising an additional single dose of the anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen includes four to ten additional dosing cycles (e.g., four additional dosing cycles, five additional dosing cycles, six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles) comprising an additional single dose of the anti-CD79b antibody drug conjugate.

[0022] In some embodiments, one or more additional dosing cycles include an additional single dose of the bispecific antibody and do not include administration of the anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen includes at least two additional dosing cycles that include an additional single dose of the bispecific antibody and do not include administration of the anti-CD79b antibody drug conjugate.In some embodiments, the dosing regimen includes two to ten additional dosing cycles that include an additional single dose of the bispecific antibody and do not include administration of the anti-CD79b antibody drug conjugate (e.g., two additional dosing cycles, three additional dosing cycles, four additional dosing cycles, five additional dosing cycles, six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles).

[0023] In some embodiments, the additional single dose of the bispecific antibody is approximately equivalent in amount to the C2D1 bispecific antibody. In some embodiments, the additional single dose of the bispecific antibody is approximately 30 mg. In some embodiments, the additional single dose of the bispecific antibody is administered or intended to be administered to the subject on approximately day 1 (±1 day) of each additional dosing cycle including the additional dose of the bispecific antibody.

[0024] In some embodiments, the dosing regimen includes six or more additional dosing cycles (e.g., six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles), wherein each of the six or more additional dosing cycles includes a single dose of the bispecific antibody, and wherein no more than four (e.g., none, no more than one, no more than two, no more than three, or no more than four; such as none, one, two, three, or four) of the six or more additional dosing cycles include administration of an anti-CD79b antibody drug conjugate.In some embodiments, the dosing regimen includes six additional dosing cycles, wherein each of the six or more additional dosing cycles includes a single dose of the bispecific antibody, and wherein no more than four (e.g., none, no more than one, no more than two, no more than three, or no more than four; e.g., none, one, two, three, or four) of the six additional dosing cycles include administration of an anti-CD79b antibody drug conjugate.

[0025] In one aspect of the present invention, a method of treating a subject having a CD20-positive cell proliferative disorder is described, comprising administering to the subject a drug conjugate of an anti-CD79b antibody and a bispecific antibody that binds to CD20 and CD3, in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises: (i) a single dose (C1D1) of the anti-CD79b antibody drug conjugate; and (ii) a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein each of the C1D1 and C1D2 bispecific antibody is administered to the subject after the C1D1 anti-CD79b antibody drug conjugate, wherein the C1D1 bispecific antibody is approximately 2.5 mg and the C1D2 bispecific antibody is approximately 10 mg;and (b) the second dosing cycle comprises: (i) a single dose (C2D1) of the CD79b antibody drug conjugate; and (ii) a single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is about 10 mg, about 16 mg, or about 30 mg.

[0026] In another aspect, the present invention provides a drug conjugate of an anti-CD79b antibody and a bispecific antibody that binds to CD20 and CD3 for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein the drug conjugate of the anti-CD79b antibody and the bispecific antibody that binds to CD20 and CD3 are administered to the subject in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises: (i) a single dose (C1D1) of the anti-CD79b antibody drug conjugate;and (ii) a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein each of the C1D1 and C1D2 bispecific antibody is intended to be administered to the subject subsequent to the C1D1 of the anti-CD79b antibody drug conjugate, wherein the C1D1 of the bispecific antibody is approximately 2.5 mg and the C1D2 of the bispecific antibody is approximately 10 mg; and (b) the second dosing cycle comprises: (i) a single dose (C2D1) of the anti-CD79b antibody drug conjugate; and (ii) a single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0027] In another aspect, the present invention provides a use of an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 for treating a subject having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody that binds to CD20 and CD3 are administered to the subject in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises: (i) a single dose (C1D1) of the anti-CD79b antibody drug conjugate;and (ii) a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein each of the C1D1 and C1D2 bispecific antibody is intended to be administered to the subject subsequent to the C1D1 of the anti-CD79b antibody drug conjugate, wherein the C1D1 of the bispecific antibody is approximately 2.5 mg and the C1D2 of the bispecific antibody is approximately 10 mg; and (b) the second dosing cycle comprises: (i) a single dose (C2D1) of the anti-CD79b antibody drug conjugate; and (ii) a single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0028] In another aspect, the present invention provides the use of an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 in the manufacture of a medicament for treating a subject having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody that binds to CD20 and CD3 are administered to the subject in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises: (i) a single dose (C1D1) of the anti-CD79b antibody drug conjugate;and (ii) a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein each of the C1D1 and C1D2 bispecific antibody is intended to be administered to the subject subsequent to the C1D1 of the anti-CD79b antibody drug conjugate, wherein the C1D1 of the bispecific antibody is approximately 2.5 mg and the C1D2 of the bispecific antibody is approximately 10 mg; and (b) the second dosing cycle comprises: (i) a single dose (C2D1) of the anti-CD79b antibody drug conjugate; and (ii) a single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0029] In some embodiments, the single dose of the C1D1 anti-CD79b antibody drug conjugate is from about 0.1 mg / kg to about 2.4 mg / kg (e.g., from about 0.1 mg / kg to about 2.2 mg / kg, from about 0.1 mg / kg to about 2.0 mg / kg, from about 0.5 mg / kg to about 2.2 mg / kg, from about 0.8 mg / kg to about 2.2 mg / kg, from about 1 mg / kg to about 2.2 mg / kg, from about 1.2 mg / kg to about 2.2 mg / kg, from about 1.4 mg / kg to about 2.2 mg / kg, from about 1.6 mg / kg to about 2.2 mg / kg, from about 1.8 mg / kg to about 2.0 mg / kg, from about 0.1 mg / kg to about 1.6 mg / kg, from about 0.5 mg / kg to about 1.6 mg / kg, or from about 1 mg / kg to about 1.8 mg / kg;for example, about 1 mg / kg, about 1.2 mg / kg, about 1.6 mg / kg, or about 1.8 mg / kg), and a single dose of the C2D1 anti-CD79b antibody drug conjugate is from about 0.1 mg / kg to about 2.4 mg / kg (e.g., from about 0.1 mg / kg to about 2.2 mg / kg, from about 0.1 mg / kg to about 2.0 mg / kg, from about 0.5 mg / kg to about 2.2 mg / kg, from about 0.8 mg / kg to about 2.2 mg / kg, from about 1 mg / kg to about 2.2 mg / kg, from about 1.2 mg / kg to about 2.2 mg / kg, from about 1.4 mg / kg to about 2.2 mg / kg, from about 1.6 mg / kg to about 2.2 mg / kg, from about 1.8 mg / kg up to about 2.0 mg / kg, from about 0.1 mg / kg to about 1.6 mg / kg, from about 0.5 mg / kg to about 1.6 mg / kg, or from about 1 mg / kg to about 1.8 mg / kg;for example, about 1 mg / kg, about 1.2 mg / kg, about 1.6 mg / kg, or about 1.8 mg / kg). In some embodiments, a single dose of the C1D1 anti-CD79b antibody drug conjugate is about 1.8 mg / kg, and a single dose of the C2D1 anti-CD79b antibody drug conjugate is about 1.8 mg / kg;

[0030] In some embodiments, the C1D1 bispecific antibody and the C1D2 bispecific antibody are administered or intended to be administered to a subject on approximately days 8 (±1 day) and 15 (±1 day), respectively, of the first dosing cycle.

[0031] In some embodiments, the C2D1 bispecific antibody is administered or intended to be administered to the subject on or about day 1 (± 1 day) of the second dosing cycle. In some embodiments, the C1D1 anti-CD79b antibody drug conjugate is administered or intended to be administered to the subject on or about day 2 (± 1 day) of the first dosing cycle, and the C2D1 anti-CD79b antibody drug conjugate is administered or intended to be administered to the subject on or about day 1 (± 1 day) of the second dosing cycle.

[0032] In some embodiments, the first and second dosing cycles are 14-day (e.g., 14 ± 3 days) dosing cycles. In some embodiments, the first and second dosing cycles are 21-day (e.g., 21 ± 3 days) dosing cycles.

[0033] In some embodiments, the dosing regimen includes one or more additional dosing cycles. In some embodiments, the dosing regimen includes six to ten additional dosing cycles (e.g., six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles).

[0034] In some embodiments, the dosing regimen includes ten additional dosing cycles. In some embodiments, the additional dosing cycles are 21-day (e.g., 21 ± 3 days) dosing cycles. In some embodiments, the dosing regimen includes one or more additional dosing cycles. In some embodiments, the dosing regimen includes six to ten additional dosing cycles. In some embodiments, the dosing regimen includes ten additional dosing cycles.

[0035] In some embodiments, the additional dosing cycles are 14-day (e.g., 14 ± 3 days) dosing cycles.

[0036] In some embodiments, one or more additional dosing cycles include an additional single dose of the bispecific antibody and an additional single dose of the anti-CD79b antibody drug conjugate. In some embodiments, the additional single dose of the anti-CD79b antibody drug conjugate is approximately equivalent in amount to the C2D1 anti-CD79b antibody drug conjugate. In some embodiments, the additional single dose of the anti-CD79b antibody conjugate is administered or intended to be administered to the subject on approximately day 1 (±1 day) of each additional dosing cycle including the additional dose of the anti-CD79b antibody drug conjugate.

[0037] In some embodiments, the additional single dose of the bispecific antibody of each additional dosing cycle including the additional dose of the CD79b antibody drug conjugate is administered or is intended to be administered after completion of the administration of the additional single dose of the CD79b antibody drug conjugate.In some embodiments, the additional single dose of the bispecific antibody of each additional dosing cycle comprising an additional dose of the anti-CD79b antibody drug conjugate is administered or is intended to be administered about 60-120 minutes (e.g., about 60-100 minutes, about 60-90 minutes, about 60-80 minutes, about 90-120 minutes, about 80-100 minutes, about 80-120 minutes, about 75-105 minutes, or about 85-95 minutes; e.g., about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes, about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes) after completion of the additional single dose of the anti-CD79b antibody drug conjugate. medicinal product.In some embodiments, the additional single dose of the bispecific antibody of each additional dosing cycle including the additional dose of the anti-CD79b antibody drug conjugate is administered or is intended to be administered approximately 90 minutes after completion of the administration of the additional single dose of the anti-CD79b antibody drug conjugate.

[0038] In some embodiments, the dosing regimen includes at least four additional dosing cycles, each comprising an additional single dose of the bispecific antibody and an additional single dose of the anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen includes four to ten additional dosing cycles (e.g., four additional dosing cycles, five additional dosing cycles, six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles), each comprising an additional single dose of the bispecific antibody and an additional single dose of the anti-CD79b antibody drug conjugate.In some embodiments, the one or more additional dosing cycles include an additional single dose of the bispecific antibody and do not include administration of the anti-CD79b antibody drug conjugate.

[0039] In some embodiments, the dosing regimen includes at least two additional dosing cycles that include an additional single dose of the bispecific antibody and do not include administration of the anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen includes two to ten additional dosing cycles (e.g., two additional dosing cycles, three additional dosing cycles, four additional dosing cycles, five additional dosing cycles, six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles) that include an additional single dose of the bispecific antibody and do not include administration of the anti-CD79b antibody drug conjugate.

[0040] In some embodiments, the additional single dose of the bispecific antibody is approximately equivalent in amount to the C2D1 bispecific antibody. In some embodiments, the additional single dose of the bispecific antibody is approximately 30 mg. In some embodiments, the additional single dose of the bispecific antibody is administered or intended to be administered to the subject on approximately day 1 (±1 day) of each additional dosing cycle including the additional dose of the bispecific antibody.

[0041] In some embodiments, the dosing regimen includes six or more additional dosing cycles (e.g., six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles), wherein each of the six or more additional dosing cycles includes a single dose of the bispecific antibody, and wherein no more than four (e.g., none, no more than one, no more than two, no more than three, or no more than four; e.g., none, one, two, three, or four) of the six or more additional dosing cycles include administration of an anti-CD79b antibody drug conjugate.In some embodiments, the dosing regimen comprises six to ten additional dosing cycles (e.g., six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles), wherein each of the six to ten additional dosing cycles comprises a single dose of the bispecific antibody, and wherein no more than four (e.g., none, no more than one, no more than two, no more than three, or no more than four; e.g., none, one, two, three, or four) of the six additional dosing cycles comprises administration of an anti-CD79b antibody drug conjugate.In some embodiments, the dosing regimen comprises ten additional dosing cycles, wherein each of the ten additional dosing cycles comprises a single dose of the bispecific antibody, and wherein no more than four (e.g., none, no more than one, no more than two, no more than three, or no more than four; e.g., none, one, two, three, or four) of the ten additional dosing cycles comprise administration of the anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen comprises ten additional dosing cycles, wherein each of the ten additional dosing cycles comprises a single dose of the bispecific antibody, and wherein each of the ten additional dosing cycles comprise administration of the anti-CD79b antibody drug conjugate.

[0042] In one aspect, the present invention describes a method of treating a subject having a CD20-positive cell proliferative disorder, comprising administering to the subject a drug conjugate of an anti-CD79b antibody and a bispecific antibody that binds to CD20 and CD3, in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein the C1D1 of the bispecific antibody is approximately 2.5 mg and the C1D2 of the bispecific antibody is approximately 10 mg; (ii) a single dose (C1D1) of the anti-CD79b antibody drug conjugate; (b) each of the second through sixth dosing cycles comprises a single dose of (C2D1-C6D1) bispecific antibody and a single dose of (C2D1-C6D1) anti-CD79b antibody drug conjugate;and (c) each of the seventh through 12th dosing cycles includes a single dose of the (C7D1-C12D1) bispecific antibody and does not include administration of an anti-CD79b antibody drug conjugate, and wherein each single dose of the C2D1-C12D1 bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0043] In another aspect, the present invention provides a drug conjugate of an anti-CD79b antibody and a bispecific antibody that binds to CD20 and CD3 for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein the drug conjugate of the anti-CD79b antibody and the bispecific antibody that binds to CD20 and CD3 is administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein C1D1 of the bispecific antibody is approximately 2.5 mg and C1D2 of the bispecific antibody is approximately 10 mg; (ii) a single dose (C1D1) of an anti-CD79b antibody drug conjugate;(b) each of the second through sixth dosing cycles includes a single dose of (C2D1-C6D1) bispecific antibody and a single dose of (C2D1-C6D1) anti-CD79b antibody drug conjugate; and (c) each of the seventh through 12th dosing cycles includes a single dose of (C7D1-C12D1) bispecific antibody and does not include administration of an anti-CD79b antibody drug conjugate, and wherein each single dose of C2D1-C12D1 bispecific antibody is about 10 mg, about 16 mg, or about 30 mg.

[0044] In another aspect, the present invention describes the use of a drug conjugate of an anti-CD79b antibody and a bispecific antibody that binds to CD20 and CD3 in the treatment of a subject having a CD20-positive cell proliferative disorder, wherein the drug conjugate of the anti-CD79b antibody and the bispecific antibody that binds to CD20 and CD3 is administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein C1D1 of the bispecific antibody is approximately 2.5 mg and C1D2 of the bispecific antibody is approximately 10 mg; (ii) a single dose (C1D1) of an anti-CD79b antibody drug conjugate;(b) each of the second through sixth dosing cycles includes a single dose of (C2D1-C6D1) bispecific antibody and a single dose of (C2D1-C6D1) anti-CD79b antibody drug conjugate; and (c) each of the seventh through 12th dosing cycles includes a single dose of (C7D1-C12D1) bispecific antibody and does not include administration of an anti-CD79b antibody drug conjugate, and wherein each single dose of C2D1-C12D1 bispecific antibody is about 10 mg, about 16 mg, or about 30 mg.

[0045] In another aspect, the present invention describes the use of a drug conjugate of an anti-CD79b antibody and a bispecific antibody that binds to CD20 and CD3 in the manufacture of a medicament for treating a subject having a CD20-positive cell proliferative disorder, wherein the drug conjugate of the anti-CD79b antibody and the bispecific antibody that binds to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein C1D1 of the bispecific antibody is approximately 2.5 mg and C1D2 of the bispecific antibody is approximately 10 mg; (ii) a single dose (C1D1) of an anti-CD79b antibody drug conjugate;(b) each of the second through sixth dosing cycles includes a single dose of (C2D1-C6D1) bispecific antibody and a single dose of (C2D1-C6D1) anti-CD79b antibody drug conjugate; and (c) each of the seventh through 12th dosing cycles includes a single dose of (C7D1-C12D1) bispecific antibody and does not include administration of an anti-CD79b antibody drug conjugate, and wherein each single dose of C2D1-C12D1 bispecific antibody is about 10 mg, about 16 mg, or about 30 mg.

[0046] In some embodiments, the C2D1-C12D1 bispecific antibodies are approximately equivalent in amount. In some embodiments, the C2D1 bispecific antibody is approximately 30 mg.

[0047] In some embodiments, the C1D1-C6D1 anti-CD79b antibody drug conjugates are approximately equivalent in amount.In some embodiments, each of the C1D1-C6D1 anti-CD79b antibody drug conjugates is from about 0.1 mg / kg to about 2.4 mg / kg (e.g., from about 0.1 mg / kg to about 2.2 mg / kg, from about 0.1 mg / kg to about 2.0 mg / kg, from about 0.5 mg / kg to about 2.2 mg / kg, from about 0.8 mg / kg to about 2.2 mg / kg, from about 1 mg / kg to about 2.2 mg / kg, from about 1.2 mg / kg to about 2.2 mg / kg, from about 1.4 mg / kg to about 2.2 mg / kg, from about 1.6 mg / kg to about 2.2 mg / kg, from about 1.8 mg / kg to about 2.0 mg / kg, from about 0.1 mg / kg to about 1.6 mg / kg, from about 0.5 mg / kg to about 1.6 mg / kg or from about 1 mg / kg to about 1.8 mg / kg; for example, about 1 mg / kg, about 1.2 mg / kg, about 1.6 mg / kg, or about 1.8 mg / kg).In some embodiments, each of the C1D1-C6D1 anti-CD79b antibody drug conjugates is approximately 1.8 mg / kg.

[0048] In some embodiments, the C1D1 bispecific antibody and the C1D2 bispecific antibody are administered or intended to be administered to the subject on or about days 8 (±1 day) and 15 (±1 day), respectively, of the first dosing cycle. In some embodiments, the C2D1-C12D1 bispecific antibody is administered or intended to be administered to the subject on or about day 1 of each dosing cycle. In some embodiments, the C1D1 anti-CD79b antibody drug conjugate is administered or intended to be administered to the subject on or about day 2 (±1 day) of the first dosing cycle, and the C2D1-C6D1 anti-CD79b antibody drug conjugate is administered or intended to be administered to the subject on or about day 1 of each dosing cycle comprising administration of the anti-CD79b antibody drug conjugate.

[0049] In some embodiments, the C2D1-C6D1 bispecific antibody is administered or intended to be administered after completion of administration of the C2D1-C6D1 anti-CD79b antibody drug conjugate.In some embodiments, the C2D1-C6D1 bispecific antibody is administered or is intended to be administered about 60-120 minutes (e.g., about 60-100 minutes, about 60-90 minutes, about 60-80 minutes, about 90-120 minutes, about 80-100 minutes, about 80-120 minutes, about 75-105 minutes, or about 85-95 minutes; e.g., about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes, about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes) after completion of administration of the C2D1-C6D1 anti-CD79b antibody drug conjugate.In some embodiments, the C2D1-C6D1 bispecific antibody is administered or intended to be administered approximately 90 minutes after completion of administration of the C2D1-C6D1 anti-CD79b antibody drug conjugate.

[0050] In some embodiments, each dosing cycle is a 14-day (e.g., 14 ± 3 days) dosing cycle. In some embodiments, each dosing cycle is a 21-day (e.g., 21 ± 3 days) dosing cycle.

[0051] In some embodiments, the dosing regimen includes an additional retreatment regimen after completion of 12 dosing cycles of the dosing regimen. In some embodiments, the additional retreatment regimen includes 12 additional dosing cycles, wherein: (a) the first additional dosing cycle includes: (i) a first dose (C13D1) of the bispecific antibody and a second dose (C13D2) of the bispecific antibody, wherein the C13D1 bispecific antibody is approximately 2.5 mg,and C13D2 of the bispecific antibody is approximately 10 mg; (ii) a single dose (C13D1) of the anti-CD79b antibody drug conjugate; (b) each of the second through sixth additional dosing cycles comprises a single dose (C14D1-C18D1) of the bispecific antibody and a single dose (C14D1-C18D1) of the anti-CD79b antibody drug conjugate; and (c) each of the seventh through 12th additional dosing cycles comprises a single dose (C19D1-C24D1) of the bispecific antibody and does not include administration of the anti-CD79b antibody drug conjugate, and wherein each single dose of C14D1-C24D1 of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg. In some embodiments, (a) the first additional dosing cycle comprises: (i) a first dose (C13D1) of the bispecific antibody that is administered or that is intended to be administered on day 8 (± 1 day) of the first additional dosing cycle,and a second dose (C13D2) of the bispecific antibody administered or intended to be administered on day 15 (± 1 day) of the first additional dosing cycle, wherein the C13D1 of the bispecific antibody is approximately 2.5 mg and the C13D2 of the bispecific antibody is approximately 10 mg; and (ii) a single dose (C13D1) of the anti-CD79b antibody drug conjugate administered or intended to be administered on day 2 (± 1 day) of the first additional dosing cycle; (b) each of the second through sixth additional dosing cycles comprises a single dose (C14D1-C18D1) of the bispecific antibody and a single dose (C14D1-C18D1) of the anti-CD79b antibody drug conjugate; and (c) each of the seventh through 12th additional dosing cycles includes a single dose of (C19D1-C24D1) bispecific antibody and does not include administration of an anti-CD79b antibody drug conjugate,and wherein the C14D1-C24D1 bispecific antibody is administered or intended to be administered on day 1 (± 1 day) of each additional dosing cycle, and the C14D1-C18D1 anti-CD79b antibody conjugate is administered or intended to be administered on day 1 (± 1 day) of each additional dosing cycle, and wherein each single dose of the C14D1-C24D1 bispecific antibody is approximately 30 mg, and each single dose of the C13D1-C18D1 anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg. In some embodiments, a waiting period is provided between the completion of the 12 dosing cycles and the start of the 12 additional dosing cycles of the additional retreatment regimen. In some embodiments, the waiting period is from about one to about eight weeks. In some embodiments, each additional dosing cycle of the additional retreatment regimen is a 14-day (e.g.,14 ± 3 days) dosing cycle. In some embodiments, each additional dosing cycle of the additional retreatment regimen is a 21-day (e.g., 21 ± 3 days) dosing cycle.

[0052] In some embodiments, the method further comprises administering to the subject one or more additional therapeutic agents. In some embodiments, the anti-CD79b antibody drug conjugate and the bispecific antibody are administered together with one or more additional therapeutic agents.

[0053] In some embodiments, the one or more additional therapeutic agents comprise one or more chemotherapeutic agents. In some embodiments, the one or more chemotherapeutic agents comprise cyclophosphamide, doxorubicin, and rituximab.

[0054] In some embodiments, one or more additional therapeutic agents is tocilizumab. In some embodiments, one or more additional therapeutic agents is a corticosteroid. In some embodiments, the corticosteroid comprises prednisone, prednisolone, methylprednisolone, and dexamethasone.

[0055] In some embodiments, the method further comprises administering rituximab, cyclophosphamide, doxorubicin, and prednisone (R-CHP) to the subject. In some embodiments, the anti-CD79b antibody drug conjugate and the bispecific antibody are administered together with rituximab, cyclophosphamide, doxorubicin, and prednisone (R-CHP).

[0056] In some embodiments, one or more additional therapeutic agents is an antihistamine. In some embodiments, the antihistamine is diphenhydramine. In some embodiments, one or more additional therapeutic agents comprises allopurinol and rasburicase. In some embodiments, one or more additional therapeutic agents is an antipyretic.

[0057] In some embodiments, one or more additional therapeutic agents is obinutuzumab. In some embodiments, obinutuzumab is administered or intended to be administered before the bispecific antibody is administered. In some embodiments, obinutuzumab is administered or intended to be administered about seven days (±1 day) before the bispecific antibody is administered. In some embodiments, obinutuzumab is administered or intended to be administered as a single dose of about 1000 mg. In some embodiments, obinutuzumab is administered in a first dose of about 1000 mg and in a second dose of about 1000 mg. In some embodiments, the first dose of obinutuzumab is administered about seven days (±1 day) before the C1D1 bispecific antibody is administered. In some embodiments, the second dose of obinutuzumab is administered about one day (±1 day) before the C1D1 bispecific antibody is administered.

[0058] In some embodiments, the anti-CD79b antibody drug conjugate is polatuzumab vedotin or anti-CD79b-MC-vc-PAB-MMAE. In some embodiments, the anti-CD79b antibody drug conjugate is polatuzumab vedotin.

[0059] In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds to CD20, comprising the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence YSWIN (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence RIFPGDGDTDYNGKFKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence NVFDGYWLVY (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence RSSKSLLHSNGITYLY (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence QMSNLVS (SEQ ID NO: 5); and (e) HVR-L3 comprising the amino acid sequence (SEQ ID NO: 6).In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds to CD20, comprising (a) a heavy chain variable domain VH comprising an amino acid sequence having at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%; e.g., 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO:7; (b) a light chain variable domain (VL) comprising an amino acid sequence having at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%; e.g., 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 8; or (c) a VH domain as in (a) and a VL domain as in (b).In some embodiments, a Fab molecule that specifically binds to CD20 comprises (a) a VH domain comprising the amino acid sequence of SEQ ID NO:7 and (b) a VL domain comprising the amino acid sequence of SEQ ID NO:8.

