Administration of combination therapy using an anti-CD20 / anti-CD3 bispecific antibody and an anti-CD79B antibody drug conjugate.

Abstract

The present invention relates to the treatment of a subject having a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder, such as non-Hodgkin's lymphoma). More specifically, the present invention relates to the treatment of a subject having a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder) by administering a combination of an anti-CD20 / anti-CD3 bispecific antibody and an anti-CD79b antibody drug conjugate.

Description

[Technical Field]

[0001] Sequence List This application includes a sequence listing submitted electronically in ASCII format (the entire listing is incorporated herein by reference). The ASCII copy, created on 26 April 2021, is named 51177-033001_Sequence_Listing_4.26.21_ST25 and has a size of 32,706 bytes.

[0002] The present invention relates to a method for treating diseases, particularly B-cell proliferative disorders, by administering an anti-CD20 / anti-CD3 bispecific antibody and anti-CD79b antibody drug conjugate, and to a method for reducing adverse effects of administering an anti-CD20 / anti-CD3 bispecific antibody and anti-CD79b antibody drug conjugate. [Background technology]

[0003] B-cell proliferative disorders are a heterogeneous group of malignancies that include both leukemia and lymphoma. Lymphomas originate from lymphoid cells and are classified into two main categories: Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL). In the United States, B-cell lymphomas account for approximately 80–85% of all non-Hodgkin lymphoma cases, and within the B-cell subset, there is considerable heterogeneity based on B-cell genotype and phenotypic expression patterns. For example, the B-cell lymphoma subset includes slow-growing, painless, and refractory diseases such as follicular lymphoma (FL) or chronic lymphocytic leukemia (CLL), as well as more aggressive subtypes, 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 NHL diagnoses, followed by follicular lymphoma (FL; 20-25% of all NHL diagnoses) and mantle cell lymphoma (MCL; 6-10% of all NHL diagnoses). B-cell chronic lymphocytic leukemia (CLL) is the most common type of leukemia in adults, with approximately 15,000 new cases diagnosed annually in the United States (American Cancer Society 2015).

[0004] Bispecific antibodies can simultaneously bind cell surface antigens of cytotoxic cells (e.g., T cells; by binding to differentiation antigen group 3 (CD3)) and cancer cells (e.g., B cells; by binding to CD20), and are intended to destroy the bound cancer cells by the bound cytotoxic cells. Glofitamab is a T cell-induced bispecific (TCB) antibody that targets CD20 expressed on B cells and the CD3 epsilon chain (CD3e) present on T cells.

[0005] However, immunotherapy with anti-D20 / anti-CD3 bispecific antibodies such as grofitamab can be limited by undesirable side effects such as cytokine toxicity (e.g., cytokine release syndrome (CRS)), fluid-related reactions (IRRs), severe tumor lysis syndrome (TLS), and central nervous system (CNS) toxicity.

[0006] Therefore, there is an unmet need in the field of treatment for CD20-positive B-cell proliferative disorders (e.g., non-Hodgkin lymphoma, NHL) for the development of effective administration methods for anti-CD20 / anti-CD3 bispecific antibodies (e.g., grofitamab) that achieve a better benefit-risk profile. [Overview of the project]

[0007] In one embodiment, the present invention is characterized by a method for treating a subject having a CD20-positive cell proliferative disorder, comprising administering to the subject having a CD20-positive cell proliferative disorder 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) and a second dose (C1D2) of the bispecific antibody, with C1D1 being approximately 2.5 mg and C1D2 being approximately 10 mg; and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody, with C2D1 being approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0008] In another embodiment, the present invention features an anti-CD79b antibody drug conjugate and a bispecific antibody conjugated to CD20 and CD3 for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugated to CD20 and bCD3 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) and a second dose (C1D2) of the bispecific antibody, with C1D1 being approximately 2.5 mg and C1D2 being approximately 10 mg; and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody, with C2D1 being approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0009] In another embodiment, the present invention is characterized by the use of an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating to CD20 and CD3 in the treatment of a subject having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugating 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) and a second dose (C1D2) of the bispecific antibody: (a) the first dosing cycle comprises a first dose (C1D1) and a second dose (C1D2) of the bispecific antibody, where C1D1 is approximately 2.5 mg and C1D2 is approximately 10 mg; (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody, where C2D1 is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0010] In another embodiment, the present invention is characterized by the use of an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating to CD20 and CD3 in the manufacture of a pharmacopoeia for the treatment of subjects having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugating to CD20 and CD3 are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) the first dosing cycle comprises a first dose (C1D1) and a second dose (C1D2) of the bispecific antibody, where C1D1 is approximately 2.5 mg and C1D2 is approximately 10 mg; (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody, where C2D1 is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0011] 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.

[0012] In some embodiments, the first administration cycle includes a single dose C1D1 of the anti-CD79b antibody drug conjugate. In some embodiments, the single dose C1D1 of the anti-CD79b antibody drug conjugate ranges from about 0.1 mg / kg to about 2.4 mg / kg (e.g., about 0.1 mg / kg to about 2.2 mg / kg, about 0.1 mg / kg to about 2.0 mg / kg, about 0.5 mg / kg to about 2.2 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.2 mg / kg, about 1.2 mg / kg to about The dosages are approximately 2.2 mg / kg, 1.4 mg / kg to 2.2 mg / kg, 1.6 mg / kg to 2.2 mg / kg, 1.8 mg / kg to 2.0 mg / kg, 0.1 mg / kg to 1.6 mg / kg, 0.5 mg / kg to 1.6 mg / kg, or 1 mg / kg to 1.8 mg / kg (for example, 1 mg / kg, 1.2 mg / kg, 1.6 mg / kg, or 1.8 mg / kg). In some embodiments, a single dose C1D1 of the anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg. In some embodiments, C1D1 of the anti-CD79b antibody drug conjugate is administered on or around day 2 of the administration cycle (±1 day). In some embodiments, a second administration cycle includes a single dose C2D1 of the anti-CD79b antibody drug conjugate. In some embodiments, a single dose C2D1 of the anti-CD79b antibody drug conjugate ranges from approximately 0.1 mg / kg to approximately 2.4 mg / kg (e.g., approximately 0.1 mg / kg to approximately 2.2 mg / kg, approximately 0.1 mg / kg to approximately 2.0 mg / kg, approximately 0.5 mg / kg to approximately 2.2 mg / kg, approximately 0.8 mg / kg to approximately 2.2 mg / kg, approximately 1 mg / kg to approximately 2.2 mg / kg, approximately 1.2 mg / kg to approximately 2.2 mg / kg, approximately 1.4 mg / kg to approximately 2.2 mg / kg, approximately 1.6 mg / kg to approximately 2.2 mg / kg, approximately 1.8 mg / kg to approximately 2.0 mg / kg, approximately 0.1 mg / kg to approximately 1.6 mg / kg, approximately 0.5 mg / kg to approximately 1.6 mg / kg, or approximately 1 mg / kg to approximately 1.8 mg / kg; for example, approximately 1 mg / kg, approximately 1.2 mg / kg, approximately 1.6 mg / kg, or approximately 1.8 mg / kg).In some embodiments, the single dose C2D1 of the anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg.

[0013] In some embodiments, the bispecific antibody C1D1 and the bispecific antibody C1D2 are administered to the subject on day 8 or around (±1 day) and day 15 or around (±1 day), respectively, of the first administration cycle. In some embodiments, the bispecific antibody C2D1 is administered to the subject on day 1 or around (±1 day) of the second administration cycle. In some embodiments, the bispecific antibody C2D1 is administered after the completion of C2D1 of the anti-CD79b antibody drug conjugate. In some embodiments, the bispecific antibody C2D1 is administered or is administered approximately 60 to 120 minutes after the completion of administration of the anti-CD79b antibody drug conjugate C2D1 (e.g., approximately 60 to 100 minutes, approximately 60 to 90 minutes, approximately 60 to 80 minutes, approximately 90 to 120 minutes, approximately 80 to 100 minutes, approximately 80 to 120 minutes, approximately 75 to 105 minutes, or approximately 85 to 95 minutes; e.g., approximately 60 minutes, approximately 70 minutes, approximately 80 minutes, approximately 85 minutes, approximately 88 minutes, approximately 90 minutes, approximately 92 minutes, approximately 95 minutes, approximately 100 minutes, approximately 110 minutes, or approximately 120 minutes). In some embodiments, the bispecific antibody C2D1 is administered or is administered approximately 90 minutes after the completion of administration of the anti-CD79b antibody drug conjugate C2D1. In some embodiments, the anti-CD79b antibody drug conjugate C1D1 is administered to the subject on day 2 of the first administration cycle or around that day (±1 day), and the anti-CD79b antibody drug conjugate C2D1 is administered to the subject on day 1 of the second administration cycle or around that day (±1 day). In some embodiments, the first and second administration cycles are each 14-day (e.g., 14±3 day) administration cycles. In some embodiments, the first and second administration cycles are each 21-day (e.g., 21±3 day) administration cycles.

[0014] In some embodiments, the dosing regimen includes one or more additional dosing cycles. In some embodiments, the dosing regimen includes 6 to 10 additional dosing cycles (e.g., 6 additional dosing cycles, 7 additional dosing cycles, 8 additional dosing cycles, 9 additional dosing cycles, or 10 additional dosing cycles). In some embodiments, the dosing regimen includes 10 additional dosing cycles. In some embodiments, each additional dosing cycle is 14 days long (e.g., 14 ± 3 days). In some embodiments, each additional dosing cycle is 21 days long (e.g., 21 ± 3 days).

[0015] 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 equal to C2D1 of the 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 to the subject on day 1 (±1 day) of each additional dosing cycle that includes the additional dose of the anti-CD79b antibody drug conjugate.

[0016] In some embodiments, the additional single dose of the bispecific antibody in each additional dosing cycle, including an additional dose of the anti-CD79b antibody drug conjugate, is administered or is administered after the administration of the additional single dose of the anti-CD79b antibody drug conjugate is completed. In some embodiments, the additional single dose of the bispecific antibody for each additional dosing cycle, including an additional dose of the anti-CD79b antibody drug conjugate, is administered or will be administered approximately 60 to 120 minutes after the completion of administration of the additional single dose of the anti-CD79b antibody drug conjugate (e.g., approximately 60 to 100 minutes, approximately 60 to 90 minutes, approximately 60 to 80 minutes, approximately 90 to 120 minutes, approximately 80 to 100 minutes, approximately 80 to 120 minutes, approximately 75 to 105 minutes, or approximately 85 to 95 minutes; e.g., approximately 60 minutes, approximately 70 minutes, approximately 80 minutes, approximately 85 minutes, approximately 88 minutes, approximately 90 minutes, approximately 92 minutes, approximately 95 minutes, approximately 100 minutes, approximately 110 minutes, or approximately 120 minutes). In some embodiments, the additional single dose of this bispecific antibody for each additional dosing cycle, including the additional dose of the anti-CD79b antibody drug conjugate, is administered approximately 90 minutes after the completion of the administration of the additional single dose of the anti-CD79b antibody drug conjugate.

[0017] In some embodiments, the dosing regimen includes at least four additional dosing cycles, each containing an additional single dose of the anti-CD79b antibody-drug conjugate. In some embodiments, the dosing regimen includes between four and 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 containing an additional single dose of the anti-CD79b antibody-drug conjugate.

[0018] In some embodiments, one or more of the additional dosing cycles include an additional single dose of the bispecific antibody and do not include the 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 the administration of the anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen includes between 2 and 10 additional dosing cycles that include an additional single dose of the bispecific antibody and do not include the administration of the anti-CD79b antibody drug conjugate (e.g., 2 additional dosing cycles, 3 additional dosing cycles, 4 additional dosing cycles, 5 additional dosing cycles, 6 additional dosing cycles, 7 additional dosing cycles, 8 additional dosing cycles, 9 additional dosing cycles, or 10 additional dosing cycles).

[0019] In some embodiments, the additional single dose of the bispecific antibody is approximately equivalent to the C2D1 dose of the 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 to the subject on or around day 1 of each additional administration cycle containing the additional dose of the bispecific antibody (±1 day).

[0020] In some embodiments, the administration regimen comprises six or more additional administration cycles (e.g., six additional administration cycles, seven additional administration cycles, eight additional administration cycles, nine additional administration cycles, or ten additional administration cycles), each of which comprises a single dose of the bispecific antibody, and four or fewer of the six or more additional administration cycles (e.g., zero, one or fewer, two or fewer, three or fewer, or four or fewer; e.g., zero, one, two, three, or four) include the administration of an anti-CD79b antibody drug conjugate.

[0021] In one embodiment, the present invention is characterized by a method for treating a subject having a CD20-positive cell proliferative disorder, comprising administering to the subject having a CD20-positive cell proliferative disorder 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 the bispecific antibody (b) The second dose (C1D2) of the subject is administered to the subject after C1D1 of the anti-CD79b antibody drug conjugate, with C1D1 being approximately 2.5 mg and C1D2 being approximately 10 mg; (b) The second administration cycle is administered to the subject (i) a single dose (C2D1) of the anti-CD79b antibody drug conjugate and (ii) a single dose (C2D1) of the bispecific antibody, with C2D1 being approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0022] In another embodiment, the present invention features an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating to CD20 and CD3 for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugating 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 of the bispecific antibody (b) The second administration cycle comprises (i) a single dose (C1D1) of the anti-CD79b antibody drug conjugate and (ii) a single dose (C2D1) of the bispecific antibody, where C1D1 and C1D2 are administered to the subject after C1D1 of the anti-CD79b antibody drug conjugate, with C1D1 being approximately 2.5 mg and C1D2 being approximately 10 mg; (b) The second administration cycle comprises (i) a single dose (C2D1) of the anti-CD79b antibody drug conjugate and (ii) a single dose (C2D1) of the bispecific antibody, where C2D1 is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0023] In another embodiment, the present invention is characterized by the use of an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating to CD20 and CD3 for the treatment of a subject having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugating 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 of the bispecific antibody ( (b) The regimen comprises (i) a single dose of the anti-CD79b antibody drug conjugate (C1D1) and a second dose of the bispecific antibody (C1D2), wherein C1D1 and C1D2 of the bispecific antibody are administered to the subject after C1D1 of the anti-CD79b antibody drug conjugate, with C1D1 being approximately 2.5 mg and C1D2 being approximately 10 mg; (b) The second administration cycle comprises (i) a single dose of the anti-CD79b antibody drug conjugate (C2D1) and (ii) a single dose of the bispecific antibody (C2D1), with C2D1 being approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0024] In another embodiment, the present invention is characterized by the use of an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating to CD20 and CD3 in the manufacture of a pharmacopoeci for the treatment of a subject having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugating 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 fraction of the bispecific antibody conjugate (b) The regimen comprises a dose 1 (C1D1) and a second dose (C1D2) of the bispecific antibody, where C1D1 and C1D2 of the bispecific antibody are administered to the subject after C1D1 of the anti-CD79b antibody drug conjugate, with C1D1 being approximately 2.5 mg and C1D2 being approximately 10 mg; (b) The second administration cycle comprises (i) a single dose (C2D1) of the anti-CD79b antibody drug conjugate and (ii) a single dose (C2D1) of the bispecific antibody, with C2D1 being approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0025] In some embodiments, a single dose C1D1 of the anti-CD79b antibody drug conjugate ranges from approximately 0.1 mg / kg to approximately 2.4 mg / kg (e.g., approximately 0.1 mg / kg to approximately 2.2 mg / kg, approximately 0.1 mg / kg to approximately 2.0 mg / kg, approximately 0.5 mg / kg to approximately 2.2 mg / kg, approximately 0.8 mg / kg to approximately 2.2 mg / kg, approximately 1 mg / kg to approximately 2.2 mg / kg, approximately 1.2 mg / kg). Approximately 2.2 mg / kg from kg, approximately 1.4 mg / kg to approximately 2.2 mg / kg, approximately 1.6 mg / kg to approximately 2.2 mg / kg, approximately 1.8 mg / kg to approximately 2.0 mg / kg, approximately 0.1 mg / kg to approximately 1.6 mg / kg, approximately 0.5 mg / kg to approximately 1.6 mg / kg, or approximately 1 mg / kg to approximately 1.8 mg / kg; for example, approximately 1 mg / kg, approximately 1.2 mg / kg, approximately 1.6 mg / kg, or approximately 1.8 mg / kg. The dosage range is approximately 0.1 mg / kg to 2.4 mg / kg (e.g., approximately 0.1 mg / kg to 2.2 mg / kg, approximately 0.1 mg / kg to 2.0 mg / kg, approximately 0.5 mg / kg to 2.2 mg / kg, approximately 0.8 mg / kg to 2.2 mg / kg, approximately 1 mg / kg to 2.2 mg / kg, approximately 1.2 mg / kg to 2. The dosages are approximately 0.2 mg / kg, 1.4 mg / kg to 2.2 mg / kg, 1.6 mg / kg to 2.2 mg / kg, 1.8 mg / kg to 2.0 mg / kg, 0.1 mg / kg to 1.6 mg / kg, 0.5 mg / kg to 1.6 mg / kg, or 1 mg / kg to 1.8 mg / kg (for example, 1 mg / kg, 1.2 mg / kg, 1.6 mg, or 1.8 mg / kg). In some embodiments, a single dose C1D1 of the anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg, and a single dose C2D1 of the anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg.

[0026] In some embodiments, the bispecific antibody C1D1 and the bispecific antibody C1D2 are administered to the subject on or around day 8 (±1 day) and on or around day 15 (±1 day), respectively, of the first administration cycle.

[0027] In some embodiments, the bispecific antibody C2D1 is administered to the subject on day 1 of the second administration cycle or around that day (±1 day). In some embodiments, the anti-CD79b antibody drug conjugate C1D1 is administered to the subject on day 2 of the first administration cycle or around that day (±1 day), and the anti-CD79b antibody drug conjugate C2D1 is administered to the subject on day 1 of the second administration cycle or around that day (±1 day).

[0028] In some embodiments, the first and second administration cycles are each 14-day (e.g., 14 ± 3 days) administration cycles. In some embodiments, the first and second administration cycles are each 21-day (e.g., 21 ± 3 days) administration cycles.

[0029] In some embodiments, the dosing regimen includes one or more additional dosing cycles. In some embodiments, the dosing regimen includes 6 to 10 additional dosing cycles (e.g., 6 additional dosing cycles, 7 additional dosing cycles, 8 additional dosing cycles, 9 additional dosing cycles, or 10 additional dosing cycles).

[0030] In some embodiments, the dosing regimen includes 10 additional dosing cycles. In some embodiments, each additional dosing cycle is 21 days (e.g., 21 ± 3 days). In some embodiments, the dosing regimen includes one or more additional dosing cycles. In some embodiments, the dosing regimen includes 6 to 10 additional dosing cycles. In some embodiments, the dosing regimen includes 10 additional dosing cycles.

[0031] In some embodiments, each additional dosing cycle is a 14-day (e.g., 14 ± 3 days) dosing cycle.

[0032] In some embodiments, one or more of the 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 to C2D1 of the anti-CD79b antibody drug conjugate. In some embodiments, the additional single dose of the anti-CD79b antibody drug conjugate is administered to the subject on day 1 (±1 day) of each additional dosing cycle that includes the additional dose of the anti-CD79b antibody drug conjugate.

[0033] In some embodiments, the additional single dose of the bispecific antibody in each additional dosing cycle, including an additional dose of the anti-CD79b antibody drug conjugate, is administered or is administered after the administration of the additional single dose of the anti-CD79b antibody drug conjugate is completed. In some embodiments, the additional single dose of the bispecific antibody for each additional dosing cycle, including an additional dose of the anti-CD79b antibody drug conjugate, is administered or will be administered approximately 60 to 120 minutes after the completion of administration of the additional single dose of the anti-CD79b antibody drug conjugate (e.g., approximately 60 to 100 minutes, approximately 60 to 90 minutes, approximately 60 to 80 minutes, approximately 90 to 120 minutes, approximately 80 to 100 minutes, approximately 80 to 120 minutes, approximately 75 to 105 minutes, or approximately 85 to 95 minutes; e.g., approximately 60 minutes, approximately 70 minutes, approximately 80 minutes, approximately 85 minutes, approximately 88 minutes, approximately 90 minutes, approximately 92 minutes, approximately 95 minutes, approximately 100 minutes, approximately 110 minutes, or approximately 120 minutes). In some embodiments, the additional single dose of this bispecific antibody for each additional dosing cycle, including the additional dose of the anti-CD79b antibody drug conjugate, is administered approximately 90 minutes after the completion of the administration of the additional single dose of the anti-CD79b antibody drug conjugate.

[0034] In some embodiments, the dosing regimen includes at least four additional dosing cycles, each containing 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, five, six, seven, eight, nine, or ten additional dosing cycles) each containing an additional single dose of the bispecific antibody and an additional single dose of the anti-CD79b antibody drug conjugate. In some embodiments, one or more of the additional dosing cycles contain an additional single dose of the bispecific antibody but do not contain an administration of the anti-CD79b antibody drug conjugate.

[0035] In some embodiments, the administration regimen includes at least two additional administration cycles, each containing an additional single dose of the bispecific antibody and not including the administration of an anti-CD79b antibody drug conjugate. In some embodiments, the administration regimen includes between 2 and 10 additional administration cycles (e.g., 2 additional administration cycles, 3 additional administration cycles, 4 additional administration cycles, 5 additional administration cycles, 6 additional administration cycles, 7 additional administration cycles, 8 additional administration cycles, 9 additional administration cycles, or 10 additional administration cycles) each containing an additional single dose of the bispecific antibody and not including the administration of an anti-CD79b antibody drug conjugate.

[0036] In some embodiments, the additional single dose of the bispecific antibody is approximately equivalent to the C2D1 dose of the 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 to the subject on or around day 1 of each additional administration cycle containing the additional dose of the bispecific antibody (±1 day).

[0037] In some embodiments, the administration regimen comprises six or more additional administration cycles (e.g., six additional administration cycles, seven additional administration cycles, eight additional administration cycles, nine additional administration cycles, or ten additional administration cycles), each of which comprises a single dose of the bispecific antibody, and four or fewer of the six or more additional administration cycles (e.g., zero, one or fewer, two or fewer, three or fewer, or four or fewer; e.g., zero, one, two, three, or four) of which comprises administration of an anti-CD79b antibody drug conjugate. In some embodiments, the administration regimen comprises 6 to 10 or more additional administration cycles (e.g., 6 additional administration cycles, 7 additional administration cycles, 8 additional administration cycles, 9 additional administration cycles, or 10 additional administration cycles), each of which comprises a single dose of the bispecific antibody, and four or fewer of the six additional administration cycles (e.g., 0, 1 or fewer, 2 or fewer, 3 or fewer, or 4 or fewer; e.g., 0, 1, 2, 3, or 4) include the administration of the anti-CD79b antibody drug conjugate. In some embodiments, the administration regimen comprises 10 additional administration cycles, each of which comprises a single dose of the bispecific antibody, and four or fewer of the ten additional administration cycles (e.g., 0, 1 or fewer, 2 or fewer, 3 or fewer, or 4 or fewer; e.g., 0, 1, 2, 3, or 4) include the administration of the anti-CD79b antibody drug conjugate. In some embodiments, the administration regimen comprises 10 additional administration cycles, each of which comprises a single dose of the bispecific antibody, and each of the 10 additional administration cycles comprises administration of an anti-CD79b antibody drug conjugate.

[0038] In one embodiment, the present invention is characterized by a method for treating a subject having a CD20-positive cell proliferative disorder, comprising administering to the subject having a CD20-positive cell proliferative disorder 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 (where C1D1 is approximately 2.5 mg and C1D2 is approximately 10 mg) (i) a single dose (C1D1) of the anti-CD79b antibody drug conjugate; (b) each of the second to sixth administration cycles includes a single dose (C2D1 to C6D1) of the bispecific antibody and a single dose (C2D1 to C6D1) of the anti-CD79b antibody drug conjugate; (c) each of the seventh to twelfth administration cycles includes a single dose (C7D1 to C12D1) of the bispecific antibody and does not include administration of the anti-CD79b antibody drug conjugate; each single dose C2D1 to C12D1 of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0039] In another embodiment, the present invention features an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating to CD20 and CD3 for use in a method of treating a subject having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugating to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose of the bispecific antibody (C1D1) and a second dose of the bispecific antibody (C1D2) (where C1D1 of the bispecific antibody is approximately 2 (i) a single dose (C1D2) of the bispecific antibody is approximately 10 mg; and (ii) a single dose (C1D1) of the anti-CD79b antibody drug conjugate is included; (b) each of the 2nd to 6th administration cycles includes a single dose (C2D1 to C6D1) of the bispecific antibody and a single dose (C2D1 to C6D1) of the anti-CD79b antibody drug conjugate; (c) each of the 7th to 12th administration cycles includes a single dose (C7D1 to C12D1) of the bispecific antibody and does not include administration of the anti-CD79b antibody drug conjugate; each single dose (C2D1 to C12D1) of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0040] In another embodiment, the present invention is characterized by the use of an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating to CD20 and CD3 in the treatment of subjects having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugating to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose of the bispecific antibody (C1D1) and a second dose of the bispecific antibody (C1D2) (where C1D1 of the bispecific antibody is approximately 2.5 mg and C1 of the bispecific antibody D2 is approximately 10 mg; and (ii) a single dose (C1D1) of the anti-CD79b antibody drug conjugate; (b) each of the second to sixth administration cycles includes a single dose (C2D1 to C6D1) of the bispecific antibody and a single dose (C2D1 to C6D1) of the anti-CD79b antibody drug conjugate; (c) each of the seventh to twelfth administration cycles includes a single dose (C7D1 to C12D1) of the bispecific antibody and does not include administration of the anti-CD79b antibody drug conjugate; each single dose (C2D1 to C12D1) of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0041] In another embodiment, the present invention is characterized by the use of an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating to CD20 and CD3 in the manufacture of a pharmacopoeia for the treatment of subjects having a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugating to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose of the bispecific antibody (C1D1) and a second dose of the bispecific antibody (C1D2) (where C1D1 of the bispecific antibody is approximately 2.5 mg, and the bispecific antibody (i) the amount of C1D2 in the body is approximately 10 mg; and (ii) a single dose (C1D1) of the anti-CD79b antibody drug conjugate; (b) each of the second to sixth administration cycles includes a single dose (C2D1 to C6D1) of the bispecific antibody and a single dose (C2D1 to C6D1) of the anti-CD79b antibody drug conjugate; (c) each of the seventh to twelfth administration cycles includes a single dose (C7D1 to C12D1) of the bispecific antibody and does not include administration of the anti-CD79b antibody drug conjugate; each single dose (C2D1 to C12D1) of the bispecific antibody is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0042] In some embodiments, the amounts of C2D1 to C12D1 of this bispecific antibody are approximately equal. In some embodiments, the amount of C2D1 of this bispecific antibody is approximately 30 mg.

[0043] In some embodiments, the C1D1-C6D1 of the anti-CD79b antibody drug conjugate are approximately equal in amount. In some embodiments, each of the C1D1-C6D1 of the anti-CD79b antibody drug conjugate is approximately 0.1 mg / kg to approximately 2.4 mg / kg (for example, approximately 0.1 mg / kg to approximately 2.2 mg / kg, approximately 0.1 mg / kg to approximately 2.0 mg / kg, approximately 0.5 mg / kg to approximately 2.2 mg / kg, approximately 0.8 mg / kg to approximately 2.2 mg / kg, approximately 1 mg / kg to approximately 2.2 mg / kg, approximately 1.2 mg / kg). The dosages range from approximately 2.2 mg / kg, 1.4 mg / kg to approximately 2.2 mg / kg, 1.6 mg / kg to approximately 2.2 mg / kg, 1.8 mg / kg to approximately 2.0 mg / kg, 0.1 mg / kg to approximately 1.6 mg / kg, 0.5 mg / kg to approximately 1.6 mg / kg, or 1 mg / kg to approximately 1.8 mg / kg; for example, approximately 1 mg / kg, approximately 1.2 mg / kg, approximately 1.6 mg / kg, or approximately 1.8 mg / kg). In some embodiments, each of the anti-CD79b antibody drug conjugates C1D1 to C6D1 is approximately 1.8 mg / kg.

[0044] In some embodiments, the bispecific antibody C1D1 and the bispecific antibody C1D2 are administered to the subject on day 8 or around (±1 day) and day 15 or around (±1 day), respectively, of the first administration cycle. In some embodiments, the bispecific antibodies C2D1 to C12D1 are administered to the subject on day 1 of each administration cycle or around. In some embodiments, the anti-CD79b antibody drug conjugate C1D1 is administered to the subject on day 2 of the first administration cycle or around (±1 day), and the anti-CD79b antibody drug conjugates C2D1 to C6D1 are administered to the subject on day 1 of each administration cycle that includes the administration of the anti-CD79b antibody drug conjugate.

[0045] In some embodiments, the C2D1-C6D1 of the bispecific antibody is administered or is administered after the completion of the C2D1-C6D1 of the anti-CD79b antibody drug conjugate. In some embodiments, the bispecific antibodies C2D1-C6D1 are administered or will be administered approximately 60-120 minutes after the completion of administration of the anti-CD79b antibody drug conjugate C2D1-C6D1 (e.g., approximately 60-100 minutes, approximately 60-90 minutes, approximately 60-80 minutes, approximately 90-120 minutes, approximately 80-100 minutes, approximately 80-120 minutes, approximately 75-105 minutes, or approximately 85-95 minutes; e.g., approximately 60 minutes, approximately 70 minutes, approximately 80 minutes, approximately 85 minutes, approximately 88 minutes, approximately 90 minutes, approximately 92 minutes, approximately 95 minutes, approximately 100 minutes, approximately 110 minutes, or approximately 120 minutes). In some embodiments, the bispecific antibodies C2D1–C6D1 are administered approximately 90 minutes after the completion of administration of the anti-CD79b antibody drug conjugate C2D1–C6D1.

[0046] In some embodiments, each administration cycle is 14 days (e.g., 14 ± 3 days). In some embodiments, each administration cycle is 21 days (e.g., 21 ± 3 days).

[0047] In some embodiments, the administration regimen includes an additional retreatment regimen after the completion of 12 administration cycles of the administration regimen. In some embodiments, the additional retreatment regimen includes 12 additional administration cycles, where (a) the first additional administration cycle includes (i) a first dose of the bispecific antibody (C13D1) and a second dose of the bispecific antibody (C13D2) (where C13D1 is approximately 2.5 mg and C13D2 is approximately 10 mg); and (ii) a single dose of the anti-CD79b antibody drug conjugate (C13D1); and (b) the second... Each of the sixth additional dosing cycles comprises a single dose of the bispecific antibody (C14D1-C18D1) and a single dose of the anti-CD79b antibody drug conjugate (C14D1-C18D1); (c) Each of the seventh-twelfth additional dosing cycles comprises a single dose of the bispecific antibody (C19D1-C24D1) and does not include administration of the anti-CD79b antibody drug conjugate; each single dose of the bispecific antibody C14D1-C24D1 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 administered or to be administered on day 8 (±1) of the first additional dosing cycle and a second dose (C13D2) of the bispecific antibody administered or to be administered on day 15 (±1) of the first additional dosing cycle (where C13D1 of the bispecific antibody is approximately 2.5 mg and C13D2 of the bispecific antibody is approximately 10 mg); and (ii) a single dose (C13D1) of the anti-CD79b antibody drug conjugate administered or to be administered on day 2 (±1) of the first additional dosing cycle; and (b) the second to sixth additional dosing cycles each comprise a single dose (C14D1 to C1) of the bispecific antibody. c) each includes a single dose of the bispecific antibody (C14D1-C18D1) and an anti-CD79b antibody drug conjugate; a) each of the 7th to 12th additional administration cycles includes a single dose of the bispecific antibody (C19D1-C24D1) and does not include administration of the anti-CD79b antibody drug conjugate; the bispecific antibody C14D1-C24D1 is administered on day 1 (±1 day) of each additional administration cycle, and the anti-CD79b antibody drug conjugate C14D1-C18D1 is administered on day 1 (±1 day) of each additional administration cycle, with each single dose of the bispecific antibody C14D1-C24D1 being approximately 30 mg and each single dose of the anti-CD79b antibody drug conjugate C13D1-C18D1 being approximately 1.8 mg / kg. In some embodiments, there is a waiting period between the completion of 12 dosing cycles of the treatment regimen and the start of 12 additional dosing cycles of the additional retreatment regimen. In some embodiments, this waiting period is between approximately one week and approximately 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.

[0048] In some embodiments, the method of the present invention further includes administering one or more additional therapeutic agents to a target. In some embodiments, the anti-CD79b antibody drug conjugate and the bispecific antibody are administered together with one or more additional therapeutic agents.

[0049] In some embodiments, one or more additional therapeutic agents include one or more chemotherapeutic agents. In some embodiments, one or more chemotherapeutic agents include cyclophosphamide, doxorubicin, and rituximab.

[0050] In some embodiments, one or more additional therapeutic agents are tocilizumab. In some embodiments, one or more additional therapeutic agents are corticosteroids. In some embodiments, the corticosteroids include prednisone, prednisolone, methylprednisolone, and dexamethasone.

[0051] In some embodiments, the method of the present invention further comprises administering rituximab, cyclophosphamide, doxorubicin, and prednisone (R-CHP) to the target. In some embodiments, the anti-CD79b antibody drug conjugate and the bispecific antibody are administered together with rituximab, cyclophosphamide, doxorubicin, and prednisone (R-CHP).

[0052] In some embodiments, one or more additional therapeutic agents are antihistamines. In some embodiments, the antihistamine is diphenhydramine. In some embodiments, one or more additional therapeutic agents include allopurinol and rasburicase. In some embodiments, one or more additional therapeutic agents are antipyretics.

[0053] In some embodiments, one or more additional therapeutic agents are obinutuzumab. In some embodiments, obinutuzumab is administered or will be administered before the administration of the bispecific antibody. In some embodiments, obinutuzumab is administered or will be administered about 7 days (±1 day) before the administration of the bispecific antibody. In some embodiments, obinutuzumab is administered or will 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 a second dose of about 1000 mg. In some embodiments, the first dose of obinutuzumab is administered about 7 days (±1 day) before the administration of C1D1 of the bispecific antibody. In some embodiments, the second dose of obinutuzumab is administered about 1 day (±1 day) before the administration of C1D1 of the bispecific antibody.

[0054] In some embodiments, the anti-CD79b antibody-drug conjugate is polatuzumab vedotin or anti-CD79b-MC-vc-PAB-MMAE.

[0055] In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds to CD20 and includes the following six hypervariable regions (HVRs): (a) HVR-H1 containing the amino acid sequence of YSWIN (SEQ ID NO: 1); (b) HVR-H2 containing the amino acid sequence of RIFPGDGDTDYNGKFKG (SEQ ID NO: 2); (c) HVR-H3 containing the amino acid sequence of NVFDGYWLVY (SEQ ID NO: 3); (d) HVR-L1 containing the amino acid sequence of RSKSLLHSNGITYLY (SEQ ID NO: 4); (e) HVR-L2 containing the amino acid sequence of QMSNLVS (SEQ ID NO: 5); and (f) HVR-L3 containing the amino acid sequence of AQNLELPYT (SEQ ID NO: 6). In some embodiments, the bispecific antibody comprises (a) a heavy-chain variable VH domain containing an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 7 (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%); (b) a variable light (VL) domain containing an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 8 (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%); or (c) at least one Fab molecule that specifically binds to CD20, comprising a VH domain as in (a) and a VL domain as in (b). In some embodiments, the Fab molecule that specifically binds to CD20 includes (a) a VH domain containing the amino acid sequence of SEQ ID NO: 7 and (b) a VL domain containing the amino acid sequence of SEQ ID NO: 8.

[0056] In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds to CD3, the molecule comprising the following six HVRs: (a) HVR-H1 comprising the amino acid sequence of TYAMN (SEQ ID NO: 9); (b) HVR-H2 comprising the amino acid sequence of RIRSKYNNYATYYADSVKG (SEQ ID NO: 10); (c) HVR-H3 comprising the amino acid sequence of HGNFGNSYVSWFAY (SEQ ID NO: 11); (d) HVR-L1 comprising the amino acid sequence of GSSTGAVTTSNYAN (SEQ ID NO: 12); (e) HVR-L2 comprising the amino acid sequence of GTNKRAP (SEQ ID NO: 13); and (f) HVR-L3 comprising the amino acid sequence of ALWYSNLWV (SEQ ID NO: 14). In some embodiments, the bispecific antibody comprises (a) a heavy-chain variable VH domain containing an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 15 (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%); (b) a variable light (VL) domain containing an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16 (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%); or (c) at least one Fab molecule that specifically binds to CD3, comprising a VH domain as in (a) and a VL domain as in (b). In some embodiments, the Fab molecule that specifically binds to CD3 includes (a) a VH domain containing the amino acid sequence of SEQ ID NO: 15 and (b) a VL domain containing the amino acid sequence of SEQ ID NO: 16.

[0057] In some embodiments, the bispecific antibody comprises a Fab molecule that specifically binds to CD3, wherein (a) the variable domain of the Fab heavy chain and the variable domain of the Fab light chain are exchanged, or (b) the constant domain of the Fab heavy chain and the constant domain of the Fab light chain are exchanged. In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds to CD20, wherein in the constant domain CL of the Fab molecule, the amino acid at position 124 is substituted with lysine (K) (Kabat numbering), the amino acid at position 123 is substituted with arginine (R) or lysine (K) (Kabat numbering), and in the constant domain CH1 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 divalent with respect to CD20 and monovalent with respect to 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.

[0058] 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; where the third Fab molecule in (c) is identical to the first Fab molecule in (a); and in the constant domain CL of the first Fab molecule in (a) and the third Fab molecule in (c), the amino acid at position 124 is substituted with lysine (K) (Kabat numbering), and the amino acid at position 123 is arginine (R). Alternatively, it is substituted with lysine (K) (numbered by Kabat); where, in the constant domain CH1 of the first Fab molecule (a) and the third Fab molecule (c), 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); where, the first Fab molecule (a) is fused at the C-terminus of the Fab heavy chain of the second Fab molecule (b) with the N-terminus of the Fab heavy chain of the second Fab molecule (b) and the third Fab molecule (c) are each fused at the C-terminus of one of the N-terminuses of a subunit of the Fc domain of (d) with the C-terminus of the Fab heavy chain.

[0059] In some embodiments, the bispecific antibody is a humanized antibody. In some embodiments, the bispecific antibody is a chimeric antibody.

[0060] In some embodiments, the bispecific antibody includes an Fc domain, which is an IgG Fc domain. In another embodiment, the IgG Fc domain is an IgG1 Fc domain. In some embodiments, the IgG Fc domain includes a mutation at amino acid residue N297 (EU numbering) resulting in the absence of glycosylation. In some embodiments, the mutation at amino acid residue N297 is a substitutional mutation. In some embodiments, the mutation at amino acid residue N297 reduces the effector function of the Fc region. In some embodiments, this mutation is an N297G or N297A mutation. In some embodiments, the bispecific antibody includes a mutation in the Fc region that reduces the effector function. In some embodiments, this mutation is a substitutional mutation. In some embodiments, the substitutional mutation is at amino acid residues 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.

[0061] In some embodiments, the bispecific antibody comprises one or more heavy chain constant domains, which are selected from a first CH1(CH11) domain, a first CH2(CH21) domain, a first CH3(CH31) domain, a second CH1(CH12) domain, a second CH2(CH22) domain, and a second CH3(CH32) domain. In some embodiments, at least one of the one or more heavy chain constant domains is paired with another heavy chain constant domain. In some embodiments, the CH31 domain and the CH32 domain each comprise a projection or cavity, and the projection or cavity of the CH31 domain can be positioned in the cavity or projection of the CH32 domain, respectively. In some embodiments, the CH31 domain and the CH32 domain are in contact at the interface between the projection and the cavity. In some embodiments, the CH21 domain and the CH22 domain each include a projection or cavity, and the projection or cavity of the CH21 domain can be positioned within the cavity or projection of the CH22 domain. In some embodiments, the CH21 domain and the CH22 domain are in contact at the interface between the projection and the cavity.

[0062] In some embodiments, the bispecific antibody is grofitamab.

[0063] In some embodiments, the bispecific antibody is administered intravenously. In some embodiments, the anti-CD79b antibody drug conjugate is administered intravenously. In some embodiments, if the bispecific antibody and the anti-CD79b antibody drug conjugate are administered on the same day, the bispecific antibody is administered after the administration of the anti-CD79b antibody drug conjugate is completed. In some embodiments, the bispecific antibody is administered or will be administered approximately 60 to 120 minutes after the completion of administration of the anti-CD79b antibody drug conjugate (e.g., approximately 60 to 100 minutes, approximately 60 to 90 minutes, approximately 60 to 80 minutes, approximately 90 to 120 minutes, approximately 80 to 100 minutes, approximately 80 to 120 minutes, approximately 75 to 105 minutes, or approximately 85 to 95 minutes; e.g., approximately 60 minutes, approximately 70 minutes, approximately 80 minutes, approximately 85 minutes, approximately 88 minutes, approximately 90 minutes, approximately 92 minutes, approximately 95 minutes, approximately 100 minutes, approximately 110 minutes, or approximately 120 minutes). In some embodiments, the bispecific antibody is administered or will be administered approximately 90 minutes after the completion of administration of the anti-CD79b antibody drug conjugate.

[0064] In one embodiment, the present invention is a method for treating a subject having a CD20-positive cell proliferative disorder, characterized by administering polatuzumab vedotin and grofitamab 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 a first dose (C1D1) of grofitamab administered on day 8 (±1 day) of the first dosing cycle and grofitamab administered on day 15 (±1 day) of the first dosing cycle. (b) The second dose of grofitamab (C1D2) is approximately 2.5 mg of grofitamab C1D1 and approximately 10 mg of grofitamab C1D2; (b) The second administration cycle is approximately 10 mg of grofitamab C2D1, administered on day 1 (±1) of the second administration cycle, and approximately 10 mg of grofitamab C2D1, administered on day 1 (±1) of the second administration cycle.

[0065] In another embodiment, the present invention features polatuzumab vedotin and grofitamab for use in a method of treating subjects having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, (a) the first dosing cycle comprising a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and (b) administered on day 15 (±1) of the first dosing cycle (b) The second dose (C1D2) of grofitamab administered, where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg; (b) The second administration cycle comprises a single dose (C2D1) of grofitamab administered on day 1 (±1) of the second administration cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1) of the second administration cycle, where C2D1 of grofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0066] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab for the treatment of subjects having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and grofitamab 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 grofitamab administered on day 8 (±1) of the first dosing cycle and an administration of grofitamab administered on day 15 (±1) of the first dosing cycle (b) The second administration cycle includes a second dose (C1D2) of grofitamab, where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg; (b) The second administration cycle includes a single dose (C2D1) of grofitamab administered on day 1 (±1) of the second administration cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1) of the second administration cycle, where C2D1 of grofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0067] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the manufacture of a pharmacopoeci for the treatment of subjects having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and grofitamab 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 grofitamab administered on day 8 (±1) of the first dosing cycle and (b) administered on day 15 (±1) of the first dosing cycle (b) The second administration cycle includes a second dose (C1D2) of grofitamab, where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg; (b) The second administration cycle includes a single dose (C2D1) of grofitamab administered on day 1 (±1) of the second administration cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1) of the second administration cycle, where C2D1 of grofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0068] In one embodiment, the present invention relates to a method for treating a subject having a CD20-positive cell proliferative disorder, comprising administering polatuzumab vedotin and grofitamab 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 polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle, and (ii) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and an additional dose administered on day 15 (±1) of the first dosing cycle. (b) The second administration cycle includes (i) a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1) of the second administration cycle and (ii) a single dose (C2D1) of grofitamab administered on day 1 (±1) of the second administration cycle, with grofitamab C2D1 being approximately 10 mg, approximately 16 mg, or approximately 30 mg, and polatuzumab vedotin C1D1 and C2D1 being approximately 1.8 mg / kg each.