[0060] In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds to CD3, comprising the following six HVRs: (a) HVR-H1 comprising the amino acid sequence TYAMN (SEQ ID NO: 9); (b) HVR-H2 comprising the amino acid sequence RIRSKYNNYATYYADSVKG (SEQ ID NO: 10); (c) HVR-H3 comprising the amino acid sequence HGNFGNSYVSWFAY (SEQ ID NO: 11); (d) HVR-L1 comprising the amino acid sequence GSSTGAVTTSNYAN (SEQ ID NO: 12); (e) HVR-L2 comprising the amino acid sequence GTNKRAP (SEQ ID NO: 13); and (e) HVR-L3 comprising the amino acid sequence ALWYSNLWV (SEQ ID NO: 14).In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds to CD3, comprising (a) a heavy chain variable domain VH comprising an amino acid sequence having at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%; e.g., 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 15; (b) a light chain variable domain (VL) comprising an amino acid sequence having at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%; e.g., 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 16; or (c) a VH domain as in (a) and a VL domain as in (b).In some embodiments, a Fab molecule that specifically binds to CD3 comprises (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 15 and (b) a VL domain comprising the amino acid sequence of SEQ ID NO: 16.

[0061] In some embodiments, the bispecific antibody comprises a Fab molecule that specifically binds to CD3, wherein (a) the variable domains of the heavy and light chains of the Fab are exchanged, or (b) the constant domains of the heavy and light chains of the Fab are exchanged. In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds to CD20, wherein in the CL constant domain of the Fab molecule, the amino acid at position 124 is substituted with lysine (K) (Kabat numbering), and the amino acid at position 123 is substituted with arginine (R) or lysine (K) (Kabat numbering), and wherein in the CH1 constant domain of the Fab molecule, the amino acid at position 147 is substituted with glutamic acid (E) (EU numbering), and the amino acid at position 213 is substituted with glutamic acid (E) (EU numbering). In some embodiments, the bispecific antibody is bivalent for CD20 and monovalent for CD3.In some embodiments, the bispecific antibody comprises two Fab molecules that specifically bind to CD20 and one Fab molecule that specifically binds to CD3.

[0062] In some embodiments, the bispecific antibody comprises (a) a first Fab molecule that specifically binds to CD20; (b) a second Fab molecule that specifically binds to CD3; (c) a third Fab molecule that specifically binds to CD20; and (d) an Fc domain consisting of first and second subunits capable of stable association; wherein the third Fab molecule of (c) is identical to the first Fab molecule of (a); wherein in the CL constant domain of the first Fab molecule of (a) and the third Fab molecule of (c), the amino acid at position 124 is substituted with lysine (K) (Kabat numbering), and the amino acid at position 123 is substituted with arginine (R) or lysine (K) (Kabat numbering); and in the constant domain CH1 of the first Fab molecule according to (a) and of the third Fab molecule according to (b), the amino acid at position 147 is substituted with glutamic acid (E) (EU numbering), and the amino acid at position 213 is substituted with glutamic acid (E) (EU numbering);and wherein the first Fab molecule according to (a) is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab molecule according to (b), and each of the second Fab molecule according to (b) and the third Fab molecule according to (c) is fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain according to (d).;

[0063] In some embodiments, the bispecific antibody is a humanized antibody. In some embodiments, the bispecific antibody is a chimeric antibody.

[0064] In some embodiments, the bispecific antibody comprises an Fc domain, wherein the Fc domain is an IgG Fc domain. In some embodiments, the IgG Fc domain is an IgG1 Fc domain. In some embodiments, the IgG Fc domain comprises a mutation at amino acid residue N297 (EU numbering) that results in the absence of glycosylation. In some embodiments, the mutation at amino acid residue N297 is a substitution mutation. In some embodiments, the mutation at amino acid residue N297 reduces the effector function of the Fc region. In some embodiments, the mutation is N297G or N297A. In some embodiments, the bispecific antibody comprises a mutation in the Fc region that reduces effector function. In some embodiments, the mutation is a substitution mutation.In some embodiments, the substitution mutation occurs at amino acid residue L234, L235, D265, and / or P329 (EU numbering). In some embodiments, the mutation is selected from the group consisting of L234A, L235A, D265A, and P329G.

[0065] In some embodiments, the bispecific antibody comprises one or more heavy chain constant domains, wherein the one or more heavy chain constant domains are selected from a first CH1 domain (CH11), a first CH2 domain (CH21), a first CH3 domain (CH31), a second CH1 domain (CH12), a second CH2 domain (CH22), and a second CH3 domain (CH32). In some embodiments, at least one of the one or more heavy chain constant domains is fused with another heavy chain constant domain. In some embodiments, each of the CH31 and CH32 domains comprises a protrusion or a cavity, and wherein the protrusion or cavity in the CH31 domain can be located in a cavity or protrusion, respectively, in the CH32 domain. In some embodiments, the CH31 and CH32 domains converge at the surface between the protrusion and the cavity.In some embodiments, each of the CH21 and CH22 domains comprises a protrusion or cavity, and wherein the protrusion or cavity in the CH21 domain may be located within the cavity or protrusion, respectively, in the CH22 domain. In some embodiments, the CH21 and CH22 domains converge at the surface between said protrusion and cavity.

[0066] In some embodiments, the bispecific antibody is glofitamab.

[0067] In some embodiments, the bispecific antibody is administered or intended to be administered intravenously. In some embodiments, the anti-CD79b antibody drug conjugate is administered or intended to be administered intravenously. In some embodiments, if the bispecific antibody and the anti-CD79b antibody drug conjugate are administered or intended to be administered on the same day, the bispecific antibody is administered or intended to be administered after completion of administration of the anti-CD79b antibody drug conjugate.In some embodiments, the bispecific antibody is administered or is intended to be administered about 60-120 minutes (e.g., about 60-100 minutes, about 60-90 minutes, about 60-80 minutes, about 90-120 minutes, about 80-100 minutes, about 80-120 minutes, about 75-105 minutes, or about 85-95 minutes; e.g., about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes, about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes) after completion of administration of the anti-CD79b antibody drug conjugate.In some embodiments, the bispecific antibody is administered or is intended to be administered approximately 90 minutes after completion of administration of the anti-CD79b antibody drug conjugate.

[0068] In one aspect, the present invention describes a method of treating a subject having a CD20-positive cell proliferative disorder, comprising administering to the subject polatuzumab vedotin and glofitamab in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab that is administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab that is administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg, C1D2 of glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises a single dose (C2D1) of glofitamab administered on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0069] In another aspect, the present invention describes polatuzumab vedotin and glofitamab for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg, C1D2 of glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises a single dose (C2D1) of glofitamab intended to be administered on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin intended to be administered on day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0070] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab for treating a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg, C1D2 of glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises a single dose (C2D1) of glofitamab intended to be administered on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin intended to be administered on day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0071] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the manufacture of a medicament for treating a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg, C1D2 of glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises a single dose (C2D1) of glofitamab intended to be administered on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin intended to be administered on day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0072] In one aspect, the present invention describes a method of treating a subject having a CD20-positive cell proliferative disorder, comprising administering to the subject polatuzumab vedotin and glofitamab in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises: (i) a single dose (C1D1) of polatuzumab vedotin, which is administered on day 2 (± 1 day) of the first dosing cycle; and (ii) a first dose (C1D1) of glofitamab administered on day 8 (± day 1) of the first dosing cycle and a second dose (C1D2) of glofitamab administered on day 15 (± day 1) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg and C1D2 of glofitamab is approximately 10 mg; and (b) the second dosing cycle comprises: (i) a single dose (C2D1) of polatuzumab vedotin administered on day 1 (± day 1) of the second dosing cycle;and (ii) a single dose (C2D1) of glofitamab administered on day 1 (±1 day) of the second dosing cycle, wherein the C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg, and the C1D1 and C2D1 of polatuzumab vedotin are each approximately 1.8 mg / kg;

[0073] In another aspect, the present invention describes polatuzumab vedotin and glofitamab for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises: (i) a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle; and (ii) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg and C1D2 of glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises: (i) a single dose (C2D1) of polatuzumab vedotin intended to be administered on day 1 (± 1 day) of the second dosing cycle; and (ii) a single dose (C2D1) of glofitamab intended to be administered on day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg, and the C1D1 and C2D1 of polatuzumab vedotin are each approximately 1.8 mg / kg.

[0074] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the treatment of a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises: (i) a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle; and (ii) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg and C1D2 of glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises: (i) a single dose (C2D1) of polatuzumab vedotin intended to be administered on day 1 (± 1 day) of the second dosing cycle; and (ii) a single dose (C2D1) of glofitamab intended to be administered on day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg, and the C1D1 and C2D1 of polatuzumab vedotin are each approximately 1.8 mg / kg.

[0075] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the manufacture of a medicament for treating a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises: (i) a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle; and (ii) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg and C1D2 of glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises: (i) a single dose (C2D1) of polatuzumab vedotin intended to be administered on day 1 (± 1 day) of the second dosing cycle; and (ii) a single dose (C2D1) of glofitamab intended to be administered on day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg, and the C1D1 and C2D1 of polatuzumab vedotin are each approximately 1.8 mg / kg.

[0076] In one aspect, the present invention describes a method of treating a subject having a CD20-positive cell proliferative disorder, comprising administering to the subject polatuzumab vedotin and glofitamab in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab that is administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of glofitamab that is administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin that is administered on day 2 (± 1 day) of the first dosing cycle, wherein the C1D1 of glofitamab is from about 1 mg to about 5 mg, and the C1D2 of glofitamab is about 10 mg;(b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, wherein C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg; and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in amount to C2D1 glofitamab, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0077] In another aspect, the present invention describes polatuzumab vedotin and glofitamab for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg, and C1D2 glofitamab is approximately 10 mg;(b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, wherein C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg; and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in the amount of C2D1 glofitamab, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0078] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the treatment of a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg, and C1D2 glofitamab is approximately 10 mg;(b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, wherein C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg; and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in the amount of C2D1 glofitamab, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0079] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the manufacture of a medicament for the treatment of a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle, wherein the C1D1 of glofitamab is approximately 2.5 mg, and The C1D2 of glofitamab is approximately 10 mg;(b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, wherein C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg; and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in the amount of C2D1 glofitamab, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0080] In one aspect, the present invention describes a method of treating a subject having a CD20-positive cell proliferative disorder, comprising administering to the subject polatuzumab vedotin and glofitamab in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab that is administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab that is administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of glofitamab is approximately 2.5 mg and the C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin that is administered on day 2 (± 1 day) of the first dosing cycle; (b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, wherein C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg;and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in amount to C2D1 glofitamab, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0081] In another aspect, the present invention describes polatuzumab vedotin and glofitamab for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg and C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin, administered on day 2 (±1 day) of the first dosing cycle;(b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, wherein C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg; and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in the amount of C2D1 glofitamab, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0082] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the treatment of a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg and C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin, administered on day 2 (±1 day) of the first dosing cycle;(b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, wherein C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg; and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in the amount of C2D1 glofitamab, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0083] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the manufacture of a medicament for treating a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg and C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin, administered on day 2 (±1 day) of the first dosing cycle;(b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, wherein C2D1 of glofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg; and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in the amount of C2D1, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0084] In one aspect, the present invention describes a method of treating a subject having a CD20-positive cell proliferative disorder, comprising administering to the subject polatuzumab vedotin and glofitamab in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab that is administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab that is administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of glofitamab is approximately 2.5 mg and the C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin that is administered on day 2 (± 1 day) of the first dosing cycle; (b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, with C2D1 of glofitamab being approximately 30 mg;and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in amount to C2D1 glofitamab, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0085] In another aspect, the present invention describes polatuzumab vedotin and glofitamab for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg and C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin, administered on day 2 (±1 day) of the first dosing cycle;(b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, with C2D1 of glofitamab being approximately 30 mg; and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in the amount of C2D1 glofitamab, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0086] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the treatment of a subject having a CD20 positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg and C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin, administered on day 2 (±1 day) of the first dosing cycle;(b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, with C2D1 of glofitamab being approximately 30 mg; and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in the amount of C2D1 glofitamab, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0087] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the manufacture of a medicament for treating a subject having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg and C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin, administered on day 2 (±1 day) of the first dosing cycle;(b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin, with C2D1 of glofitamab being approximately 30 mg; and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C3D1-C12D1 glofitamab is approximately equal in the amount of C2D1, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0088] In some embodiments, the dosing cycles are 14-day (e.g., 14 ± 3 days) dosing cycles. In some embodiments, the dosing cycles are 21-day (e.g., 21 ± 3 days) dosing cycles.

[0089] In some embodiments, glofitamab is administered intravenously. In some embodiments, polatuzumab vedotin is administered intravenously. In some embodiments, if glofitamab and polatuzumab vedotin are administered or intended to be administered on the same day, glofitamab is administered or intended to be administered after completion of administration of polatuzumab vedotin.In some embodiments, glofitamab is administered or intended to be administered about 60-120 minutes (e.g., about 60-100 minutes, about 60-90 minutes, about 60-80 minutes, about 90-120 minutes, about 80-100 minutes, about 80-120 minutes, about 75-105 minutes, or about 85-95 minutes; e.g., about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes, about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes) after completion of administration of polatuzumab. In some embodiments, glofitamab is administered or intended to be administered about 90 minutes after completion of administration of polatuzumab.

[0090] In some embodiments, the method further comprises administering obinutuzumab to the subject. In some embodiments, polatuzumab vedotin and glofitamab are administered together with obinutuzumab. In some embodiments, obinutuzumab is administered or intended to be administered before glofitamab. In some embodiments, obinutuzumab is administered or intended to be administered approximately seven days (±1 day) before glofitamab. In some embodiments, obinutuzumab is administered or intended to be administered as a single dose of approximately 1000 mg.

[0091] In some embodiments, the CD20-positive cell proliferative disorder is a B-cell proliferative disorder. In some embodiments, the B-cell proliferative disorder is non-Hodgkin's lymphoma (NHL) or central nervous system lymphoma (CNS). In some embodiments, the NHL is relapsed and / or refractory. In some embodiments, the NHL is diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), high-grade B-cell lymphoma, primary mediastinal (thymic) large B-cell lymphoma (PMLBCL), diffuse B-cell lymphoma, or small lymphocytic lymphoma.

[0092] In some embodiments, the NHL is DLBCL. In some embodiments, the DLBCL is relapsed or refractory DLBCL.

[0093] In some embodiments, the NHL is FL. In some embodiments, the FL is relapsed or refractory FL. In some embodiments, the FL is transformed FL.

[0094] In some embodiments, the NHL is MCL. In some embodiments, the MCL is relapsed or refractory MCL.

[0095] In some embodiments, the CD20-positive cell proliferative disorder is not chronic lymphoid leukemia (CLL), acute lymphoblastic leukemia (ALL), Richter's transformation, Burkitt's lymphoma, or lymphoplasmacytic lymphoma.

[0096] In one aspect, the present invention describes a method of treating a population of subjects having R / R NHL, comprising administering to the subjects an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of the bispecific antibody that is administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of the bispecific antibody that is administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin that is administered on day 2 (± 1 day) of the first dosing cycle, wherein C1D1 of the bispecific antibody is approximately 2.5 mg, and C1D2 bispecific antibody is approximately 10 mg;and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody administered on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (± 1 day) of the second dosing cycle, wherein C2D1 of the bispecific antibody is approximately 30 mg, and wherein C1D1 and C1D2 of the anti-CD79b antibody drug conjugate are each approximately 1.8 mg / kg;

[0097] In another aspect, the present invention provides an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 for use in a method of treating a population of subjects having R / R NHL, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody that binds to CD20 and CD3 are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of the bispecific antibody intended to be administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of the bispecific antibody intended to be administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle, with C1D1 bispecific antibody being approximately 2.5 mg,and C1D2 of the bispecific antibody is approximately 10 mg; and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody intended for administration on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin intended for administration on day 1 (± 1 day) of the second dosing cycle, wherein C2D1 of the bispecific antibody is approximately 30 mg, and C1D1 and C1D2 of the anti-CD79b antibody drug conjugate are each approximately 1.8 mg / kg.

[0098] In another aspect, the present invention provides the use of an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 in the treatment of a population of subjects having R / R NHL, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody that binds to CD20 and CD3 are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of the bispecific antibody intended to be administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of the bispecific antibody intended to be administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle, with the C1D1 bispecific antibody being approximately 2.5 mg,and C1D2 of the bispecific antibody is approximately 10 mg; and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody intended for administration on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin intended for administration on day 1 (± 1 day) of the second dosing cycle, wherein C2D1 of the bispecific antibody is approximately 30 mg, and C1D1 and C1D2 of the anti-CD79b antibody drug conjugate are each approximately 1.8 mg / kg.

[0099] In another aspect, the present invention describes the use of an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 in the manufacture of a medicament for the treatment of a population of subjects having R / R NHL, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody that binds to CD20 and CD3 are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of the bispecific antibody intended for administration on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of the bispecific antibody intended for administration on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin intended for administration on day 2 (± 1 day) of the first dosing cycle, with C1D1 bispecific antibody being approximately 2.5 mg,and C1D2 of the bispecific antibody is approximately 10 mg; and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody intended for administration on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin intended for administration on day 1 (± 1 day) of the second dosing cycle, wherein C2D1 of the bispecific antibody is approximately 30 mg, and C1D1 and C1D2 of the anti-CD79b antibody drug conjugate are each approximately 1.8 mg / kg.

[0100] In one aspect, the present invention describes a method of treating a population of subjects having R / R NHL, comprising administering to the subjects an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3, in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of the bispecific antibody that is administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of the bispecific antibody that is administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of the bispecific antibody is approximately 2.5 mg, and the C1D2 of the bispecific antibody is approximately 10 mg; and (ii) a single dose (C1D1) of the anti-CD79b antibody drug conjugate administered on day 2 (±1 day) of the first dosing cycle;(b) each of the second through sixth dosing cycles contains a single dose of (C2D1-C6D1) bispecific antibody and a single dose of (C2D1-C6D1) anti-CD79b antibody drug conjugate; and (c) each of the seventh through 12th dosing cycles comprises a single dose of the (C7D1-C12D1) bispecific antibody and does not include administration of the anti-CD79b antibody drug conjugate, and wherein each single dose of the C2D1-C12D1 bispecific antibody is administered on day 1 (± 1 day) of each dosing cycle, and each single dose of the C2D1-C6D1 anti-CD79b antibody drug conjugate is administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of the C2D1-C12D1 bispecific antibody is approximately 30 mg, and each single dose of the C1D1-C6D1 anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg.

[0101] In another aspect, the present invention provides an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 for use in a method of treating a population of subjects having R / R NHL, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody that binds to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of the bispecific antibody intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of the bispecific antibody intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of the bispecific antibody is approximately 2.5 mg, and C1D2 of the bispecific antibody is approximately 10 mg;and (ii) a single dose (C1D1) of the anti-CD79b antibody drug conjugate to be administered on day 2 (± 1 day) of the first dosing cycle; (b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of the bispecific antibody and a single dose (C2D1-C6D1) of the anti-CD79b antibody drug conjugate;and (c) each of the seventh through 12th dosing cycles comprises a single dose of (C7D1-C12D1) bispecific antibody and does not include administration of an anti-CD79b antibody drug conjugate, and wherein each single dose of the C2D1-C12D1 bispecific antibody is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of the C2D1-C6D1 anti-CD79b antibody drug conjugate is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of the C2D1-C12D1 bispecific antibody is approximately 30 mg, and each single dose of the C1D1-C6D1 anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg.

[0102] In another aspect, the present invention describes the use of an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 for the treatment of a population of subjects having R / R NHL, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody that binds to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of the bispecific antibody intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of the bispecific antibody intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of the bispecific antibody is approximately 2.5 mg, and C1D2 of the bispecific antibody is approximately 10 mg;and (ii) a single dose (C1D1) of the anti-CD79b antibody drug conjugate to be administered on day 2 (± 1 day) of the first dosing cycle; (b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of the bispecific antibody and a single dose (C2D1-C6D1) of the anti-CD79b antibody drug conjugate;and (c) each of the seventh through 12th dosing cycles comprises a single dose of (C7D1-C12D1) bispecific antibody and does not include administration of an anti-CD79b antibody drug conjugate, and wherein each single dose of the C2D1-C12D1 bispecific antibody is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of the C2D1-C6D1 anti-CD79b antibody drug conjugate is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of the C2D1-C12D1 bispecific antibody is approximately 30 mg, and each single dose of the C1D1-C6D1 anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg.

[0103] In another aspect, the present invention describes the use of an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 in the manufacture of a medicament for the treatment of a population of subjects having R / R NHL, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody that binds to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of the bispecific antibody intended for administration on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of the bispecific antibody intended for administration on day 15 (± 1 day) of the first dosing cycle, wherein C1D1 of the bispecific antibody is approximately 2.5 mg, and C1D2 of the bispecific antibodies are approximately 10 mg;and (ii) a single dose (C1D1) of the anti-CD79b antibody drug conjugate to be administered on day 2 (± 1 day) of the first dosing cycle; (b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of the bispecific antibody and a single dose (C2D1-C6D1) of the anti-CD79b antibody drug conjugate;and (c) each of the seventh through 12th dosing cycles comprises a single dose of (C7D1-C12D1) bispecific antibody and does not include administration of an anti-CD79b antibody drug conjugate, and wherein each single dose of the C2D1-C12D1 bispecific antibody is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of the C2D1-C6D1 anti-CD79b antibody drug conjugate is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of the C2D1-C12D1 bispecific antibody is approximately 30 mg, and each single dose of the C1D1-C6D1 anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg.

[0104] In one aspect, the present invention describes a method of treating a population of subjects having R / R NHL, comprising administering to the subjects polatuzumab vedotin and glofitamab in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab that is administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of glofitamab that is administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin that is administered on day 2 (± 1 day) of the first dosing cycle, wherein the C1D1 of glofitamab is approximately 2.5 mg and the C1D2 of glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises a single dose (C2D1) of glofitamab administered on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (± 1 day) of the second dosing cycle, wherein C2D1 of glofitamab is approximately 30 mg, and wherein C1D1 and C1D2 of polatuzumab vedotin are each approximately 1.8 mg / kg.

[0105] In another aspect, the present invention describes polatuzumab vedotin and glofitamab for use in a method of treating a population of subjects having R / R NHL, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg, and C1D2 of glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises a single dose (C2D1) of glofitamab intended to be administered on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin intended to be administered on day 1 (± 1 day) of the second dosing cycle, wherein C2D1 of glofitamab is approximately 30 mg, and C1D1 and C1D2 of polatuzumab vedotin are each approximately 1.8 mg / kg;

[0106] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the treatment of a population of subjects having R / R NHL, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle, wherein the C1D1 of glofitamab is approximately 2.5 mg and the C1D2 of glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises a single dose (C2D1) of glofitamab intended to be administered on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin intended to be administered on day 1 (± 1 day) of the second dosing cycle, wherein C2D1 of glofitamab is approximately 30 mg, and C1D1 and C1D2 of polatuzumab vedotin are each approximately 1.8 mg / kg;

[0107] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the manufacture of a medicament for the treatment of a population of subjects having R / R NHL, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin intended to be administered on day 2 (± 1 day) of the first dosing cycle, wherein C1D1 of glofitamab is approximately 2.5 mg, and C1D2 glofitamab is approximately 10 mg;and (b) the second dosing cycle comprises a single dose (C2D1) of glofitamab intended to be administered on day 1 (± 1 day) of the second dosing cycle and a single dose (C2D1) of polatuzumab vedotin intended to be administered on day 1 (± 1 day) of the second dosing cycle, wherein C2D1 of glofitamab is approximately 30 mg, and C1D1 and C1D2 of polatuzumab vedotin are each approximately 1.8 mg / kg;

[0108] In one aspect, the present invention describes a method of treating a population of subjects having R / R NHL, comprising administering to the subjects polatuzumab vedotin and glofitamab in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab that is administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab that is administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of glofitamab is approximately 2.5 mg and the C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin that is administered on day 2 (± 1 day) of the first dosing cycle; (b) each of the second through sixth dosing cycles consists of a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin;and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C2D1-C12D1 glofitamab is approximately 30 mg, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0109] In another aspect, the present invention describes polatuzumab vedotin and glofitamab for use in a method of treating a population of subjects having R / R NHL, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of glofitamab is approximately 2.5 mg and the C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin, administered on day 2 (±1 day) of the first dosing cycle; (b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin;and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1-C12D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C2D1-C12D1 glofitamab is approximately 30 mg, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0110] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the treatment of a population of subjects having R / R NHL, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of glofitamab is approximately 2.5 mg and the C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin, administered on day 2 (±1 day) of the first dosing cycle; (b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin;and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C2D1-C12D1 glofitamab is approximately 30 mg, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0111] In another aspect, the present invention describes the use of polatuzumab vedotin and glofitamab in the manufacture of a medicament for the treatment of a population of subjects having R / R NHL, wherein polatuzumab vedotin and glofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) a first dose (C1D1) of glofitamab intended to be administered on day 8 (± 1 day) of the first dosing cycle, and a second dose (C1D2) of glofitamab intended to be administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of glofitamab is approximately 2.5 mg and the C1D2 of glofitamab is approximately 10 mg; and (ii) a single dose (C1D1) of polatuzumab vedotin, administered on day 2 (±1 day) of the first dosing cycle; (b) each of the second through sixth dosing cycles comprises a single dose (C2D1-C6D1) of glofitamab and a single dose (C2D1-C6D1) of polatuzumab vedotin;and (c) each of the seventh through 12th dosing cycles includes a single dose (C7D1) of glofitamab and does not include administration of polatuzumab vedotin, and wherein each single dose of C2D1-C12D1 glofitamab is intended to be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1-C6D1 polatuzumab vedotin is intended to be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of C2D1-C12D1 glofitamab is approximately 30 mg, and each single dose of C1D1-C6D1 polatuzumab vedotin is approximately 1.8 mg / kg.