[0069] In another embodiment, the present invention features polatuzumab vedotin and grofitamab for use in a method of treating subjects having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and grofitamab 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 administered on day 2 (±1) of the first dosing cycle, and (ii) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and on day 15 (± (b) The second administration cycle includes (i) a single dose (C2D1) of grofitamab administered on day 1 (±1 day), where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg; (b) the second administration cycle includes (i) a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1 day) of the second administration cycle and (ii) a single dose (C2D1) of grofitamab administered on day 1 (±1 day) of the second administration cycle, where C2D1 of grofitamab is approximately 10 mg, approximately 16 mg or approximately 30 mg, and C1D1 and C2D1 of polatuzumab vedotin are approximately 1.8 mg / kg, respectively.

[0070] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the treatment of subjects having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and grofitamab 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 administered on day 2 (±1) of the first dosing cycle, and (ii) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and on day 15 (±1) of the first dosing cycle. (b) The second administration cycle includes (i) a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1 day) of the second administration cycle, and (ii) a single dose (C2D1) of grofitamab administered on day 1 (±1 day) of the second administration cycle, with grofitamab C2D1 being approximately 10 mg, approximately 16 mg, or approximately 30 mg, and polatuzumab vedotin C1D1 and C2D1 being approximately 1.8 mg / kg each.

[0071] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the manufacture of a pharmacopoeci for the treatment of subjects having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and grofitamab 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 administered on day 2 (±1) of the first dosing cycle and (ii) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and on day 15 (±1) of the first dosing cycle. (b) The second administration cycle includes (i) a single dose (C2D1) of grofitamab administered on day 1 (±1 day) of the second administration cycle, where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg; (b) the second administration cycle includes (i) a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1 day) of the second administration cycle, and (ii) a single dose (C2D1) of grofitamab administered on day 1 (±1 day) of the second administration cycle, where C2D1 of grofitamab is approximately 10 mg, approximately 16 mg or approximately 30 mg, and C1D1 and C2D1 of polatuzumab vedotin are each approximately 1.8 mg / kg.

[0072] In one embodiment, the present invention relates to a method for treating a subject having a CD20-positive cell proliferative disorder, comprising administering polatuzumab vedotin and grofitamab to the subject in a dosing regimen comprising 12 dosing cycles, wherein (a) the first dosing cycle comprises a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle, a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle, wherein C1D1 of grofitamab is between approximately 1 mg and approximately 5 mg, and C1D2 of grofitamab is approximately 10 mg; and (b) the second to sixth dosing cycles each comprise grofitamab (c) Each of the 7th to 12th dosing cycles includes a single dose of grofitamab (C2D1 to C6D1) and a single dose of polatuzumab vedotin (C2D1 to C6D1), where C2D1 of grofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg; (c) Each of the 7th to 12th dosing cycles includes a single dose of grofitamab (C7D1 to C12D1) and does not include administration of polatuzumab vedotin; each of the single doses of grofitamab C 2D1 to C12D1 are administered on day 1 (±1 day) of each dosing cycle, and each single dose of polatuzumab vedotin C2D1 to C6D1 is administered on day 1 (±1 day) of each dosing cycle. Here, each single dose of grofitamab C3D1 to C12D1 is approximately equivalent to grofitamab C2D1, and each single dose of polatuzumab vedotin C1D1 to C6D1 is approximately 1.8 mg / kg.

[0073] In another embodiment, the present invention features polatuzumab vedotin and grofitamab for use in a method of treating subjects having CD20-positive cell proliferative disorders, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle, a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle, where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg; (b) the second to sixth dosing cycles Each of these includes a single dose of grofitamab (C2D1-C6D1) and a single dose of polatuzumab vedotin (C2D1-C6D1), where C2D1 of grofitamab is approximately 10 mg, 16 mg, or 30 mg; (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1-C12D1) and does not include administration of polatuzumab vedotin; grofitamab Each single dose C2D1 to C12D1 of polatuzumab vedotin is administered on day 1 (±1 day) of each dosing cycle, each single dose C2D1 to C6D1 of polatuzumab vedotin is administered on day 1 (±1 day) of each dosing cycle, each single dose C3D1 to C12D1 of grofitamab is approximately equivalent to C2D1 of grofitamab, and each single dose C1D1 to C6D1 of polatuzumab vedotin is approximately 1.8 mg / kg.

[0074] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the treatment of subjects having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein (a) the first dosing cycle comprises a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle, a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle, wherein C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg, and (b) the second to sixth dosing cycles each comprise grofitamab (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C2D1 to C6D1) and a single dose of polatuzumab vedotin (C2D1 to C6D1), where C2D1 of grofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg; (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1 to C12D1) and does not include administration of polatuzumab vedotin; each of the single doses of grofitamab The doses C2D1 to C12D1 are administered on day 1 (±1 day) of each dosing cycle, and each single dose of polatuzumab vedotin C2D1 to C6D1 is administered on day 1 (±1 day) of each dosing cycle. Here, each single dose of grofitamab C3D1 to C12D1 is approximately equivalent to grofitamab C2D1, and each single dose of polatuzumab vedotin C1D1 to C6D1 is approximately 1.8 mg / kg.

[0075] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the manufacture of a pharmacopoeia for the treatment of subjects having CD20-positive cell proliferative disorders, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle, a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle, and a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle, where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg; (b) the second to sixth dosing cycles Each of the following cycles includes a single dose of grofitamab (C2D1-C6D1) and a single dose of polatuzumab vedotin (C2D1-C6D1), where C2D1 of grofitamab is approximately 10 mg, 16 mg, or 30 mg; (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1-C12D1) and does not include administration of polatuzumab vedotin; grofitamab Each single dose of bu (C2D1-C12D1) is administered on day 1 (±1 day) of each dosing cycle, each single dose of polatuzumab vedotin (C2D1-C6D1) is administered on day 1 (±1 day) of each dosing cycle, each single dose of grofitamab (C3D1-C12D1) is approximately equivalent to grofitamab (C2D1), and each single dose of polatuzumab vedotin (C1D1-C6D1) is approximately 1.8 mg / kg.

[0076] In one embodiment, the present invention relates to a method for treating a subject having a CD20-positive cell proliferative disorder, comprising administering polatuzumab vedotin and grofitamab to the subject in a dosing regimen comprising 12 dosing cycles, wherein (a) the first dosing cycle comprises (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg) and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle; and (b) the second to sixth dosing cycles each comprise grofitamab vedotin. (c) Each of the following administration cycles includes a single dose of grofitamab (C2D1-C6D1) and a single dose of polatuzumab vedotin (C2D1-C6D1), where C2D1 of grofitamab is approximately 10 mg, 16 mg, or 30 mg; (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1-C12D1) and does not include administration of polatuzumab vedotin; each of the single doses of grofitamab C2D1 to C12D1 are administered on day 1 (±1 day) of each dosing cycle, and each single dose of polatuzumab vedotin C2D1 to C6D1 is administered on day 1 (±1 day) of each dosing cycle. Here, each single dose of grofitamab C3D1 to C12D1 is approximately equivalent to grofitamab C2D1, and each single dose of polatuzumab vedotin C1D1 to C6D1 is approximately 1.8 mg / kg.

[0077] In another embodiment, the present invention features polatuzumab vedotin and grofitamab for use in a method of treating subjects having CD20-positive cell proliferative disorders, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg); and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle; and (b) the second to sixth dosing cycles Each administration cycle includes a single dose of grofitamab (C2D1-C6D1) and a single dose of polatuzumab vedotin (C2D1-C6D1), where C2D1 of grofitamab is approximately 10 mg, approximately 16 mg, or approximately 30 mg; (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1-C12D1) and does not include administration of polatuzumab vedotin; grofi Each single dose of tamab, C2D1 to C12D1, is administered on day 1 (±1 day) of each dosing cycle; each single dose of polatuzumab vedotin, C2D1 to C6D1, is administered on day 1 (±1 day) of each dosing cycle; each single dose of grofitamab, C3D1 to C12D1, is approximately equivalent to grofitamab C2D1; and each single dose of polatuzumab vedotin, C1D1 to C6D1, is approximately 1.8 mg / kg.

[0078] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the treatment of subjects having CD20-positive cell proliferative disorders, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein (a) the first dosing cycle comprises (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg); and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle; and (b) the second to sixth dosing cycles Each of the cycles includes a single dose of grofitamab (C2D1-C6D1) and a single dose of polatuzumab vedotin (C2D1-C6D1), where C2D1 of grofitamab is approximately 10 mg, 16 mg, or 30 mg; (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1-C12D1) and does not include administration of polatuzumab vedotin; each of the single doses of grofitamab Each dose C2D1 to C12D1 is administered on day 1 (±1 day) of each dosing cycle, and each single dose of polatuzumab vedotin C2D1 to C6D1 is administered on day 1 (±1 day) of each dosing cycle. Here, each single dose of grofitamab C3D1 to C12D1 is approximately equivalent to grofitamab C2D1, and each single dose of polatuzumab vedotin C1D1 to C6D1 is approximately 1.8 mg / kg.

[0079] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the manufacture of a pharmacopoeia for the treatment of subjects having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, wherein (a) the first dosing cycle comprises (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg); and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle; and (b) the second to sixth dosing cycles Each administration cycle includes a single dose of grofitamab (C2D1-C6D1) and a single dose of polatuzumab vedotin (C2D1-C6D1), where C2D1 of grofitamab is approximately 10 mg, 16 mg, or 30 mg; (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1-C12D1) and does not include administration of polatuzumab vedotin; Each single dose C2D1 to C12D1 is administered on day 1 (±1 day) of each dosing cycle, and each single dose of polatuzumab vedotin C2D1 to C6D1 is administered on day 1 (±1 day) of each dosing cycle. Here, each single dose of grofitamab C3D1 to C12D1 is approximately equivalent to grofitamab C2D1, and each single dose of polatuzumab vedotin C1D1 to C6D1 is approximately 1.8 mg / kg.

[0080] In one embodiment, the present invention relates to a method for treating a subject having a CD20-positive cell proliferative disorder, comprising administering polatuzumab vedotin and grofitamab to the subject in a dosing regimen comprising 12 dosing cycles, wherein (a) the first dosing cycle comprises (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg) and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle; and (b) the second to sixth dosing cycles Each cycle includes a single dose of grofitamab (C2D1-C6D1) and a single dose of polatuzumab vedotin (C2D1-C6D1), where C2D1 of grofitamab is approximately 30 mg; (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1-C12D1) and does not include administration of polatuzumab vedotin; each single dose of grofitamab C2D Dose 1-C12D1 is administered on day 1 (±1 day) of each dosing cycle, and each single dose of polatuzumab vedotin C2D1-C6D1 is administered on day 1 (±1 day) of each dosing cycle. Here, each single dose of grofitamab C3D1-C12D1 is approximately equivalent to grofitamab C2D1, and each single dose of polatuzumab vedotin C1D1-C6D1 is approximately 1.8 mg / kg.

[0081] In another embodiment, the present invention features polatuzumab vedotin and grofitamab for use in a method of treating subjects having CD20-positive cell proliferative disorders, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg); and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle; (b) the Each of the 2nd to 6th administration cycles includes a single dose of grofitamab (C2D1 to C6D1) and a single dose of polatuzumab vedotin (C2D1 to C6D1), where C2D1 of grofitamab is approximately 30 mg; (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1 to C12D1) and does not include administration of polatuzumab vedotin; Each single dose C2D1 to C12D1 is administered on day 1 (±1 day) of each dosing cycle, each single dose C2D1 to C6D1 of polatuzumab vedotin is administered on day 1 (±1 day) of each dosing cycle, each single dose C3D1 to C12D1 of grofitamab is approximately equivalent to C2D1 of grofitamab, and each single dose C1D1 to C6D1 of polatuzumab vedotin is approximately 1.8 mg / kg.

[0082] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the treatment of subjects having CD20-positive cell proliferative disorders, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg); and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle; and (b) the second to sixth... Each administration cycle includes a single dose of grofitamab (C2D1-C6D1) and a single dose of polatuzumab vedotin (C2D1-C6D1), where grofitamab C2D1 is approximately 30 mg; (c) Each of the 7th-12th administration cycles includes a single dose of grofitamab (C7D1-C12D1) and does not include administration of polatuzumab vedotin; each single dose of grofitamab C 2D1 to C12D1 are administered on day 1 (±1 day) of each dosing cycle, and each single dose of polatuzumab vedotin C2D1 to C6D1 is administered on day 1 (±1 day) of each dosing cycle. Here, each single dose of grofitamab C3D1 to C12D1 is approximately equivalent to grofitamab C2D1, and each single dose of polatuzumab vedotin C1D1 to C6D1 is approximately 1.8 mg / kg.

[0083] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the manufacture of a pharmacopoeia for the treatment of subjects having a CD20-positive cell proliferative disorder, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg); and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle; (b) the second to the second Each of the 6 administration cycles includes a single dose of grofitamab (C2D1-C6D1) and a single dose of polatuzumab vedotin (C2D1-C6D1), where C2D1 of grofitamab is approximately 30 mg; (c) Each of the 7th-12th administration cycles includes a single dose of grofitamab (C7D1-C12D1) and does not include administration of polatuzumab vedotin; each single dose of grofitamab C2D1 to C12D1 are administered on day 1 (±1 day) of each dosing cycle, and each single dose of polatuzumab vedotin C2D1 to C6D1 is administered on day 1 (±1 day) of each dosing cycle. Here, each single dose of grofitamab C3D1 to C12D1 is approximately equivalent to grofitamab C2D1, and each single dose of polatuzumab vedotin C1D1 to C6D1 is approximately 1.8 mg / kg.

[0084] In some embodiments, the administration cycle is 14 days (e.g., 14 ± 3 days). In some embodiments, the administration cycle is 21 days (e.g., 21 ± 3 days).

[0085] In some embodiments, grofitamab is administered intravenously. In some embodiments, polatuzumab vedotin is administered intravenously. In some embodiments, if grofitamab and polatuzumab vedotin are administered on the same day or if they are administered on the same day, grofitamab is administered after the administration of polatuzumab vedotin is completed. In some embodiments, grofitamab is administered or will be administered approximately 60 to 120 minutes after the completion of polatuzumab administration (e.g., approximately 60 to 100 minutes, approximately 60 to 90 minutes, approximately 60 to 80 minutes, approximately 90 to 120 minutes, approximately 80 to 100 minutes, approximately 80 to 120 minutes, approximately 75 to 105 minutes, or approximately 85 to 95 minutes; e.g., approximately 60 minutes, approximately 70 minutes, approximately 80 minutes, approximately 85 minutes, approximately 88 minutes, approximately 90 minutes, approximately 92 minutes, approximately 95 minutes, approximately 100 minutes, approximately 110 minutes, or approximately 120 minutes). In some embodiments, grofitamab is administered or will be administered approximately 90 minutes after the completion of polatuzumab vedotin administration.

[0086] In some embodiments, the method further includes administering obinutuzumab to the subject. In some embodiments, polatuzumab vedotin and grofitamab are administered together with obinutuzumab. In some embodiments, obinutuzumab is administered before or to administer grofitamab. In some embodiments, obinutuzumab is administered about 7 days (±1 day) before or to administer grofitamab. In some embodiments, obinutuzumab is administered as a single dose of about 1000 mg.

[0087] 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 lymphoma (NHL) or central nervous system lymphoma (CNSL). 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.

[0088] In some embodiments, NHL is DLBCL. In some embodiments, DLBCL is relapsed or refractory DLBCL.

[0089] In some embodiments, NHL is FL. In some embodiments, FL is relapsed or refractory FL. In some embodiments, FL is transformed FL.

[0090] In some embodiments, NHL is MCL. In some embodiments, MCL is relapsed or refractory MCL.

[0091] In some embodiments, CD20-positive cell proliferative disorders are not chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), Richter transformation, Burkitt lymphoma, or lymphoplasmacytic lymphoma.

[0092] In one embodiment, the present invention relates to a method for treating a population of subjects having R / R NHL, comprising administering to the subjects, in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, the method comprising: (a) the first dosing cycle comprising: a first dose (C1D1) of the bispecific antibody administered on day 8 (±1) of the first dosing cycle; a second dose (C1D2) of the bispecific antibody administered on day 15 (±1) of the first dosing cycle; and a third dose administered on day 2 (±1) of the first dosing cycle. (b) The second administration cycle includes a single dose (C1D1) of the bispecific antibody, where C1D1 is approximately 2.5 mg and C1D2 is approximately 10 mg; (b) The second administration cycle includes a single dose (C2D1) of the bispecific antibody administered on day 1 (±1 day) of the second administration cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1 day) of the second administration cycle, where 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.

[0093] In another embodiment, the present invention features an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating 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 conjugating to CD20 and CD3 are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, (a) the first dosing cycle comprising a first dose (C1D1) of the bispecific antibody administered on day 8 (±1) of the first dosing cycle, a second dose (C1D2) of the bispecific antibody administered on day 15 (±1) of the first dosing cycle, and the first dosing cycle (b) The second administration cycle includes a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1 day) of the cycle, where C1D1 of the bispecific antibody is approximately 2.5 mg and C1D2 of the bispecific antibody is approximately 10 mg; (b) The second administration cycle includes a single dose (C2D1) of the bispecific antibody administered on day 1 (±1 day) of the second administration cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1 day) of the second administration cycle, where 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.

[0094] In another embodiment, the present invention is characterized by the use of an anti-CD79b antibody drug conjugate and a bispecific antibody conjugated to CD20 and CD3 for treating a target population having R / R NHL, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugated to CD20 and CD3 are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, (a) the first dosing cycle comprising a first dose (C1D1) of the bispecific antibody administered on day 8 (±1) of the first dosing cycle, a second dose (C1D2) of the bispecific antibody administered on day 15 (±1) of the first dosing cycle, and the first dosing cycle (b) The second administration cycle includes a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1 day) of the cycle, where C1D1 of the bispecific antibody is approximately 2.5 mg and C1D2 of the bispecific antibody is approximately 10 mg; (b) The second administration cycle includes a single dose (C2D1) of the bispecific antibody administered on day 1 (±1 day) of the second administration cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1 day) of the second administration cycle, where 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.

[0095] In another embodiment, the present invention is characterized by the use of an anti-CD79b antibody drug conjugate and a bispecific antibody conjugated to CD20 and CD3 in the manufacture of a pharmacopoeci for the treatment of a target population having R / R NHL, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugated to CD20 and CD3 are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, (a) the first dosing cycle comprising a first dose (C1D1) of the bispecific antibody administered on day 8 (±1) of the first dosing cycle, a second dose (C1D2) of the bispecific antibody administered on day 15 (±1) of the first dosing cycle, and the first (b) The second administration cycle includes a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1 day) of the administration cycle, where C1D1 of the bispecific antibody is approximately 2.5 mg and C1D2 of the bispecific antibody is approximately 10 mg; (b) The second administration cycle includes a single dose (C2D1) of the bispecific antibody administered on day 1 (±1 day) of the second administration cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1 day) of the second administration cycle, where 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.

[0096] In one embodiment, the present invention relates to a method for treating a population of subjects having R / R NHL, comprising administering to the subjects, in a dosing regimen comprising 12 dosing cycles, the method comprising: (a) the first dosing cycle comprising: (i) a first dose (C1D1) of the bispecific antibody administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of the bispecific antibody administered on day 15 (±1) of the first dosing cycle (where 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 administered on day 2 (±1) of the first dosing cycle; and (b) the second Each of the six administration cycles includes a single dose of the bispecific antibody (C2D1-C6D1) and a single dose of the anti-CD79b antibody drug conjugate (C2D1-C6D1). (c) Each of the seventh to twelfth administration cycles includes a single dose of the bispecific antibody (C7D1-C12D1) and does not include the administration of the anti-CD79b antibody drug conjugate; each single dose of the bispecific antibody C2D1-C12D1 is administered on day 1 (±1 day) of each administration cycle, and each single dose of the anti-CD79b antibody drug conjugate C2D1-C6D1 is administered on day 1 (±1 day) of each administration cycle. Each single dose of the bispecific antibody C2D1-C12D1 is approximately 30 mg, and each single dose of the anti-CD79b antibody drug conjugate C1D1-C6D1 is approximately 1.8 mg / kg.

[0097] In another embodiment, the present invention features an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating 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 conjugating to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose (C1D1) of the bispecific antibody administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of the bispecific antibody administered on day 15 (±1) of the first dosing cycle (where C1D1 is approximately 2.5 mg and C1D2 is approximately 10 mg), and (ii) the anti-CD79b antibody drug conjugate administered on day 2 (±1) of the first dosing cycle. (b) Each of the 2nd to 6th administration cycles includes a single dose (C1D1) of the bispecific antibody and a single dose (C2D1 to C6D1) of the anti-CD79b antibody drug conjugate; (c) Each of the 7th to 12th administration cycles includes a single dose (C7D1 to C12D1) of the bispecific antibody and does not include administration of the anti-CD79b antibody drug conjugate; Each single dose of the bispecific antibody C2D1-C12D1 is administered on day 1 (±1 day) of each administration cycle, and each single dose of the anti-CD79b antibody drug conjugate C2D1-C6D1 is administered on day 1 (±1 day) of each administration cycle. Each single dose of this bispecific antibody C2D1-C12D1 is approximately 30 mg, and each single dose of the anti-CD79b antibody drug conjugate C1D1-C6D1 is approximately 1.8 mg / kg.