[0112] In some embodiments, the complete response rate is at least 20% (e.g., at least 25%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%; such as in the range of 20-100%, in the range of 40-100%, in the range of 60-100%, in the range of 80-100%, in the range of 20-80%, in the range of 20-60%, in the range of 20-40%, in the range of 40-80%, in the range of 40-60%, in the range of 30-50%, or in the range of 35-45%; such as about 20%, about 25%, about 30%, about 35%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 50%, about 60%, or more). In some embodiments, the complete response rate is at least 40%. In particular embodiments, the complete response rate in a population of subjects having R / R NHL is about 42%.In some embodiments, the overall response rate is at least 30% (e.g., at least 35%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%; e.g., in the range of 30-100%, in the range of 50-100%, in the range of 70-100%, in the range of 30-90%, in the range of 30-70%, in the range of 30-50%, in the range of 40-80%, in the range of 40-60%, in the range of 45-55%, or in the range of 35-45%; e.g., about 30%, about 35%, about 40%, about 45%, about 48%, about 49%, about 50%, about 51%, about 52%, about 55%, about 60%, about 70%, or more). In some embodiments, the overall response rate is at least 50%. In particular embodiments, the overall response rate in a population of subjects with R / R NHL is about 50%.

[0113] In some embodiments, the B-cell proliferative disorder is R / R MCL. In some embodiments, the R / R NHL is R / R MCL. In some embodiments, the complete response rate is at least 60% (e.g., at least 60%, at least 70%, at least 80%, or at least 90%; e.g., in the range of 60-100%, in the range of 70-100%, in the range of 80-100%, in the range of 90-100%, in the range of 60-90%, in the range of 60-80%, in the range of 60-70%, in the range of 70-90%, in the range of 80-90%, in the range of 80-100%, or in the range of 90-100%; e.g., about 60%, about 70%, about 75%, about 80%, about 83%, about 85%, about 87%, about 90%, about 95%, about 97%, about 98%, about 99%, or more). In some embodiments, the complete response rate is at least 80%.In particular embodiments, the complete response rate in a population of subjects having R / R NHL is at least about 85%. In particular embodiments, the complete response rate in a population of subjects having R / R LCL is at least about 85%. In particular embodiments, the complete response rate in a population of subjects having R / R LCL is approximately 100%.In some embodiments, the overall response rate is at least 60% (e.g., at least 60%, at least 70%, at least 80%, or at least 90%; e.g., in the range of 60-100%, in the range of 70-100%, in the range of 80-100%, in the range of 90-100%, in the range of 60-90%, in the range of 60-80%, in the range of 60-70%, in the range of 70-90%, in the range of 80-90%, in the range of 80-100%, or in the range of 90-100%; e.g., about 60%, about 70%, about 75%, about 80%, about 83%, about 85%, about 87%, about 90%, about 95%, about 97%, about 98%, about 99%, or more). In particular embodiments, the overall response rate in a population of subjects having R / R NHL is at least about 85%. In particular embodiments, the overall response rate in a population of subjects having R / R LCL is at least about 85%.In specific embodiments, the overall response rate in a population of subjects having R / R LCM is approximately 100%. In some embodiments, the overall response rate is at least 80%.

[0114] In some embodiments, the B-cell proliferative disorder is R / R DLBCL. In some embodiments, R / R NHL is R / R MCL. In some embodiments, R / R NHL is R / R DLBCL.In some embodiments, the complete response rate is at least 60% (e.g., at least 60%, at least 70%, at least 80%, or at least 90%; e.g., in the range of 60-100%, in the range of 70-100%, in the range of 80-100%, in the range of 90-100%, in the range of 60-90%, in the range of 60-80%, in the range of 60-70%, in the range of 60-65%, in the range of 65-75%, or in the range of 75-85%; e.g., about 60%, about 61%, about 62%, about 65%, about 68%, about 69%, about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 76%, about 77%, about 80%, about 85%, about 90%, or more). In some embodiments, the complete response rate is at least 65%. In some embodiments, the complete response rate is at least 70%.In particular embodiments, the complete response rate in a population of subjects having R / R DLBCL is approximately 60%. In particular embodiments, the complete response rate in a population of subjects having R / R DLBCL is approximately 65%. In particular embodiments, the complete response rate in a population of subjects having R / R DLBCL is approximately 70%. In particular embodiments, the complete response rate in a population of subjects having R / R DLBCL is approximately 75%.In some embodiments, the overall response rate is at least 60% (e.g., at least 60%, at least 70%, at least 80%, or at least 90%; e.g., in the range of 60-100%, in the range of 70-100%, in the range of 80-100%, in the range of 90-100%, in the range of 60-90%, in the range of 60-80%, in the range of 60-70%, in the range of 60-65%, in the range of 65-75%, in the range of 70-90%, or in the range of 75-85%; e.g., about 60%, about 63%, about 64%, about 65%, about 66%, about 67%, about 70%, about 73%, about 74%, about 75%, about 76%, about 77%, about 80%, about 83%, about 84%, about 85%, about 86%, about 87%, about 90%, or more). In some embodiments, the overall response rate is at least 70%.In some embodiments, the overall response rate is at least 80%. In particular embodiments, the overall response rate in a population of subjects having R / R DLBCL is approximately 65%. In particular embodiments, the overall response rate in a population of subjects having R / R DLBCL is approximately 73%. In particular embodiments, the overall response rate in a population of subjects having R / R DLBCL is approximately 75%. In particular embodiments, the overall response rate in a population of subjects having R / R DLBCL is approximately 85%.

[0115] In some embodiments, the B-cell proliferative disorder is R / R DLBCL. In some embodiments, R / R NHL is R / R DLBCL. In some embodiments, the complete response rate is at least 35% (e.g., at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more; e.g., in the range of 30-100%, in the range of 50-100%, in the range of 70-100%, in the range of 35-90%, in the range of 45-90%, in the range of 35-70%, in the range of 35-50%, in the range of 40-80%, in the range of 40-60%, in the range of 45-55%, or in the range of 35-45%; e.g., about 35%, about 40%, about 45%, about 48%, about 49%, about 50%, about 51%, about 52%, about 55%, about 60%, approximately 70%, approximately 75%, approximately 80%, approximately 85%, or more).In some embodiments, the complete response rate is at least 45%. In some embodiments, the complete response rate is at least 55%. In some embodiments, the complete response rate is at least 75%. In some embodiments, the complete response rate is at least 85%. In some embodiments, the complete response rate is at least 90%. In particular embodiments, the complete response rate in a population of subjects having R / R DLBCL is approximately 46%. In particular embodiments, the complete response rate in a population of subjects having R / R DLBCL is approximately 52%. In particular embodiments, the complete response rate in a population of subjects having R / R DLBCL is approximately 86%.In some embodiments, the overall response rate is at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more; e.g., in the range of 85-100%, in the range of 87-100%, in the range of 90-100%, in the range of 95-100%, in the range of 85-97%, in the range of 85-95%, in the range of 85-90%, in the range of 85-87%, in the range of 90-95%, or in the range of 93-97%; e.g., about 85%, approximately 86%, approximately 87%, approximately 88%, approximately 89%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, approximately 99%, or more).In some embodiments, the overall response rate is at least 85%. In some embodiments, the overall response rate is at least 90%. In specific embodiments, the overall response rate in a population of subjects with R / R DLBCL is approximately 86%.

[0116] In some embodiments, the complete response rate exceeds the reference complete response rate in a control population of subjects who received a combination therapy comprising an anti-CD20 / CD3 bispecific antibody and an anti-PD-L1 antagonist antibody and does not contain an anti-CD79b antibody drug conjugate. In some embodiments, the objective response rate exceeds the reference objective response rate in a control population of subjects who received a combination therapy comprising an anti-CD20 / CD3 bispecific antibody and an anti-PD-L1 antagonist antibody and does not contain an anti-CD79b antibody drug conjugate. In some embodiments, the complete response rate exceeds the reference complete response rate in a control population of patients who received a combination therapy comprising glofitamab and atezolizumab and does not contain polatuzumab vedotin.In some embodiments, the objective response rate exceeds the control objective response rate in a control population of subjects receiving combination therapy containing glofitamab and atezolizumab and not containing polatuzumab vedotin.

[0117] In some embodiments, the subject is a human. In some embodiments, each subject in the subject population is a human. In some embodiments, each subject in the control subject population is a human. In some embodiments, the subject or subject population has received at least two prior systemic therapies (e.g., two, three, four, five, six, or more prior systemic therapies). In some embodiments, the subject or subject population is not eligible for autologous stem cell transplantation (AST).

[0118] BRIEF DESCRIPTION OF GRAPHIC MATERIALS

[0119] The application file contains at least one graphic representation executed in color. Copies of the publication of this patent or this application with color representations will be provided by the Office upon request and payment of the required fee.

[0120] Figures 1A-1P are diagrams showing the configurations of exemplary CD20 / CD3 bispecific antibodies.

[0121] Figure 2 is a schematic diagram showing the structure of glofitamab.

[0122] Figure 3 shows a schematic representation of the study design described in Example 1. Atezo = atezolizumab; CRM = continuous reassessment method; DLBCL = diffuse large cell lymphoma; ECOG = Eastern Cooperative Oncology Group; EWOC = dose escalation with overdose control; FL = follicular lymphoma; Pola = polatuzumab vedotin; R / R = relapsed and / or refractory; SCT = Society for Clinical Trials; GSV = glofitamab.

[0123] Figures 4A-4B show the study design flowcharts for the atezolizumab arm (Figure 4A), the polatuzumab dose escalation phase (Figure 4B), and the polatuzumab expansion phase (Figure 4C), as described in Example 1. Atezolizumab = atezolizumab; CR = complete response; ED = dose escalation; DLBCL = diffuse large cell lymphoma; EoS = end of study; F / U = follow-up; Glofit = glyfitamab; NHL = non-Hodgkin lymphoma; Pola = polatuzumab vedotin; PD = disease progression; PR = partial response; PC = patients; RP2D = recommended phase II dose; R / R = relapsed and / or refractory; SD = stable disease.

[0124] Figure 5A and Figure 5B are diagrams showing the timing of dose administration in the study described in Example 1 for the atezolizumab group (Figure 5A) and for the polatuzumab vedotin group (Figure 5B). Atezo = atezolizumab; Glofit = glofitamab; Gpt = pre-treatment with GAZVAYA® (pre-treatment with obinutuzumab); Pola = polatuzumab vedotin.

[0125] Figure 6 shows a bar chart depicting the incidence of adverse events (AEs) with an incidence of ≥10% or NCI-CTCAE Grade 5 severity for patients with assessed safety in Cohorts 1 and 2 treated with glofitamab + polatuzumab vedotin. Color indicates the severity of the AEs. The left panel represents all AEs in the study. The right panel represents only AEs related to the study treatment (e.g., glofitamab or polatuzumab vedotin).

[0126] Figure 7 presents updated efficacy data at the end of the clinical period for dose-escalation cohorts 1 and 2 and the expansion cohort in RP2D; Glofit = glofitamab. Polatuzumab = polatuzumab vedotin.

[0127] DETAILED DESCRIPTION OF THE ESSENCE OF THE INVENTION

[0128] The present invention provides methods of treating a subject having a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., non-Hodgkin's lymphoma (NHL) (e.g., relapsed and / or refractory NHL, diffuse large B-cell lymphoma (DLBCL) (e.g., relapsed and / or refractory DLBCL), follicular lymphoma (FL) (e.g., relapsed and / or refractory FL or transformed FL), mantle cell lymphoma (MCL) (e.g., relapsed or refractory MCL)) or central nervous system lymphoma (CNS)), comprising administering to the subject an anti-CD79b antibody drug conjugate and / or an anti-CD20 / CD3 bispecific antibody, e.g., as a fractionated dosage regimen with increasing dose.The method includes at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle includes a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein C1D1 of the bispecific antibody is approximately 2.5 mg, and C1D2 of the bispecific antibody is approximately 10 mg; and (b) the second dosing cycle includes a single dose (C2D1) of the bispecific antibody, wherein C2D1 of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0129] The present invention is based, in part, on the discovery that a fractionated, dose-escalating dosing regimen that involves administering a bispecific antibody that binds to CD20 and CD3 (e.g., glofitamab) over multiple dosing cycles (e.g., wherein the first dosing cycle is a stepwise, fractionated dosing cycle) can very effectively treat subjects with a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder) and have an acceptable safety profile (e.g., with respect to cytokine release syndrome).

[0130] I. General Methods

[0131] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology, which are within the skill in the art. Such techniques are explained in detail in such references as "Molecular Cloning: A Laboratory Manual," second edition (Sambrook et al., 1989); "Oligonucleotide Synthesis" (M.J. Gait, ed., 1984); "Animal Cell Culture" (R.I. Freshney, ed., 1987); "Methods in Enzymology" (Academic Press, Inc.); "Current Protocols in Molecular Biology" (F.M. Ausubel et al., eds., 1987, and periodic updates); "PCR: The Polymerase Chain Reaction", (Mullis et al., ed., 1994); "A Practical Guide to Molecular Cloning" (Perbal Bernard V., 1988); "Phage Display: A Laboratory Manual" (Barbas et al., 2001).

[0132] II. Definitions

[0133] In this document, terms are used as generally accepted in the art unless otherwise defined below.

[0134] The term "cluster of differentiation 20" or "CD20" as used herein refers to any native CD20 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. CD20 (also known as B-lymphocyte antigen CD20, B-lymphocyte surface antigen B1, Leu-16, Bp35, BM5, and LF5; the human protein is characterized in the UniProt database under entry P11836) is a hydrophobic transmembrane protein with a molecular mass of approximately 35 kDa expressed on pre-B and mature B lymphocytes (Valentine, M. A. et al., J. Biol. Chem. 264 (1989) 11282–11287; Tedder, T. F., et al., Proc. Natl. Acad. Sci. USA 85 (1988) 208–212; Stamenkovic, I., et al., J. Exp. Med. 167 (1988) 1975–1980; Einfeld, D. A., et al., EMBO J. 7 (1988) 711-717; Tedder, T. F., et al., J. Immunol. 142 (1989) 2560-2568).The corresponding human gene is transmembrane 4-domain subfamily A member 1, also known as MS4A1. This gene encodes a member of the transmembrane 4A gene family. Members of this growing protein family are characterized by common structural features and similar intron / exon splice junctions and exhibit unique expression profiles among hematopoietic cells and non-lymphoid tissues. This gene encodes a B-lymphocyte surface molecule that plays a role in the development and differentiation of B cells into plasma cells. This family member is localized on 11q12, among a cluster of family members. This term encompasses "full-length" unprocessed CD20, as well as any form of CD20 produced by cellular processing. The term also includes naturally occurring CD20 variants, such as splice variants or allelic variants. Alternative splicing of this gene results in two transcript variants that encode a single protein.In one embodiment, CD20 is human CD20.

[0135] The terms "anti-CD20 antibody" and "antibody that binds to CD20" refer to an antibody that is capable of binding CD20 with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent for targeting CD20. In one embodiment, the extent of binding of the anti-CD20 antibody to an unrelated, non-CD20 protein is less than about 10% of the binding of the antibody to CD20, as determined, for example, by radioimmunoassay (RIA). In certain embodiments, the antibody that binds to CD20 has a dissociation constant (K Д ) ≤ 1 μM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM (e.g., 10 -8 M or less, for example, from 10 -8 M to 10 -13 M, for example, from 10 -9 M to 10 -13M). In certain embodiments, the anti-CD20 antibody binds to an epitope of CD20 that is conserved across CD20 from different species.

[0136] The “anti-CD20 type II antibody” refers to an anti-CD20 antibody that has the binding properties and biological activity of anti-CD20 type II antibodies as described in Cragg et al., Blood 103 (2004) 2738–2743; Cragg et al., Blood 101 (2003) 1045–1052, Klein et al., mAbs 5 (2013), 22–33, and summarized in Table 1 below.

[0137]

[0138]

[0139] Examples of the anti-CD20 type II antibodies include, for example, obinutuzumab (GA101), tositumumab (B1), humanized B-Lyl IgG1 antibody (chimeric humanized IgG1 antibody as disclosed in WO 2005 / 044859), 11B8 IgG1 (as disclosed in WO 2004 / 035607), and AT80 IgG1.

[0140] Examples of anti-CD20 type I antibodies include, for example, rituximab, ofatumumab, veltuzumab, ocaratuzumab, ocrelizumab, PRO131921, ublituximab, HI47 IgG3 (ECACC, hybridoma), 2C6 IgG1 (as disclosed in WO 2005 / 103081), 2F2 IgG1 (as disclosed in WO 2004 / 035607 and WO 2005 / 103081), and 2H7 IgG1 (as disclosed in WO 2004 / 056312).

[0141] "CD3" refers to any native CD3 from any vertebrate source, including mammals such as primates (e.g., humans), non-human primates (e.g., cynomolgus macaques), and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses "full-length" unprocessed CD3, as well as any form of CD3 produced by cellular processing. This term also includes naturally occurring CD3 variants, such as splice variants or allelic variants. In one embodiment, the CD3 is human CD3, in particular the epsilon subunit of human CD3 (CD3s). The amino acid sequence of human CD3s is listed under UniProt (www.uniprot.org) accession number P07766 (version 144) or NCBI (www.ncbi.nlm.nih.gov / ) RefSeq NP 000724.1. The amino acid sequence of cynomolgus macaque [Macaca fascicularis] CD3s is listed in NCBI GenBank No. BAB71849.1.

[0142] The terms "bispecific antibody to CD20 / CD3" and "bispecific antibody binding to CD20 and CD3" refer to a bispecific antibody capable of binding CD20 and CD3 with sufficient affinity such that the antibody can be used as a diagnostic and / or therapeutic agent for targeting CD20 and / or CD3. In one embodiment, the degree of binding of the bispecific antibody that binds to CD20 and CD3 to an unbound non-CD3 protein and / or a non-CD20 protein is less than about 10% of the degree of binding of the antibody to CD3 and / or CD20, measured, for example, by radioimmunoassay (RIA). In certain aspects, a bispecific antibody that binds to CD20 and CD3 has a dissociation constant (Kd) of ≤ 1 μM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM (e.g., 10 -8 M or less, for example, from 10 -8 M to 10 -13 M, for example, from 10 -9 M to 10-13 M). In some embodiments, a bispecific antibody that binds to CD20 and CD3 binds to an epitope of CD3 that is conserved among CD3 species and / or an epitope of CD20 that is conserved among CD20 species. One example of a CD20 / CD3 bispecific antibody is glofitamab (proposed INR: WHO Drug List 121, Vol. 33, No. 2, 2019, p. 276, also known as CD20-TCB, RO7082859, or RG6026; CAS No.: 2229047-91-8).

[0143] The term “cluster of differentiation 79b” or “CD79b” as used herein refers to any native CD79b from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses “full-length” unprocessed CD79b, as well as any form of CD79b produced by cellular processing. This term also includes naturally occurring CD79b variants, including, for example, splice variants or allelic variants. CD79b includes, for example, the human CD79b protein (NCBI RefSeq #NP_000617), which is 229 amino acids long.

[0144] The terms "anti-CD79b antibody" and "antibody that binds to CD79b" refer to an antibody that is capable of binding CD79b with sufficient affinity such that the antibody can be used as a diagnostic and / or therapeutic agent for targeting CD79b. In one embodiment, the degree of binding of the anti-CD79b antibody to an unrelated, non-CD79b protein is less than about 10% of the binding of the antibody to CD79b, as determined, for example, by radioimmunoassay (RIA). In certain embodiments, an antibody that binds to CD79b has a dissociation constant (Kd) of ≤ 1 μM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM (e.g., 10 -8 M or less, for example, from 10 -8 M to 10 -13 M, for example, from 10 -9 M to 10 -13 M). In certain embodiments, the anti-CD79b antibody binds to an epitope of CD79b that is conserved across CD79b from different species.

[0145] As used herein, the term "cytokine release" or "cytokine surge" is synonymous with the terms "cytokine storm" or "cytokine release syndrome" (abbreviated "CRS") and refers to an increase in the levels of cytokines, in particular tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-2 (IL-2) and / or interleukin-8 (IL-8), in the blood of a subject during or shortly after (e.g., within 1 day) administration of a therapeutic agent, resulting in adverse symptoms. Cytokine release is defined as a supraphysiological response following administration of any immune therapy that results in the activation or recruitment of endogenous or administered T cells and / or other immune effector cells. Symptoms can be progressive, are always accompanied by fever at onset, and may include hypotension, capillary leakage (hypoxia), and end organ dysfunction (Lee et al., 2019).In some cases, such as after CAR-T cell infusion, CRS may occur several days after infusion as the CAR-T cells expand. With subsequent infusions, the incidence and severity typically decrease. Symptoms can range from mild discomfort to death and include fever, chills, dizziness, hypertension, hypotension, dyspnea, anxiety, sweating, flushing, skin rash, tachycardia, tachypnea, headache, tumor pain, nausea, vomiting, and / or organ failure.

[0146] The term "amino acid mutation" as used herein is intended to include amino acid substitutions, deletions, insertions, and modifications. Any combination of substitutions, deletions, insertions, and modifications can be made to obtain the final construct, provided that the final construct has the desired characteristics, such as reduced binding to the Fc receptor. Deletions and insertions in the amino acid sequence include amino- and / or carboxyl-terminal deletions and insertions of amino acids. Specific amino acid mutations are amino acid substitutions. For the purpose of altering, for example, the binding characteristics of the Fc region, non-conservative amino acid substitutions, i.e., the replacement of one amino acid with another amino acid having different structural and / or chemical properties, are particularly preferred.Amino acid substitutions include substitution of unnatural amino acids or naturally occurring amino acid derivatives of the twenty standard amino acids (e.g., 4-hydroxyproline, 3-methylhistidine, ornithine, homoserine, 5-hydroxylysine). Amino acid mutations can be created using genetic or chemical methods well known in the art. Genetic methods may include site-directed mutagenesis, PCR, gene synthesis, etc. It is understood that methods for altering amino acid side chain groups by methods other than genetic engineering, such as chemical modification, may also be applicable. Various designations may be used herein to refer to the same amino acid mutation. For example, a substitution of proline at position 329 of the Fc region with glycine may be designated as 329G, G329, G. 329 , P329G or Pro329Gly.

[0147] "Affinity" refers to the strength of the net non-covalent interactions between one binding site of a molecule (e.g., a receptor) and its binding partner (e.g., a ligand). Unless otherwise specified, in this document, the term "binding affinity" refers to the intrinsic binding affinity of a molecule, reflecting the interaction between the members of a binding pair (e.g., a receptor and a ligand) at a 1:1 ratio. The affinity of a molecule X for its partner Y can generally be expressed by the dissociation constant (K Д ), which is the ratio of the dissociation and association constants (k дис and k асс(respectively). Thus, equivalent affinities may involve different rate constants, provided that the ratio between the rate constants remains the same. Affinity can be measured using well-established methods known in the art. A specific method for measuring affinity is surface plasmon resonance (SPR).

[0148] An affinity matured antibody refers to an antibody with one or more changes in one or more hypervariable regions (HVRs) compared to a parent antibody that does not contain such changes, wherein such changes result in an improvement in the affinity of the antibody for the antigen.

[0149] As used herein, the term "antigen-binding fragment" refers to a molecule that specifically binds to an antigenic determinant. In one embodiment, the antigen-binding fragment is capable of directing a compound to which it is attached (e.g., a cytokine or a second antigen-binding fragment) to a target site, such as a specific type of tumor cell bearing the antigenic determinant. Antigen-binding fragments include antibodies and fragments thereof, as further defined herein. Preferred antigen-binding fragments include an antigen-binding domain of an antibody, comprising the variable region of the antibody heavy chain and the variable region of the antibody light chain. In certain embodiments, antigen-binding fragments may comprise antibody constant regions, as further defined herein and known in the art.Usable heavy chain constant regions belong to any of five isotypes: α, δ, ε, γ, or μ. Usable light chain constant regions include any of two isotypes: κ and λ.

[0150] By the terms “binds,” “specifically binds,” or “is specific for,” it is meant that the binding is selective for the antigen and can be separated from unwanted or nonspecific interactions. The ability of an antigen-binding fragment to bind to a specific antigenic determinant can be measured either by enzyme-linked immunosorbent assay (ELISA) or other techniques known to those skilled in the art, such as surface plasmon resonance (BIAcore analysis) (Liljeblad et al., Glyco J. 17, 323-329 (2000)), and conventional binding assays (Heeley, Endocr Res. 28, 217-229 (2002)). In one embodiment, the degree of binding of the antigen-binding fragment to the unrelated protein is less than about 10% of the binding of the antigen-binding fragment to the antigen, as measured, for example, by SPR.In certain embodiments, an antigen-binding fragment that binds to an antigen, or an antigen-binding molecule comprising the antigen-binding fragment, has a dissociation constant (K. Д ) ≤ 1 μM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM (e.g., 10 -8 M or less, for example, from 10 -8 M -13 up to 10 -9 M, for example, from 10 -9 M to 10 -13 M).