[0098] In another embodiment, the present invention is characterized by the use of an anti-CD79b antibody drug conjugate and a bispecific antibody conjugating to CD20 and CD3 for treating a population of subjects having R / R NHL, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugating to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose (C1D1) of the bispecific antibody administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of the bispecific antibody administered on day 15 (±1) of the first dosing cycle (where C1D1 is approximately 2.5 mg and C1D2 is approximately 10 mg), and (ii) a single dose of the anti-CD79b antibody drug conjugate administered on day 2 (±1) of the first dosing cycle. (b) Each of the 2nd to 6th administration cycles includes a single dose of the bispecific antibody (C2D1 to C6D1) and a single dose of the anti-CD79b antibody drug conjugate (C2D1 to C6D1), and (c) Each of the 7th to 12th administration cycles includes a single dose of the bispecific antibody (C7D1 to C12D1) and does not include administration of the anti-CD79b antibody drug conjugate; the bispecific antibody Each single dose C2D1 to C12D1 of the bispecific antibody is administered on day 1 (±1 day) of each administration cycle, and each single dose C2D1 to C6D1 of the anti-CD79b antibody drug conjugate is administered on day 1 (±1 day) of each administration cycle. Each single dose C2D1 to C12D1 of this bispecific antibody is approximately 30 mg, and each single dose C1D1 to C6D1 of the anti-CD79b antibody drug conjugate is approximately 1.8 mg / kg.

[0099] In another embodiment, the present invention is characterized by the use of an anti-CD79b antibody drug conjugate and a bispecific antibody conjugated to CD20 and CD3 in the manufacture of a pharmacopoeci for the treatment of a target population having R / R NHL, wherein the anti-CD79b antibody drug conjugate and the bispecific antibody conjugated to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose (C1D1) of the bispecific antibody administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of the bispecific antibody administered on day 15 (±1) of the first dosing cycle (where C1D1 is approximately 2.5 mg and C1D2 is approximately 10 mg), and (ii) the anti-CD79b antibody drug conjugate administered on day 2 (±1) of the first dosing cycle (b) each of the 2nd to 6th administration cycles includes a single dose (C1D1) of the bispecific antibody and a single dose (C2D1 to C6D1) of the anti-CD79b antibody drug conjugate; (c) each of the 7th to 12th administration cycles includes a single dose (C7D1 to C12D1) of the bispecific antibody and does not include administration of the anti-CD79b antibody drug conjugate; this bispecific Each single dose of the antibody, C2D1 to C12D1, is administered on day 1 (±1 day) of each dosing cycle, and each single dose of the anti-CD79b antibody drug conjugate, C2D1 to C6D1, is administered on day 1 (±1 day) of each dosing cycle. Each single dose of this bispecific antibody, C2D1 to C12D1, is approximately 30 mg, and each single dose of the anti-CD79b antibody drug conjugate, C1D1 to C6D1, is approximately 1.8 mg / kg.

[0100] In one embodiment, the present invention is a method for treating a population of subjects having R / R NHL, comprising administering polatuzumab vedotin and grofitamab to the subjects 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 grofitamab administered on day 8 (±1) of the first dosing cycle, a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle, and polatuzumab administered on day 2 (±1) of the first dosing cycle. (b) The second administration cycle includes a single dose of buvedotin (C1D1), where grofitamab C1D1 is approximately 2.5 mg and grofitamab C1D2 is approximately 10 mg; (b) The second administration cycle includes a single dose of grofitamab (C2D1) administered on day 1 (±1) of the second administration cycle and a single dose of polatuzumab vedotin (C2D1) administered on day 1 (±1) of the second administration cycle, where grofitamab C2D1 is approximately 30 mg and polatuzumab vedotin C1D1 and C1D2 are each approximately 1.8 mg / kg.

[0101] In another embodiment, the present invention features polatuzumab vedotin and grofitamab for use in a method of treating a target population having R / R NHL, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, (a) the first dosing cycle comprising a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle, a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle, and (b) administered on day 2 (±1) of the first dosing cycle (b) The second administration cycle includes a single dose (C1D1) of polatuzumab vedotin, where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg; (b) The second administration cycle includes a single dose (C2D1) of grofitamab administered on day 1 (±1) of the second administration cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1) of the second administration cycle, where C2D1 of grofitamab is approximately 30 mg and C1D1 and C1D2 of polatuzumab vedotin are each approximately 1.8 mg / kg.

[0102] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in a method for treating a population of subjects having R / R NHL, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, (a) the first dosing cycle comprising a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle, a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle, and (b) administered on day 2 (±1) of the first dosing cycle (b) The second administration cycle includes a single dose of polatuzumab vedotin (C1D1), where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg; (b) The second administration cycle includes a single dose of grofitamab (C2D1) administered on day 1 (±1) of the second administration cycle and a single dose of polatuzumab vedotin (C2D1) administered on day 1 (±1) of the second administration cycle, where C2D1 of grofitamab is approximately 30 mg and C1D1 and C1D2 of polatuzumab vedotin are each approximately 1.8 mg / kg.

[0103] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the manufacture of a pharmacopoeci for the treatment of a target population having R / R NHL, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, (a) the first dosing cycle comprising: a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle; a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle; and (b) administered on day 2 (±1) of the first dosing cycle. (b) The second administration cycle includes a single dose (C1D1) of polatuzumab vedotin administered on day 1 (±1) of the second administration cycle and a single dose (C2D1) of polatuzumab vedotin administered on day 1 (±1) of the second administration cycle, with a C2D1 of grofitamab being approximately 30 mg and C1D1 and C1D2 of polatuzumab vedotin being approximately 1.8 mg / kg each.

[0104] In one embodiment, the present invention is a method for treating a population of subjects having R / R NHL, comprising administering polatuzumab vedotin and grofitamab to the subjects in a dosing regimen comprising 12 dosing cycles, wherein (a) the first dosing cycle comprises (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg) and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle; and (b) the second to sixth Each of the following administration cycles includes a single dose of grofitamab (C2D1-C6D1) and a single dose of polatuzumab vedotin (C2D1-C6D1); (c) Each of the 7th-12th administration cycles includes a single dose of grofitamab (C7D1-C12D1) and does not include administration of polatuzumab vedotin; each single dose of grofitamab C2D1-C12D1 is administered on day 1 (±1 day) of each administration cycle, and each single dose of polatuzumab vedotin C2D1-C6D1 is administered on day 1 (±1 day) of each administration cycle, where each single dose of grofitamab C2D1-C12D1 is approximately 30 mg / kg and each single dose of polatuzumab vedotin C1D1-C6D1 is approximately 1.8 mg / kg.

[0105] In another embodiment, the present invention features polatuzumab vedotin and grofitamab for use in a method of treating a population of subjects having R / R NHL, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg); and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle. (b) Each of the 2nd to 6th administration cycles includes a single dose of grofitamab (C2D1 to C6D1) and a single dose of polatuzumab vedotin (C2D1 to C6D1); (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1 to C12D1) and no administration of polatuzumab vedotin; each single dose of grofitamab C2D1 to C12D1 is administered on day 1 (±1 day) of each administration cycle, and each single dose of polatuzumab vedotin C2D1 to C6D1 is administered on day 1 (±1 day) of each administration cycle, with each single dose of grofitamab C2D1 to C12D1 being approximately 30 mg and each single dose of polatuzumab vedotin C1D1 to C6D1 being approximately 1.8 mg / kg.

[0106] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the treatment of a target population having R / R NHL, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg); and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle; b) Each of the 2nd to 6th administration cycles includes a single dose of grofitamab (C2D1 to C6D1) and a single dose of polatuzumab vedotin (C2D1 to C6D1); (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1) and does not include administration of polatuzumab vedotin; each single dose of grofitamab C2D1 to C12D1 is administered on day 1 (±1 day) of each administration cycle, and each single dose of polatuzumab vedotin C2D1 to C6D1 is administered on day 1 (±1 day) of each administration cycle, where each single dose of grofitamab C2D1 to C12D1 is approximately 30 mg / kg and each single dose of polatuzumab vedotin C1D1 to C6D1 is approximately 1.8 mg / kg.

[0107] In another embodiment, the present invention is characterized by the use of polatuzumab vedotin and grofitamab in the manufacture of a pharmacopoeci for the treatment of a target population having R / R NHL, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising 12 dosing cycles, (a) the first dosing cycle comprising (i) a first dose (C1D1) of grofitamab administered on day 8 (±1) of the first dosing cycle and a second dose (C1D2) of grofitamab administered on day 15 (±1) of the first dosing cycle (where C1D1 of grofitamab is approximately 2.5 mg and C1D2 of grofitamab is approximately 10 mg), and (ii) a single dose (C1D1) of polatuzumab vedotin administered on day 2 (±1) of the first dosing cycle. (b) Each of the 2nd to 6th administration cycles includes a single dose of grofitamab (C2D1 to C6D1) and a single dose of polatuzumab vedotin (C2D1 to C6D1); (c) Each of the 7th to 12th administration cycles includes a single dose of grofitamab (C7D1) and does not include administration of polatuzumab vedotin; each single dose of grofitamab C2D1 to C12D1 is administered on day 1 (±1 day) of each administration cycle, and each single dose of polatuzumab vedotin C2D1 to C6D1 is administered on day 1 (±1 day) of each administration cycle, with each single dose of grofitamab C2D1 to C12D1 being approximately 30 mg / kg and each single dose of polatuzumab vedotin C1D1 to C6D1 being approximately 1.8 mg / kg.

[0108] 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%; for example, between 20-100%, between 40-100%, between 60-100%, between 80-100%, between 20-80%, between 20-60%, between 20-40%, between 40-80%, between 40-60%, between 30-50%, or between 35-45%; for example, 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 certain embodiments, the complete response rate in the target population having R / R NHL is approximately 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., between 30-100%, between 50-100%, between 70-100%, between 30-90%, between 30-70%, between 30-50%, between 40-80%, between 40-60%, between 45-55%, or between 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 certain embodiments, the overall response rate in a population of subjects with R / R NHL is approximately 50%.

[0109] In some embodiments, the B-cell proliferative disorder is R / R MCL. In some embodiments, 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., between 60-100%, between 70-100%, between 80-100%, between 90-100%, between 60-90%, between 60-80%, between 60-70%, between 70-90%, between 80-90%, between 80-100%, or between 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 certain embodiments, the complete response rate in the population of subjects with R / R NHL is at least about 85%. In certain embodiments, the complete response rate in the population of subjects with R / R MCL is at least about 85%. In certain embodiments, the complete response rate in the population of subjects with R / R MCL is about 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., between 60-100%, between 70-100%, between 80-100%, between 90-100%, between 60-90%, between 60-80%, between 60-70%, between 70-90%, between 80-90%, between 80-100%, or between 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 higher). In certain embodiments, the overall response rate in the population of subjects having R / R NHL is at least about 85%. In certain embodiments, the overall response rate in the target population with R / R MCL is at least about 85%. In certain embodiments, the overall response rate in the target population with R / R MCL is about 100%. In some embodiments, the overall response rate is at least 80%.

[0110] 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., between 60-100%, between 70-100%, between 80-100%, between 90-100%, between 60-90%, between 60-80%, between 60-70%, between 60-65%, between 65-75%, or 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 certain embodiments, the complete response rate in the target population with R / R DLBCL is approximately 60%. In certain embodiments, the complete response rate in the target population with R / R DLBCL is approximately 65%. In certain embodiments, the complete response rate in the target population with R / R DLBCL is approximately 70%. In certain embodiments, the complete response rate in the target population with 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., between 60-100%, between 70-100%, between 80-100%, between 90-100%, between 60-90%, between 60-80%, between 60-70%, between 60-65%, between 65-75%, between 70-90%, or between 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 higher). In some embodiments, the overall response rate is at least 70%. In some embodiments, the overall response rate is at least 80%.In certain embodiments, the overall response rate in a population of subjects with R / R DLBCL is approximately 65%. In certain embodiments, the overall response rate in a population of subjects with R / R DLBCL is approximately 73%. In certain embodiments, the overall response rate in a population of subjects with R / R DLBCL is approximately 75%. In certain embodiments, the overall response rate in a population of subjects with R / R DLBCL is approximately 85%.

[0111] In some embodiments, 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., between 30-100%, between 50-100%, between 70-100%, between 35-90%, between 45-90%, between 35-70%, between 35-50%, between 40-80%, between 40-60%, between 45-55%, or between 35-45%; e.g., about 35%, about 40%, about 45%, about 48%, about 49%, about 50%, about 51%, about 52%, about 55%, about 60%, about 70%, about 75%, about 80%, about 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 certain embodiments, the complete response rate in the population of subjects with R / R DLBCL is approximately 46%. In certain embodiments, the complete response rate in the population of subjects with R / R DLBCL is approximately 52%. In certain embodiments, the complete response rate in the population of subjects with 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 higher; e.g., 85-100%). The overall response rates are between 87% and 100%, between 90% and 100%, between 95% and 100%, between 85% and 97%, between 85% and 95%, between 85% and 90%, between 85% and 87%, between 90% and 95%, or between 93% and 97%; for example, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or higher). In some embodiments, the overall response rate is at least 85%. In some embodiments, the overall response rate is at least 90%. In certain embodiments, the overall response rate in a population of subjects with R / R DLBCL is about 86%.

[0112] In some embodiments, the complete response rate is higher than the baseline complete response rate in a reference population of subjects treated with a combination therapy comprising an anti-CD20 / anti-CD3 bispecific antibody and an anti-PD-L1 antagonist antibody, but without an anti-CD79b antibody drug conjugate. In some embodiments, the objective response rate is higher than the baseline objective response rate in a reference population of subjects treated with a combination therapy comprising an anti-CD20 / anti-CD3 bispecific antibody and an anti-PD-L1 antagonist antibody, but without an anti-CD79b antibody drug conjugate. In some embodiments, the complete response rate is higher than the baseline complete response rate in a reference population of subjects treated with a combination therapy comprising grofitamab and atezolizumab, but without polatuzumab vedotin. In some embodiments, the objective response rate is higher than the baseline objective response rate in a reference population of subjects treated with a combination therapy comprising grofitamab and atezolizumab, but without polatuzumab vedotin.

[0113] In some embodiments, the subjects are human. In some embodiments, each subject in the subject population is human. In some embodiments, each subject in the reference population of subjects is human. In some embodiments, the subjects or subject population have received at least two prior systemic therapies (e.g., two, three, four, five, six or more prior systemic therapies). In some embodiments, the subjects or subject population are ineligible for autologous stem cell transplantation (SCT).

[0114] This application file includes at least one drawing prepared in color. A copy of this patent or patent application accompanied by the color drawing will be provided by the Patent Office upon request and payment of the necessary fees. [Brief explanation of the drawing]

[0115] [Figure 1A-1F] Figures 1A-1F are schematic diagrams showing the composition of exemplary anti-CD20 / anti-CD3 bispecific antibodies. [Figure 1G-1N] Figure 1G-1N is a schematic diagram showing the composition of an exemplary anti-CD20 / anti-CD3 bispecific antibody. [Figure 2] This is a schematic diagram showing the structure of grofitamab. [Figure 3] This is a schematic diagram illustrating the overview of the trial design described in Example 1. Atezo = Atezolizumab; CRM = Continuous reassessment method; DLBCL = Diffuse large B-cell lymphoma; ECOG = Eastern Cooperative Oncology Group; EWOC = Escalation by overdose control; FL = Follicular lymphoma; Pola = Polatuzumab vedotin; R / R = Relapsed and / or refractory; SCT = Society for Clinical Trials; TCB = Grofitamab. [Figure 4A-4C]This is a schematic diagram showing the study design for the atezolizumab group (arm) (Figure 4A), the dose escalation phase of the polatuzumab group (Figure 4B), and the expansion phase of the polatuzumab group (Figure 4C) as described in Example 1. Atezo = Atezolizumab; CR = Complete response; DE = Dose escalation; DLBCL = Diffuse large B-cell lymphoma; EoS = End of study; F / U = Follow-up; Glofit = Glofitamab; NHL = Non-Hodgkin lymphoma; Pola = Polatuzumab vedotin; PD = Progressive disease; PR = Partial response; Pts = Patients; RP2D = Recommended Phase II dose; R / R = Relapsed and / or refractory; SD = Stable condition [Figure 5A-5B] This is a schematic diagram showing the timing of administration for the atezolizumab group (Figure 5A) and the polatuzumab vedotin group (Figure 5B) in the study described in Example 1. Atezo = Atezolizumab; Glofit = Glofitamab; Gpt = Prior treatment with Gazyva® (prior treatment with obinutuzumab); Pola = Polatuzumab vedotin. [Figure 6] This table lists the frequency of adverse events (AEs) with an incidence of ≥10% or grade 5 according to NCI-CTCAE for safety evaluation patients in cohorts 1 and 2 who had previously received treatment with grofitamab + polatuzumab vedotin. The color indicates the grade of the AE. All AEs during the study are listed on the left. Only AEs considered to be related to the study treatment (e.g., grofitamab or polatuzumab vedotin) are listed on the right. [Figure 7] This shows the latest efficacy data at the clinical cutoff date for dose escalation cohorts 1 and 2, as well as the expanded cohort, in RP2D. Glofit = grofitamab. Pola = polatuzumab vedotin. [Modes for carrying out the invention]

[0116] The present invention provides a method for treating a subject having a CD20-positive cell proliferative disorder (e.g., B-cell proliferative disorder (e.g., non-Hodgkin 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), or mantle cell lymphoma (MCL) (e.g., relapsed or refractory MCL)), or central nervous system lymphoma (CNSL))), comprising administering an anti-CD79b antibody drug conjugate and / or an anti-CD20 / anti-CD3 bispecific antibody to the subject, for example, in a divided dose escalation regimen. The method comprises at least a first administration cycle and a second administration cycle, wherein (a) the first administration cycle comprises a first dose (C1D1) and a second dose (C1D2) of the bispecific antibody, with C1D1 being approximately 2.5 mg and C1D2 being approximately 10 mg; and (b) the second administration cycle comprises a single dose (C2D1) of the bispecific antibody, with C2D1 being approximately 10 mg, approximately 16 mg, or approximately 30 mg.

[0117] The present invention is in part based on the discovery that a divided dose escalation regimen involving the administration of a bispecific antibody (e.g., grofitamab) that binds to CD20 and CD3 over multiple administration cycles (e.g., the first administration cycle being a step-up divided dose cycle) can very effectively treat subjects with CD20-positive cell proliferative disorders (e.g., B-cell proliferative disorders) and an acceptable safety profile (e.g., with respect to cytokine release syndrome).

[0118] I. General Techniques Unless otherwise stated, the implementation of this invention utilizes prior art in molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology, which are within the scope of the art of those skilled in the art. Such techniques are well described in the following literature, for example: "Molecular Cloning: A Laboratory Manual", 2nd edition (Sambrook et al., 1989); "Oligonucleotide Synthesis" (MJ Gait, ed., 1984); "Animal Cell Culture" (RI Freshney, ed., 1987); "Methods in Enzymology" (Academic Press, Inc.); "Current Protocols in Molecular Biology" (FMAusubel 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).

[0119] II. Definition The terms used herein shall be used as commonly used in the art unless otherwise defined below.

[0120] As used herein, the terms “differentiation antigen group 20” or “CD20” refer to any natural CD20 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise indicated. CD20 (also known as B lymphocyte antigen CD20, B lymphocyte surface antigen B1, Leu-16, Bp35, BM5, and LF5; this human protein is characterized in UniProt database entry P11836) is a hydrophobic transmembrane protein with a molecular weight of approximately 35 kD expressed in pre-B lymphocytes and mature B lymphocytes (Valentine, MA et al., J. Biol. Chem. 264 (1989) 11282-11287; Tedder, TF, et al., Proc. Natl. Acad. Sci. USA 85 (1988) 208-212; Stamenkovic, I., et al., J. Exp. Med. 167 (1988) 1975-1980; Einfeld, DA, et al., EMBO J.7(1988)711-717; Tedder, TF, 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 nascent protein family are characterized by common structural features and similar intron / exon splice boundaries, and exhibit unique expression patterns 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 to 11q12 within the cluster of family members. The term encompasses not only "full-length," raw CD20 but also any form of CD20 resulting from intracellular processing. The term also encompasses naturally occurring variants of CD20, such as splice variants or allele variants. Alternative splicing of this gene results in two transcriptional variants encoding the same protein. In one embodiment, CD20 is human CD20.

[0121] The terms “anti-CD20 antibody” and “CD20-binding antibody” refer to an antibody capable of binding to CD20 with sufficient affinity to be useful as a diagnostic and / or therapeutic agent in targeting CD20. In some embodiments, the degree of binding of an anti-CD20 antibody to unrelated non-CD20 proteins is less than about 10% of the binding of an antibody to CD20 as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the antibody that binds to CD20 has a degree 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, 10 -8 ~10 -13 M, for example 10 -9 M~10 -13 The dissociation constant (K) of M D ) has. In certain embodiments, the anti-CD20 antibody binds to CD20 epitopes that are conserved among different species of CD20.

[0122] "Type II anti-CD20 antibody" refers to an anti-CD20 antibody that possesses the binding characteristics and biological activity of a type II anti-CD20 antibody, 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), 22-333, and summarized in Table 1 below. Table 1. Characteristics of Type I and Type II anti-CD20 antibodies TIFF0007836317000001.tif70170

[0123] Examples of type II anti-CD20 antibodies include, for example, obinutuzumab (GA101), tositumomab (B1), humanized B-Ly1 antibody IgG1 (a chimeric humanized IgG1 antibody disclosed in International Publication No. 2005 / 044859), 11B8 IgG1 (disclosed in International Publication No. 2004 / 035607), and AT80 IgG1.

[0124] Examples of type I anti-CD20 antibodies include, for example, rituximab, ofatumumab, vertuzumab, okalatuzumab, ocrelizumab, PRO131921, ubrituximab, HI47 IgG3 (ECACC, hybridoma), 2C6 IgG1 (disclosed in International Publication No. 2005 / 103081), 2F2 IgG1 (disclosed in International Publication No. 2004 / 035607 and International Publication No. 2005 / 103081), and 2H7 IgG1 (disclosed in International Publication No. 2004 / 056312).

[0125] Unless otherwise specified, “CD3” refers to any natural CD3 from any vertebrate source, including mammals such as primates (e.g., humans), non-human primates (e.g., cynomolgus monkeys), and rodents (e.g., mice and rats). This term encompasses not only “full-length,” raw CD3, but also any form of CD3 resulting from intracellular processing. This term also encompasses naturally occurring variants of CD3, such as splice variants or allele variants. In some embodiments, CD3 refers to human CD3, in particular the epsilon subunit of human CD3 (CD3ε). The amino acid sequence of human CD3ε is shown in 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 monkey [Macaca fascicularis] CD3ε is shown in NCBI GenBank number BAB71849.1.

[0126] The terms "anti-CD20 antibody / anti-CD3 bispecific antibody" and "bispecific antibody that binds to CD20 and CD3" refer to bispecific antibodies that can bind to both CD20 and CD3 with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent in targeting CD20 and / or CD3. In certain embodiments, the degree of binding of a bispecific antibody that binds to CD20 and CD3 to unrelated non-CD3 proteins and / or non-CD20 proteins is, for example, less than about 10% of the binding of the antibody to CD3 and / or CD20, measured by, for example, radioimmunoassay (RIA). In certain embodiments, the bispecific antibody that binds to CD20 and CD3 has a dissociation constant (K D ) 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, e.g., 10 -8 M to 10 -13 M, e.g., 10 -9 M to 10 -13 M). In certain embodiments, the bispecific antibody that binds to CD20 and CD3 binds to an epitope of CD3 that is conserved among different species of CD3 and / or an epitope of CD20 that is conserved among different species of CD20. An example of an anti-CD20 / anti-CD3 bispecific antibody is glofitamab (Proposed International Nonproprietary Name: List 121 WHO Drug Information, Vol. 33, No. 2, 2019, page 276; also known as CD20-TCB, RO7082859 or RG6026; CAS number: 2229047-91-8).

[0127] As used herein, the terms “differentiation antigen group 79b” or “CD79b” refer to any natural CD79b from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. The term encompasses “full-length,” untreated CD79b, and any form of CD79b resulting from intracellular processing. The term also encompasses naturally occurring variants of CD79b, including, for example, splice variants or allele variants. CD79b includes, for example, the human CD79b protein with a length of 229 amino acids (NCBI RefSeq number NP_000617).

[0128] The terms “anti-CD79b antibody” and “antibody that binds to CD79b” refer to an antibody that can bind to CD79b with sufficient affinity to be useful as a diagnostic and / or therapeutic agent in targeting CD79b. In some embodiments, the degree of binding of an anti-CD79b antibody to an unrelated non-CD79b protein is less than about 10% of the binding of the antibody to CD79b as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the antibody that binds to CD79b has a viscosity 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, 10 -8 M~10 -13 M, for example 10 -9 M~10 -13 It has a dissociation constant (KD) of M). In certain embodiments, the anti-CD79b antibody binds to CD79b epitopes that are conserved among different species of CD79b.

[0129] As used herein, the terms “cytokine release” or “cytokine release” are synonymous with “cytokine storm” or “cytokine release syndrome” (abbreviated as “CRS”), and refer to the elevated levels of cytokines in the target blood, particularly 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), during or immediately after administration of a therapeutic agent (e.g., within 1 day after administration), and the resulting adverse symptoms. Cytokines are defined as the hyperphysiological response following the administration of any immunotherapy resulting in the activation or involvement of endogenous or infused T cells and / or other immune effector cells. Symptoms may be progressive and always include fever at onset, and may include hypotension, capillary leakage (hypoxia), and terminal organ dysfunction (Lee et al. 2019). In some cases, for example after CAR-T cell administration, CRS may develop several days after administration as the CAR-T cells proliferate. The incidence and severity are typically reduced with subsequent fluid administration. Symptoms can range from symptomatic discomfort to fatal events and may include fever, chills, dizziness, hypertension, hypotension, dyspnea, restlessness, sweating, flushing, skin rash, tachycardia, tachypnea, headache, tumor pain, nausea, vomiting, and / or organ failure.

[0130] As used herein, the term "amino acid mutation" encompasses substitutions, deletions, insertions, and modifications of amino acids. Substitutions, deletions, insertions, and modifications can be combined in any way to obtain the final construct, provided that the final construct has a desired characteristic, such as reduced binding to the Fc receptor. Deletions and insertions of amino acid sequences include deletions and insertions of the amino-terminus and / or carboxy-terminus of amino acids. A specific amino acid mutation is an amino acid substitution. For example, to alter the binding properties of the Fc region, non-conservative amino acid substitutions, i.e., replacing one amino acid with another amino acid having different structural and / or chemical properties, are particularly preferred. Amino acid substitutions include substitutions with non-natural amino acids or with natural amino acid derivatives of 20 standard amino acids (e.g., 4-hydroxyproline, 3-methylhistidine, ornithine, homoserine, 5-hydroxylysine). Amino acid mutations can be induced using genetic or chemical methods well known in the art. Genetic methods may include site-directed mutagenesis, PCR, gene synthesis, etc. Methods other than genetic engineering, such as chemical modification to alter the side chain groups of amino acids, may also be useful. In this specification, various notations are used to indicate the same amino acid mutation. For example, the substitution of proline to glycine at position 329 of the Fc region is expressed as 329G, G329, G 329 It may be indicated as P329G or Pro329Gly.

[0131] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., a receptor) and its binding partner (e.g., a ligand). As used herein, "binding affinity" refers to the intrinsic binding affinity that reflects the 1:1 interaction between members of a binding pair (e.g., receptor and ligand), unless otherwise specified. The affinity of molecule X for its partner Y is generally expressed by the dissociation constant (K). D ) is expressed as the dissociation rate constant and the association rate constant (where k is the relative position of the dissociation rate constant and the association rate constant, respectively). off , k onThis is the ratio of the rate constants. Therefore, as long as the ratio of these rate constants is the same, equivalent affinities can include different rate constants. Affinity can be measured by established methods known in the art. A specific method for measuring affinity is surface plasmon resonance (SPR).

[0132] An "affinity-mature" antibody refers to an antibody that has one or more modifications in one or more hypervariable regions (HVRs) compared to a parent antibody that does not have such modifications, and such modifications result in an improved affinity of the antibody to the antigen.

[0133] As used herein, the term “antigen-binding moiety” refers to a polypeptide molecule that specifically binds to an antigenic determinant. In some embodiments, the antigen-binding moiety can direct the entity to which it binds (e.g., a cytokine or a second antigen-binding moiety) to a target site, for example, to a specific type of tumor cell or tumor stroma containing an antigenic determinant. Antigen-binding moieties include antibodies and fragments thereof, as further defined herein. Preferred antigen-binding moieties include the antigen-binding domain of an antibody, including an antibody heavy chain variable region and an antibody light chain variable region. In certain embodiments, the antigen-binding moiety may include an antibody constant region, as further defined herein and known in the art. Useful heavy chain constant regions include any of five isotypes: α, δ, ε, γ, or μ. Useful light chain constant regions include any of two isotypes: κ and λ.

[0134] "To bind," "to bind specifically," or "specific to" means that the binding is selective to the antigen and can be distinguished from undesirable or nonspecific interactions. The ability of an antigen-binding moiety to bind to a specific antigenic determinant can also be measured by enzyme-linked immunosorbent assay (ELISA) or other techniques well known to those skilled in the art, such as surface plasmon resonance (analysis using a BIAcore instrument) (Liljeblad et al., Glyco J. 17, 323-329 (2000)) and classical binding assays (Heeley, Endocr Res. 28, 217-229 (2002)). In some embodiments, the degree of binding of an antigen-binding moiety to an unrelated protein is less than about 10% of the binding of the antigen-binding moiety to the antigen, as measured, for example, by SPR. In certain embodiments, the antigen-binding portion that binds to the antigen, or the antigen-binding molecule containing the antigen-binding portion, has a minimum size 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, 10 -8 M to 10 -13 M, for example 10 -9 M to 10 -13 The dissociation constant (K) of M D ) has.

[0135] "Decreased binding," for example, decreased binding to the Fc receptor, refers to a decrease in the affinity of the corresponding interaction, as measured, for example, by SPR. For clarity, this term also includes a decrease in affinity to zero (or below the detection limit of the analytical method), i.e., complete termination of the interaction. Conversely, "increased binding" refers to an increase in the binding affinity of the corresponding interaction.

[0136] As used herein, the term “antigen-binding molecule” refers in its broadest sense to a molecule that specifically binds to an antigenic determinant. Examples of antigen-binding molecules include immunoglobulins and their derivatives, such as fragments.

[0137] As used herein, the term “antigenic determinant” is synonymous with “antigen” and “epitope” and refers to a site on a polypeptide macromolecule (e.g., a continuous stretch of amino acids or a conformational configuration consisting of different regions of discontinuous amino acids) to which an antigen-binding moiety binds to form an antigen-antigen complex. Useful antigenic determinants can be found, for example, on the surface of tumor cells, on the surface of virus-infected cells, on the surface of other diseased cells, free in serum, and / or in the extracellular matrix (ECM). Unless otherwise specified, the term “antigen” as used herein refers to the native form of a protein from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). In certain embodiments, the antigen is a human protein. When this term refers to a particular protein as used herein, it includes “full-length,” raw protein, and any form of protein obtained from intracellular processing. This term also includes naturally occurring variants of a protein, such as splice variants or allele variants. An exemplary human protein useful as an antigen is CD3, particularly the epsilon subunit of CD3 (see UniProt number P07766 (version 130), NCBI RefSeq number NP_000724.1 for the human sequence; or UniProt number Q95LI5 (version 49), NCBI GenBank number BAB71849.1 for the cynomolgus monkey [Macaca fascicularis] sequence). In certain embodiments, the T cell activation bispecific molecules described herein bind to CD3 or target cell antigen epitopes that are conserved among various species of CD3 or target antigens.

[0138] As used herein, the term "polypeptide" refers to a molecule composed of monomers (amino acids) linked in a linear chain by amide bonds (also known as peptide bonds). The term "polypeptide" refers to a chain of two or more amino acids, not to a specific length of the product. Therefore, any other term used to refer to a peptide, dipeptide, tripeptide, oligopeptide, "protein," "amino acid chain," or a chain of two or more amino acids falls within the definition of "polypeptide," and the term "polypeptide" can be used in place of or interchangeably with these terms. The term "polypeptide" is also intended to refer to the products of post-expression modifications of a polypeptide, such modifications including, but not limited to, glycosylation, acetylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, protein cleavage, or modification with non-natural amino acids. Polypeptides may be obtained from natural biological sources or produced by recombinant techniques, but are not necessarily translated from a specified nucleic acid sequence. Polypeptides may be produced by any method, including chemical synthesis. The polypeptides of the present invention may be of a size consisting of 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, 1000 or more, or 2000 or more amino acids. Polypeptides may have a clearly defined three-dimensional structure, but do not necessarily have such a structure. Polypeptides having a clearly defined three-dimensional structure are said to be "folded," while polypeptides that do not have a clearly defined three-dimensional structure and can adopt many different conformations are said to be "unfolded."

[0139] An "isolated" polypeptide or variant or derivative thereof means a polypeptide that is not found in its natural environment. Purification is not particularly necessary. For example, an isolated polypeptide can be taken from its natural or natural environment. Recombinant production polypeptides and proteins expressed in host cells are considered isolated for the purposes of this invention, as are natural or recombinant polypeptides that have been isolated, fragmented, or partially or substantially purified by any appropriate technique.

[0140] The “amino acid sequence identity percentage (%)” for 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 of the reference polypeptide, after the sequences have been aligned and gaps introduced where necessary to obtain the maximum sequence identity percentage, and no conservative substitutions are considered as part of the sequence identity. Alignment for the purpose of determining the amino acid sequence identity percentage can be achieved in various ways within the scope of the art using commonly available computer software, such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning the sequences, including any algorithm necessary to achieve the maximum alignment over the entire length of the sequences being compared. However, as used herein, the amino acid sequence identity % values ​​are generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was created by Genentech, Inc., and its source code, along with user documentation, has been filed with the U.S. Copyright Office, Washington DC, 20559, where it is registered under U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is generally available from Genentech, Inc. in South San Francisco, California, or can be compiled from its source code. The ALIGN-2 program needs to 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 remain unchanged. In situations where ALIGN-2 is used for amino acid sequence comparison, the amino acid sequence identity % of a given amino acid sequence A to, with, or relative to a given amino acid sequence B (or, this can be described as a given amino acid sequence A having or containing a specific amino acid sequence identity % to, with, or relative to a given amino acid sequence B) is calculated as follows: 100 x fraction X / Y In the formula, X is the number of amino acid residues to which the sequence alignment program ALIGN-2 has scored a perfect match in the alignment of A and B, and Y is the total number of amino acid residues in B. It should be understood that if the length of amino acid sequence A is not equal to the length of amino acid sequence B, the amino acid sequence identity % of A to B is not equal to the amino acid sequence identity % of B to A. Unless otherwise noted, all amino acid sequence identity % values ​​used herein are obtained using the ALIGN-2 computer program as described in the preceding paragraph.

[0141] The term "antibody" as used herein is used in its broadest sense and is not limited to any particular form, but encompasses a variety of antibody structures, including monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, as long as they exhibit the desired antigen-binding activity.

[0142] The terms “full-length antibody,” “intact antibody,” and “whole antibody” are used interchangeably herein to refer to antibodies having a structure substantially similar to that of a natural antibody or antibodies having a heavy chain containing an Fc region as defined herein.

[0143] An "antibody fragment" refers to a molecule other than an intact antibody that binds to an antigen to which an intact antibody binds, and contains a portion of the intact antibody. 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. As used herein, the term "antibody fragment" also includes single-domain antibodies.

[0144] The term "immunoglobulin molecule" refers to a protein that has the structure of a naturally occurring antibody. For example, IgG class immunoglobulins are heterotetrameric glycoproteins with a weight of approximately 150,000 daltons, composed of two disulfide-bonded light chains and two heavy chains. From the N-terminus to the C-terminus, each heavy chain has a variable region (VH), also called a variable heavy chain domain or heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3), also called heavy chain constant domains. Similarly, from the N-terminus to the C-terminus, each light chain has a variable region (VL), also called a variable light chain domain or light chain variable domain, followed by a constant light (CL) domain, also called a light chain constant domain. The heavy chain of an immunoglobulin is assigned to one of five classes called α(IgA), δ(IgD), ε(IgE), γ(IgG), or μ(IgM), and some of these classes are further divided into subclasses such as γ1(IgG1), γ2(IgG2), γ3(IgG3), γ4(IgG4), α1(IgA1), and α2(IgA2). The light chain of an immunoglobulin is assigned to one of two types called kappa (κ) or lambda (λ) based on the amino acid sequence of its constant domain. An immunoglobulin essentially consists of two Fab molecules and one Fc domain linked via the hinge region of the immunoglobulin.

[0145] The term "antigen-binding domain" refers to a portion of an antibody that specifically binds to part or all of an antigen and includes a region complementary to part or all of the antigen. The antigen-binding domain may be provided, for example, by one or more antibody variable domains (also called antibody variable regions). Preferably, the antigen-binding domain includes an antibody light chain variable region (VL) and an antibody heavy chain variable region (VH).

[0146] The term "variable region" or "variable domain" refers to a domain in the antibody heavy chain or antibody light chain involved in the binding of an antibody to an antigen. The variable domains (VH and VL, respectively) of the heavy and light chains of natural antibodies generally have similar structures, with each domain containing four conserved framework regions (FRs) and three hypervariable regions (HVRs). See, for example, Kindt et al., Kuby Immunology, 6th edition, WH Freeman and Co., p. 91 (2007). A single VH or VL domain may be sufficient to confer antigen-binding specificity.

[0147] A "human antibody" is an antibody that has an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human or human cell, or an antibody derived from a non-human source that utilizes the human antibody repertoire, or a sequence that encodes another human antibody. This definition of a human antibody explicitly excludes humanized antibodies that contain non-human antigen-binding residues.

[0148] A “humanized” antibody refers to a chimeric antibody containing amino acid residues from a non-human HVR and amino acid residues from a human FR. In certain embodiments, the humanized antibody comprises substantially all of at least one, typically two, variable domains, where all or substantially all of the HVR (e.g., CDR) corresponds to that of a non-human antibody, and all or substantially all of the FR corresponds to that of a human antibody. In some cases, the humanized antibody may also contain at least a portion of the antibody constant region derived from a human antibody. The “humanized form” of an antibody, e.g., a non-human antibody, refers to a humanized antibody.

[0149] As used herein, the terms “hypervariable region” or “HVR” refer to each region of an antibody variable domain that is sequence-hypervariable (“complementarity-determining region” or “CDR”) and / or forms a structurally defined loop (“hypervariable loop”) and / or contains an antigen contact residue (“antigen contact”). Generally, an antibody contains six HVRs: three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). Examples of HVRs as used herein include: (a) Hypervariable loops formed 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)); (b) CDRs generated 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 edition. Public Health Service, National Institutes of Health, Bethesda, MD (1991)); (c) Antigen contacts occurring 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 (d) A combination of (a), (b) and / or (c) containing HVR amino acid residues 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).

[0150] Unless otherwise indicated, HVR residues and other residues within the variable domain (e.g., FR residues) are numbered herein according to Kabat et al. cited above.

[0151] "Framework" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The variable domain FR generally consists of four FR domains: FR1, FR2, FR3, and FR4. Therefore, the HVR sequence and FR sequence generally appear in the following order in VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0152] The "Human Consensus Framework" is a framework representing the most commonly present amino acid residues in the selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is performed from subgroups of variable domain sequences. Generally, these sequence subgroups are those described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, NIH Publication 91-3242, Bethesda MD (1991), vols. 1-3. In some embodiments, for the VL, the subgroup is subgroup Kappa I, as described in Kabat et al. above. In some embodiments, for the VH, the subgroup is subgroup III, as described in Kabat et al. above.

[0153] In this specification, “acceptor human framework” means a framework comprising the amino acid sequence of a light chain variable domain (VL) framework or a heavy chain variable domain (VH) framework derived from a human immunoglobulin framework or a human consensus framework, as defined below. An acceptor human framework “derived” from a human immunoglobulin framework or a human consensus framework may contain the same amino acid sequence or may contain variations of the amino acid sequence. In some embodiments, the number of amino acid variations 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 VL acceptor human framework is sequence-identical to the VL human immunoglobulin framework sequence or the human consensus framework sequence.

[0154] The "class" of an antibody refers to the type of constant domain or constant region in its heavy chain. There are five main classes of antibodies: IgA, IgD, IgE, IgG, and IgM. Some of these are further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0155] As used herein, the terms IgG “isotype” or “subclass” mean any subclass of immunoglobulin determined by the chemical and antigenic properties of its constant region.

[0156] In this specification, the terms “Fc domain” or “Fc region” are used to refer to the C-terminal region of an immunoglobulin heavy chain, including at least a portion of the constant region. This term includes both the native sequence Fc region and the variant Fc region. While the boundaries of the Fc region of an IgG heavy chain may vary subtly, the Fc region of a human IgG heavy chain is typically defined as extending from Cys226 or Pro230 to the carboxyl terminus of the heavy chain. However, antibodies produced by host cells may undergo post-translational cleavage of one or more amino acids, particularly one or two, from the C-terminus of the heavy chain. Thus, antibodies produced by host cells by the expression of a specific nucleic acid molecule encoding a full-length heavy chain may contain either a full-length heavy chain or a cleaved variant of the full-length heavy chain (also referred to herein as a “cleaved variant heavy chain”). This is also true when the last two C-terminal amino acids of the heavy chain are glycine (G446) and lysine (K447, EU numbering). Therefore, the C-terminal lysine (Lys447) or C-terminal glycine (Gly446) and lysine (K447) in the Fc domain may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc domain or constant domain follows the EU numbering system, also known as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991 (see also above). As used herein, the “subunit” of the Fc domain refers to one of the two polypeptides that form the dimeric Fc domain, i.e., the polypeptide containing the C-terminal constant domain of the immunoglobulin heavy chain and possessing stable self-associating ability. For example, the subunit of the IgG Fc domain includes the IgG CH2 and IgG CH3 constant domains.

[0157] "Modifications that promote the association of the first and second subunits of the Fc domain" are manipulations of the peptide backbone or post-translational modifications of the Fc domain subunits that reduce or prevent the formation of homodimers by association between a polypeptide containing an Fc domain subunit and an identical polypeptide. When used herein, association-promoting modifications include, in particular, distinct modifications performed on each of two Fc domain subunits that are desirable to associate (i.e., the first and second subunits of the Fc domain), and these modifications are complementary to each other in order to promote the association of the two Fc domain subunits. For example, association-promoting modifications can alter the structure or charge of one or both of these Fc domain subunits to favor their association sterically or electrostatically, respectively. Thus, (hetero)dimerization occurs between a polypeptide containing a first Fc domain subunit and a polypeptide containing a second Fc domain subunit, and these subunits may not be identical in the sense that the further components fused to each of these subunits (e.g., antigen-binding moieties) may not be the same. In some embodiments, the modification that promotes association includes amino acid mutations, specifically amino acid substitutions, within the Fc domain. In certain embodiments, the modification that promotes association includes separate amino acid mutations, specifically amino acid substitutions, in each of the two subunits of the Fc domain.

[0158] "Activated Fc receptors" are Fc receptors that, following binding via the Fc region of an antibody, trigger a signaling event that stimulates receptor-hosting cells to exert effector function. Activated Fc receptors include FcγRIIIa (CD16a), FcγRI (CD64), FcγRIIa (CD32), and FcαRI (CD89).

[0159] The term "effector function" as used in relation to antibodies refers to the biological activity resulting from the antibody's Fc region, which varies depending on the antibody isotype. Examples of antibody effector functions include C1q binding and complement-dependent cell-mediated cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent phagocytosis (ADCP), cytokine secretion, immune complex-mediated antigen uptake by antigen-presenting cells, downregulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0160] As used herein, the term "effector cells" refers to a population of lymphocytes that present effector partial receptors (e.g., cytokine receptors) and / or Fc receptors on their surface, and through these, bind to the effector portion (e.g., cytokine) and / or Fc region of antibodies, thereby contributing to the destruction of target cells (e.g., tumor cells). Effector cells mediate, for example, cytotoxic or phagocytic effects. Effector cells include CD8 + cytotoxic T cells, CD4 + This includes, but is not limited to, helper T cells, γδ T cells, NK cells, lymphocyte-activated killer (LAK) cells, and effector T cells such as macrophages / monocytes.