[0151] "Decreased binding," such as decreased binding to an Fc receptor, refers to a decrease in affinity for the corresponding interaction, as determined, for example, by SPR. For clarity, this term also includes a decrease in affinity to zero (or below the detection limit of the assay), i.e., complete elimination of the interaction. Conversely, "increased binding" refers to an increase in binding affinity for the corresponding interaction.

[0152] As used in this document, the term "antigen-binding molecule" refers in its broadest sense to a molecule that specifically binds an antigenic determinant. Examples of antigen-binding molecules include immunoglobulins and their derivatives, such as fragments.

[0153] As used herein, the term "antigenic determinant" is synonymous with the terms "antigen" and "epitope" and refers to a site (e.g., a continuous stretch of amino acids or a conformational configuration consisting of different regions of non-contiguous amino acids) on a polypeptide macromolecule to which an antigen-binding moiety binds to form an antigen-binding moiety-antigen complex. Suitable antigenic determinants may be found, for example, on the surface of tumor cells, on the surfaces of virus-infected cells, on the surfaces of other diseased cells, in free form in serum and / or in the extracellular matrix (ECM).Proteins useful as antigens (e.g., CD3) herein may be any native form of protein from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. In a specific embodiment, the antigen is a human protein. When referring to a specific protein herein, the term includes the "full-length" unprocessed protein, as well as any form of the protein that results from cellular processing. This term also includes naturally occurring protein variants, such as splice variants or allelic variants. A typical human protein useful as an antigen is CD3, in particular the epsilon subunit of CD3 (see UniProt #P07766 (version 130), NCBI RefSeq #NP 000724.1, for the human sequence; or UniProt #Q95LI5 (version 49), NCBI GenBank #BAB71849.1, with respect to the cynomolgus macaque (Macaca fascicularis) sequence). In some embodiments, the bispecific antigen-binding T cell activating molecule described herein binds to a CD3 epitope or target cell antigen that is conserved among CD3 or target cell antigens from different species.

[0154] As used herein, the term "polypeptide" refers to a molecule composed of monomers (amino acids) linked linearly by amide bonds (also known as peptide bonds). The term "polypeptide" refers to any chain of two or more amino acids and does not refer to a specific length of the product. Thus, peptides, dipeptides, tripeptides, oligopeptides, "protein," "amino acid chain," or any other term used to refer to a chain of two or more amino acids are included within the definition of "polypeptide," and the term "polypeptide" may be used instead of or interchangeably with any of these terms. The term "polypeptide" is also intended to refer to the products of post-expression modifications of a polypeptide, including, but not limited to, glycosylation, acetylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, or modification of unnatural amino acids.The polypeptide may be obtained from a natural biological source or produced by recombinant technology, but is not necessarily translated from the specified nucleic acid sequence. It can be obtained by any method, including chemical synthesis. The size of the polypeptide of the present invention can be approximately 3 or more, 5 or more, 10 or more, 20 or more, 25 or more, 50 or more, 75 or more, 100 or more, 200 or more, 500 or more, more, 1000 or more, or 2000 or more amino acids. Polypeptides may have a defined three-dimensional structure, although they do not necessarily have such a structure. Polypeptides with a defined three-dimensional structure are called folded, and polypeptides that do not have a defined three-dimensional structure but can adopt a large number of different conformations are called unfolded.

[0155] An "isolated" polypeptide, or variant, or derivative thereof, is defined as a polypeptide that is not in its natural environment. There is no specific requirement for the level of purification. For example, an isolated polypeptide may be removed from its native or natural environment. Recombinantly produced polypeptides or proteins expressed in host cells are considered isolated for the purposes of the present invention, including both native and recombinant polypeptides that have been separated, fractionated, or partially or completely purified by any suitable method.

[0156] "Percentage (%) of amino acid sequence identity" relative to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve maximum sequence identity, but without taking into account any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining the percentage of amino acid sequence identity can be performed by various methods known in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software.Those skilled in the art can determine suitable parameters for sequence alignment, including any algorithms necessary to achieve maximal alignment over the entire length of the sequences being compared. For the purposes of this document, % amino acid sequence identity values ​​are obtained using the ALIGN-2 sequence comparison software. The ALIGN-2 sequence comparison software was developed by Genentech, Inc., and the source code has been filed with the user documentation with the U.S. Copyright Office, Washington, D.C. 20559, where it is registered under U.S. Copyright Registration Number TXU510087. The ALIGN-2 software is freely available from Genentech, Inc., South San Francisco, California, or can be compiled from the source code.The ALIGN-2 program should be compiled for use on UNIX operating systems, including Digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and are not variable. When ALIGN-2 is used to compare amino acid sequences, the % amino acid sequence identity of a given amino acid sequence A with or compared to a given amino acid sequence B (which can alternatively be formulated as a given amino acid sequence A having or containing a specified % amino acid sequence identity with or compared to a given amino acid sequence B) is calculated as follows:

[0157] 100 times the X / Y ratio,

[0158] where X is the number of amino acid residues scored by the ALIGN-2 sequence alignment program as identical matches in the program alignment of A and B, and where Y is the total number of amino acid residues in B. It should be understood that where the length of the amino acid sequence of A is not equal to the length of the amino acid sequence of B, the % amino acid sequence identity of A to B will not be equal to the % amino acid sequence identity of B to A. Unless otherwise specifically stated, all % amino acid sequence identity values ​​used in this document are obtained as described in the preceding paragraph using the ALIGN-2 computer program.

[0159] The term "antibody" is used herein in the broadest sense and includes various antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, provided that they exhibit the necessary antigen-binding activity.

[0160] The terms "full-length antibody," "intact antibody," and "complete antibody" are used interchangeably herein to refer to an antibody having a structure substantially similar to the structure of a native antibody or having heavy chains that contain an Fc region as defined herein.

[0161] "Antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody that binds the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2, diabodies, linear antibodies, single-chain antibody molecules (e.g., scFv), and multispecific antibodies formed from antibody fragments. The term "antibody fragment" as used herein also encompasses single-domain antibodies.

[0162] The term "immunoglobulin molecule" refers to a protein with the structure of a naturally occurring antibody. For example, IgG immunoglobulins are heterotetrameric glycoproteins weighing approximately 150,000 daltons, consisting of two light chains and two heavy chains linked by disulfide bonds. From the N- to the C-terminus, each heavy chain contains a variable region (VH), also called the variable heavy domain or heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3), also called the heavy chain constant region. Similarly, in the direction from the N-terminus to the C-terminus, each light chain contains a variable region (VL), also called the variable light domain or light chain variable domain, followed by a light chain constant domain (CL), also called the light chain constant region.The immunoglobulin heavy chain can be classified into one of five classes, called α (IgA), δ (IgD), ε (IgE), γ (IgG), or μ (IgM), some of which can be further divided into subclasses, such as γ1 (IgG1), γ2 (IgG2), γ3 (IgG3), γ4 (IgG4), α1 (IgA1), and α2 (IgA2). The immunoglobulin light chain can be classified into one of two types, called kappa (κ) and lambda (λ), based on the amino acid sequence of its constant domain. Immunoglobulin consists primarily of two Fab molecules and an Fc domain linked via the immunoglobulin hinge region.

[0163] The term "antigen-binding domain" refers to the portion of an antibody that contains a region that specifically binds to and is complementary to part or all of an antigen. An antigen-binding domain can be formed, for example, by one or more variable domains of an antibody (also referred to as variable regions of an antibody). Preferably, the antigen-binding domain comprises the variable region of the antibody's light chain (VL) and the variable region of the antibody's heavy chain (VH).

[0164] The term "variable region" or "variable domain" refers to the domain of the heavy or light chain of an antibody that is involved in the binding of the antibody to an antigen. The variable domains of the heavy chain and light chain (VH and VL, respectively) of a native antibody typically have similar structures, with each domain containing four conserved framework regions (FRs) and three hypervariable regions (HVRs). (See, e.g., Kindt et al. Kuby Immunology, 6 thed., WH Freeman and Co., p. 91 (2007). A single VH or VL domain may be sufficient to confer antigen-binding specificity.

[0165] A "human antibody" is an antibody that has an amino acid sequence corresponding to that of an antibody produced by a human being or a human cell, or that is derived from a non-human source and utilizes human antibody repertoires or other human antibody-encoding sequences. This definition of a human antibody explicitly excludes a humanized antibody containing non-human antigen-binding residues.

[0166] A "humanized" antibody refers to a chimeric antibody comprising amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody comprises substantially all of at least one, and typically two, variable domains in which all or substantially all of the HVRs (e.g., CDRs) correspond to those of a non-human antibody, and all or substantially all of the FRs correspond to those of a human antibody. A humanized antibody may optionally comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody, for example, a non-human antibody, refers to an antibody that has undergone humanization.

[0167] The term "hypervariable region" or "HVR" as used herein refers to each of the regions of the variable domain of an antibody that are hypervariable in sequence ("complementarity determining regions" or "CDRs"), and / or form structurally defined loops ("hypervariable loops"), and / or contain antigen-contacting residues ("antigenic contacts"). In general, antibodies contain six HVRs: three in VH (H1, H2, H3) and three in VL (L1, L2, L3). Illustrative HVRs in this document include:

[0168] (a) hypervariable loops located at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987));

[0169] (b) CDRs are located at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991));

[0170] (c) antigenic contacts are located at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3) (MacCallum et al. J. Mol. Biol. 262: 732-745 (1996)); and

[0171] (d) combinations of (a), (b) and / or (c) comprising amino acid residues HVR 46-56 (L2), 47-56 (L2), 48-56 (L2), 49-56 (L2), 26-35 (H1), 26-35b (H1), 49-65 (H2), 93-102 (H3) and 94-102 (H3).

[0172] Unless otherwise noted, HVR residues and other residues in the variable domain (e.g., FR residues) are numbered in this document according to Kabat et al., see above.

[0173] "Framework region" or "FR" refers to the residues of the variable domain other than the hypervariable region (HVR) residues. The FR of a variable domain typically consists of four FR domains: FR1, FR2, FR3, and FR4. Accordingly, the HVR and FR sequences are typically found in the VH (or VL) in the following order: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0174] A "human consensus framework" is a framework region that represents the most frequently occurring amino acid residues in a set of human immunoglobulin VL or VH framework sequences. Typically, a set of human immunoglobulin VL or VH sequences is derived from a subset of variable domain sequences. In general, the subset of sequences is a subset according to Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda MD (1991), vols. 1-3. In one embodiment, for VL, the subgroup is subgroup kappa I according to Kabat et al., supra. In one embodiment, for VH, the subgroup is subgroup III according to Kabat et al., supra.

[0175] For the purposes of this document, an "acceptor human framework" is a framework region comprising an amino acid sequence of a light chain variable domain (VL) framework region or a heavy chain variable domain (VH) framework region derived from a human immunoglobulin framework region or a human consensus framework region, as defined below. A human acceptor framework region "derived from" a human immunoglobulin framework region or a human consensus framework region may comprise the same amino acid sequence or may comprise changes in the amino acid sequence. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less.In some embodiments, the human VL acceptor framework is identical in sequence to a human immunoglobulin VL framework sequence or a human consensus framework sequence.

[0176] Antibody "class" refers to the type of constant domain, or constant region, contained in its heavy chain. There are five main classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and some can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different immunoglobulin classes are called α, δ, ε, γ, and μ, respectively.

[0177] As used herein, the term "isotype" or "subclass" of IgG means any of the subclasses of immunoglobulins defined by the chemical and antigenic characteristics of their constant regions.

[0178] As used herein, the term "Fc domain" or "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain, which contains at least a portion of the constant region. This term includes native-sequence Fc regions and variant Fc regions. Although the boundaries of the IgG heavy chain Fc region may vary slightly, the human IgG heavy chain Fc region, by definition, typically extends from Cys226 or Pro230 to the carboxyl terminus of the heavy chain. Antibodies produced by host cells may undergo post-translational cleavage of one or more, in particular one or two, amino acids from the C-terminus of the heavy chain.Therefore, an antibody produced by a host cell by expressing a specific nucleic acid molecule encoding a full-length heavy chain may comprise the full-length heavy chain or may comprise a cleaved version of the full-length heavy chain (also referred to herein as a "cleaved version of the heavy chain"). This may be the case when the two terminal amino acids of the C-terminus of the heavy chain are glycine (G446) and lysine (K447, EU numbering). Therefore, the C-terminal lysine (Lys447) or the C-terminal glycine (Gly446) and lysine (K447) of the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region follows the EU numbering system, also referred to as the EU index, described in Kabat et al., Sequences of Proteins of Immunological Interest, 5. thEd. Public Health Service, National Institutes of Health, Bethesda, MD, 1991 (see also above). As used herein, the term "subunit" of the Fc domain refers to one of the two polypeptides that form a dimeric Fc domain, i.e., a polypeptide containing the C-terminal constant regions of the immunoglobulin heavy chain capable of stable self-association. For example, the Fc domain subunit of IgG contains the CH2 and CH3 constant domains of IgG.

[0179] "A modification that promotes the association of the first and second Fc domain subunits" is a manipulation of the peptide backbone or post-translational modifications of an Fc domain subunit that reduces or prevents the association of a polypeptide containing the Fc domain subunit with an identical polypeptide to form a homodimer. As used herein, an association-promoting modification specifically includes separate modifications made to each of the Fc domain subunits whose association is required (i.e., the first and second Fc domain subunits), wherein the modifications are complementary to each other so as to promote the association of the two Fc domain subunits. For example, an association-promoting modification may alter the structure or charge of one or both Fc domain subunits so as to make their association sterically or electrostatically favorable, respectively.Thus, (hetero)dimerization occurs between a polypeptide comprising a first Fc domain subunit and a polypeptide comprising a second Fc domain subunit, which may be non-identical in the sense that additional components fused to each subunit (e.g., antigen-binding fragments) are not identical. In some embodiments, the modification that promotes association comprises an amino acid mutation in the Fc domain, in particular an amino acid substitution. In a specific embodiment, the modification that promotes association comprises a separate amino acid mutation, in particular an amino acid substitution, in each of the two Fc domain subunits.

[0180] An "activating Fc receptor" is an Fc receptor that, upon interaction with the Fc region of an antibody, triggers signaling events that stimulate the receptor-bearing cell to perform effector functions. Activating Fc receptors include FcγRIIIa (CD16a), FcγRI (CD64), FcγRIIa (CD32), and FcαRI (CD89).

[0181] The term "effector functions," when used in relation to antibodies, refers to those biological activities that can be attributed to the Fc region of an antibody, which vary depending on the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), cytokine secretion, immune complex-mediated antigen uptake by antigen-presenting cells, downregulation of cell surface receptors (e.g., the B cell receptor), and B cell activation.

[0182] As used herein, the term "effector cells" refers to a population of lymphocytes that have receptors on their surface for effector molecules, such as cytokine receptors, and / or Fc receptors, through which they bind effector molecules, such as cytokines, and / or the Fc region of an antibody, and promote the killing of target cells, such as tumor cells. Effector cells can, for example, exert cytotoxic or phagocytic activity. Effector cells include, in particular, effector T cells, such as CD8 + cytotoxic T cells, CD4 + helper T cells, γδ T cells, NK cells, lymphokine-activated killer (LAK) cells, and macrophages / monocytes.

[0183] For the purposes of this document, the terms "engineer," "engineered," and "engineering" are intended to include any manipulation of the peptide backbone or post-translational modifications of a naturally occurring or recombinant polypeptide or fragment thereof. Engineering includes modifications of the amino acid sequence, glycosylation profile, or side chain groups of individual amino acids, as well as combinations of these approaches. The term "engineering," particularly with the prefix "glyco-," as well as the term "glycosylation engineering," includes metabolic engineering of the cellular glycosylation machinery, including genetic manipulation of oligosaccharide synthetic pathways to achieve altered glycosylation of glycoproteins expressed in cells. Furthermore, glycosylation engineering includes studying the effects of mutations and the cellular environment on glycosylation.In one embodiment, glycosylation engineering involves altering the activity of glycosyltransferases. In a specific embodiment, engineering results in altered activity of glucosaminyltransferases and / or fucosyltransferases. Engineering of glycosylation can be used to generate a "GnTIII-enhanced host cell" (e.g., a host cell that has been manipulated to express increased levels of one or more polypeptides having β(1,4)-N-acetylglucosaminyltransferase III (GnTIII) activity), a "ManII-enhanced host cell" (e.g., a host cell that has increased levels of one or more polypeptides having α-mannosidase II (ManII) activity), or a "α(1,6) fucosyltransferase-depleted host cell" (e.g., a host cell that has decreased levels of α(1,6) fucosyltransferase).

[0184] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells," which include initially transformed cells and the progeny obtained from them, regardless of the number of passages. Progeny may not be completely identical to the original cell in nucleic acid content and may contain mutations. Mutant progeny that have the same function or biological activity for which the initially transformed cells are screened or selected are included herein. A host cell is any type of cellular system that can be used to produce the proteins used in the present invention.In one embodiment, the host cell is engineered to produce an antibody with modified oligosaccharides. In some embodiments, the host cells have been manipulated to express increased levels of one or more polypeptides having β(1,4)-N-acetylglucosaminyltransferase III (GnTIII) activity. In some embodiments, the host cells have been further manipulated to express increased levels of one or more polypeptides having α-mannosidase II (ManII) activity. Host cells include cultured cells, such as cultured mammalian cells such as CHO cells, BHK cells, NS0 cells, SP2 / 0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER cells.C6 or hybridoma cells, yeast cells, insect cells and plant cells, to name just a few, but also cells belonging to a transgenic animal, transgenic plant or cultured plant or animal tissue.

[0185] In this document, the term "polypeptide having GnTIII activity" refers to a polypeptide capable of catalyzing the addition of an N-acetylglucosamine (GlcNAc) residue in a β-1,4-linkage to a β-linked mannoside of the trimannosyl backbone of N-linked oligosaccharides. These include fusion polypeptides that exhibit an enzymatic activity similar to, but not necessarily identical to, that of β(1,4)-N-acetylglucosaminyltransferase III, also known as β-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC 2.4.1.144), according to the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (NC-IUBMB), as measured in a specified biological assay, with or without dose dependence. Where a dose dependence exists, it need not be identical to the dose dependence of GnTIII, but rather should be substantially similar to the dose dependence for a given activity compared to GnTIII (i.e.,(the candidate polypeptide will exhibit greater activity or no more than about 25 times less activity, preferably no more than about ten times less activity, and most preferably no more than about three times less activity compared to GnTIII). In some embodiments, the polypeptide having GnTIII activity is a fusion polypeptide comprising a GnTIII catalytic domain and a Golgi localization domain of a heterologous Golgi-resident polypeptide. In particular, the Golgi localization domain is a mannosidase II or GnTI localization domain, more particularly a mannosidase II localization domain. Alternatively, the Golgi localization domain is selected from the group consisting of: a mannosidase I localization domain, a GnTII localization domain, and an α1,6-core fucosyltransferase localization domain.Methods for producing such fusion polypeptides and their use to produce antibodies with enhanced effector functions are disclosed in WO2004 / 065540, US Provisional Patent Application No. 60 / 495142, and US Patent Application Publication No. 2004 / 0241817, the entire contents of which are expressly incorporated herein by reference.

[0186] In this document, the term "Golgi localization domain" refers to the amino acid sequence of a Golgi-resident polypeptide that is responsible for anchoring the polypeptide to its location within the Golgi complex. Localization domains typically contain the amino-terminal "tails" of the enzyme.

[0187] As used herein, the term “polypeptide with ManII activity” refers to polypeptides that are capable of catalyzing the hydrolysis of terminal 1,3- and 1,6-linked α-D-mannose residues in the branched GlcNAcMan5GlcNAc2mannose intermediate in N-linked oligosaccharides. These include polypeptides that exhibit enzymatic activity similar to, but not necessarily identical to, that of Golgi α-mannosidase II, also known as mannosyl-oligosaccharide 1,3-1,6-α-mannosidase II (EC 3.2.1.114), according to the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (NC-IUBMB).

[0188] Antibody-dependent cell-mediated cytotoxicity (ADCC) is an immune mechanism that results in the lysis of antibody-coated target cells by immune effector cells. Target cells are those to which antibodies or their fragments containing the Fc region specifically bind, generally via the protein portion N-terminal to the Fc region. As used herein, the term "increased / decreased ADCC" is defined as an increase / decrease in the number of target cells lysed in a given time at a given concentration of antibody in the medium surrounding the target cells via the ADCC mechanism defined above, and / or as a decrease / increase in the concentration of antibody in the medium surrounding the target cells required to ensure lysis of a given number of target cells in a given time via the ADCC mechanism.The increase / decrease in ADCC is determined relative to the ADCC mediated by the same antibody produced by the same type of host cells, using the same standard methods of production, purification, formulation, and storage (which are well known to those skilled in the art), but which was not engineered. For example, an increase in ADCC mediated by an antibody produced by host cells engineered to have an altered glycosylation pattern (e.g., to express glycosyltransferase, GnTIII, or other glycosyltransferases) using the methods described herein, compared to ADCC mediated by the same antibody produced by the same type of non-engineered host cells.

[0189] An antibody with increased / decreased antibody-dependent cell-mediated cytotoxicity (ADCC) is defined as an antibody with increased / decreased ADCC, as determined by any suitable method known to those skilled in the art. One accepted in vitro ADCC assay is as follows:

[0190] 1) The assay uses target cells that are known to express the target antigen recognized by the antigen-binding region of the antibody;

[0191] 2) The effector cells used in the assay are human peripheral blood mononuclear cells (PBMCs) isolated from the blood of a randomly selected healthy donor;

[0192] 3) The analysis is carried out in accordance with the following protocol:

[0193] i) PBMCs are isolated using standard density centrifugation procedures and suspended at 5×10 6 cells / ml in RPMI cell culture medium;

[0194] ii) Target cells are grown by standard tissue culture methods, harvested from exponential growth phase with viability above 90%, washed in RPMI cell culture medium, labeled with 100 microcuries 51 Cr, washed twice in cell culture medium and resuspended in cell culture medium at a density of 105 cells / ml;

[0195] iii) 100 microliters of the prepared target cell suspension mentioned above is transferred to each well of a 96-well microtiter plate;

[0196] iv) The antibody is serially diluted from 4000 ng / mL to 0.04 ng / mL in the cell culture medium, and 50 microliters of the resulting antibody solutions are added to the target cells in a 96-well microtiter plate, tested in triplicate at different antibody concentrations, covering the entire concentration range specified above;

[0197] v) For the maximum release (MR) control, 50 microliters of 2% (v / v) aqueous non-ionic detergent solution (Nonidet, Sigma, St. Louis) are added to 3 additional wells of the plate containing labeled target cells instead of the antibody solution (item iv above);

[0198] vi) For the spontaneous release (SR) control, 50 microliters of RPMI cell culture medium instead of the antibody solution (item iv above) are added to 3 additional wells of the plate containing labeled target cells;

[0199] vii) Then the 96-well microtiter plate is centrifuged at 50 x g for 1 minute and incubated for 1 hour at 4°C;

[0200] viii) 50 microliters of the PBMC suspension (item i above) is added to each well to obtain an effector gap cell ratio of 25:1, and the plates are placed in an incubator with a 5% CO2 atmosphere at 37°C for 4 hours;

[0201] ix) The cell-free supernatant from each well is collected and the experimentally released radioactivity (ER) is quantified using a gamma counter;

[0202] x) the percentage of specific lysis is calculated for each antibody concentration using the formula (ER-MR) / (MR-SR) × 100, where ER is the quantitative expression of the mean radioactivity (see item ix above) for the antibody concentration, MR is the quantitative expression of the mean radioactivity (see item ix above) for the MR controls (see item v above), and SR is the quantitative expression of the mean radioactivity (see item ix above) for the SR controls (see item vi above);

[0203] 4) “increased / decreased AZCC” is defined as

[0204] increase / decrease in the maximum percentage of specific lysis observed within the antibody concentration range tested above, and / or increase / decrease in the antibody concentration required to achieve one-half of the maximum percentage of specific lysis observed within the antibody concentration range tested above. The increase / decrease in ADCC is determined relative to the ADCC measured using the above assay, mediated by the same antibody produced by the same type of host cells, using the same standard methods of production, purification, formulation, and storage that are well known to those skilled in the art, but which was not designed.

[0205] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical and / or bind to the same epitope, with the exception of possible variant antibodies, such as those containing mutations that occur naturally or arise during the production of the monoclonal antibody preparation, such variants being generally present in small amounts. Unlike polyclonal antibody preparations, which typically contain different antibodies directed against different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is directed against a single determinant on the antigen.Thus, the modifier "monoclonal" indicates the characteristic of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be interpreted as requiring the production of the antibody by any particular method. For example, monoclonal antibodies for use in accordance with the present invention can be produced by a variety of techniques, including, but not limited to, hybridoma technology, recombinant DNA technology, phage display technology, and methods using transgenic animals containing all or part of the human immunoglobulin loci, with such methods and other typical methods for producing monoclonal antibodies described herein.

[0206] "Naked antibody" means an antibody that is not conjugated to a heterologous component (e.g., a cytotoxic component) or a radiolabel. A naked antibody may be present in a pharmaceutical formulation.

[0207] "Native antibodies" refer to naturally occurring immunoglobulin molecules with varying structures. For example, native IgG-based antibodies are heterotetrameric glycoproteins weighing approximately 150,000 daltons, consisting of two identical light chains and two identical heavy chains linked by a disulfide bond. From the N-terminus to the C-terminus, each heavy chain has a variable region (VH), also called the variable heavy domain or heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3). Similarly, from the N-terminus to the C-terminus, each light chain has a variable region (VL), also called the variable light domain or light chain variable domain, followed by a light chain constant domain (CL). An antibody light chain can be classified into one of two types, called kappa (κ) and lambda (λ), based on the amino acid sequence of its constant domain.

[0208] In the context of this document, the terms "first," "second," "third," etc., in relation to antigen-binding fragments or domains are used to facilitate distinction when more than one of each type of fragment or domain is present. The use of these terms does not imply a specific order or orientation unless explicitly stated otherwise.

[0209] The terms "polyspecific" and "bispecific" mean that an antigen-binding molecule is capable of specifically binding to at least two different antigenic determinants. Typically, a bispecific antigen-binding molecule comprises two antigen-binding sites, each specific for a different antigenic determinant. In certain embodiments, a bispecific antigen-binding molecule is capable of simultaneously binding two antigenic determinants, particularly two antigenic determinants expressed on two different cells.

[0210] The term "valence" or "valence," as used herein, refers to the presence of a specified number of antigen-binding sites in an antigen-binding molecule. Therefore, the term "univalent antigen binding" refers to the presence of one (and no more than one) antigen-binding site specific for an antigen in an antigen-binding molecule.

[0211] "Antigen-binding site" refers to the site, i.e., one or more amino acid residues, of an antigen-binding molecule that mediates interaction with the antigen. For example, the antigen-binding site of an antibody contains amino acid residues from the complementarity-determining regions (CDRs). A native immunoglobulin molecule typically has two antigen-binding sites, while an Fab molecule typically has one.

[0212] As used herein, the term "activating T cell antigen" refers to an antigenic determinant expressed by T lymphocytes, particularly cytotoxic T lymphocytes, that is capable of inducing or enhancing T cell activation following interaction with an antigen-binding molecule. Specifically, interaction of an antigen-binding molecule with an activating T cell antigen can induce T cell activation by initiating the signaling cascade of the T cell receptor complex. A typical T cell activating antigen is CD3. In a specific embodiment, the activating T cell antigen is CD3, in particular the epsilon subunit of CD3 (see UniProt No. P07766 (version 130), NCBI RefSeq No. NP 000724.1, for the human sequence; or UniProt No. Q95LI5 (version 49), NCBI GenBank No. BAB71849.1, for the cynomolgus monkey [Macaca fascicularis] sequence).

[0213] As used herein, the term "T cell activation" refers to one or more cellular responses of a T lymphocyte, in particular a cytotoxic T lymphocyte, selected from: proliferation, differentiation, cytokine secretion, release of cytotoxic effector molecules, cytotoxic activity, and expression of activation markers. The T cell activating therapeutic agents used in the present invention are capable of inducing T cell activation. Suitable assays for determining T cell activation are known in the art and are described herein.

[0214] The term "target cell antigen" as used herein refers to an antigenic determinant located on the surface of a target cell, for example, a cell in a tumor, such as a cancer cell or a tumor stromal cell. In a specific embodiment, the target cell antigen is CD20, in particular human CD20 (see UniProt Accession No. P11836).

[0215] In the context of this document, “B cell antigen” refers to an antigenic determinant presented on the surface of a B lymphocyte, particularly a malignant B lymphocyte (in this case, the antigen is also called “malignant B cell surface antigen”).

[0216] As used in this document, the term "T cell antigen" refers to an antigenic determinant presented on the surface of a T lymphocyte, particularly a cytotoxic T lymphocyte.

[0217] “Fab molecule” refers to a protein consisting of the VH and CH1 domain of the heavy chain (“Fab heavy chain”) and the VL and CL domain of the light chain (“Fab light chain”) of immunoglobulin.

[0218] A chimeric antigen receptor or CAR is a genetically engineered receptor protein comprising an antigen-binding moiety, such as a single-chain variable fragment (scFv) of a targeting antibody, a transmembrane domain, an intracellular T-cell activating signaling domain (e.g., T-cell receptor zeta chains of the CD3 protein), and optionally one or more intracellular costimulatory domains (e.g., CD28, CD27, CD137 (4-1BB), Ox40). CARs mediate antigen recognition, T-cell activation, and, in the case of second-generation CARs, costimulation to enhance T-cell functionality and persistence. For a review, see, e.g., Jackson et al., Nat Rev Clin Oncol. (2016) 13, 370–383.

[0219] By “fused” is meant that the components (e.g., a Fab molecule and an Fc domain subunit) are linked by peptide bonds, either directly or through one or more peptide linkers.

[0220] The “effective amount” of an agent refers to the amount required to produce a physiological change in the cell or tissue into which it is administered.

[0221] A "therapeutically effective amount" of a drug, such as a pharmaceutical composition, refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result. A therapeutically effective amount of a drug, for example, eliminates, reduces, slows, minimizes, or prevents adverse effects of a disease.

[0222] A "therapeutic agent" means an active component, such as a pharmaceutical composition, that is administered to a subject in an attempt to modify the natural course of a disease in the subject being treated, and may be administered either prophylactically or during the progression of clinical pathology. An "immunotherapeutic agent" means a therapeutic agent that is administered to a subject in an attempt to restore or enhance the subject's immune response, such as to a tumor.

[0223] The term "pharmaceutical composition" refers to a preparation which is in a form that ensures the effectiveness of the biological activity of the active ingredient contained therein and which does not contain additional components that are unacceptably toxic to the subject to whom the composition is to be administered.

[0224] "Pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical composition other than the active ingredient that is nontoxic to the subject. A pharmaceutically acceptable carrier includes, but is not limited to, a buffer, excipient, stabilizer, or preservative.

[0225] The term "package insert" or "instructions for use" is used to refer to the instructions customarily included in the commercial packages of therapeutic products that contain information on the indications, usage, dosage, administration, combination therapy, contraindications and / or warnings regarding the use of such therapeutic products.

[0226] The term "combination therapy" as referred to herein encompasses combined administration (in which two or more therapeutic agents are included in one or different formulations) and separate administration, in which case the administration of the antibody of the present invention may occur before, simultaneously and / or after the administration of an additional therapeutic agent or agents, preferably an antibody or antibodies.

[0227] By a "crossover" Fab molecule (also called "Crossfab"), it is meant a Fab molecule in which the variable domains or the constant domains of the heavy and light chains of the Fab are exchanged (i.e., substituted for each other), i.e., the crossover Fab molecule comprises a peptide chain composed of the light chain variable domain VL and the heavy chain constant domain CH1 (VL-CH1, in the direction from the N- to the C-terminus), and a peptide chain composed of the heavy chain variable domain VH and the light chain constant domain CL (VH-CL, in the direction from the N- to the C-terminus). For clarity, in a crossover Fab molecule in which the variable domains of the Fab light chain and Fab heavy chain are exchanged, the peptide chain containing the heavy chain constant domain 1 CH1 is referred to herein as the "heavy chain" of the (crossover) Fab molecule.Meanwhile, in the crossover Fab molecule in which the constant domains of the Fab light chain and the Fab heavy chain are exchanged, the peptide chain containing the variable domain of the heavy chain VH is referred to in this document as the "heavy chain" of the (crossover) Fab molecule.

[0228] In contrast, a “standard” Fab molecule means that the Fab molecule has its natural format, i.e., it contains a heavy chain consisting of the variable and constant domains of the heavy chain (VH-CH1, in the direction from N- to C-terminus), and a light chain consisting of the variable and constant domains of the light chain (VL-CL, in the direction from N- to C-terminus).

[0229] The term "polynucleotide" refers to an isolated nucleic acid molecule or construct, such as messenger RNA (mRNA), viral RNA, or plasmid DNA (pDNA). A polynucleotide may contain a conventional phosphodiester linkage or a non-conventional linkage (e.g., an amide linkage such as that found in peptide nucleic acids (PNA)). The term "nucleic acid molecule" refers to any one or more nucleic acid segments, such as DNA or RNA fragments, present in a polynucleotide.

[0230] An "isolated" nucleic acid molecule or polynucleotide means a nucleic acid molecule, DNA, or RNA, that has been removed from its native environment. For example, a recombinant polynucleotide encoding a polypeptide contained in a vector is considered isolated in the context of the present invention. Additional examples of isolated polynucleotides include recombinant polynucleotides contained in heterologous host cells or purified (partially or substantially) polynucleotides in solution. An isolated polynucleotide includes a polynucleotide molecule contained in cells that normally contain the polynucleotide molecule, but in which the polynucleotide molecule is present extrachromosomally or in a chromosomal location that differs from its natural chromosomal location. Isolated RNA molecules include in vivo or in vitro RNA transcripts of the present invention, as well as positive and negative strand forms and double-stranded forms.Isolated polynucleotides or nucleic acids according to the present invention further include such molecules produced synthetically. Additionally, a polynucleotide or nucleic acid may comprise or comprise a regulatory element, such as a promoter, a ribosome binding site, or a transcription terminator.

[0231] A nucleic acid or polynucleotide having a nucleotide sequence that is at least, for example, 95% "identical" to a reference sequence according to the present invention means that the nucleotide sequence of the polynucleotide is identical to the reference sequence, except that the polynucleotide sequence may include up to five point mutations for every 100 nucleotides of the reference nucleotide sequence. In other words, to obtain a polynucleotide having a nucleotide sequence that is at least 95% identical to the reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or replaced with other nucleotides, or a number of nucleotides representing up to 5% of the total number of nucleotides in the reference sequence may be inserted into the reference sequence.These changes in the reference sequence may be at the 5' or 3' terminal positions of the reference nucleotide sequence or anywhere between these terminal positions, located individually between residues in the reference sequence or one or more contiguous groups in the reference sequence. In practice, the determination that any particular polynucleotide sequence is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence of the present invention can be carried out in a conventional manner using known computer programs, such as those discussed above for polypeptides (e.g., ALIGN-2).

[0232] The term "expression cassette" refers to a polynucleotide created recombinantly or synthetically with a number of defined nucleic acid elements that enable the transcription of a specific nucleic acid in a target cell. A recombinant expression cassette can be included in a plasmid, chromosome, mitochondrial DNA, plastid DNA, virus, or nucleic acid fragment. Typically, a recombinant expression cassette, as part of an expression vector, contains, among other sequences, a nucleic acid sequence intended for transcription and a promoter. In certain embodiments, an expression cassette of the invention comprises polynucleotide sequences that encode the bispecific antigen-binding molecules of the invention or fragments thereof.

[0233] The terms "vector" or "expression vector" are synonymous with "expression construct" and refer to a DNA molecule that is used to introduce and direct the expression of a specific gene to which it is operably linked in a target cell. This term includes a vector in the form of a self-replicating nucleic acid structure, as well as a vector incorporated into the genome of the host cell into which it is introduced. The expression vector of the present invention comprises an expression cassette. Expression vectors allow the transcription of large amounts of stable mRNA. Once the expression vector is in the target cell, the cellular transcriptional and / or translational machinery produces the ribonucleic acid molecule or protein encoded by the gene. In one embodiment, the expression vector of the invention comprises an expression cassette that contains polynucleotide sequences that encode the bispecific antigen-binding molecules of the invention or fragments thereof.

[0234] The term "approximately" as used herein refers to the typical error range for the corresponding value and is well known to those skilled in the art. In this document, the use of "approximately" with respect to a quantity or parameter includes (and describes) embodiments that directly relate to this quantity or parameter.

[0235] B-cell proliferative disorder refers to a disease in which the patient's B-cell count is elevated compared to that of a healthy subject, and in particular, in which the elevated B-cell count is a hallmark of the disease. "CD20-positive B-cell proliferative disorder" refers to a B-cell proliferative disorder in which B cells, particularly malignant B cells (in addition to normal B cells), express CD20.

[0236] Typical diseases associated with B cell proliferation include non-Hodgkin lymphoma (NHL), diffuse large B cell lymphoma (DLBCL; such as relapsed or refractory DLBCL not otherwise specified (NOS), high-grade B cell lymphoma (HGBCL; such as HGBCL NOS, double-lesion HGBCL, and triple-lesion HGBCL), primary mediastinal large B cell lymphoma (PMBCL), and DLBCL derived from FL (transformed FL; trFL)); follicular lymphoma (FL), including FL classes 1-3b; mantle cell lymphoma (MCL); marginal zone lymphoma (MZL), including splenic, nodal, or extranodal MZL. In one embodiment, the CD20-positive B-cell proliferative disorder is relapsed or refractory NHL (e.g., relapsed or refractory DLBCL, relapsed or refractory FL, relapsed or refractory MCL).

[0237] "Refractory disease" means the absence of a complete remission during first-line therapy. In one embodiment, refractory disease is defined as the absence of response or relapse within 6 months after previous therapy. In one embodiment, refractory disease is characterized by one or more of the following: progressive disease (PD) as the best response to first-line therapy, stable disease (SD) as the best response after at least 4 cycles of first-line therapy (e.g., 4 cycles of rituximab, cyclophosphamide, doxorubicin hydrochloride (hydroxydaunorubicin), vincristine sulfate (Oncovin), and prednisone, also abbreviated as R-CHOP), or partial response (PR) as the best response after at least 6 cycles and biopsy-proven residual disease or disease progression after a partial response. "Disease relapse" means a complete remission during first-line therapy.In one embodiment, disease recurrence is confirmed by biopsy. In one embodiment, patients have relapsed or failed to respond to at least two prior systemic treatment regimens (including at least one regimen containing anthracyclines and at least one regimen containing anti-CD20 therapy).

[0238] An "individual" or "subject" is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). Preferably, the individual or subject is a human. In one case, each subject in the subject population is a human. In one case, each subject in the reference subject population is a human.

[0239] A subject who is “not suitable for transplantation” or “not eligible for autologous stem cell transplantation (ASCT)” is a subject who is not eligible for, is not recommended to receive, is unable to receive, or refuses autologous SCT. Examples of preferred subject characteristics include age ≤ 65 years, Karnofsky performance status (KPS; Karnofsky et al. Cancer. 1948; 1(4): 634–656) > 60, forced expiratory volume in 1 second (FEV1) > 60% of predicted, diffusing capacity of the lungs (DLCO) > 60% of predicted, left ventricular ejection fraction > 45%, normal heart rhythm, serum bilirubin ≤ 2 mg / 100 mL, alanine aminotransferase (ALT) / aspartate aminotransferase (AST) ≤ 2 × normal, serum creatinine ≤ 1.5 mg / 100 mL, creatinine clearance > 60 mL / min., absence of a second active malignant tumor, absence of pregnancy and uncontrolled infections (including dental) (Hamadani M et al. Bone Marrow Transplant. 2010; 45:1259-68).

[0240] As used herein, the term "treatment" (and its grammatical variants, such as "treat" or "treating") refers to a clinical intervention with the aim of altering the natural course of a disease in the individual being treated, and can be carried out both prophylactically and during the course of clinical pathology. Desirable treatment effects include, but are not limited to, preventing the onset or recurrence of the disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, preventing metastasis, reducing the rate of disease progression, reducing the intensity or temporary alleviation of the painful condition, and remission or improving the prognosis. In some embodiments, the methods of the present invention are used to delay the development of a disease or slow the progression of a disease.

[0241] As used herein, "slowing down the progression" of a disorder or disease means delaying, inhibiting, slowing, arresting, stabilizing, and / or postponing the development of the disease or disorder (e.g., a CD20-positive B-cell proliferative disorder, such as NHL, such as DLBCL). This slowing may vary in duration, depending on the disease history and / or the individual being treated. It is understood by those skilled in the art that a substantial or significant slowing may, in practice, include preventing the development of the disease in a subject. For example, in advanced cancer, the development of metastases to the central nervous system (CNS) may be slowed.

[0242] The term "reduce" or "inhibit" refers to the ability to produce an overall reduction of, for example, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or more. For clarity, this term also includes a reduction to zero (or below the detection limit of the assay), i.e., complete elimination or abolition.In some embodiments, reducing or inhibiting may mean reducing or inhibiting adverse events such as cytokine-mediated toxicity (e.g., cytokine release syndrome (CRS)), infusion-related reactions (IRR), macrophage activation syndrome (MAS), neurological toxicity, severe tumor lysis syndrome (STS), neutropenia, thrombocytopenia, liver enzyme elevations, and / or central nervous system (CNS) toxicity following treatment with an anti-CD20 / CD3 bispecific antibody using a stepped dosing regimen of the present invention, compared to an unchanged, pre-determined dosing regimen with a target dose of the bispecific antibody.In other embodiments, the terms "reduce" or "inhibit" may refer to an effector function of an antibody that is mediated by the Fc region of the antibody, wherein such effector functions include, but are not limited to, complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC), and antibody-dependent cellular phagocytosis (ADCP). In other embodiments, the reduction or inhibition may refer to the symptoms of the CD20-positive B-cell proliferative disorder being treated (e.g., NHL (e.g., DLBCL), FL (e.g., relapsed and / or refractory FL or transformed FL), MCL, high-grade B-cell lymphoma, or PMLBCL), the presence or size of metastases, or the size of the primary tumor.

[0243] As used herein, "administering" refers to a method of administering a dose of a compound (e.g., a CD20 / CD3 bispecific antibody) or a composition (e.g., a pharmaceutical composition comprising a CD20 / CD3 bispecific antibody) to a subject. The compounds and / or compositions used in the methods described herein can be administered intravenously (e.g., by intravenous infusion).

[0244] A "fixed" or "basic" dose of a therapeutic agent (e.g., a bispecific antibody) in this document refers to the dose administered to a patient regardless of their weight or body surface area (BSA). Therefore, the fixed or basic dose is not expressed as mg / kg or mg / m, but as an absolute amount of therapeutic agent (e.g., mg).

[0245] The "target dose" in this document refers to the dose of the CD20 / CD3 bispecific antibody that achieves a therapeutic effect, i.e., the desired clinical efficacy. For glofitamab, the potential target dose has been determined to be 16 mg or 30 mg.

[0246] “Fixed or set dosing with a target dose” and “treatment regimen without step dosing” mean a dosing schedule that uses the same dose in the first and second cycles and optionally in any subsequent treatment cycle, as opposed to step dosing, which uses lower doses in the first few treatment cycles and achieves the target dose only in the second or subsequent treatment cycle.

[0247] The terms "treatment cycle" or "cycle" as used herein (abbreviated: "C") mean a course of one or more doses of an anti-CD20 / CD3 bispecific antibody that is repeated on a regular schedule, optionally with rest periods (no treatment) in between. In one aspect of the invention, the first treatment cycle includes first and second doses of the anti-CD20 / CD3 bispecific antibody, followed by a rest period. In one such embodiment, the first treatment cycle includes a first dose of the anti-CD20 / CD3 bispecific antibody on day 1 of the first cycle and a second dose of the anti-CD20 / CD3 bispecific antibody on day 8 of the first cycle, followed by 12 days of rest. In one embodiment, the second and all subsequent cycles include one dose of the anti-CD20 / CD3 bispecific antibody, which is administered on the first day of the cycle, followed by 20 days of rest. In one embodiment, one treatment cycle is 21 days.In another embodiment, one treatment cycle is 14 days. A treatment cycle comprising one or more doses of the CD20 / CD3 bispecific antibody may further comprise one or more doses of one or more other therapeutic agents, for example, an anti-CD20 antibody, in particular obinutuzumab. The treatment schedule according to the invention may comprise 2 or more treatment cycles, or 3, 4, 5, 6, 7, 8, 9, 10, 11, in particular 12 treatment cycles.

[0248] “Individual response” or “response” may be assessed by any endpoint that indicates benefit to the subject, including, but not limited to, (1) inhibition to some extent of disease progression (e.g., progression of a CD20-positive B-cell proliferative disorder such as non-Hodgkin's lymphoma (NHL)), including slowing and stopping; (2) reduction in tumor size; (3) inhibition (i.e., reducing, slowing, or stopping) of cancer cell infiltration into adjacent peripheral organs and / or tissues; (4) inhibition (i.e.,(5) alleviating, to some extent, one or more symptoms associated with a CD20-positive B-cell proliferative disorder, such as a B-cell proliferative disorder; (6) prolonging or increasing the duration of survival, including overall survival and progression-free survival; and / or (7) reducing mortality at a specified time point after treatment.

[0249] As used herein, “complete response” or “CR” refers to the disappearance of all target lesions. In one embodiment, standard response criteria in NHL are used to determine CR. (Lugano Classification, Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27): 3059-3067.)

[0250] In this document, "partial response" or "PR" means a reduction in the sum of the longest diameters (SLD) of the target lesions by at least 30%, using the baseline SLD as the reference, or a reduction in the product of the diameters (PMD) of the target lesions by at least 50%, using the baseline PLD as the reference.

[0251] "Long-term response" refers to a long-term effect of tumor growth reduction after the end of treatment. For example, the tumor size may remain the same or become smaller compared to the size at the beginning of the administration phase. In some embodiments, the long-term response has a duration of at least the same as the duration of treatment, at least 1.5x, 2.0x, 2.5x, or 3.0x the duration of treatment.

[0252] An "effective response" of a subject or a subject's "responsiveness" to treatment with a medicinal product and similar expressions refer to a clinical or therapeutic benefit for a subject at risk for or suffering from a disease or disorder, such as cancer. In one embodiment, such benefit includes one or more of: prolongation of survival (including overall survival and progression-free survival); obtaining an objective response (including a complete response or partial response); or improvement in the signs or symptoms of cancer.

[0253] "Duration of complete response" (DCR) is defined as the time elapsed from the initial onset of documented CR to documented disease progression or death from any cause, whichever occurs first. In one embodiment, DCR is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27): 3059-3067.).

[0254] “Duration of objective response” (DOR) is defined as the first occurrence of a documented objective response before disease progression, relapse, or death from any cause. In one embodiment, DOR is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27): 3059-3067.).

[0255] “Progression-free survival” (PFS) is defined as the time from the first treatment with a CD20 / CD3 bispecific antibody to the first event of disease progression or death from any cause, whichever occurs first. In one embodiment, PFS is estimated based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27): 3059-3067.).

[0256] “Overall survival” (OS) is defined as the time from the first treatment with CD20 / CD3 bispecific antibody to the date of death from any cause.

[0257] “Time to first overall response” (TOOR) is defined as the time from the start of treatment to the first documented response. In one embodiment, TOOR is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27): 3059-3067.).

[0258] “Time to first complete response” (TFR) is defined as the time from the start of treatment to the first documented complete response. In one embodiment, TFR is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27): 3059-3067.).

[0259] In this document, the “objective response rate” or “overall response rate” (ORR) is defined as the sum of the partial response (PR) and complete response (CR) rates. In one embodiment, the ORR is estimated based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27): 3059-3067).

[0260] In this document, “stable disease” or “SD” refers to neither a reduction in target lesions sufficient to confirm PR nor an increase sufficient to confirm PD, with the lowest SND value since the start of treatment being taken as the starting point.

[0261] In this document, "progressive disease" or "PD" means an increase in the SND of target lesions by at least 20%, using the lowest SND as the reference, or an increase in the SPD of target lesions by at least 50%, using the lowest SPD as the reference, recorded since the start of treatment, or the presence of one or more new lesions.

[0262] As used herein, an “infusion-related reaction,” “IRR,” or “infusion-related adverse event” is an adverse event that occurs in a patient or subject during or within 24 hours after administration of a medicinal product (e.g., a CD20 / CD3 bispecific antibody, such as glofitamab, or an anti-CD79b antibody-drug conjugate, such as polatuzumab vedotin). IRRs can be classified into severity grades 1-5 in accordance with, for example, NCI CTCAE v.4.

[0263] The term "PD-1 axis binding antagonist" refers to a molecule that inhibits the interaction of a PD-1 axis binding partner with one or more of its binding partners so as to eliminate T cell dysfunction resulting from signaling in the PD-1 signaling axis, leading to the restoration or enhancement of T cell function (e.g., proliferation, cytokine production, target cell killing). As used herein, a PD-1 axis binding antagonist includes a PD-1 binding antagonist, a PD-L1 binding antagonist, and a PD-L2 binding antagonist.

[0264] The term "PD-1-binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signal transduction due to the interaction of PD-1 with one or more of its binding partners, such as PD-L1, PD-L2. In some embodiments, the PD-1-binding antagonist is a molecule that inhibits the binding of PD-1 to one or more of its binding partners. In a particular aspect, the PD-1-binding antagonist inhibits the binding of PD-1 to PD-L1 and / or PD-L2. For example, PD-1-binding antagonists include anti-PD-1 antibodies, their antigen-binding fragments, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, suppress, or disrupt signal transduction resulting from the interaction of PD-1 with PD-L1 and / or PD-L2.In one embodiment, a PD-1-binding antagonist reduces a negative costimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-1 to render a dysfunctional T cell less dysfunctional (e.g., increasing the effector response to antigen recognition). In some embodiments, the PD-1-binding antagonist is an anti-PD-1 antibody. In a specific aspect, the PD-1-binding antagonist is MDX-1106 (nivolumab). In another specific aspect, the PD-1-binding antagonist is pembrolizumab (formerly lambrolizumab (MK-3475)). In another specific aspect, the PD-1-binding antagonist is AMP-224. In some embodiments, the PD-1-binding antagonist is MDX-1106 (nivolumab).In some embodiments, the PD-1-binding antagonist is MK-3475 (pembrolizumab). In some embodiments, the PD-1-binding antagonist is MED1-0680. In some cases, the PD-1-binding antagonist is PDR001 (spartalizumab). In some cases, the PD-1-binding antagonist is REGN2810 (cemiplimab). In some cases, the PD-1-binding antagonist is BGB-108. In other cases, the PD-1-binding antagonist is prolgolimab, camrelizumab, sintilimab, tislelizumab, or toripalimab.