[0161] As used herein, the terms “engineer,” “engineered,” and “engineering” are understood to include the manipulation or post-translational modification of the peptide backbone of natural or recombinant polypeptides or fragments thereof. Engineering includes the modification of amino acid sequences, the modification of glycosylation patterns, or the modification of side chain groups of individual amino acids, and combinations thereof. Engineering, particularly those prefixed “glyco,” and “glycosylation engineering,” include metabolic manipulation of cellular glycosylation mechanisms, including genetic manipulation of oligosaccharide synthesis pathways to alter the glycosylation of glycoproteins expressed in cells. Furthermore, glycosylation engineering also includes the effects of mutation and the cellular environment on glycosylation. In some embodiments, glycosylation engineering is a change in glycosyltransferase activity. In certain embodiments, the manipulation results in a change in glucosaminyltransferase activity and / or fucosyltransferase activity. Glycosylation operations can be used to obtain "host cells with increased GnTIII activity" (e.g., host cells manipulated to increase the expression level of one or more polypeptides having β(1,4)-N-acetylglucosaminyltransferase III (GnTIII) activity), "host cells with increased ManII activity" (e.g., host cells manipulated to increase the expression level of one or more polypeptides having α-mannosidase II (ManII) activity), or "host cells with decreased α(1,6) fucosyltransferase activity" (e.g., host cells manipulated to decrease the expression level of α(1,6) fucosyltransferase).

[0162] The terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells into which exogenous nucleic acids have been introduced and their offspring. Host cells include “transformers” and “transformed cells,” which include primary transformed cells and offspring derived from primary transformed cells, regardless of passage number. Offspring may not have exactly the same nucleic acid content as the parent cells and may contain mutations. Offspring of mutants having the same function or biological activity as those screened or selected in the original transformed cells are included herein. Host cells are any type of cell line available for producing the proteins used in the present invention. In some embodiments, host cells are engineered to enable the production of antibodies having modified oligosaccharides. In certain embodiments, host cells are engineered to increase the expression level of one or more polypeptides having β(1,4)-N-acetylglucosaminyltransferase III (GnTIII) activity. In certain embodiments, host cells are further engineered to increase the expression level of one or more polypeptides having α-mannosidase II (ManII) activity. Host cells include mammalian cultured cells such as cultured cells, for example, to name just a few, CHO cells, BHK cells, NS0 cells, SP2 / 0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells, or hybridoma cells, yeast cells, insect cells, and plant cells, but also include cells contained in transgenic animals, transgenic plants, or cultured plants or animal tissues.

[0163] As used herein, the term “GnTIII activity-possessing polypeptide” refers to a polypeptide capable of catalyzing the addition of an N-acetylglucosamine (GlcNAc) residue to the β-1,4 linkage of a trimannosyl core of an N-linked oligosaccharide to a β-linked mannoside. This includes fusion polypeptides that exhibit enzymatic activity similar to, but not necessarily identical to, β(1,4)-N-acetylglucosaminyltransferase III, also known as β-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyl-transferase (EC 2.4.1.144) by the International Union for Biochemistry and Molecular Biology Nomenclature (NC-IUBMB), when measured in specific biological assays with or without dose-dependent effects. If dose-dependency exists, the enzyme activity does not need to be identical to that of GnTIII; rather, the dose-dependency at a given activity needs to be substantially similar to that of GnTIII (i.e., the candidate polypeptide exhibits greater activity than GnTIII, or about 25 times or less, preferably about 10 times or less, and most preferably about 3 times or less). In certain embodiments, the polypeptide having GnTIII activity is a fusion polypeptide comprising the catalytic domain of GnTIII and the Golgi localization domain of a heterologous Golgi commensal polypeptide. In particular, the Golgi localization domain is the localization domain of mannosidase II or GnTI, and especially the localization domain of mannosidase II. Alternatively, the Golgi localization domain is selected from the group consisting of the localization domain of mannosidase I, the localization domain of GnTII, and the localization domain of α1,6 corefucosyltransferase. Methods for producing such fusion polypeptides and using them to produce antibodies with enhanced effector function are disclosed in International Publication No. 2004 / 065540, U.S. Provisional Patent Application Publication No. 60 / 495142, and U.S. Patent Application Publication No. 2004 / 0241817 (all of which are expressly incorporated herein by reference).

[0164] As used herein, the term "Golgi localization domain" refers to the amino acid sequence of a Golgi resident polypeptide that is responsible for fixing the Golgi resident polypeptide to a specific location within the Golgi complex. Generally, the localization domain includes the amino-terminus "tail" of the enzyme.

[0165] As used herein, the term “ManII-active polypeptide” refers to polypeptides capable of catalyzing the hydrolysis of terminal 1,3- and 1,6-linked α-D-mannose residues of the branched GlcNAcMan5GlcNAc2 mannose intermediate of N-linked oligosaccharides. This includes fusion polypeptides that exhibit enzymatic activity similar to, but not necessarily identical to, Golgi α-mannosidase II, also known as mannosyl-oligosaccharide 1,3-1,6-α-mannosidase II (EC 3.2.1.114) by the International Union for Biochemistry and Molecular Biology Nomenclature (NC-IUBMB).

[0166] Antibody-dependent cell-mediated cytotoxicity (ADCC) is an immune mechanism that leads to the lysis of antibody-coated target cells by immune effector cells. These target cells are those to which an antibody or fragment thereof, containing an Fc region, specifically binds, typically via a protein portion that is the N-terminus of the Fc region. As used herein, the term "increase / decrease in ADCC" is defined as either an increase / decrease in the number of target cells lysed within a given time at a given concentration of antibody in the culture medium surrounding the target cells by the ADCC mechanism, and / or a decrease / increase in the concentration of antibody in the culture medium surrounding the target cells required to achieve the lysis of a given number of target cells within a given time by the ADCC mechanism. The increase / decrease in ADCC is compared to ADCC mediated by the same antibody, which is produced by the same type of host cells using the same standard production, purification, formulation, and storage methods (known to those skilled in the art), but is not engineered. For example, an increase in ADCC mediated by antibodies produced by host cells engineered to alter the glycosylation pattern (e.g., to express glycosyltransferase, GnTIII, or other glycosyltransferase) by the methods described herein has been compared to ADCC mediated by the same antibodies produced by non-engineered host cells of the same type.

[0167] "Antibodies with increased / decreased antibody-dependent cytotoxicity (ADCC)" refers to antibodies in which ADCC is increased / decreased as measured by any appropriate method known to those skilled in the art. One commonly accepted in vitro ADCC assay is as follows: 1) This assay uses target cells known to express the target antigen recognized by the antigen-binding region of the antibody; 2) This assay uses human peripheral blood mononuclear cells (PBMCs) isolated from the blood of randomly selected healthy donors as effector cells; 3) This assay is performed according to the following protocol: i) Isolate the PBMCs using a standard density centrifugation procedure, and measure 5 × 10⁻⁶ 6Suspend cells in RPMI cell culture medium at a concentration of cells / ml; ii) Target cells are grown using standard tissue culture methods, harvested from the exponential growth phase with a viability of over 90%, washed in RPMI cell culture medium, and heated to 100 microcuries. 51 Labeled with Cr, washed twice with cell culture medium, 10 5 Resuspend the cells in the cell culture medium at a density of cells / ml; iii) Transfer 100 microliters of the above final target cell suspension to each well of a 96-well microtiter plate; iv) The antibody is sequentially diluted in cell culture medium from 4000 ng / ml to 0.04 ng / ml, and 50 microliters of the resulting antibody solution are added to target cells in a 96-well microtiter plate. A total of three antibody concentrations covering the entire concentration range described above are tested in a series; v) As a maximum release (MR) control, replace the antibody solution (iv above) in three additional wells of the plate containing labeled target cells with 50 microliters of a 2% (V / ) aqueous solution of a nonionic surfactant (nonidet, Sigma-St. Louis); vi) As a spontaneous release (SR) control, replace the antibody solution (iv) above with 50 microliters of RPMI cell culture medium in three additional wells of the plate containing labeled target cells; vii) Then, centrifuge the 96-well microtiter plate at 50 × g for 1 minute and incubate at 4°C for 1 hour; viii) Add 50 microliters of PBMC suspension (i above) to each well to achieve an effector:target cell ratio of 25:1, and incubate the plate in a 37°C incubator under a 5% CO2 atmosphere for 4 hours; ix) Collect the cell-free supernatant from each well and quantify the experimentally released radioactivity (ER) using a gamma counter; x) Calculate the specific solubility for each antibody concentration according to the formula (ER-MR) / (MR-SR) × 100 [wherein ER is the average radioactivity quantified for that antibody concentration (see ix above), MR is the average radioactivity quantified for the MR control (see V above) (see ix above), and SR is the average radioactivity quantified for the SR control (see vi above) (see ix above)]; 4) “Increased / decreased ADCC” is defined as either an increase / decrease in the maximum specific lysis rate observed within the antibody concentration range tested above, and / or a decrease / increase in the antibody concentration required to achieve half of the maximum specific lysis rate observed within the antibody concentration range tested above. The increase / decrease in ADCC is compared to the ADCC measured in the above assay, which is mediated by the same antibody produced by the same type of host cells using the same standard production, purification, formulation and storage methods (known to those skilled in the art), but which has not been engineered.

[0168] As used herein, the term “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous population of antibodies; that is, the individual antibodies constituting that population are identical and / or bind to the same epitope, except for possible variant antibodies (e.g., those containing spontaneous mutations or those arising during the manufacture of the monoclonal antibody preparation; such variants are usually present in small amounts). In contrast to polyclonal antibody preparations, which typically contain various antibodies against various determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is against a single determinant on an antigen. Therefore, the modifier “monoclonal” indicates the characteristic of an antibody obtained from a substantially homogeneous population of antibodies and should not be interpreted as requiring the production of the antibody by some specific method. For example, monoclonal antibodies used in accordance with the present invention can be produced by a variety of techniques, including but not limited to hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of a human immunoglobulin locus, and these methods and other exemplary methods for producing monoclonal antibodies are described herein.

[0169] A "naked antibody" refers to an antibody that is not conjugated with a heterogeneous portion (e.g., a cytotoxic portion) or a radioactive label. Naked antibodies may be present in pharmaceutical formulations.

[0170] "Natural antibodies" refer to naturally occurring immunoglobulin molecules with various structures. For example, a natural IgG antibody is a heterotetrameric glycoprotein with approximately 150,000 daltons, composed of two identical disulfide-linked light chains and two identical heavy chains. From the N-terminus to the C-terminus, each heavy chain has a variable region (VH) (also called a variable heavy chain 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 a variable light domain or light chain variable domain) followed by one constant light (CL) domain. Based on the amino acid sequence of its constant domain, the light chains of an antibody can be assigned to one of two types: kappa (κ) or lambda (λ).

[0171] As used herein, terms such as “first,” “second,” and “third” relating to antigen-binding moieties or domains are used for convenience to distinguish between more than one of each type of moiety or domain. Unless otherwise explicitly stated, such use of terminology is not intended to confer any particular order or direction.

[0172] The terms "multispecificity" and "bispecificity" mean that an antigen-binding molecule can specifically bind to at least two distinct antigenic determinants. Typically, a bispecific antigen-binding molecule contains two antigen-binding sites, each of which is specific to a different antigenic determinant. In certain embodiments, a bispecific antigen-binding molecule can simultaneously bind to two antigenic determinants, particularly two antigenic determinants expressed on two different cells.

[0173] As used herein, the terms "valent" or "valency" indicate the presence of a specific number of antigen-binding sites within an antigen-binding molecule. Therefore, the term "monovalent binding to an antigen" indicates the presence of one (but not more than one) antigen-binding sites specific to that antigen within the antigen-binding molecule.

[0174] The "antigen-binding site" refers to the part of an antigen-binding molecule that provides interaction with an antigen, i.e., one or more amino acid residues. For example, the antigen-binding site of an antibody contains amino acid residues from the complementarity-determining region (CDR). Natural immunoglobulin molecules typically have two antigen-binding sites, while Fab molecules typically have a single antigen-binding site.

[0175] As used herein, “activated T cell antigen” refers to an antigenic determinant expressed by T lymphocytes, particularly cytotoxic T lymphocytes, which can induce or enhance T cell activation upon interaction with antigen-binding molecules. Specifically, the interaction of an antigen-binding molecule with an activated T cell antigen can induce T cell activation by triggering a signaling cascade of the T cell receptor complex. An exemplary activated T cell antigen is CD3. In certain embodiments, the activated T cell antigen is CD3, particularly the epsilon subunit of CD3 (see UniProt number P07766 (version 130), NCBI RefSeq number Np_000724.1 for the human sequence; or UniProt number Q95LI5 (version 49), NCBI GenBank number BAB71849.1 for the cynomolgus monkey [Macaca fascicularis] sequence).

[0176] As used herein, “T cell activation” refers to one or more cellular responses of T lymphocytes, particularly cytotoxic T lymphocytes, selected from proliferation, differentiation, cytokine secretion, release of cytotoxic effector molecules, cytotoxic activity, and expression of activation markers. The T cell activation therapeutic agents used in the present invention can induce T cell activation. Suitable assays for measuring T cell activation are known in the art described herein.

[0177] As used herein, “target cell antigen” refers to an antigenic determinant presented on the surface of target cells, such as cancer cells or cells within a tumor, including tumor stroma cells. In certain embodiments, the target cell antigen is CD20, particularly human CD20 (see UniProt number P11836).

[0178] As used herein, “B cell antigen” refers to the antigenic determinants presented on the surface of B lymphocytes, particularly malignant B lymphocytes (in which case the antigen is also called “malignant B cell surface antigen”).

[0179] As used herein, "T cell antigen" refers to the antigenic determinants presented on the surface of T lymphocytes, particularly cytotoxic T lymphocytes.

[0180] A "Fab molecule" refers to a protein consisting of the VH and CH1 domains of the immunoglobulin heavy chain ("Fab heavy chain") and the VL and CL domains of the light chain ("Fab light chain").

[0181] A "chimeric antigen receptor" or "CAR" refers to a genetically engineered receptor protein that includes an antigen-binding moiety, e.g., a single-chain variable fragment (scFv) of a target antibody, a transmembrane domain, an intracellular T cell activation signaling domain (e.g., the CD3 zeta chain of the T cell receptor), 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 that enhances T cell functionality and persistence. For a review, see, for example, Jackson et al., Nat Rev Clin Oncol. (2016) 13, 370-383.

[0182] "Fused" means that the constituent elements (e.g., the Fab molecule and the Fc domain subunit) are linked by peptide bonds, either directly or via one or more peptide linkers.

[0183] The "effective dose" of a drug is the amount necessary to produce a physiological change in the cells or tissues to which it is administered.

[0184] The "therapeutically effective amount" of an agent, such as a pharmaceutical composition, refers to an effective amount for a dosage and period required to obtain a desired therapeutic or prophylactic result. The therapeutically effective amount of an agent, for example, removes, reduces, delays, minimizes, or prevents side effects of a disease.

[0185] The "therapeutic agent" means, for example, the active ingredient of a pharmaceutical composition, which is administered to a subject to change the natural course of a disease in the subject being treated and can be carried out for prevention or during the course of clinical pathology. The "immunotherapeutic agent" refers to a therapeutic agent administered to a subject for the purpose of restoring or enhancing the subject's immune response to a tumor.

[0186] The term "pharmaceutical composition" refers to a preparation in a form that enables the biological activity of the active ingredient contained therein and does not contain additional ingredients that are excessively toxic to the subject to whom the composition is administered.

[0187] A "pharmaceutically acceptable carrier" is a component other than the active ingredient in a pharmaceutical composition and refers to a component that is non-toxic to the subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0188] The term "package insert" or "instructions for use" is used to refer to the instructions normally included in the commercial package of a therapeutic product that contains information about indications, usage, dosage, administration, combination therapy, contraindications, and / or warnings regarding the use of the therapeutic product.

[0189] The term "combination therapy" as described herein includes co-administration (when two or more therapeutic agents are included in the same or separate formulations) and separate administration, and in the case of separate administration, the administration of the antibody reported herein is carried out before, simultaneously with, and / or after the administration of one or more additional therapeutic agents, preferably one or more antibodies.

[0190] A "crossover" Fab molecule (also called a "Crossfab") refers to a Fab molecule in which the variable domain or constant domain of the Fab heavy chain is exchanged with (i.e., replaced by) the variable domain or constant domain of the Fab light chain. Specifically, a crossover Fab molecule includes a peptide chain composed of a light chain variable domain VL and a heavy chain constant domain 1 CH1 (VL-CH1, N-terminus to C-terminus), and a peptide chain composed of a heavy chain variable domain VH and a light chain constant domain CL (VH-CL, N-terminus to C-terminus). For clarity, in a crossover Fab molecule in which the variable domain of the Fab light chain and the variable domain of the 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. Conversely, in a crossover Fab molecule in which the constant domain of the Fab light chain and the constant domain of the Fab heavy chain are exchanged, the peptide chain containing the heavy chain variable domain VH is referred to herein as the "heavy chain" of the (crossover) Fab molecule.

[0191] In contrast, a "conventional" Fab molecule refers to a Fab molecule in its natural format, that is, a Fab molecule that includes a heavy chain composed of a variable domain and a constant domain (VH-CH1, from the N-terminus to the C-terminus) and a light chain composed of a variable domain and a constant domain (VL-CL, from the N-terminus to the C-terminus).

[0192] The term "polynucleotide" refers to an isolated nucleic acid molecule or construct, such as messenger RNA (mRNA), viral RNA, or plasmid DNA (pDNA). Polynucleotides may contain common phosphodiester bonds or less common bonds (e.g., amide bonds, as seen in peptide nucleic acids (PNAs)). The term "nucleic acid molecule" refers to any one or more nucleic acid segments present in a polynucleotide, such as DNA or RNA fragments.

[0193] "Isolated" nucleic acid molecules or polynucleotides are nucleic acid molecules, DNA, or RNA that have been removed from their natural environment. For example, recombinant polynucleotides encoding polypeptides contained in a vector are considered isolated for the purposes of this invention. Further examples of isolated polynucleotides include recombinant polynucleotides maintained within heterologous host cells, or (partially or substantially) purified polynucleotides in solution. Isolated polynucleotides include polynucleotide molecules found in cells that normally contain polynucleotide molecules, but where the polynucleotide molecules are located outside the chromosome or at a chromosomal location different from their original location on the chromosome. Isolated RNA molecules include RNA transcripts of this invention in vivo or in vitro, as well as positive-strand, negative-strand, and double-stranded forms. Isolated polynucleotides or nucleic acids according to the present invention further include such molecules produced by synthesis. Polynucleotides or nucleic acids may or may not contain regulatory elements such as promoters, ribosome-binding sites, or transcription terminators.

[0194] A nucleic acid or polynucleotide having a nucleotide sequence that is at least, for example, 95% "identical" to the reference nucleotide sequence of the present invention means that the nucleotide sequence of the polynucleotide is identical to the reference nucleotide sequence, except that the nucleotide sequence of the polynucleotide may contain up to 5 point mutations per 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 substituted with other nucleotides, or up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. Such modifications to the reference sequence may be individually scattered among residues in the reference sequence at the 5' or 3' terminal positions of the reference nucleotide sequence or at any position between these terminal positions, or they may be scattered as one or more consecutive groups within the reference sequence. In practice, whether 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 determined using conventionally known computer programs, such as those described above for polypeptides (e.g., ALIGN-2).

[0195] The term "expression cassette" refers to a polynucleotide produced recombinantly or synthetically using a set of specific nucleic acid elements that enable the transcription of a particular nucleic acid within a target cell. Recombinant expression cassettes can be incorporated into plasmids, chromosomes, mitochondrial DNA, plastid DNA, viruses, or nucleic acid fragments. Typically, the recombinant expression cassette portion of an expression vector includes, among several sequences, the nucleic acid sequence to be transcribed and a promoter. In certain embodiments, the expression cassette of the present invention includes a polynucleotide sequence or fragment thereof encoding the bispecific antigen-binding molecule of the present invention.

[0196] The term “vector” or “expression vector” is synonymous with “expression construct” and refers to a DNA molecule used to introduce a specific gene into a target cell, enabling it to bind operably, and to induce its expression. This term includes vectors as self-replicating nucleic acid structures and vectors that have been incorporated into the genome of the host cell into which they have been introduced. The expression vector of the present invention comprises an expression cassette. The expression vector enables the transcription of a large amount of stable mRNA. Once the expression vector enters the target cell, the ribonucleic acid molecule or protein encoded by the gene is produced by the cellular transcription and / or translation mechanism. In one embodiment, the expression vector of the present invention comprises an expression cassette containing a polynucleotide sequence encoding the bispecific antigen-binding molecule or a fragment thereof of the present invention.

[0197] As used herein, the term “about” refers to the normal range of error of the corresponding value, as would be readily understood by those skilled in the art. References to a value or parameter “about” herein include (and describe) embodiments relating to that value or parameter itself.

[0198] "B-cell proliferative disorders" refer to diseases in which the number of B cells in a patient is increased compared to the number of B cells in a healthy control group, and in particular, diseases in which the increase in the number of B cells is the cause or characteristic of the disease. "CD20-positive B-cell proliferative disorders" refer to B-cell proliferative disorders in which B cells, (in addition to normal B cells) particularly malignant B cells, express CD20.

[0199] Exemplary B-cell proliferative disorders include non-Hodgkin lymphoma (NHL), diffuse large B-cell lymphoma (DLBCL; e.g., relapsed or refractory DLBCL not otherwise specified (NOS)), high-grade B-cell lymphoma (HGBCL; e.g., HGBCL NOS, double-hit HGBCL, and triple-hit HGBCL), primary mediastinal large B-cell lymphoma (PMBCL), and DLBCL arising from FL (transformed FL; trFL); follicular lymphoma (FL) (including grade 1-3b FL); mantle cell lymphoma (MCL); and marginal zone lymphoma (MZL) (including splenic MZL, nodal MZL, or extranodal MZL). In one embodiment, a CD20-positive B-cell proliferative disorder is relapsed or refractory NHL (e.g., relapsed or refractory DLBCL, relapsed or refractory FL, or relapsed or refractory MCL).