[0265] Other examples of PD-1 axis binding antagonists include cemiplimab, prolgolimab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostarlimab, retifanlimab, spartalizumab, sasanlimab, penpulimab, CS1003, HLX10, SCT-I10A, SHR-1316, CS1001, envafolimab, TQB2450, ZKAB001, LP-002, zimberelimab, balstilimab, genolimzumab, BI 754091, cetrelimab, YBL-006, BAT1306, HX008, CX-072, IMC-001, KL-A167, budigalimab, AMG 404, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, APL-502, cosibelimab, lodapolimab, GS-4224, INCB086550, FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, MAX-10181, RC98, BION-004, AM0001, SV201, ENUM 244С8, ENUM 388D4, AUNP-012, STI-1110, ADG104, AK-103, LBL-006, hAb21, AVA-004, PDL-GEX, INCB090244, KD036, KY1003, LYN192, MT-6035, VXM10, YBL-007, ABSK041, GB7003, JS-003, HS-636.

[0266] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signal transduction resulting from the interaction of PD-L1 with one or more of its binding partners, such as PD-1 or B7-1. In some embodiments, the PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partners. In a particular aspect, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1 and / or B7-1. In some embodiments, PD-L1-binding antagonists include PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, suppress, or interfere with signaling resulting from the interaction of PD-L1 with one or more of its binding partners, such as PD-1 and B7-1.In one embodiment, a PD-L1-binding antagonist reduces a negative costimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-L1 to render a dysfunctional T cell less dysfunctional (e.g., increasing the effector response to antigen recognition). In some embodiments, the PD-L1-binding antagonist is an anti-PD-L1 antibody. In some specific embodiments, the anti-PD-L1 antibody is atezolizumab (CAS Registry Number: 1422185-06-5), also known as MPDL3280A, and described herein. In another specific embodiment, the anti-PD-L1 antibody is MDX-1105, described herein. In another specific aspect, the anti-PD-L1 antibody is MEDI4736, as described herein.

[0267] In this document, the term “atezolizumab” refers to the PD-L1 antagonist antibody with the International Nonproprietary Name (INN) of Schedule 112 (WHO Drug Information, Vol.28, No.4, 2014, p.488) or CAS Registration Number 1380723-44-3.

[0268] The term "PD-L2-binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signal transduction resulting from the interaction of PD-L2 with one or more of its binding partners, such as PD-1. In some embodiments, the PD-L2-binding antagonist is a molecule that inhibits the binding of PD-L2 to one or more of its binding partners. In a particular aspect, the PD-L2-binding antagonist inhibits the binding of PD-L2 to PD-1. In some embodiments, PD-L2-binding antagonists include PD-L2 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, suppress, or interfere with signal transduction resulting from the interaction of PD-L2 with one or more of its binding partners, such as PD-1.In one embodiment, a PD-L2-binding antagonist reduces the negative costimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-L2 to render a dysfunctional T cell less dysfunctional (e.g., increasing the effector response to antigen recognition). In some embodiments, the PD-L2-binding antagonist is an immunoadhesin.

[0269] As used herein, the term “chemotherapeutic agent” refers to a compound useful for the treatment of cancer such as a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., relapsed or refractory B-cell proliferative disorder), e.g., non-Hodgkin's lymphoma (NHL; e.g., diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL; e.g., grade 1 FL, grade 2 FL, grade 3 FL (e.g., grade 3a FL, grade 3b FL), or transformed FL), mantle cell lymphoma (MCL), or marginal zone lymphoma (MZL)), e.g., relapsed or refractory NHL (e.g., relapsed or refractory DLBCL, relapsed or refractory FL, relapsed or refractory MCL or marginal zone lymphoma (MZL). Examples of chemotherapy agents include EGFR inhibitors (including small molecule inhibitors (e.g., erlotinib (TARCEVA®, Genentech / OSI Pharm.);PD 183805 (CI 1033, 2-propenamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-, dihydrochloride, Pfizer Inc.); ZD1839, gefitinib (IRESSA®) 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca); ZM 105180 ((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-(1-methyl-piperidin-4-yl)-pyrimido[5,4-c1]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166 ((R)-4-[4-[(l-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide); EKB-569 (N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolinyl]-4-(dimethylamino)-2-butynamide) (Wyeth); AG1478 (Pfizer); AG1571 (SU 5271; Pfizer);and dual EGFR / HER2 tyrosine kinase inhibitors such as lapatinib (TYKERB®, GSK572016, or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[[2-methylsulfonyl)ethyl]amino]methyl]-2-furanyl]-4-quinazolinamine]); tyrosine kinase inhibitor (e.g., EGFR inhibitor; small molecule HER2 tyrosine kinase inhibitor, e.g., TAK 165 (Takeda); CP-724,714, an oral selective ErbB2 receptor tyrosine kinase inhibitor (Pfizer and OSI); dual HER inhibitors, e.g., EKB-569 (Wyeth), which preferentially binds EGFR but inhibits both HER2 and EGFR-overexpressing cells; PKI-166 (Novartis); pan-HER inhibitors, such as canertinib (CI-1033, Pharmacia); Raf-1 inhibitors, such as the antisense agent ISIS-5132 (ISIS Pharmaceuticals), which inhibits Raf-1 signaling; non-HER-targeting tyrosine kinase inhibitors, such as imatinib mesylate (GLEEVEC®, Glaxo SmithKline); multitargeted tyrosine kinase inhibitors such as sunitinib (SUTENT®, Pfizer);VEGF receptor tyrosine kinase inhibitors such as vatalanib (PTK787 / ZK222584, Novartis / Schering AG); extracellular regulated kinase I MAPK inhibitor CI-1040 (Pharmacia); quinazolines such as PD 153035, 4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyrimidopyrimidines; pyrrolopyrimidines such as CGP 59326, CGP 60261 and CGP 62706; pyrazolopyrimidines, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidines; curcumin (diferuloylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tyrphostins containing nitrothiophene molecules; PD-0183805 (Warner-Lamber); antisense molecules (e.g., binding to HER-encoding nucleic acid); quinoxalines (U.S. Patent No. 5,804,396); triphostins (U.S. Patent No. 5,804,396); ZD6474 (Astra Zeneca); PTK-787 (Novartis / Schering AG); pan-HER inhibitors such as CI-1033 (Pfizer); Affinitac (ISIS 3521; Isis / Lilly); PKI 166 (Novartis); GW2016 (Glaxo SmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering AG);INC-1C11 (Imclone); rapamycin (sirolimus, RAPAMUNE®); proteasome inhibitors such as bortezomib (VELCADE®, Millennium Pharm.); disulfiram; epigallocatechin gallate; salinosporamide A; carfilzomib; 17-AAG (geldanamycin); radicicol; lactate dehydrogenase A (LDH-A); fulvestrant (FASLODEX®, AstraZeneca); letrozole (FEMARA®, Novartis), finasunate (VATALANIB®, Novartis); oxaliplatin (ELOXATIN®, Sanofi); 5-fluorouracil (5-FU); leucovorin; lonafamib (SCH 66336); sorafenib (NEXAVAR®, Bayer Labs); AG1478, alkylating agents such as thiotepa and cyclophosphamide CYTOXAN®; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carbochone, meturedopa, and uredopa; ethyleneimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylomelamin; acetogenins (particularly bulatacin and bulatacinone); camptothecin (including topotecan and irinotecan); bryostatin; callistatin;CC-1065 (including its synthetic analogs adozelesin, carzelesin and bizelesin); cryptophycins (in particular, cryptophycin-1 and cryptophycin-8); adrenocorticosteroids (including prednisone and prednisolone); cyproterone acetate; 5α-reductase inhibitors, including finasteride and dutasteride); vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat dolastatin; aldesleukin, talc duocarmycin (including synthetic analogs - KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictin; spongistatin; Nitrogen mustards such as chlorambucil, chlomaphazine, chlorphosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novambiquin, phenesterine, prednimustine, trofosfamide, uracil mustard; Nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; Antibiotics such as enedine antibiotics (e.g. calicheamicin, in particular calicheamicin γ1 and calicheamicin ω1); Dynemicin, including dynemicin A; Bisphosphonates such as clodronate; Esperamicin;and the neocarzinostatin chromophore and related chromophores of the enedin antibiotics), aclacinomycins, actinomycin, authramycin, azazerine, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycin, dactinomycin, detorubicin, 6-diazo-5-oxo-L-norleucine, morpholinodoxorubicin, cyanomorpholinodoxorubicin, 2-pyrrolinodoxorubicin, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine;Androgens such as calusterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; Antiadrenal drugs such as aminoglutethimide, mitotane, trilostane; Folic acid replenisher such as frolinic acid; Aceglatone; Aldophosphamide glycoside; Aminolevulinic acid; Eniluracil; Amsacrine; Bestrabucil; Bisantrene; Edatraxate; Defofamine; Demecolcine; Diazicon; Elfomitin; Elliptinium acetate; Epothilone; Etoglucide; Gallium nitrate; Hydroxyurea; Lentinan; Lonidainine; Maytansinoids such as maytansine and ansamitocins; Mitoguazone; Mitoxantrone; Mopidaminol; Nitraerin; pentostatin; fenamet; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products); razoxan; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triazicon; 2,2',2''-trichlorotriethylamine; trichothecenes (including T-2 toxin, verracurin A, roridin A, and anhydrin); urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol;pipobroman; gacytosin; arabinoside (Ara-C); thiotepa; chloranbucil; GEMZAR® (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; etoposide (VP-16); ifosfamide; mitoxantrone; novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine (XELODA®); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; pharmaceutically acceptable salts, acids, prodrugs, and derivatives of any of the foregoing.

[0270] Chemotherapeutic agents also include (i) antihormonal agents that regulate or inhibit the action of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including NOLVADEX®; tamoxifen citrate), raloxifene, droloxifene, iodoxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and FARESTON® (toremifine citrate); (ii) aromatase inhibitors, which inhibit the aromatase enzyme, which regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazoles, aminoglutethimide, MEGASE® (megestrol acetate), AROMASIN® (exemestane; Pfizer), formestani, fadrozole, RIVISOR® (vorozole), FEMARA® (letrozole; Novartis) and ARIMIDEX® (anastrozole; AstraZeneca); (iii) antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, goserelin;buserelin, tripterelin, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, all-trans retinoic acid, fenretinide, and troxacitabine (1,3-dioxolane nucleoside cytosine analogue); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, in particular those that inhibit the expression of genes in signaling pathways involved in impaired cell proliferation, such as, for example, PKC-alpha, Ralf and H-Ras; (vii) ribozymes, such as VEGF expression inhibitors (e.g., ANGIOZYME®) and HER2 expression inhibitors; (viii) vaccines, such as gene therapy vaccines, for example ALLOVECTIN®, LEUVECTIN® and VAXID®;(ix) growth inhibitors, including vincristine (e.g., vincristine and vinblastine), NAVELBINE® (vinorelbine), taxanes (e.g., paclitaxel, nab-paclitaxel, and docetaxel), topoisomerase II inhibitors (e.g., doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin), DNA alkylating agents (e.g., tamoxifen, dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C); and (x) pharmaceutically acceptable salts, acids, prodrugs, and derivatives of any of the foregoing.

[0271] The term "cytotoxic agent" as used herein refers to a substance that inhibits or interferes with the function of cells and / or results in the death or destruction of cells. Cytotoxic agents include, but are not limited to, radioactive isotopes (e.g., At 211 , I 131 , I 125 , Y 90 , Re 186 , Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212and radioactive isotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitory agents; enzymes and fragments thereof, such as nucleolytic enzymes; antibiotics; toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof; and various antineoplastic or anticancer agents described below.

[0272] III. Therapeutic agents for use in the methods of the present invention

[0273] A. Bispecific antibodies to CD20 / CD3

[0274] The present invention provides novel dosages of anti-CD20 / CD3 bispecific antibodies. In one embodiment, the antibody is a monoclonal antibody. In one embodiment, the anti-CD20 / CD3 bispecific antibody is a polyclonal antibody. In one embodiment, the anti-CD20 / CD3 bispecific antibody is a human antibody. In one embodiment, the anti-CD20 / CD3 bispecific antibody is a humanized antibody. In one embodiment, the anti-CD20 / CD3 bispecific antibody is a chimeric antibody. In one embodiment, the anti-CD20 / CD3 bispecific antibody is a full-length antibody. In one embodiment, the anti-CD20 / CD3 bispecific antibody is an IgG class antibody, in particular an IgG1 subclass antibody. In one embodiment, the anti-CD20 / CD3 bispecific antibody is a recombinant antibody.

[0275] In some embodiments, the anti-CD20 / CD3 bispecific antibody comprises an antibody fragment. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab')2, Fv, and scFv fragments and other fragments described below. For a review of certain antibody fragments, see Hudson et al. Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, e.g., Pluckthün, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York), pp. 269-315 (1994); see also WO 93 / 16185; and U.S. Patent Nos. 5,571,894 and 5,587,458. For a discussion of Fab and F(ab')2 fragments containing salvage receptor binding epitope residues and having an increased half-life in vivo, see U.S. Patent No. 5,869,046. In one embodiment, the antibody fragment is a Fab fragment or an scFv fragment.

[0276] Diabodies are antibody fragments with two antigen-binding sites, which may be bivalent or bispecific. See, e.g., EP 404097; WO 1993 / 01161; Hudson et al., Nat. Med. 9:129–134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444–6448 (1993). Tribodies and tetrabodies are also described in Hudson et al., Nat. Med. 9:129–134 (2003).

[0277] Single-domain antibodies are antibody fragments comprising all or part of a heavy chain variable domain or all or part of a light chain variable domain of an antibody. In certain embodiments, the single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U.S. Patent No. 6,248,516 B1).

[0278] Antibody fragments can be produced by a variety of methods, including, but not limited to, proteolytic cleavage of an intact antibody, as well as production using recombinant host cells (e.g., E. coli or phage), as described herein.

[0279] In certain embodiments, the CD20 / CD3 bispecific antibody is a chimeric antibody. Some chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). In one example, the chimeric antibody comprises a non-human variable region (e.g., a variable region derived from an antibody of a mouse, rat, hamster, rabbit, or non-human primate, such as a monkey) and a human constant region. In a further example, the chimeric antibody is a "class-switched" antibody, the class or subclass of which has been altered compared to the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0280] In certain embodiments, the CD20 / CD3 bispecific antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce immunogenicity in humans while maintaining the specificity and affinity of the parent non-human antibody. Typically, a humanized antibody comprises one or more variable domains in which the HVRs, such as CDRs (or fragments thereof), are derived from an antibody of non-human origin, and the FRs (or fragments thereof) are derived from human antibody sequences. The humanized antibody optionally also comprises at least a portion of a human constant region. In some embodiments, some FR residues in the humanized antibody are substituted with the corresponding residues of a non-human antibody (e.g., the antibody from which the HVR residues are derived), for example, to restore or improve the specificity or affinity of the antibody.

[0281] Humanized antibodies and methods for producing them are discussed, for example, in Almagro and Fransson, Front. Biosci. 13:1619–1633 (2008), and further described, for example, in Riechmann et al., Nature 332:323–329 (1988); Queen et al., Proc. Nat'l Acad. Sci. USA 86:10029–10033 (1989); U.S. Pat. Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods 36:25–34 (2005) (which describes grafting of a specificity determining region (SDR)); Padlan, Mol. Immunol. 28:489–498 (1991) (described “surface modification”); Dall'Acqua et al., Methods 36:43–60 (2005) (described “FR shuffling”); and Osbourn et al., Methods 36:61–68 (2005) and Klimka et al., Br. J. Cancer, 83:252–260 (2000) (described a “directed selection” approach to FR shuffling).

[0282] Human framework regions that can be used for humanization include, but are not limited to, framework regions selected by the "best match" method (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from the consensus sequence of human antibodies of a particular subgroup of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol, 151:2623 (1993)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and framework regions obtained by screening FR libraries (see, e.g., Baca et al., J. Biol. Chem. 272:10678–10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611–22618 (1996)).

[0283] In certain embodiments, the CD20 / CD3 bispecific antibody is a human antibody. Human antibodies can be produced using various techniques known in the art. Human antibodies are described generally in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20: 450-459 (2008).

[0284] Human antibodies can be produced by administering an immunogen to a transgenic animal modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigen stimulation. Such animals typically contain all or part of the human immunoglobulin loci that replace the endogenous immunoglobulin loci or that are present extrachromosomally or randomly integrated into the animal's chromosomes. Typically, such transgenic mice have inactivated endogenous immunoglobulin loci. For a review of methods for producing human antibodies using transgenic animals, see Lonberg, Nat. Biotech. 23:1117–1125 (2005).See also, for example, U.S. Patent Nos. 6,075,181 and 6,150,584, which describe XENOMOUSE™ technology; U.S. Patent No. 5,770,429, which describes HuMab® technology; U.S. Patent No. 7,041,870, which describes K-M MOUSE® technology; and U.S. Patent Application Publication No. US 2007 / 0061900, which describes VelociMouse® technology. Human variable regions from intact antibodies generated by such animals can be further modified, for example, by combining with different human constant regions.

[0285] Human antibodies can be produced using hybridoma-based methods. Human myeloma and mouse and human heteromyeloma cell lines for the production of human monoclonal antibodies have been described. (See, e.g., Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991).) Human antibodies produced by human B-cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103: 3557-3562 (2006). Additional methods include those described, for example, in U.S. Patent No. 7,189,826 (which describes the production of human IgM monoclonal antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) (which describes human-human hybridomas).The method for producing human hybridoma (trioma method) is also described in Vollmers and Brandlein, Histology and Histopathology, 20(3):927-937 (2005), and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27(3): 185-91 (2005).

[0286] Human antibodies can also be generated by isolating Fv clone variable domain sequences from human-derived phage display libraries. These variable domain sequences can then be fused with the desired human constant domain. Methods for selecting human antibodies from antibody libraries are described below.

[0287] Binding domains comprising a CD20 / CD3 bispecific antibody can be isolated by screening combinatorial libraries for binding molecules with the desired activity or activities. For example, a number of methods for generating phage display libraries and screening such libraries for antibodies having the desired binding characteristics are known in the art. Such methods are discussed, for example, by Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, 2001) and are further described, for example, in McCafferty et al., Nature 348:552-554; Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Marks and Bradbury, in Methods in Molecular Biology 248:161-175 (Lo, ed., Human Press, Totowa, NJ, 2003); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellowes, Proc. Natl. Acad. Sci.USA 101(34): 12467-12472 (2004); и Lee et al., J. Immunol. Methods 284(1-2): 119-132(2004).

[0288] In some phage display-based methods, VH and VL gene repertoires are separately cloned by polymerase chain reaction (PCR) and randomly recombined into phage libraries, which can then be screened for antigen-binding phage as described by Winter et al., Ann. Rev. Immunol., 12: 433–455 (1994). Typically, phage display antibody fragments as single-chain Fv fragments (scFv) or Fab fragments. Libraries from immunized sources allow the generation of antibodies with high affinity for the immunogen without the need for hybridoma construction. Alternatively, a naive repertoire (e.g., from humans) can be cloned to produce a single source of antibodies to a wide range of non-self as well as self antigens without any immunization, as described in Griffiths et al., EMBO J, 12: 725-734 (1993).Finally, naive libraries can also be produced synthetically by cloning unrearranged V gene segments from stem cells and using PCR primers containing random sequence to encode highly variable CDR3 regions and performing rearrangement in vitro, as described by Hoogenboom and Winter, J. Mol. Biol., 227: 381–388 (1992). Patent publications that describe human antibody phage libraries include, for example, U.S. Patent No. 5,750,373 and U.S. Patent Publication Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360.

[0289] Antibodies or antibody fragments isolated from human antibody libraries are referred to herein as human antibodies or human antibody fragments.

[0290] Technologies for producing bispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs having different specificities (see Milstein et al. Nature 305: 537, (1983), WO 93 / 08829 and Traunecker et al. EMBO J. 10: 3655, (1991)), and knob-into-hole engineering (see, for example, U.S. Patent No. 5,731,168). Multispecific antibodies can also be produced by engineering using the electrostatic interaction effect to produce Fc-heterodimeric antibody molecules (WO 2009 / 089004 A1); cross-linking two or more antibodies or fragments (see, e.g., U.S. Patent No. 4,676,980, and Brennan et al., Science, 229: 81 (1985)); using leucine zippers to produce bispecific antibodies (see, e.g., Kostelny et al., J. Immunol., 148(5): 1547-1553 (1992)); using the diabody method to produce bispecific antibody fragments (see, e.g., Hollinger et al., Proc.Natl. Acad. Sci. USA, 90:6444–6448 (1993)); and the use of single-chain Fv (scFv) dimers (see, e.g., Gruber et al., J. Immunol., 152:5368 (1994)); and the production of trispecific antibodies, as described, e.g., in Tutt et al. J. Immunol. 147:60 (1991).

[0291] Engineered antibodies with three or more functional antigen-binding sites, including "octopus antibodies", are also included herein (see, for example, US 2006 / 0025576 A1).

[0292] The bispecific antibody to CD20 / CD3 herein also includes a "dual-action FAb" or "DAF" having an antigen-binding site that binds to two different antigens (see, for example, US 2008 / 0069820).

[0293] This document also includes “cross-mab” antibodies (see, for example, WO 2009080251, WO 2009080252, WO 2009080253, WO 2009080254).

[0294] Another technique for producing bispecific antibody fragments is the “bispecific T-cell-engaging activator” or BiTE® approach (see, e.g., WO 2004 / 106381, WO 2005 / 061547, WO 2007 / 042261, and WO 2008 / 119567). This approach utilizes two antibody variable domains located on a single polypeptide. For example, a single polypeptide chain comprises two single-chain Fv fragments (scFv), each having a heavy-chain variable domain (VH) and a light-chain variable domain (VL), separated by a polypeptide linker of sufficient length to provide an intramolecular linkage between the two domains. This single polypeptide further comprises a polypeptide spacer sequence between the two scFv fragments.Each scFv recognizes its own epitope, and these epitopes can be specific to different cell types, so that cells of two different cell types are brought into proximity or bind when each scFv interacts with its cognate epitope. In one embodiment of this approach, an scFv that recognizes a cell surface antigen expressed by an immune cell, such as the CD3 polypeptide on a T cell, is linked to another scFv that recognizes a cell surface antigen expressed by a target cell, such as a malignant or tumor cell.

[0295] Since the T-cell-attracting bispecific activator is a single polypeptide, it can be expressed using any prokaryotic or eukaryotic cell expression system known in the art, such as a CHO cell line. However, special purification techniques may be required to separate monomeric T-cell-attracting bispecific activators from other multimeric species that may have biological activity different from the intended activity of the monomer (see, for example, EP 1 691 833). In one exemplary purification scheme, a solution containing the secreted polypeptides is first subjected to metal affinity chromatography, and the polypeptides are eluted with an imidazole concentration gradient. This eluate is then purified using anion exchange chromatography, where the polypeptides are eluted with a sodium chloride concentration gradient.Finally, this eluate is subjected to size exclusion chromatography to separate monomers from multimers.

[0296] In some embodiments, the CD20 / CD3 bispecific antibody may be further modified to contain additional non-protein components known in the art and readily available. Moieties suitable for derivatization of the CD20 / CD3 bispecific antibody include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymer, polyamino acids (homopolymers or random copolymers), dextran or poly(n-vinylpyrrolidone)polyethylene glycol, propylene glycol homopolymers, oxide / ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerin), polyvinyl alcohol, and mixtures thereof.Polyethylene glycol propionaldehyde may offer manufacturing advantages due to its stability in water. The polymer can have any molecular weight and can be branched or unbranched. The number of polymers attached to the antibody can vary, and if more than one polymer is attached, they may be the same or different molecules. In general, the number and / or type of polymers used for derivatization can be determined based on considerations that include, but are not limited to, the specific properties or functions of the antibody that need to be improved, whether the antibody derivative will be used therapeutically under certain conditions, etc.

[0297] The CD20 / CD3 bispecific antibody may also be conjugated to one or more cytotoxic agents, such as chemotherapeutic agents or drugs, growth inhibitors, toxins (e.g., protein toxins, enzymatically active toxins of bacterial, fungal, plant or animal origin or fragments thereof) or radioactive isotopes.

[0298] In one embodiment, the anti-CD20 / CD3 bispecific antibody comprises an antibody drug conjugate (ADC) in which the antibody is conjugated to one or more drugs, including, but not limited to, a maytansinoid (see U.S. Patent Nos. 5,208,020, 5,416,064 and European Patent EP 0 425 235 B1); an auristatin such as the drug moieties monomethyl auristatin DE and DF (MMAE and MMAF) (see U.S. Patent Nos. 5,635,483, 5,780,588 and 7,498,298); dolastatin; calicheamicin or a derivative thereof (see U.S. Patent Nos. 5,712,374, 5,714,586, 5,739,116, 5,767,285, 5,770,701, 5,770,710, 5,773,001, and 5,877,296; Hinman et al., Cancer Res. 53:3336-3342 (1993); and Lode et al., Cancer Res. 58:2925-2928 (1998)); an anthracycline such as daunomycin or doxorubicin (see Kratz et al., Current Med. Chem. 13:477–523 (2006); Jeffrey et al., Bioorganic & Med. Chem. Letters 16:358–362 (2006); Torgov et al., Bioconj. Chem. 16:717–721 (2005); Nagy et al., Proc. Natl. Acad. Sci. USA 97:829–834 (2000); Dubowchik et al., Bioorg.& Med. Chem. Letters 12:1529–1532 (2002); King et al., J. Med. Chem. 45:4336–4343 (2002); and U.S. Patent No. 6,630,579); methotrexate; vindesine; taxanes such as docetaxel, paclitaxel, larotaxel, tesetaxel, and ortataxel; trichothecene; and CC1065.

[0299] In another embodiment, the CD20 / CD3 bispecific antibody is conjugated to an enzymatically active toxin or fragment thereof, including, but not limited to, diphtheria toxin A chain, non-binding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modecin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII and PAP-S), Momordica charantia inhibitor, curcin, crotin, Saponaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin and tricothecenes.

[0300] In another embodiment, the CD20 / CD3 bispecific antibody is conjugated to a radioactive atom to form a radioconjugate. A number of radioactive isotopes are available for the preparation of radioconjugates. Examples include At 211 , I 131 , I 125 , Y 90 , Re 186 , Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212 and radioactive isotopes of Lu. If the radioconjugate is used for detection purposes, it may contain a radioactive atom for scintigraphic studies, such as Tc 99m or I 123 , or a spin label for nuclear magnetic resonance (NMR) (also known as magnetic resonance imaging, MRI), such as iodine-123, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese, or iron.

[0301] CD20 / CD3 bispecific antibody-cytotoxic agent conjugates can be prepared using a variety of bifunctional protein binding agents, such as N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP), succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), iminothiolane (IT), bifunctional imidoester derivatives (such as dimethyl adipimidate HCl), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis(p-azidobenzoyl)-hexanediamine), bis-diazonium derivatives (such as bis(p-diazonium (benzoyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, a ricin-based immunotoxin can be prepared as described in Vitetta et al., Science 238:1098 (1987).Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugating a radionuclide to an antibody. See WO 94 / 11026. The linker can be a "cleavable linker" that facilitates release of the cytotoxic drug within the cell. For example, an acid-labile linker, a peptidase-sensitive linker, a photolabile linker, a dimethyl linker, or a disulfide-containing linker can be used (Chari et al., Cancer Res. 52:127-131 (1992); U.S. Patent No. 5,208,020).

[0302] In one embodiment, the CD20 / CD3 bispecific antibody is indicated for the treatment of cancer. In one embodiment, the cancer is a B-cell proliferative disorder. In one embodiment, the cancer is a CD20-positive B-cell proliferative disorder. In one embodiment, the cancer is non-Hodgkin's lymphoma (NHL). In one embodiment, the NHL is diffuse large B-cell lymphoma (DLBCL), high-grade B-cell lymphoma (HGBCL), DLBCL arising from FL [transformed FL; trFL], primary mediastinal large B-cell lymphoma (PMBCL), or marginal zone lymphoma (LMZ). LMZ can be classified into splenic, nodal, and extranodal LMZ. In one embodiment, the NHL is mantle cell lymphoma (MCL). In one embodiment, the NHL is follicular lymphoma (FL) grade 1-3a.In one embodiment, the CD20-positive B-cell proliferative disorder is a relapsed or refractory B-cell proliferative disorder. In one embodiment, the relapsed or refractory B-cell proliferative disorder is relapsed or refractory NHL (e.g., relapsed or refractory DLBCL, relapsed or refractory FL, relapsed or refractory MCL).

[0303] In one embodiment, the CD20 / CD3 bispecific antibody specifically binds to CD3ε.

[0304] In one embodiment, the anti-CD20 / CD3 bispecific antibody may compete for binding with the H2C antibody (PCT Publication No. WO 2008 / 119567), the V9 antibody (Rodrigues et al., Int J Cancer Suppl. 7, 45-50 (1992) and U.S. Patent No. 6,054,297), the FN18 antibody (Nooij et al., Eur J Immunol. 19, 981-984 (1986)), the SP34 antibody (Pessano et al., EMBO J. 4, 337-340 (1985)), the OKT3 antibody (Kung et al., Science 206, 347-349 (1979)), the WT31 antibody (Spits et al., J Immunol. 135, 1922 (1985)), UCHT1 antibody (Burns et al., J Immunol. 129, 1451-1457 (1982)), 7D6 antibody (Coulie et al., Eur J Immunol. 21, 1703-1709 (1991)) or Leu-4 antibody.In some embodiments, the CD20 / CD3 bispecific antibody may also include an antigen-binding fragment that specifically binds to CD3, as described in WO 2005 / 040220, WO 2005 / 118635, WO 2007 / 042261, WO 2008 / 119567, WO 2008 / 119565, WO 2012 / 162067, WO 2013 / 158856, WO 2013 / 188693, WO 2013 / 186613, WO 2014 / 110601, WO 2014 / 145806, WO 2014 / 191113, WO 2014 / 047231, WO 2015 / 095392, WO 2015 / 181098, WO 2015 / 001085, WO 2015 / 104346, WO 2015 / 172800, WO 2016 / 020444 or WO 2016 / 014974.

[0305] In some embodiments, the CD20 / CD3 bispecific antibody may comprise an antibody or antigen-binding fragment of rituximab, obinutuzumab, ocrelizumab, ofatumumab, ocaratuzumab, veltuzumab, and ublituximab.

[0306] In one embodiment, the CD20 / CD3 bispecific antibody is XmAb®13676. In one embodiment, the CD20 / CD3 bispecific antibody is REGN1979. In one embodiment, the CD20 / CD3 bispecific antibody is FBTA05 (Lymphomun). In one embodiment, the CD20 / CD3 bispecific antibody is glofitamab.

[0307] In some embodiments, the CD20 / CD3 bispecific antibody may be a generic, biosimilar, or non-comparable biological version of an antibody named herein.

[0308] In one embodiment, the CD20 / CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD20, comprising a heavy chain variable region comprising:

[0309] (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 1;

[0310] (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 2;

[0311] (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 3;

[0312] and a light chain variable region comprising:

[0313] (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 4;

[0314] (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 5, and

[0315] (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 6.

[0316] In one embodiment, the anti-CD20 / CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD20, comprising a heavy chain variable region sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 7, and a light chain variable region sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 8. In another embodiment, the anti-CD20 / CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD20, comprising the heavy chain variable region sequence of SEQ ID NO: 7, and light chain variable region sequence SEQ ID NO: 8.

[0317] In one embodiment, the CD20 / CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD3, comprising a heavy chain variable region comprising:

[0318] (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 9;

[0319] (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 10;

[0320] (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 11; and

[0321] and a light chain variable region comprising:

[0322] (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 12;

[0323] (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 13, and

[0324] (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 14.

[0325] In one embodiment, the anti-CD20 / CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD3, comprising a heavy chain variable region sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 15, and a light chain variable region sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 16. In another embodiment, the anti-CD20 / CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD3, comprising a heavy chain variable region sequence of SEQ ID NO: 15 and the light chain variable region sequence SEQ ID NO: 16.

[0326] In one embodiment, the CD20 / CD3 bispecific antibody comprises:

[0327] a) at least one antigen-binding domain that specifically binds to CD20, comprising a heavy chain variable region comprising:

[0328] (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 1;

[0329] (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 2;

[0330] (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 3;

[0331] and a light chain variable region comprising:

[0332] (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 4;

[0333] (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 5;

[0334] (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 6, and

[0335] b) at least one antigen-binding domain that specifically binds to CD3, comprising a heavy chain variable region comprising:

[0336] (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 9;

[0337] (ii)HVR-H2, comprising the amino acid sequence of SEQ ID NO: 10;

[0338] (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 11; and a light chain variable region comprising:

[0339] (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 12;

[0340] (ii)HVR-L2 comprising the amino acid sequence of SEQ ID NO: 13, and

[0341] (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 14.

[0342] In one embodiment, the CD20 / CD3 bispecific antibody comprises:

[0343] (i) at least one antigen-binding domain that specifically binds to CD20, comprising the heavy chain variable region sequence of SEQ ID NO: 7 and the light chain variable region sequence of SEQ ID NO: 8, and

[0344] (ii)no at least one antigen-binding domain that specifically binds to CD3, comprising the heavy chain variable region sequence of SEQ ID NO: 15 and the light chain variable region sequence of SEQ ID NO: 16.

[0345] In one embodiment, the antigen-binding domain that specifically binds to CD3 of the anti-CD20 / CD3 bispecific antibody is an antibody fragment, in particular a Fab molecule or an scFv molecule, more particularly a Fab molecule. In a specific embodiment, the antigen-binding domain that specifically binds to CD3 of the anti-CD20 / CD3 bispecific antibody is a crossover Fab molecule in which the variable domains or constant domains of the heavy and light chains of the Fab are exchanged (i.e., substituted for each other).

[0346] In one embodiment, the anti-CD20 / CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD20 and one antigen-binding domain that specifically binds to CD3. In one embodiment, the anti-CD20 / CD3 bispecific antibody comprises a first antigen-binding domain that specifically binds to CD3 and second and third antigen-binding domains that specifically bind to CD20. In one embodiment, the first antigen-binding domain is a crossover Fab molecule, and the second and third antigen-binding domains are conventional Fab molecules. In one embodiment, the anti-CD20 / CD3 bispecific antibody further comprises an Fc domain. The anti-CD20 / CD3 bispecific antibody may comprise modifications of the Fc region and / or antigen-binding domains described herein.In one embodiment, the CD20 / CD3 bispecific antibody comprises an IgG1 Fc domain comprising one or more amino acid substitutions that reduce Fc receptor binding and / or effector function. In one embodiment, the CD20 / CD3 bispecific antibody comprises an IgG1 Fc domain comprising the amino acid substitutions L234A, L235A, and P329G (EU numbering).

[0347] In one embodiment, the CD20 / CD3 bispecific antibody comprises:

[0348] (i) an antigen-binding domain that specifically binds to CD3, which is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first subunit of the Fc domain;

[0349] (ii) a first antigen-binding domain that specifically binds to CD20, which is fused at the C-terminus of a Fab heavy chain to the N-terminus of a Fab heavy chain antigen-binding domain that specifically binds to CD3; and

[0350] (iii) a second antigen-binding domain that specifically binds to CD20, which is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second subunit of the Fc domain.

[0351] In one specific embodiment, the CD20 / CD3 bispecific antibody comprises:

[0352] a) a first Fab molecule that specifically binds to CD3, in particular CD3 epsilon; and wherein the variable domains VL and VH of the Fab light chain and the Fab heavy chain are substituted for each other;

[0353] b) a second Fab molecule and a third Fab molecule that specifically bind to CD20, wherein in the CL constant domain of the second Fab molecule and the third Fab molecule, the amino acid at position 124 is substituted with lysine (K) (Kabat numbering), and the amino acid at position 123 is substituted with lysine (K) or arginine (R), in particular arginine (R) (Kabat numbering), and wherein in the CH1 constant domain of the second Fab molecule and the third Fab molecule, the amino acid at position 147 is substituted with glutamic acid (E) (EU numbering), and the amino acid at position 213 is substituted with glutamic acid (E) (EU numbering); and

[0354] c) Fc domain, consisting of the first and second subunits capable of stable association.

[0355] In one embodiment, the CD20 / CD3 bispecific antibody comprises two antigen-binding domains that specifically bind to CD20 and one antigen-binding domain that specifically binds to CD3. In one embodiment, the CD20 / CD3 bispecific antibody is bivalent for CD20 and monovalent for CD3.

[0356] In one embodiment, the first Fab molecule according to a) is fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the Fc domain subunits according to c), the second Fab molecule according to b) is fused at the C-terminus of the Fab heavy chain to the N-terminus of the heavy chain of the first Fab molecule according to a), and the third Fab molecule according to b) is fused at the C-terminus of the Fab heavy chain to the N-terminus of the other Fc domain subunit according to c). In one embodiment, the first Fab molecule according to a) comprises a heavy chain variable region that is at least 95%, 96%, 97%, 98% or 99% identical to the sequence of SEQ ID NO: 15, and a light chain variable region that is at least 95%, 96%, 97%, 98% or 99% identical to the sequence of SEQ ID NO: 16.

[0357] In yet another further embodiment, the first Fab molecule according to a) comprises the heavy chain variable region sequence of SEQ ID NO: 15 and the light chain variable region sequence of SEQ ID NO: 16.

[0358] In one embodiment, the second Fab molecule and the third Fab molecule according to b) comprise a heavy chain variable region that is at least 95%, 96%, 97%, 98% or 99% identical to the sequence of SEQ ID NO: 7, and a light chain variable region that is at least 95%, 96%, 97%, 98% or 99% identical to the sequence of SEQ ID NO: 8.

[0359] In one embodiment, the second Fab molecule and the third Fab molecule according to b) comprise the heavy chain variable region sequence of SEQ ID NO: 7 and the light chain variable region sequence of SEQ ID NO: 8.

[0360] In a specific embodiment, the anti-CD20 / CD3 bispecific antibody comprises a polypeptide that is at least 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 17, a polypeptide that is at least 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 18, a polypeptide that is at least 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 19, and a polypeptide that is at least 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 20. In a further specific embodiment, the bispecific antibody comprises the polypeptide sequence of SEQ ID NO: 17, the polypeptide sequence of SEQ ID NO: 18, the polypeptide sequence of SEQ ID NO: 19 and the polypeptide sequence of SEQ ID NO: 20.In a further specific embodiment, the bispecific antibody comprises one polypeptide chain comprising SEQ ID NO: 17, one polypeptide chain comprising SEQ ID NO: 18, two polypeptide chains comprising SEQ ID NO: 19, and one polypeptide chain comprising SEQ ID NO: 20.

[0361] In particular, the CD20 / CD3 bispecific antibody is described in PCT Publication No. WO 2016 / 020309 and European Patent Application Nos. EP 15188093 and EP 16169160 (each of which is incorporated herein by reference in its entirety).

[0362] Glofitamab

[0363] In one embodiment, the CD20 / CD3 bispecific antibody useful in the methods described herein is glofitamab. Glofitamab (proposed INN: WHO Drug List 121, Vol. 33, No. 2, 2019, p. 276, also known as CD20-TCB, RO7082859, or RG6026; CAS No.: 2229047-91-8) is a novel T-cell-capture bispecific (TCB) full-length antibody with a 2:1 molecular configuration for bivalent binding to CD20 on B cells and monovalent binding to CD3, in particular the epsilon chain of CD3 (CD3e), on T cells. Its CD3-binding region is fused head-to-tail to one of the CD20-binding regions via a flexible linker. This structure provides glofitamab with superiority in vitro compared to other 1:1 CD20-CD3 bispecific antibodies and leads to significant antitumor efficacy in preclinical models of DLBCL.The bivalence of CD20 preserves this potential in the presence of competing anti-CD20 antibodies, enabling pre- or co-therapy with these agents. Glofitamab contains an engineered heterodimeric Fc region with completely abolished binding to FcgRs and C1q. By simultaneously binding to human tumor cells expressing CD20 and to the CD3e T-cell receptor (TCR) complex on T cells, it induces tumor cell lysis, as well as T-cell activation, proliferation, and cytokine release. Glofitamab-mediated B-cell lysis is CD20-specific and does not occur in the absence of CD20 expression or with simultaneous binding (cross-linking) of T cells to CD20-expressing cells.In addition to killing, T cells undergo activation due to CD3 cross-linking, which is manifested by an increase in T-cell activation markers (CD25 and CD69), the release of cytokines (IFNγ, TNFα, IL-2, IL-6, IL-10), cytotoxic granules (granzyme B), and T-cell proliferation. The structural diagram of the glofitamab molecule is shown in Fig. 2. The sequences of glofitamab are summarized in Table 2.

[0364]

[0365]

[0366] B. Anti-CD79b antibody drug conjugates

[0367] Anti-CD79b antibody drug conjugates useful in the methods described herein (e.g., for treating a CD20-positive cell proliferative disorder, such as a B-cell proliferative disorder (e.g., NHL (e.g., relapsed and / or refractory NHL, DLBCL (e.g., relapsed and / or refractory DLBCL), FL (e.g., relapsed and / or refractory FL or transformed FL), or MCL (e.g., relapsed or refractory MCL)), or LCNS) include any of the anti-CD79b antibody drug conjugates described in U.S. Patent No. 8,088,378, which is incorporated herein by reference in its entirety.In some cases, the anti-CD79b antibody drug conjugate comprises a CD79b binding domain comprising at least one, two, three, four, five, or six hypervariable regions (HVRs) selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (e) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26.In some cases, the anti-CD79b antibody drug conjugate comprises a CD79b binding domain comprising all six of the following HVRs: (a) HVR-H1 comprising the amino acid sequence GYTFSSYWIE (SEQ ID NO: 21); (b) HVR-H2 comprising the amino acid sequence GEILPGGGDTNYNEIFKG (SEQ ID NO: 22); (c) HVR-H3 comprising the amino acid sequence TRRVPIRLDY (SEQ ID NO: 23); (d) HVR-L1 comprising the amino acid sequence KASQSVDYEGDSFLN (SEQ ID NO: 24); (e) HVR-L2 comprising the amino acid sequence AASNLES (SEQ ID NO: 25); and (e) HVR-L3 comprising the amino acid sequence QQSNEDPLT (SEQ ID NO: 26).In some cases, the anti-CD79b antibody drug conjugate comprises at least one (e.g., 1, 2, 3, or 4) of the heavy chain regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 29-32, respectively, and / or at least one (e.g., 1, 2, 3, or 4) of the light chain regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 33-36, respectively.In some cases, the anti-CD79b antibody drug conjugate comprises (a) a heavy chain variable domain (VH) comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 27 or that sequence; (b) a light chain variable domain (VL) comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 28 or that sequence; or (c) the VH domain of (a) and the VL domain of (b). Thus, in some cases, the first binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 27 and a VL domain comprising the amino acid sequence of SEQ ID NO: 28.

[0368] In some cases, the anti-CD79b antibody drug conjugate comprises (a) a heavy chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 37 or that sequence; (b) a light chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 38 or that sequence; or (c) the VH domain of (a) and the VL domain of (b). Thus, in some cases, the first binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 37 and a VL domain comprising the amino acid sequence of SEQ ID NO: 38.

[0369] The sequences of the CD79b antibody from polatuzumab vedotin are shown below in Table 3.

[0370]

[0371] In some cases, the CD79b antibody binds to a toxin such as monomethyl auristatin E (MMAE, i.e. vedotin). In some cases, the anti-CD79b antibody drug conjugate is polatuzumab vedotin (immunoglobulin G1-kappa auristatin E conjugate, anti-[Homo sapiens CD79b (immunoglobulin-associated CD79 beta)], humanized monoclonal antibody conjugated to auristatin E; gamma1 heavy chain (1-447) [humanized VH (Homo sapiens IGHV3-23*04 (76.50%)-(IGHD)-IGHJ4*01) [8.8.10] (1-117) -Homo sapiens IGHG1*03 (CH1 R120>K (214)(118-215), loop (216-230), CH2 (231-340), CH3 (341-445), CHS (446-447)) (118-447)], (220-218')-Kappa light chain disulfide (1'-218') [humanized V-KAPPA (Homo sapiens IGKV1 -39*01 (85.90%) -IGKJ1*01) [10.3.9] (l'-111') -Homo sapiens IGKC*01 (112'-218')]; dimer (226-226'':229-229'')-bisdisulfide; conjugated, on average, 3-4 cysteinyls, to monomethyl auristatin E (MMAE) via a cleavable linker of the maleimidocaproyl-valyl-citrullinyl-p-aminobenzyloxycarbonyl (mc-val-cit-PABC) type; also known as RG-7596 or RO5541077-000)), as defined in the List of 110 International Nonproprietary Names (INN) (WHO Drug Formation, Vol. 27, No. 4, 2016, p. 443). Polatuzumab vedotin is also referred to as IUPHAR / BPS number 8404, KEGG number D10761, or CAS registry number 1313206-42-6. Polatuzumab vedotin-piiq is also interchangeably referred to as "polatuzumab vedotin," "huMA79bv28-MC-vc-PAB-MMAE," or "DCDS4501A." In some embodiments, an anti-CD79b antibody (e.g., an anti-CD79b ADC) comprises the heavy chain sequence of SEQ ID NO: 37 and the light chain sequence of SEQ ID NO: 38.

[0372] In some cases, the anti-CD79b antibody drug conjugate has the formula:

[0373]

[0374] wherein Ab is an anti-CD79b antibody comprising (i) a hypervariable region-H1 (HVR-H1) comprising the amino acid sequence of SEQ ID NO: 21; (ii) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (vi) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is from 1 to 8.

[0375] In some embodiments, the antibody drug conjugate comprises an anti-CD79b antibody comprising (a) a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22, and (iii) HVR-H3 comprising an amino acid sequence selected from SEQ ID NO: 23; and (b) a VL domain comprising at least one, at least two, or all three VL HVR sequences selected from (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24, (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 25, and (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26.In some embodiments, the antibody drug immunoconjugate comprises an anti-CD79b antibody that comprises at least one of: (i) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23 and / or (ii) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24. In some embodiments, the antibody drug conjugate comprises an anti-CD79b antibody that comprises (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (d) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (e) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (e) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26.

[0376] In some embodiments, the antibody drug conjugate comprises at least one of: an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23 and / or an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24. In some embodiments, the antibody drug conjugate comprises an anti-CD79b antibody that comprises (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (d) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (e) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (e) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26.

[0377] In some embodiments, the anti-CD79b antibody drug conjugate comprises a humanized anti-CD79b antibody. In some embodiments, the anti-CD79b antibody comprises an HVR according to any of the embodiments provided herein and further comprises a human acceptor framework, such as a human immunoglobulin framework or a human consensus framework. In some embodiments, the human acceptor framework is a VL kappa 1 (VLK1) framework and / or a VH VHIII framework.In some embodiments, a humanized anti-CD79b antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (e) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26.In some embodiments, a humanized anti-CD79b antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (e) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26.

[0378] In some embodiments, an antibody drug conjugate (e.g., an anti-CD79b antibody drug conjugate) comprises an anti-CD79 antibody that comprises a heavy chain variable domain (VH) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 27. In some embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 27 comprises substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an anti-CD79b antibody comprising this sequence retains the ability to bind to CD79b. In some embodiments, a total of 1-10 amino acids were substituted, inserted, and / or deleted in SEQ ID NO: 27.In some embodiments, a total of 1-5 amino acids were substituted, inserted, and / or deleted in SEQ ID NO: 27. In some embodiments, the substitutions, insertions, or deletions occur in regions outside of the HVRs (i.e., in the FRs, e.g., SEQ ID NOs: 29-32). In some embodiments, an antibody drug conjugate (e.g., an anti-CD79b antibody conjugate) comprises the VH sequence of SEQ ID NO: 27, including post-translational modifications of that sequence. In some embodiments, the VH comprises one, two, or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23.

[0379] In some embodiments, an antibody drug conjugate (e.g., an anti-CD79b antibody drug conjugate) comprises an anti-CD79b antibody that comprises a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 28. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 28 comprises substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but the CD79b antibody drug conjugate comprising this sequence retains the ability to bind to CD79b. In certain embodiments, a total of 1-10 amino acids were substituted, inserted, and / or deleted in SEQ ID NO: 28.In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deletion in SEQ ID NO: 28. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside of the HVRs (i.e., in the FRs, e.g., SEQ ID NOs: 33-36). In some embodiments, the anti-CD79b antibody drug conjugate comprises an anti-CD79b antibody that comprises the VL sequence of SEQ ID NO: 28, including post-translational modifications of that sequence. In some embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26.In some embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26.

[0380] In some embodiments, an antibody drug conjugate (e.g., an anti-CD79b antibody drug conjugate) comprises an anti-CD79b antibody that comprises a VH as in any of the embodiments provided herein and a VL as in any of the embodiments provided herein. In some embodiments, the antibody drug conjugate comprises an anti-CD79b antibody that comprises the VH and VL sequences of SEQ ID NO: 27 and SEQ ID NO: 28, respectively, including post-translational modifications of these sequences.

[0381] In some embodiments, an antibody drug conjugate (e.g., an anti-CD79b antibody drug conjugate) comprises an anti-CD79b antibody that binds to the same epitope as the anti-CD79b antibody described herein. For example, in some embodiments, an antibody drug conjugate (e.g., an anti-CD79b antibody drug conjugate) comprises an anti-CD79b antibody that binds to the same epitope as the anti-CD79b antibody, comprising the VH sequence of SEQ ID NO: 27 and the VL sequence of SEQ ID NO: 28.

[0382] For example, in some embodiments, an antibody drug conjugate comprises an anti-CD79b antibody that is a monoclonal antibody, a chimeric antibody, a humanized antibody, or a human antibody. For example, in some embodiments, the antibody drug conjugate comprises an antigen-binding fragment of an anti-CD79b antibody described herein, such as an Fv, Fab, Fab', scFv, diabody, or F(ab')2 fragment. For example, in some embodiments, the antibody drug conjugate comprises substantially a full-length anti-CD79b antibody, such as an IgG1 antibody or another antibody class or isotype, as described elsewhere herein. Anti-CD79b antibody drug conjugates can be produced using recombinant methods and compositions, such as described in U.S. Patent No. 4,816,567.

[0383] In some cases, the anti-CD79b antibody drug conjugates of any of the embodiments may include any feature, alone or in combination, described below.

[0384] B. Antibody formats

[0385] 1. Bispecific antibody to CD20 / CD3

[0386] The components of the CD20 / CD3 bispecific antibody can be fused to each other in various configurations. Typical configurations are illustrated in Figure 1.