[0200] "Refractory disease" refers to cases where complete remission is not achieved with first-line treatment. In some embodiments, refractory disease is defined as a lack of response to previous treatment or a relapse within six months of previous treatment. In some embodiments, refractory disease is characterized by one or more of the following: progressive disease (PD) as the best response to first-line treatment; stable state (SD) as the best response after at least four cycles of first-line treatment (e.g., four 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 six cycles; and residual lesions or disease progression as demonstrated by biopsy after partial response. "Relapsing disease" is defined as complete remission to first-line treatment. In some embodiments, disease relapse is demonstrated by biopsy. In one embodiment, the patient relapsed or did not respond to at least two prior systemic treatment regimens, including at least one prior regimen containing anthracyclines and at least one prior regimen containing anti-CD20 targeted therapy.

[0201] The “individual” or “subject” is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cattle, 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 human. In some cases, each subject in the population of subjects is human. In some cases, each subject in the standard population of subjects is human.

[0202] A "transplant-ineligible" subject or a "subject ineligible for autologous stem cell transplantation (SCT)" is a subject who does not meet the eligibility requirements for autologous stem cell transplantation, a subject for whom such transplantation is not recommended, a subject who cannot receive such transplantation, or a subject who refuses such transplantation. Examples of desirable patient characteristics include age ≤65 years, Karnofsky Performance Status (KPS; Karnofsky et al. 1948; 1(4):634-656) >60, forced expiratory volume in one second (FEV1) >60% of predicted value, pulmonary diffusion capacity (DLCO) >60% of predicted value, left ventricular ejection fraction >45%, normal heart rhythm, serum bilirubin ≤2 mg / 100 mL, alanine aminotransferase (ALT) / aspartate aminotransferase (AST) <2 × normal value, serum creatinine ≤1.5 mg / 100 mL, creatinine clearance >60 mL / min, absence of secondary active malignancies, non-pregnancy, and absence of uncontrolled infections (including dental infections) (Hamadani M et al. Bone Marrow Transplant. 2010; 45:1259-68).

[0203] As used herein, “treatment” (and its grammatical variations, “treat” or “treating”) refers to a clinical intervention to alter the natural course of a disease in the subject being treated, and may be performed for preventive purposes or during the course of a clinicopathological disease. Desired effects of treatment include, but are not limited to, preventing the onset or recurrence of the disease, reducing symptoms, attenuating any direct or indirect pathological consequences of the disease, preventing metastasis, slowing the rate of disease progression, achieving remission or mitigation of symptoms, and achieving recovery or improving prognosis. In some embodiments, the methods of the present invention are used to delay the onset of a disease or to slow the progression of a disease.

[0204] As used herein, “delaying the progression” of a disorder or disease means delaying, hindering, slowing, slowing, inhibiting, and / or prolonging the progression of a disease or disorder (e.g., CD20-positive B-cell proliferative disorders, e.g., NHL, e.g., DLBCL). This delay may be of varying duration depending on the medical history and / or the individual being treated. As will be apparent to those skilled in the art, a sufficient or substantial delay may substantially encompass the prevention of the individual not developing the disease. For example, in late-stage cancer, the development of central nervous system (CNS) metastases may be delayed.

[0205] "Reduce" or "inhibit" means the ability to cause 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 reduction to zero (or below the detection limit of the analytical method), i.e., complete disappearance or elimination. In certain embodiments, "reduce" or "inhibit" refers to a reduction or inhibition of an undesirable event following treatment with the anti-CD20 / anti-CD3 bispecific antibody using the step-up dosing regimen of the present invention, such as cytokine-induced toxicity (e.g., cytokine release syndrome (CRS)), infusion-related reactions (IRR), macrophage activation syndrome (MAS), neurotoxicity, severe tumor lysis syndrome (TLS), neutropenia, thrombocytopenia, elevated liver enzymes, and / or central nervous system (CNS) toxicity, compared to a certain preset dosing with a target dose of the bispecific antibody. In other embodiments, reducing or inhibiting may mean the effector function of an antibody mediated by the antibody Fc region, such effector function specifically includes complement-dependent cell cytotoxicity (CDC), antibody-dependent cell cytotoxicity (ADCC), and antibody-dependent cell phagocytosis (ADCP). In other embodiments, reducing or inhibiting may mean the symptoms, presence or size of metastases, or size of the primary tumor 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).

[0206] As used herein, “adminstering” refers to a method of administering a certain dosage of a compound (e.g., an anti-CD20 / anti-CD3 bispecific antibody) or composition (e.g., a pharmaceutical composition, e.g., a pharmaceutical composition containing an anti-CD20 / anti-CD3 bispecific antibody) to a subject. Compounds and / or compositions used in the methods described herein may be administered intravenously (e.g., by intravenous infusion).

[0207] The "fixed" or "flat" dose of a therapeutic agent (e.g., bispecific antibody) in this specification refers to the dose administered to a patient regardless of the patient's body weight or body surface area (BSA). Thus, this fixed dose or flat dose is defined as the absolute amount of the therapeutic agent (e.g., mg), rather than as a mg / kg dose or mg / m 2 dose.

[0208] As used herein, "target dose" refers to the dose of an anti-CD20 / anti-CD3 bispecific antibody that achieves a therapeutic effect, i.e., the desired clinical effect. It has been determined that the potential target doses of glofitamab are 16 mg or 30 mg.

[0209] "Fixed dosing or preset dosing at the target dose" and "treatment regimens without a step-up dosing regimen" refer to a dosing schedule in which the same dose is used in the first and second cycles, and optionally subsequent treatment cycles, in contrast to step-up dosing, in which a low dose is used in the first few treatment cycles and the target dose is reached only in treatment cycles after the second treatment cycle.

[0210] As used herein, the term “treatment cycle” or “cycle” (abbreviated as “C”) means one or more courses of administration of anti-CD20 / anti-CD3 bispecific antibody repeated on a regular schedule, with optionally selected rest periods in between. In one embodiment of the present invention, a first treatment cycle comprises first and second administrations of anti-CD20 / anti-CD3 bispecific antibody and a subsequent rest period. In such one embodiment, the first treatment cycle comprises a first dose of anti-CD20 / anti-CD3 bispecific antibody on day 1 of the first cycle, a second dose of anti-CD20 / anti-CD3 bispecific antibody on day 8 of the first cycle, and a subsequent 12-day rest period. In one embodiment, subsequent cycles comprise one dose of anti-CD20 / anti-CD3 bispecific antibody on day 1 of that cycle and a subsequent 20-day rest period. In one embodiment, one treatment cycle consists of 21 days. In another embodiment, one treatment cycle consists of 14 days. A treatment cycle comprising one or more doses of an anti-CD20 / anti-CD3 bispecific antibody may further comprise one or more doses of one or more other therapeutic agents, such as an anti-CD20 antibody, particularly obinutuzumab. The treatment schedule according to the present invention may consist of two or more treatment cycles, or three, four, five, six, seven, eight, nine, ten, eleven, and especially twelve treatment cycles.

[0211] An “individual response” or “response” can be evaluated using any endpoint that demonstrates the effect on the subject, and endpoints include, but are not limited to, (1) some degree of inhibition (including slowing and complete cessation) of disease progression (e.g., progression of CD20-positive B-cell proliferative disorders, e.g., non-Hodgkin lymphoma); (2) reduction in tumor size; (3) inhibition (i.e., reduction, slowing or complete cessation) of cancer cell invasion into adjacent peripheral organs and / or tissues; (4) inhibition (i.e., reduction, slowing or complete cessation) of metastasis; (5) some degree of alleviation of one or more symptoms associated with CD20-positive B-cell proliferative disorders, e.g., B-cell proliferative disorders; (6) increased or prolonged survival, including overall survival and progression-free survival; and / or (7) a reduction in mortality at any point in time after treatment.

[0212] As used herein, “complete response” or “CR” refers to the disappearance of all target lesions. In one embodiment, standard NHL response criteria are evaluated for CR determination (Lugano Classification, Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27): 3059-3067).

[0213] As used herein, “partial response” or “PR” means a reduction of at least 30% of the total longest diameter (SLD) of the target lesion relative to baseline SLD, or a reduction of at least 50% of the product of diameters (SPD) of the target lesion relative to baseline SPD.

[0214] "Sustained response" refers to a sustained effect on the reduction of tumor growth after discontinuation of treatment. For example, tumor size may remain the same as or smaller than its size at the start of the treatment phase. In some implementations, the sustained response is at least the same length as the treatment period, or at least 1.5x, 2.0x, 2.5x, or 3.0x the length of the treatment period.

[0215] The terms "effective response" or "responsiveness" and similar phrases for a subject to a treatment using medicine refer to a clinical or therapeutic effect conferred upon a subject who is at risk of or suffering from a disease or disorder such as cancer. In some embodiments, such an effect includes one or more of the following: extending survival (including overall survival and progression-free survival); achieving an objective response (including complete response or partial response); or improving the signs or symptoms of cancer.

[0216] The "duration of complete response" (DOCR) is defined as the period from the first recorded complete response (CR) to the earlier of recorded disease progression or death from any cause. In one embodiment, DOCR is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27):3059-3067).

[0217] The "objective response period" (DOR) is defined as the time from the first recorded objective response to disease progression, relapse, or death from any cause. In one embodiment, the DOR is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27):3059-3067).

[0218] Progression-free survival (PFS) is defined as the period from the first treatment with an anti-CD20 / anti-CD3 bispecific antibody until disease progression or death from any cause occurs first. In one embodiment, PFS is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27):3059-3067).

[0219] Overall survival (OS) is defined as the period from the first treatment with an anti-CD20 / anti-CD3 bispecific antibody to the date of death from any cause.

[0220] The "time to first complete response" (TFOR) is defined as the time from the start of treatment until the first response is recorded. In one embodiment, the TFOR is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27):3059-3067).

[0221] The "time to first complete response" (TFCR) is defined as the time from the start of treatment until the first response is recorded. In one embodiment, the TFCR is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27):3059-3067).

[0222] As used herein, “objective response rate” or “overall response rate” (ORR) is defined as the sum of the partial response (PR) rate and the complete response (CR) rate. In one embodiment, the ORR is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27):3059-3067).

[0223] As used herein, “stable state” or “SD” refers to a state in which, based on the lowest SLD since the start of treatment, there is neither sufficient contraction to meet the criteria for PR of the target lesion nor sufficient increase to meet the criteria for PD.

[0224] As used herein, “progressive disease” or “PD” means that, since the initiation of treatment, the SLD of the target lesion has increased by at least 20% from the lowest SLD, or the SPD of the target lesion has increased by at least 50% from the lowest SPD, or the presence of one or more new lesions.

[0225] As used herein, “infusion-related reaction,” “IRR,” or “infusion-related adverse event” refers to an adverse event that occurs in a patient or subject during or within 24 hours after administration of a drug (e.g., an anti-CD20 / anti-CD3 bispecific antibody, e.g., grofitamab; or an anti-CD79b antibody drug conjugate, e.g., polatuzumab vedotin). IRRs can be graded from 1 to 5, for example, according to NCI CTCAE v.4.

[0226] The term "PD-1 axis-binding antagonist" refers to a molecule that inhibits the interaction between a PD-1 axis-binding partner and one or more of its binding partners in order to eliminate T cell dysfunction caused by signaling on the PD-1 signaling axis, thereby restoring or enhancing T cell function (e.g., proliferation, cytokine production, target cell death). As used herein, PD-1 axis-binding antagonists include PD-1 binding antagonists, PD-L1 binding antagonists, and PD-L2 binding antagonists.

[0227] The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signaling resulting from the interaction of PD-1 with one or more binding partners, such as PD-L1 and PD-L2. In some embodiments, a PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to one or more of its binding partners. In certain embodiments, a 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, immune adhexins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, suppress, or interfere with signaling resulting from the interaction of PD-1 with PD-L1 and / or PD-L2. In some embodiments, a PD-1-binding antagonist reduces negative costimulatory signals mediated by or through cell surface proteins expressed in response to PD-1-mediated signaling in T lymphocytes, thereby mitigating dysfunction in dysfunctional T cells (e.g., enhancing the effector response to antigen recognition). In some embodiments, the PD-1-binding antagonist is an anti-PD-1 antibody. In certain embodiments, the PD-1-binding antagonist is MDX-1106 (nivolumab). In other specific embodiments, the PD-1-binding antagonist is pembrolizumab (formerly lambrolizumab (MK-3475)). In other specific embodiments, 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 conjugated antagonist is MED1-0680. In some examples, the PD-1 conjugated antagonist is PDR001 (spartalizumab). In some examples, the PD-1 conjugated antagonist is REGN 2810 (semiprimab). In some examples, the PD-1 conjugated antagonist is BGB-108. In other examples, the PD-1 conjugated antagonist is prorugolimab, camrelizumab, cintilimab, tislerizumab, or tripalimab.

[0228] Further examples of PD-1 axially coupled antagonists include semiprimab, prorugolimab, camrelizumab, cintilimab, tislerizumab, tripalimab, dostalimab, retifanlimab, spartalizumab, sasanlimab, penprimab, CS1003, HLX10, SCT-I10A, SHR-1316, CS1001, emvafolimab, TQB2450, ZKAB001, LP-002, zimbererimab, valstilimab, genolimusumab, BI754091, cetrelimab, YBL-006, BAT1306, HX008, CX-072, IMC-001, KL-A167, buzicalimab, and AMG. 404, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, APL-502, Kosiberimab, Rhodapolimab, GS-4224, INCB086550, FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, MAX-10181, RC98, BION-004, AM0001, CB201, ENUM 244C8, ENUM Examples include 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, and HS-636.

[0229] The term "PD-L1-binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signaling 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, a PD-L1-binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partner. In certain embodiments, a PD-L1-binding antagonist inhibits the binding of PD-L1 to PD-1 and / or B7-1. In some embodiments, a PD-L1-binding antagonist includes anti-PD-L1 antibodies, their antigen-binding fragments, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, suppresses, or interferes with signaling 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, a PD-L1-binding antagonist reduces negative costimulatory signals mediated by or through cell surface proteins expressed in response to PD-1-mediated signaling in T lymphocytes, thereby mitigating dysfunction in dysfunctional T cells (e.g., enhancing the effector response to antigen recognition). In some embodiments, the PD-L1-binding antagonist is an anti-PD-L1 antibody. In certain embodiments, the anti-PD-L1 antibody is atezolizumab (CAS registry number: 1422185-06-5), also known as MPDL3280A, as described herein. In another particular embodiment, the anti-PD-L1 antibody is MDX-1105 as described herein. In yet another particular embodiment, the anti-PD-L1 antibody is MEDI4736 as described herein.

[0230] As used herein, the term "atezolizumab" refers to an anti-PD-L1 antagonist antibody having the International Name Name (INN) List 112 (WHO Drug Information, Vol. 28, No. 4, 2014, p. 488) or CAS Registry Number 1380723-44-3.

[0231] The term "PD-L2-binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signaling resulting from the interaction of PD-L2 with one or more of its binding partners, such as PD-1. In some embodiments, a PD-L2-binding antagonist is a molecule that inhibits the binding of PD-L2 to one or more of its binding partners. In certain embodiments, a PD-L2-binding antagonist inhibits the binding of PD-L2 to PD-1. In some embodiments, a PD-L2 antagonist includes anti-PD-L2 antibodies, their antigen-binding fragments, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, suppresses, or interferes with signaling resulting from the interaction of PD-L2 with one or more of its binding partners, such as PD-1. In some embodiments, a PD-L2-binding antagonist reduces negative costimulatory signals mediated by or through cell surface proteins expressed in response to PD-L2-mediated T lymphocyte-mediated signaling, thereby mitigating dysfunction in dysfunctional T cells (e.g., enhancing the effector response to antigen recognition). In some embodiments, the PD-L2-binding antagonist is an immunoadhesin.

[0232] As used herein, the term "chemotherapeutic agent" refers to compounds useful in the treatment of cancer, such as CD20-positive cell proliferative disorders (e.g., B-cell proliferative disorders (e.g., relapsed or refractory B-cell proliferative disorders), e.g., non-Hodgkin 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 chemotherapeutic 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); 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-fluorophenyl)-N2-(1-methyl-piperidine-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidine-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-butinamide); EKB-569 (N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolinyl]-4-(dimethylamino)-2-butenamide) (Weiss); AG1478 (Pfizer); AG1571 (SU 5271; Pfizer); and dual EGFR / HER2 tyrosine kinase inhibitors, e.g., lapatinib (TYKERB®, GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2methylsulfonyl)ethyl]amino]methyl]-2-furanyl]-4-quinazolinamine)); tyrosine kinase inhibitors (e.g., EGFR inhibitors; small molecule HER2 tyrosine kinase inhibitors, e.g., TAK165 (Takeda); oral selective inhibitors of ErbB2 receptor tyrosine kinase, CP-724, 714 (Pfizer and OSI); dual HER inhibitors, e.g., EKB-569 (available from Wyeth) (which preferentially binds to EGFR but also binds to both HER2-overexpressing and EGFR-overexpressing cells);PKI-166 (Novartis); pan-HER inhibitors, e.g., canertinib (CI-1033; Pharmacia); Raf-1 inhibitors, e.g., antisense agents that inhibit Raf-1 signaling, ISIS-5132 (ISIS Pharmaceuticals); non-HER target tyrosine kinase inhibitors, e.g., imatinib mesylate (Gleebeck®, GlaxoSmithKline); multi-target tyrosine kinase inhibitors, e.g., sunitinib (Sutent®, Pfizer); VEGF receptor tyrosine kinase inhibitors, e.g., batalanib (PTK787 / ZK222584, Novartis / Schöling); MAPK extracellular regulatory kinase I inhibitor CI-1040 (Pharmacia); quinazolines, e.g., PD 153035, 4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyridopyrimidines; pyrrolopyrimidines, e.g., CGP 59326, CGP 60261 and CGP 62706; pyrazolopyrimidine, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine; curcumin (diferloylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tilphostin containing nitrothiophene moiety; PD-0183805 (Warner-Lambert Corporation); antisense molecules (e.g., those encoding nucleic acids that code for HER); quinoxaline (US Patent No. 5804396); tryphostin (US Patent No. 5804396); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering); pan-HER inhibitors, e.g., CI-1033 (Pfizer); Affinitac (ISIS 3521; Isis / Lilly); PKI 166 (Novartis); GW2016 (GlaxoSmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering); INC-1C11 (Imukuron); and Rapamycin (Sirolimus, RapaMune®); Proteasome inhibitors, e.g., Bortezomib (VELCADE®, Millennium Pharmaceuticals); Disulfiram; Epigallocatechin gallate; Salinosporamide A; Carfilzomib;17-AAG (Geldanamycin); Radicicol; Lactate dehydrogenase A (LDH-A); Fulvestrant (Feslodex®, AstraZeneca); Letrozole (Femara®, Novartis); Finasunate (Vatalanib®, Novartis); Oxaliplatin (Eloxatin®, Sanofi); 5-FU (5-Fluorouracil); Leucovorin; Lonafarnib (SCH66336); Sorafenib (Nexavar®, Bayer / Labo); AG1478; Alkylating agents, e.g., thiotepa and cytoxane®; Cyclophosphamide; Alkyl sulfonates, e.g., busulfan, improsulfan and pigosulfan; Aziridines, e.g., benzodopa, carbocone, meturedopa and Uredopa; ethyleneimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylomellamine; acetogenins (especially bratacin and bratacinone); camptothecin (including topotecan and irinotecan); briostatin; calistatin; CC-1065 (including its adzeresin, calzelsin, and bizeresin synthetic analogs); cryptophycin (especially cryptophycin 1 and cryptophycin 8); corticosteroids (including prednisone and prednisolone); cyproterone acetate; 5α-reductase including finasteride and dutasteride; vorinostat, romidepsin, panobinostat, valproic acid, mosetinostat, dorastatin, aldesleukin, talc duocalmycin (talc duocarmycin (including synthetic analogs KW-2189 and CB1-TM1); eryuterobin; pancratistatin; sarcodictyin; spongestatin;Nitrogen mustards, such as chlorambucil, chlomaphazine, chlorophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melphalan, nobembichin, fenestrine, prednimustine, trophosphamide, uracil mustard; nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics such as engine antibiotics (e.g., Kalicare) Mycin, especially calicheamicin γ1 and calicheamicin ω1); dynemycin (including dynemycin A); bisphosphonate preparations, e.g., clodronate; esperamicin; and neocartinostatin chromophores and related pigment proteins (endiin antibiotic chromophores), acrasinomycin, actinomycin, anthramycin, azaserin, kakutinomycin, carabicin, kaminomycin, cardinophilin, chromomycin, dactinomycin, detrubicin, 6- Diazo-5-oxo-L-norleucine, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcelomycin, mitomycins such as mitomycin C, mycophenolic acid, nogaramycin, olibomycin, peplomycin, porphyromycin, puromycin, queramycin, rodorubicin, streptonigrin, streptoz Syn, tubercidine, ubenimex, dinostatin, zolubicin; antimetabolites, e.g., methotrexate and 5-fluorouracil (5-FU); folate analogs, e.g., denopterin, methotrexate, pteropterin, trimethrexate; purine analogs, e.g., fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs, e.g., ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine;Androgens, e.g., carsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; antiadrenergic drugs, e.g., aminoglutethimide, mitotane, trilostane; folic acid supplements, e.g., frolinic acid; acegraton; aldofamide glycoside; aminolevulinic acid; enyluracil; amsacrine; bestrabusil; bisantrene; edatraxate; defofamine; demecoltin; diaziquan; elfomithine; eriptinium acetate; epotilone; etogluside; gallium nitrate; hydroxyurea; lentinan; lonidainine; mytansinoids, e.g., mytansine and ansamitosine; mitogwazone; mitoxantrone; mopidamnol; nitraerine; pentostatin; Fenamet; Pirarubicin; Rosoxantrone; Podophyllic acid; 2-Ethylhydrazide; Procarbazine; PSK® Polysaccharide Complex (JHS Natural Products Co.); Lazoxane; Rhizoxin; Schizophyllan; Spirogermanium; Tenuazonic acid; Triadicone; 2,2',2''-Trichlorotriethylamine; Trichothecenes (especially T-2 toxin, Beraclin A, Loridine A and Anguidin); Urethane; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitolacto This includes ru; pipobromane; gacytosine; arabinoside ("Ara-C"); thiotepa; chlorambucil; 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; and any pharmaceutically acceptable salts, acids, prodrugs, and derivatives of the above.