[0387] In specific embodiments, the antigen-binding fragments contained in the CD20 / CD3 bispecific antibody are Fab molecules. In such embodiments, the first, second, third, etc. antigen-binding fragment may be referred to herein as the first, second, third, etc. Fab molecule, respectively. Furthermore, in specific embodiments, the CD20 / CD3 bispecific antibody comprises an Fc domain consisting of first and second subunits capable of stable association.

[0388] In some embodiments, the first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first or second Fc domain subunit.

[0389] In one such embodiment, the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first Fab molecule. In a particular such embodiment, the bispecific antibody to CD20 / CD3 consists essentially of first and second Fab molecules, an Fc domain consisting of first and second subunits, and, optionally, one or more peptide linkers, wherein the first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first or second subunit of the Fc domain, and the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first Fab molecule. Such a configuration is schematically depicted in Figures 1G and 1M. Optionally, the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule can be further fused to each other.

[0390] In another embodiment, the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first or second subunit of the Fc domain. In such a specific embodiment, the antibody consists essentially of first and second Fab molecules, an Fc domain consisting of the first and second subunits, and, optionally, one or more peptide linkers, wherein each of the first and second Fab molecules is fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain. Such a configuration is schematically depicted in Figures 1A and 1D. The first and second Fab molecules can be fused to the Fc domain directly or via a peptide linker. In a specific embodiment, each of the first and second Fab molecules is fused to the Fc domain via an immunoglobulin hinge region. In a specific embodiment, the hinge region of the immunoglobulin is a human IgG1 hinge region, particularly when the Fc domain is an IgG1 Fc domain.

[0391] In other embodiments, the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first or second Fc domain subunit. In one such embodiment, the first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab molecule. In such a specific embodiment, the antibody consists essentially of the first and second Fab molecules, an Fc domain consisting of the first and second subunits, and, optionally, one or more peptide linkers, wherein the first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab molecule, and the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first or second Fc domain subunit. Such a configuration is schematically depicted in Figures 1I and 1H. Optionally, the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule may be further fused to each other.

[0392] Fab molecules can be fused to the Fc domain or to each other directly or via a peptide linker containing one or more amino acids, typically about 2-20 amino acids. Peptide linkers are known in the art and are described herein. Suitable non-immunogenic peptide linkers include, for example, peptide linkers (G4S) n , (SG4) n , (G4S) n or G4(SG4) n "n" is generally an integer from 1 to 10, typically from 2 to 4. In one embodiment, the length of said peptide linker is at least 5 amino acids, in one embodiment from 5 to 100, in a further embodiment from 10 to 50 amino acids. In one embodiment, said peptide linker is (GxS) n or (GxS) n G m, where G=glycine, S=serine, and (x=3, n=3, 4, 5, or 6, and m=0, 1, 2, or 3) or (x=4, n=2, 3, 4, or 5, and m=0, 1, 2, or 3), in one embodiment x=4 and n=2 or 3, in a further embodiment x=4 and n=2. In one embodiment, said peptide linker is (G4S)2. A particularly suitable peptide linker for fusing the Fab light chains of the first and second Fab molecules to each other is (G4S)2. An exemplary peptide linker suitable for joining the Fab heavy chains of the first and second Fab fragments comprises the sequence (D)-(G4S)2. Another such suitable linker comprises the sequence (G4S)4. In addition, the linkers may comprise an immunoglobulin hinge region (or a portion thereof). In particular, if a Fab molecule is fused to the N-terminus of an Fc domain subunit, it may be fused via the immunoglobulin hinge region or a portion thereof, with or without an additional peptide linker.

[0393] An antibody with a single antigen-binding fragment (such as a Fab molecule) capable of specifically binding to a target cell antigen (e.g., as shown in Fig. 1A, 1D, 1G, 1I, 1M, or 1H) is particularly useful in cases where internalization of the target cell antigen can be expected after binding to a high-affinity antigen-binding fragment. In such cases, the presence of more than one antigen-binding fragment specific for the target cell antigen may enhance the internalization of the target cell antigen, thereby reducing its availability.

[0394] In many other cases, it is advantageous for an antibody to comprise two or more antigen-binding fragments (such as Fab molecules) specific for a target cell antigen (see examples illustrated in Figures 1B, 1C, 1E, 1F, 1K, 1K, 1P, or 1P), for example, to optimize targeting to a target site or to allow cross-linking of target cell antigens.

[0395] Accordingly, in certain embodiments, the CD20 / CD3 bispecific antibody comprises two CD20-binding moieties, such as two Fab molecules targeting CD20. In one embodiment, the two Fab molecules targeting CD20 are conventional Fab molecules. In one embodiment, the two Fab molecules targeting CD20 contain the same amino acid sequences of the heavy and light chains and have the same domain arrangement (i.e., conventional or crossover).

[0396] In alternative embodiments, the CD20 / CD3 bispecific antibody comprises two CD3-binding moieties, such as two Fab molecules targeting CD3. In one embodiment, the two Fab molecules targeting CD3 are crossover Fab molecules (a Fab molecule in which the variable domains VH and VL or the constant domains CL and CH1 of the heavy and light chains of the Fab are substituted for each other). In one such embodiment, the two Fab molecules targeting CD3 comprise the same amino acid sequences of the heavy and light chains and have the same domain arrangement (i.e., conventional or crossover).

[0397] In one embodiment, the third Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first or second subunit of the Fc domain.

[0398] In a specific embodiment, each of the second and third Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the Fc domain subunits, and the first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab molecule. In such a specific embodiment, the antibody consists essentially of the first, second and third Fab molecules, an Fc domain consisting of the first and second subunits, and, optionally, one or more peptide linkers, wherein the first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab molecule, the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first Fc domain subunit, and the third Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second Fc domain subunit.Such a configuration is schematically depicted in Figure 1B and Figure 1D (embodiments in which the third Fab molecule is a conventional Fab molecule and is identical to the second Fab molecule), as well as in Figure 1K and Figure 1P (embodiments in which the third Fab molecule is a crossover Fab molecule and is preferably identical to the first Fab molecule). The second and third Fab molecules can be fused to the Fc domain directly or via a peptide linker. In a specific embodiment, each of the second and third Fab molecules is fused to the Fc domain via an immunoglobulin hinge region. In a specific embodiment, the immunoglobulin hinge region is a human IgG1 hinge region, in particular when the Fc domain is an IgG1 Fc domain. Optionally, the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule can be further fused to each other.

[0399] In another embodiment, each of the second and third Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the Fc domain subunits, and the first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab molecule. In such a specific embodiment, the antibody consists essentially of the first, second and third Fab molecules, an Fc domain consisting of the first and second subunits, and, optionally, one or more peptide linkers, wherein the first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab molecule, the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first Fc domain subunit, and the third Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second Fc domain subunit.Such a configuration is schematically depicted in Figure 1B and Figure 1E (embodiments in which the third Fab molecule is a conventional Fab molecule and is identical to the second Fab molecule), as well as in Figure 1L and Figure 1P (embodiments in which the third Fab molecule is a crossover Fab molecule and is identical to the first Fab molecule). The first and third Fab molecules can be fused to the Fc domain directly or via a peptide linker. In a specific embodiment, each of the second and third Fab molecules is fused to the Fc domain via an immunoglobulin hinge region. In a specific embodiment, the immunoglobulin hinge region is a human IgG1 hinge region, in particular when the Fc domain is an IgG1 Fc domain. Optionally, the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule can be further fused to each other.

[0400] In antibody configurations in which a Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of each of the Fc domain subunits via the immunoglobulin hinge region, two Fab molecules, the hinge regions, and the Fc domain predominantly form an immunoglobulin molecule. In a specific embodiment, the immunoglobulin molecule is an IgG class immunoglobulin. In an even more specific embodiment, the immunoglobulin is an IgG1 subclass immunoglobulin. In another embodiment, the immunoglobulin is an IgG4 subclass immunoglobulin. In a further specific embodiment, the immunoglobulin is a human immunoglobulin. In other embodiments, the immunoglobulin is a chimeric immunoglobulin or a humanized immunoglobulin.

[0401] In some of the antibodies, the Fab chain of a first Fab molecule and the Fab light chain of a second Fab molecule are fused to each other, optionally via a peptide linker. Depending on the configuration of the first and second Fab molecules, the Fab light chain of the first molecule may be fused at the C-terminus to the Fab light chain of the second Fab molecule, or the Fab light chain of the second molecule may be fused at the C-terminus to the Fab light chain of the first Fab molecule. Fusion of the Fab light chains of the first and second Fab molecules further reduces mispairing of mismatched Fab heavy and light chains and also reduces the number of plasmids required for expression of some of the antibodies.

[0402] In certain embodiments, the antibody comprises a polypeptide in which the Fab light chain variable region of a first Fab molecule shares a carboxy-terminal peptide bond with a Fab heavy chain constant region of the first Fab molecule (i.e., the first Fab molecule comprises a crossover Fab heavy chain in which the heavy chain variable region is replaced by a light chain variable region), which in turn shares a carboxy-terminal peptide bond with an Fc domain subunit (VL (1) -CH1 (1) -CH2-CH3(-CH4)), and a polypeptide in which the Fab heavy chain of the second Fab molecule shares a common carboxy-terminal peptide bond with a subunit of the Fc domain (VH (2) -CH1 (2) -CH2-CH3(-CH4)). In some embodiments, the antibody further comprises a polypeptide in which the Fab heavy chain variable region of the first Fab molecule shares a common carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule (VH (1) -CL(1) ), and the Fab light chain polypeptide of the second Fab molecule (VL (2) -CL (2) ). In certain embodiments, the polypeptides are covalently linked, such as by a disulfide bond.

[0403] In certain embodiments, the antibody comprises a polypeptide in which the Fab heavy chain variable region of a first Fab molecule shares a carboxy-terminal peptide bond with a Fab light chain constant region of the first Fab molecule (i.e., the first Fab molecule comprises a crossover Fab heavy chain in which the heavy chain constant region is replaced by a light chain constant region), which in turn shares a carboxy-terminal peptide bond with an Fc domain subunit (VH (1) -CL (1) -CH2-CH3(-CH4)), and a polypeptide in which the Fab heavy chain of the second Fab molecule shares a common carboxy-terminal peptide bond with a subunit of the Fc domain (VH (2) -CH1 (2)-CH2-CH3(-CH4)). In some embodiments, the antibody further comprises a polypeptide in which the variable region of the Fab light chain of the first Fab molecule shares a common carboxy-terminal peptide bond with the constant region of the Fab heavy chain of the first Fab molecule (VL (1) -CH1 (1) ), and the Fab light chain polypeptide of the second Fab molecule (VL (2) -CL (2) ). In certain embodiments, the polypeptides are covalently linked, such as by a disulfide bond.

[0404] In some embodiments, the antibody comprises a polypeptide in which the variable region of a light chain of a first Fab molecule shares a carboxy-terminal peptide bond with the constant region of a heavy chain of a Fab of the first Fab molecule (i.e., the first Fab molecule comprises a crossover heavy chain of a Fab in which the variable region of the heavy chain is replaced by a variable region of the light chain), which in turn shares a carboxy-terminal peptide bond with the heavy chain of a second Fab molecule, which in turn shares a carboxy-terminal peptide bond with an Fc domain subunit (VL (1) -CH1 (1) -VH (2) -CH1 (2)-CH2-CH3(-CH4)). In other embodiments, the antibody comprises a polypeptide in which the Fab heavy chain of a second Fab molecule shares a carboxy-terminal peptide bond with the variable region of the Fab light chain of a first Fab molecule, which in turn shares a carboxy-terminal peptide bond with the constant region of the Fab heavy chain of the first Fab molecule (i.e., the first Fab molecule comprises a crossover Fab heavy chain in which the variable region of the heavy chain is replaced by the variable region of the light chain), which in turn shares a carboxy-terminal peptide bond with an Fc domain subunit (VH (2) -CH1 (2) -VL (1) -CH1 (1) -CH2-СН3(-СН4)).

[0405] In some of these embodiments, the antibody further comprises a Fab crossover light chain polypeptide of the first Fab molecule, wherein the Fab heavy chain variable region of the first Fab molecule shares a common carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule (VH (1) -CL (1) ), and the Fab light chain polypeptide of the second Fab molecule (VL (2) -CL (2) ). In other of these embodiments, the antibody further comprises a polypeptide in which the Fab heavy chain variable region of the first Fab molecule shares a common carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule, which in turn shares a common carboxy-terminal peptide bond with the Fab light chain polypeptide of a second Fab molecule (VH (1) -CL (1) -VL (2) -CL (2)), or a polypeptide in which the Fab light chain polypeptide of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain variable region of the first Fab molecule, which in turn shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule (VL (2) -CL (2) -VH (1) -CL (1) ), depending on the situation.

[0406] The antibody according to these embodiments may further comprise (i) an Fc domain subunit polypeptide (CH2-CH3(-CH4)) or (ii) a polypeptide in which the Fab heavy chain of a third Fab molecule shares a carboxy-terminal peptide bond with an Fc domain subunit (VH (3) -CH1 (3) -CH2-CH3(-CH4)), and the Fab light chain polypeptide of the third Fab molecule (VL (3) -CL (3) ). In certain embodiments, the polypeptides are covalently linked, such as by a disulfide bond.

[0407] In some embodiments, the antibody comprises a polypeptide in which the variable region of a Fab heavy chain of a second Fab molecule shares a carboxy-terminal peptide bond with a constant region of a Fab light chain of a first Fab molecule (i.e., the first Fab molecule comprises a crossover Fab heavy chain in which the constant region of the heavy chain is replaced by a constant region of the light chain), which in turn shares a carboxy-terminal peptide bond with a Fab heavy chain of a second Fab molecule, which in turn shares a carboxy-terminal peptide bond with a subunit of the Fc domain (VH (1) -CL (1) -VH (2) -CH1 (2)-CH2-CH3(-CH4)). In other embodiments, the antibody comprises a polypeptide in which the Fab heavy chain of a second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain variable region of a first Fab molecule, which in turn shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule (i.e., the first Fab molecule comprises a crossover Fab heavy chain in which the heavy chain constant region is replaced by the light chain constant region), which in turn shares a carboxy-terminal peptide bond with an Fc domain subunit (VH (2) -CH1 (2) -VH (1) -CL (1) -CH2-CH3(-CH4)).

[0408] In some of these embodiments, the antibody further comprises a Fab crossover light chain polypeptide of the first Fab molecule, wherein the Fab light chain variable region of the first Fab molecule shares a common carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule (VL (1) -CH1 (1) ), and the Fab light chain polypeptide of the second Fab molecule (VL (2) -CL (2) ). In other embodiments, the antibody further comprises a polypeptide in which the Fab light chain variable region of the first Fab molecule shares a common carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule, which in turn shares a common carboxy-terminal peptide bond with the Fab light chain polypeptide of a second Fab molecule (VL (1) -CH1 (1) -VL (2) -CL (2)), or a polypeptide in which the Fab light chain polypeptide of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain variable region of the first Fab molecule, which in turn shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule (VL (2) -CL (2) -VH (1) -CL (1) ), depending on the situation.

[0409] The antibody according to these embodiments may further comprise (i) an Fc domain subunit polypeptide (CH2-CH3-CH4)) or (ii) a polypeptide in which the Fab heavy chain of a third Fab molecule shares a carboxy-terminal peptide bond with an Fc domain subunit (VH (3) -CH1 (3) -CH2-CH3(-CH4)), and the Fab light chain polypeptide of the third Fab molecule (VL (3) -CL (3) ). In certain embodiments, the polypeptides are covalently linked, such as by a disulfide bond.

[0410] In certain embodiments, the antibody comprises a polypeptide in which the Fab heavy chain of a first Fab molecule shares a carboxy-terminal peptide bond with the variable region of a Fab light chain of a second Fab molecule, which in turn shares a carboxy-terminal peptide bond with the constant region of the Fab heavy chain of the second Fab molecule (i.e., the second Fab molecule comprises a crossover Fab heavy chain in which the variable region of the heavy chain is replaced by the variable region of the light chain) (VH (1) -CH1 (1) -VL (2) -CH1 (2) ). In some embodiments, the antibody further comprises a polypeptide in which the Fab heavy chain variable region of the second Fab molecule shares a common carboxy-terminal peptide bond with the Fab light chain constant region of the second Fab molecule (VH (2) -CL (2) ), and the Fab light chain polypeptide of the first Fab molecule (VL (1) -CL (1) ).

[0411] In certain embodiments, the antibody comprises a polypeptide in which the variable region of a light chain of a second Fab molecule shares a carboxy-terminal peptide bond with the constant region of a heavy chain of a Fab of the second Fab molecule (i.e., the second Fab molecule comprises a crossover heavy chain Fab in which the variable region of the heavy chain is replaced by a variable region of the light chain), which in turn shares a carboxy-terminal peptide bond with the heavy chain of a first Fab molecule (VL (2) -CH1 (2) -VH (1) -CH1 (1) ). In some embodiments, the antibody further comprises a polypeptide in which the Fab heavy chain variable region of the second Fab molecule shares a common carboxy-terminal peptide bond with the Fab light chain constant region of the second Fab molecule (VH (2) -CL (2) ), and the Fab light chain polypeptide of the first Fab molecule (VL (1) -CL (1) ).

[0412] In certain embodiments, the antibody comprises a polypeptide in which the variable region of a Fab heavy chain of a second Fab molecule shares a carboxy-terminal peptide bond with a constant region of a Fab light chain of the second Fab molecule (i.e., the second Fab molecule comprises a crossover Fab heavy chain in which the constant region of the heavy chain is replaced by a constant region of the light chain), which in turn shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule (VH (2) -CL (2) -VH (1) -CH1 (1) ). In some embodiments, the antibody further comprises a polypeptide in which the variable region of the Fab light chain of the second Fab molecule shares a common carboxy-terminal peptide bond with the constant region of the Fab heavy chain of the second Fab molecule (VL (2) -CH1 (2) ), and the Fab light chain polypeptide of the first Fab molecule (VL (1) -CL (1) ).

[0413] In certain embodiments, the antibody comprises a polypeptide in which the Fab heavy chain of a third Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of a first Fab molecule, which in turn shares a carboxy-terminal peptide bond with the variable region of the Fab light chain of a second Fab molecule, which in turn shares a carboxy-terminal peptide bond with the constant region of the Fab heavy chain of the second Fab molecule (i.e., the second Fab molecule comprises a crossover Fab heavy chain in which the variable region of the heavy chain is replaced by the variable region of the light chain) (VH (3) -CH1 (3) -VH (1) -CH1 (1) -VL (2) -CH1 (2) ). In some embodiments, the antibody further comprises a polypeptide in which the Fab heavy chain variable region of the second Fab molecule shares a common carboxy-terminal peptide bond with the Fab light chain constant region of the second Fab molecule (VH (2) -CL(2) ), and the Fab light chain polypeptide of the first Fab molecule (VL (1) -CL (1) ). In some embodiments, the antibody further comprises a Fab light chain polypeptide of a third Fab molecule (VL (3) -CL (3) ).

[0414] In certain embodiments, the antibody comprises a polypeptide in which the Fab heavy chain of a third Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of a first Fab molecule, which in turn shares a carboxy-terminal peptide bond with the variable region of the Fab heavy chain of a second Fab molecule, which in turn shares a carboxy-terminal peptide bond with the constant region of the Fab light chain of the second Fab molecule (i.e., the second Fab molecule comprises a crossover Fab heavy chain in which the constant region of the heavy chain is replaced by the constant region of the light chain) (VH (3) -CH1 (3) -VH (1) -CH1 (1) -VH (2) -CL (2)). In some embodiments, the antibody further comprises a polypeptide in which the variable region of the Fab light chain of the second Fab molecule shares a common carboxy-terminal peptide bond with the constant region of the Fab heavy chain of the second Fab molecule (VL (2) -СН1 (2) ), and the Fab light chain polypeptide of the first Fab molecule (VL (1) -CL (1) ). In some embodiments, the antibody further comprises a Fab light chain polypeptide of a third Fab molecule (VL (3) -CL (3) ).

[0415] In certain embodiments, the antibody comprises a polypeptide in which the variable region of a light chain of a second Fab molecule shares a carboxy-terminal peptide bond with the constant region of a heavy chain of a second Fab molecule (i.e., the second Fab molecule comprises a crossover heavy chain of a Fab in which the variable region of the heavy chain is replaced by a variable region of the light chain), which in turn shares a carboxy-terminal peptide bond with the heavy chain of a first Fab molecule, which in turn shares a carboxy-terminal peptide bond with the heavy chain of a third Fab molecule (VL (2) -CH1 (2) -VH (1) -CH1 (1) -VH (3) -CH1 (3) ). In some embodiments, the antibody further comprises a polypeptide in which the Fab heavy chain variable region of the second Fab molecule shares a common carboxy-terminal peptide bond with the Fab light chain constant region of the second Fab molecule (VH (2) -CL(2) ), and the Fab light chain polypeptide of the first Fab molecule (VL (1) -CL (1) ). In some embodiments, the antibody further comprises a Fab light chain polypeptide of a third Fab molecule (VL (3) -CL (3) ).

[0416] In certain embodiments, the antibody comprises a polypeptide in which the variable region of a Fab heavy chain of a second Fab molecule shares a carboxy-terminal peptide bond with a constant region of a Fab light chain of the second Fab molecule (i.e., the second Fab molecule comprises a crossover Fab heavy chain in which the constant region of the heavy chain is replaced by a constant region of the light chain), which in turn shares a carboxy-terminal peptide bond with a Fab heavy chain of a first Fab molecule, which in turn shares a carboxy-terminal peptide bond with a Fab heavy chain of a third Fab molecule (VH (2) -CL (2) -VH (1) -CH1 (1) -VH (3) -CH1 (3)). In some embodiments, the antibody further comprises a polypeptide in which the variable region of the Fab light chain of the second Fab molecule shares a common carboxy-terminal peptide bond with the constant r...

Claims

1. The use of polatuzumab and glofitamab for the treatment of a subject or population of subjects having a CD20-positive cell proliferative disorder, wherein polatuzumab and glofitamab are administered in a dosing regimen comprising a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle consists of a first dose (C1D1) of 2.5 mg and a second dose (C1D2) of 10 mg glofitamab, and a single dose of 1.8 mg / kg polatuzumab; and (b) the second dosing cycle consists of a single dose (C2D1) of 30 mg glofitamab and a single dose of 1.8 mg / kg polatuzumab, where C1D1 of polatuzumab is intended for administration on day 2 of the first dosing cycle, where C1D1 and C1D2 of glofitamab are intended to be administered to the subject on days 8 and 15 of the first dosing cycle, respectively, where C2D1 glofitamab is intended to be administered to the subject on day 1 of the second dosing cycle, where C2D1 glofitamab is intended to be administered after completion of C2D1 polatuzumab administration, and where dosing cycles represent 21-day dosing cycles.

2. Use according to claim 1, wherein the dosing regimen includes from six to ten additional dosing cycles.

3. The use according to claim 2, wherein the one or more additional dosing cycles comprise an additional single dose of 30 mg glofitamab and an additional single dose of 1.8 mg / kg polatuzumab.

4. The use according to claim 2 or 3, wherein the additional single dose of polatuzumab is intended to be administered to the subject on day 1 of each additional dosing cycle that includes the additional dose of polatuzumab.

5. The use according to any one of claims 2-4, wherein the additional single dose of glofitamab of each additional dosing cycle including the additional dose of polatuzumab is intended to be administered after completion of the additional single dose of polatuzumab.

6. The use according to any one of claims 2 to 5, wherein the dosing regimen comprises ten additional dosing cycles, wherein each of the ten additional dosing cycles comprises a single dose of 30 mg glofitamab, and wherein four of the ten additional dosing cycles comprise administration of a single dose of 1.8 mg / kg polatuzumab.

7. The use according to any one of claims 1-6, wherein polatuzumab and glofitamab are administered with one or more additional therapeutic agents selected from one or more chemotherapeutic agents, tocilizumab, a corticosteroid, an antihistamine, allopurinol, rasburicase, an antipyretic agent, and obinutuzumab.

8. The use according to claim 7, wherein the corticosteroid comprises prednisone, prednisolone, methylprednisolone and dexamethasone.

9. Use according to item 7 or 8, wherein polatuzumab and glofitamab are administered with rituximab, cyclophosphamide, doxorubicin and prednisone (R-CHP).

10. The use according to any of paragraphs 7-9, wherein obinutuzumab is intended to be administered before the administration of glofitamab.

11. The use according to claim 10, wherein obinutuzumab is intended to be administered seven days before the administration of glofitamab.

12. Use according to claim 10 or 11, wherein obinutuzumab is intended to be administered as a single dose of 1000 mg.

13. The use according to any of the preceding claims, wherein the B-cell proliferative disorder is non-Hodgkin's lymphoma (NHL) or central nervous system lymphoma (CNS).

14. The use according to claim 13, wherein the B-cell proliferative disorder is recurrent and / or refractory.

15. The use according to claim 13 or 14, wherein the NHL is diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), high-grade B-cell lymphoma, primary mediastinal (thymic) large B-cell lymphoma (PMLBCL), diffuse B-cell lymphoma, or small lymphocytic lymphoma.

16. The use according to any one of claims 1-15, wherein the subject or population of subjects has received at least two prior systemic therapies.

17. The use according to any one of claims 1-16, wherein the subject or population of subjects does not meet the requirements for autologous stem cell transplantation (ASCT).

18. The use according to any one of claims 1-17, wherein the complete response rate in the subject population is at least 20%, or wherein the overall response rate in the subject population is at least 30%.

Images