[0233] Chemotherapy agents also include (i) anti-hormone agents that act to modulate or inhibit the hormonal effects on tumors, such as anti-estrogens and selective estrogen receptor modulators (SERMs), (tamoxifen (Nolvadex®; including tamoxifen citrate), raloxifene, droloxifene, iodoxifene, 4-hydroxytamoxifen, trioxyfen, keoxyfen, LY117018, onapristone, and Fareston® (citric acid (ii) including toremifine); (ii) aromatase inhibitors that inhibit aromatase, an enzyme that regulates estrogen production in the adrenal gland, such as 4(5)-imidazole, aminoglutethimide, Megase® (megestrol acetate), Aromasin® (exemestane; Pfizer), Formestany, Fadrozol, Rivisor® (borozol), Femara® (letrozole; Novartis), and Arimidex. (Registered Trademark) (Anastrozole; AltraZeneca); (iii) Antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, trypterelin, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, all trans-retionic acids, fenretinide, and troxacitabine (1,3-dioxolane nucleoside cytosine analog); (iv) Protein kinase inhibitors; (v) Lipid kinase inhibitors; (vi) Antisense oligonucleotides, particularly those that inhibit gene expression in signaling pathways involved in abnormal cell proliferation, e.g., PKC-alpha, Ralf, and H-Ras; (vii) Ribozymes such as VEGF expression inhibitors (e.g., Angiozyme®) and HER2 expression inhibitors; (viii) Gene therapy vaccines, e.g., vaccines such as ALLOVECTIN®, leubectin®, and VAXID®;(ix) Growth inhibitors comprising vinca (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), and DNA alkylating agents (e.g., tamoxigen, dacarbazine, mechloretamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C); and (x) any pharmaceutically acceptable salts, acids, prodrugs, and derivatives of any of the above.

[0234] As used herein, the term “cytotoxic agent” refers to a substance that inhibits or prevents cellular function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioactive isotopes (e.g., 211 At, 131 I, 125 I, 90 Y, 186 Re, 188 Re, 153 Sm, 212 Bi, 32 P, 212 Radioactive isotopes of Pb and Lu; chemotherapeutic agents or drugs (e.g., methotrexate, adriamycin, or vinca alkaloids (vincristine, vinblastine, or etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin, or other inserts); growth inhibitors; enzymes and their fragments, e.g., nucleases; antibiotics; toxins such as low molecular weight toxins or enzymatically active toxins of bacterial, fungal, plant, or animal origin (including their fragments and / or variants); and various antitumor or anticancer agents disclosed below.

[0235] III. Therapeutic agents for use in the methods of the present invention A. Anti-CD20 / anti-CD3 bispecific antibody The present invention provides novel doses of anti-CD20 / anti-CD3 bispecific antibodies. In some embodiments, the antibody is a monoclonal antibody. In certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody is polyclonal. In some embodiments, the anti-CD20 / anti-CD3 bispecific antibody is a human antibody. In certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody is a humanized antibody. In certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody is a chimeric antibody. In some embodiments, the anti-CD20 / anti-CD3 bispecific antibody is a full-length antibody. In some embodiments, the anti-CD20 bispecific antibody is an IgG class antibody, particularly an IgG1 subclass antibody. In certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody is a recombinant antibody.

[0236] In certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody comprises an antibody fragment. Examples of antibody fragments include, but are not limited to, the Fab fragment, Fab' fragment, Fab'-SH fragment, F(ab')2 fragment, Fv fragment, and scFv fragment, as well as other fragments described below. For a review of specific antibody fragments, see Hudson et al. Nat. Med. 9:129-134 (2003). For a review of the scFv fragment, see, for example, Plueckthun, The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York), pp. 269-315 (1994). See also International Publication No. 93 / 16185, and U.S. Patents No. 5571894 and 5587458. For a description of the Fab and F(ab')2 fragments containing salvage receptor-binding epitope residues and having increased in vivo half-lives, see U.S. Patent No. 5,869,046. In some embodiments, the antibody fragment is either the Fab fragment or the scFv fragment.

[0237] A diabody is an antibody fragment having two antigen-binding sites that can be bivalent or bispecific. See, for example, European Patent No. 404097, International Publication No. 1993 / 01161, Hudson et al. Nat. Med. 9:129-134 (2003); and Hollinger et al. Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al. Nat. Med. 9:129-134 (2003).

[0238] A single-domain antibody is an antibody fragment that contains all or part of the heavy chain variable domain or all or part of the light chain variable domain of an antibody. In certain embodiments, the single-domain antibody is a human single-domain antibody (see, for example, Domantis, Waltham, Massachusetts; U.S. Patent No. 6,248,516).

[0239] Antibody fragments can be prepared by a variety of techniques, including, but not limited to, proteolysis of intact antibodies or production by recombinant host cells (e.g., Escherichia coli or phages), as described herein.

[0240] In certain embodiments, an anti-CD20 / anti-CD3 bispecific antibody is a chimeric antibody. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4816567; and in Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984). In one example, a chimeric antibody contains a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate (such as a monkey)) and a human constant region. In further examples, a chimeric antibody is a “class-switched” antibody in which the class or subclass is modified from that of the parent antibody. A chimeric antibody contains its antigen-binding fragment.

[0241] In some embodiments, the anti-CD20 / anti-CD3 bispecific antibody is a humanized antibody. Non-human antibodies are typically humanized to reduce immunogenicity against humans while retaining the specificity and affinity of the parent non-human antibody. Humanized antibodies typically include one or more variable domains in which the HVR, e.g., CDR (or a portion thereof), is derived from a non-human antibody and the FR (or a portion thereof) is derived from a human antibody sequence. Humanized antibodies also optionally include at least a portion of the human constant region. In some embodiments, some FR residues of the humanized antibody are replaced with corresponding residues from a non-human antibody (e.g., an antibody from which the HVR residues are derived) to restore or improve antibody specificity or affinity, for example.

[0242] Humanized antibodies and their production methods are reviewed in Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and further, see, for example, Riechmann et al., Nature 332:323-329 (1988), Queen et al., Proc. Nat'l Acad. Sci. USA 86:10029-10033 (1989), U.S. Patent Nos. 5821337, 7527791, 6982321 and 7087409, and Kashmiri et al., Methods. This is explained in 36:25-34 (2005) (describes the specificity determination region (SDR) transplantation method), Padlan, Mol.Immunol.28:489-498 (1991) (describes resurfacing), Dall'Acqua et al., Methods 36:43-60 (2005) (describes "FR shuffling"), as well as Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br.J.Cancer,83:252-260 (2000) (describes the "guided selection" method of FR shuffling).

[0243] Human framework regions that can be used for humanization are not limited to, but include framework regions selected using the "best fit" method (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from human antibody consensus sequences of specific subgroups of heavy chain variable regions or light 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 (somatic mutant) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and framework regions derived from screening of FR libraries (see, e.g., Baca et al., J. Biol. Chem. See also 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996).

[0244] In some embodiments, the anti-CD20 / anti-CD3 bispecific antibody is a human antibody. Human antibodies can be manufactured using various techniques known in the art. Human antibodies are commonly described in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5:368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20:450-459 (2008).

[0245] Human antibodies may also be prepared by administering an immunogen to transgenic animals modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigen challenge. Such animals typically contain all or part of a human immunoglobulin locus that replaces, or is located extrachromosomally, or is randomly incorporated into the animal's chromosomes, replacing the endogenous immunoglobulin locus. In such transgenic mice, the endogenous immunoglobulin locus is generally inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125 (2005). Also, see, for example, xeno mice. TM See also U.S. Patent Nos. 6,075,181 and 6,150,584, which describe the technology; U.S. Patent No. 5,770,429, which describes the HuMab® technology; U.S. Patent No. 7,041,870, which describes the KM Mouse® technology; and U.S. Patent Application Publication 2007 / 0061900, which describes the VelociMouse® technology. Human variable regions from intact antibodies produced by such animals may be further modified, for example, by combining them with different human constant regions.

[0246] Human antibodies can also be produced by hybridoma-based methods. Human myeloma cell lines and mouse-human xenomyeloma cell lines for producing human monoclonal antibodies have been described. (See, for example, 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)). In addition, human antibodies produced via human B-cell hybridoma technology are described in Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006). Additional methods include, for example, U.S. Patent No. 7189826 (describes the production of monoclonal human IgM antibodies derived from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) (describes human-human hybridomas). Human hybridoma technology (trioma technology) 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).

[0247] Human antibodies can also be produced by isolating Fv clone variable domain sequences selected from human-derived phage display libraries. These variable domain sequences may then be combined with desired human constant domains. Techniques for selecting human antibodies from antibody libraries are described below.

[0248] The binding domains contained in anti-CD20 / anti-CD3 bispecific antibodies can be isolated by screening a combinatorial library for binding sites with desired activity. For example, various methods are known in the art for creating phage display libraries and screening the library for antibodies with desired binding properties. Such methods are discussed in, for example, Hoogenboom et al. Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, 2001), and furthermore, 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. This is explained in al., J.Mol.Biol.340(5):1073-1093(2004);Fellouse, Proc.Natl.Acad.Sci.USA 101(34):12467-12472(2004); and Lee et al., J.Immunol.Methods 284(1-2):119-132(2004).

[0249] In certain phage display methods, the repertoire of VH and VL genes are separately cloned by polymerase chain reaction (PCR), randomly recombined within a phage library, and then screened for antigen-binding phages as described in Winter et al., Ann. Rev. Immunol., 12:433-455 (1994). The phages typically display antibody fragments as either single-chain Fv (scFv) fragments or Fab fragments. Libraries from immunogens provide high-affinity antibodies against immunogens without the need to construct hybridomas. Alternatively, naive repertoires can be cloned (e.g., from humans) without immunization to provide a single source of antibodies against a wide range of non-self and autoantigens, as described by Griffiths et al., EMBO J, 12:725-734 (1993). Finally, naive libraries can also be constructed synthetically by cloning an unrearranged V gene segment from stem cells, encoding a highly variable CDR3 region using PCR primers containing random sequences, and achieving rearrangement in vitro, as described in Hoogenboom and Winter, J. Mol. Biol., 227:381-388 (1992). Examples of patent publications describing human antibody phage libraries include U.S. Patent No. 5,750,373, and U.S. Patent Application Publications 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360.

[0250] Antibodies or antibody fragments isolated from a human antibody library are considered human antibodies or human antibody fragments in this specification.

[0251] Techniques for producing bispecific antibodies include, but are not limited to, the recombinant co-expression of two immunoglobulin heavy-light chain pairs with different specificities (see Milstein and Cuello, Nature 305: 537 (1983)), International Publication No. 93 / 08829, and Traunecker et al., EMBO J.10:3655 (1991)) and the "knob-into-hole" operation (see, for example, U.S. Patent No. 5731168). Furthermore, multispecific antibodies can be produced using the following methods: manipulating the electrostatic steering effect to create antibody Fc heterodimer molecules (International Publication No. 2009 / 089004); crosslinking two or more antibodies or fragments (see, e.g., U.S. Patent No. 4676980 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 "diabody" techniques to produce bispecific antibody fragments (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and using single-chain Fv(sFv) dimers (see, e.g., Gruber et al., J. Immunol., See 152:5368 (1994); and it can also be prepared by the preparation of a triplicate antibody as described, for example, Tutt et al. J. Immunol. 147:60 (1991).

[0252] Modified antibodies having three or more functional antigen-binding sites, including "octopus antibodies," are also included herein (see, for example, U.S. Patent Application Publication 2006 / 0025576).

[0253] Furthermore, anti-CD20 / anti-CD3 bispecific antibodies as used herein include "dual-acting FAb" or "DAF" that contain antigen-binding sites that bind to two different antigens (see, for example, U.S. Patent Application Publication No. 2008 / 0069820).

[0254] Crossumab antibodies are also included in this specification (see, for example, International Publication Nos. 2009080251, 2009080252, 2009080253, and 2009080254).

[0255] Another technique for producing bispecific antibody fragments is the "bispecific T cell engager" or BiTE® method (see, for example, International Publications 2004 / 106381, 2005 / 061547, 2007 / 042261, and WO2008 / 119567). This method utilizes two antibody variable domains located on a single polypeptide. For example, a single polypeptide chain contains two single-chain Fv(scFv) fragments, each having variable heavy (VH) and variable light (VL) domains separated by a polypeptide linker of sufficient length to allow intramolecular association between the two domains. This single polypeptide further contains a polypeptide spacer sequence between the two scFv fragments. Each scFv recognizes a different epitope, and these epitopes may be specific to different cell types. When each scFv binds to its homogeneous epitope, cells of two different cell types will be brought closer together or linked. A particular embodiment of this technique involves an scFv that recognizes a cell surface antigen expressed by immune cells (e.g., CD3 polypeptide on T cells) bound to another scFv that recognizes a cell surface antigen expressed by a target cell, such as a malignant or tumor cell.

[0256] Since bispecific T cell-inducing antibodies are single polypeptides, they can be expressed using any prokaryotic or eukaryotic cell expression system known in the art, such as the CHO cell line. However, specific purification techniques (see, for example, European Patent No. 1691833) may be required to separate bispecific T cell-inducing antibodies of monomers from other multimer species, as these antibodies may possess biological activities other than those intended for the monomer. In one exemplary purification scheme, a solution containing the secreted polypeptide is first subjected to metal affinity chromatography to elute the polypeptide using an imidazole concentration gradient. This eluate is further purified using anion exchange chromatography to elute the polypeptide using a sodium chloride concentration gradient. Finally, this eluate is subjected to size exclusion chromatography to separate the monomer from the multimer species.

[0257] In certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody may be further modified to include additional non-protein moieties known and readily available in the art. Suitable moieties for derivatization of the anti-CD20 / anti-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, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymers, polyamino acids (either homopolymers or random copolymers), and dextran or poly(n-vinylpyrrolidone) polyethylene glycol, polypropylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may be advantageous in production due to its stability in water. The polymer may have any molecular weight and may be branched or unbranched. The number of polymers attached to the antibody may vary, and if more than one polymer is attached, they may be the same molecule or different molecules. In general, the number and / or types of polymers used for derivatization are not limited but can be determined based on considerations including the specific properties or functions of the antibody being improved, and whether the antibody derivative will be used for therapeutic purposes under specified conditions.

[0258] Anti-CD20 / anti-CD3 bispecific antibodies may also be conjugated with one or more cytotoxic agents, such as chemotherapeutic agents or drugs, growth inhibitors, toxins (e.g., protein toxins, enzyme-active toxins of bacterial, fungal, plant or animal origin, or fragments thereof), or radioisotopes.

[0259] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is an antibody that is a meitansinoid (see U.S. Patent No. 5208020, No. 5416064 and European Patent No. 0425235); auristatin such as monomethyl auristatin drug portions DE and DF (MMAE and MMAF) (see U.S. Patents No. 5635483, No. 5780588 and No. 7498298); drastatin; calicheamycin or its derivatives (see U.S. Patents No. 5712374, No. 5714586, No. 5739116, No. 5767285, No. 5770701, No. 5770710, No. 5773001 and No. 5877296; Hinman et al., Cancer See Res.53:3336-3342 (1993); and Lode et al., Cancer Res.58:2925-2928 (1998); anthracyclines such as daunomycin or doxorubicin (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. 6630579); including methotrexate; vindesine; taxanes such as docetaxel, paclitaxel, larotaxel, tesetaxel and ortataxel; trichothecenes; and antibody-drug conjugates (ADCs) conjugated to one or more drugs, including but not limited to CC1065.

[0260] In another embodiment, the anti-CD20 / anti-CD3 bispecific antibody is conjugated to an enzyme-active toxin or fragment thereof, including but not limited to diphtheria A chain, unbound active fragment of diphtheria toxin, exotoxin A chain (derived from Pseudomonas aeruginosa), lysine A chain, abrin A chain, modesine A chain, alpha-sarcin, tallow tree protein, dianthin protein, pokeweed protein (PAPI, PAPII, and PAP-S), bitter melon inhibitor, curcin, crocin, saponna inhibitor, geronin, mitogenin, restrictosin, phenomycin, enomycin, and trichothecenes.

[0261] In another embodiment, an anti-CD20 / anti-CD3 bispecific antibody is conjugated to a radioactive atom to form a radioactive conjugate. Various radioisotopes are available for the production of radioactive conjugates. For example, At 211 , I 131 , I 125 , Y 90 Re 186 Re 188 Sm 153 , Bi 212 , P 32 Pb 212 Examples include radioactive isotopes of Lu. When a radioactive conjugate is used for detection, it is a radioactive atom for scintigraphy, such as Tc. 99m or I 123 or spin labels for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging or MRI), which may again contain iodine-123, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese, or iron.

[0262] Conjugates of anti-CD20 / anti-CD3 bispecific antibodies and cytotoxic agents can be prepared using a variety of bifunctional protein coupling agents, such as N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), iminothiolane (IT), difunctional derivatives of imide esters (e.g., dimethyladipimidate HCl), active esters (e.g., disuccinimidylsberate), aldehydes (e.g., glutaraldehyde), bis-azide compounds (e.g., bis(p-azidobenzoyl)hexanediamine), bis-diazonium derivatives (e.g., bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (e.g., toluene 2,6-diisocyanate), and bis-active fluorine compounds (e.g., 1,5-difluoro-2,4-dinitrobenzene). For example, lysine immunotoxins 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 the conjugate of radioactive nucleotides to antibodies. See International Publication No. 94 / 11026. The linker may be a "cleavage linker" that facilitates the release of cytotoxic drugs within cells. For example, acid-unstable linkers, peptidase-sensitive linkers, photo-unstable linkers, dimethyl linkers, or disulfide-containing linkers can be used (Chari et al., Cancer Res. 52:127-131 (1992); U.S. Patent No. 5208020).

[0263] In certain embodiments, anti-CD20 / anti-CD3 bispecific antibodies are used to treat cancer. In some embodiments, cancer is a B-cell proliferative disorder. In some embodiments, cancer is a CD20-positive B-cell proliferative disorder. In some embodiments, cancer is non-Hodgkin lymphoma (NHL). In some embodiments, 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 (MZL). MZL can be classified into splenic MZL, nodal MZL, and extranodal MZL. In some embodiments, NHL is mantle cell lymphoma (MCL). In some embodiments, NHL is follicular lymphoma (FL) of grade 1-3a. In one embodiment, CD20-positive B-cell proliferative disorder is a relapsed or refractory B-cell proliferative disorder. In one embodiment, a relapsed or refractory positive B-cell proliferative disorder is a relapsed or refractory NHL (e.g., relapsed or refractory DLBCL, relapsed or refractory FL, or relapsed or refractory MCL).

[0264] In one embodiment, an anti-CD20 / anti-CD3 bispecific antibody specifically binds to CD3e.

[0265] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibodies are used for binding to the following antibodies: antibody H2C (International Publication No. 2008 / 119567), antibody V9 (Rodrigues et al., Int J Cancer Suppl. 7, 45-50 (1992) and U.S. Patent No. 6054297), antibody FN18 (Nooij et al., Eur J Immunol. 19, 981-984 (1986)), antibody SP34 (Pessano et al., EMBO J. 4, 337-340 (1985)), antibody OKT3 (Kung et al., Science 206, 347-349 (1979)), antibody WT31 (Spits et al., J Immunol. 135, 1922 (1985)), antibody UCHT1 (Burns et al., J It may compete with Immunol. 129, 1451-1457 (1982), antibody 7D6 (Coulie et al., Eur J Immunol. 21, 1703-1709 (1991)), or antibody Leu-4. Furthermore, in some embodiments, the anti-CD20 / anti-CD3 bispecific antibody is included in International Publication Nos. 2005 / 040220, 2005 / 118635, 2007 / 042261, 2008 / 119567, 2008 / 119565, 2012 / 162067, 2013 / 158856, 2013 / 188693, 2013 / 186613, 2014 / 110601, and International Publication Nos. As described in Publication No. 2014 / 145806, International Publication No. 2014 / 191113, International Publication No. 2014 / 047231, International Publication No. 2015 / 095392, International Publication No. 2015 / 181098, International Publication No. 2015 / 001085, International Publication No. 2015 / 104346, International Publication No. 2015 / 172800, International Publication No. 2016 / 020444, or International Publication No. 2016 / 014974, the material may include an antigen-binding moiety that specifically binds to CD3.

[0266] In some embodiments, the anti-CD20 / anti-CD3 bispecific antibody may contain an antibody or antigen-binding moiety from rituximab, obinutuzumab, ocrelizumab, ofatumumab, okalatuzumab, vertuzumab, and ubrituximab.

[0267] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is XmAb(registered trademark) 13676. In another embodiment, the anti-CD20 / anti-CD3 bispecific antibody is REGN1979. In another embodiment, the anti-CD20 / anti-CD3 bispecific antibody is FBTA05 (Lymphomun). In another embodiment, the anti-CD20 / anti-CD3 bispecific antibody is grofitamab.

[0268] In some embodiments, the anti-CD20 / anti-CD3 bispecific antibody may include generic, biosimilar, or biononequal versions of the antibodies listed herein.

[0269] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD20, and the antigen-binding domain is (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 1 (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 2 (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 3 A heavy chain variable region including, (i) HVR-L1 containing the amino acid sequence of SEQ ID NO: 4 (ii) HVR-L2 containing the amino acid sequence of Sequence ID No. 5 (iii) HVR-L3 containing the amino acid sequence of SEQ ID NO: 6 Includes a light chain variable region.

[0270] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD20, wherein the antigen-binding domain comprises 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 the sequence of SEQ ID NO: 8. In a further embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain that comprises the heavy chain variable region sequence of SEQ ID NO: 7 and the light chain variable region sequence of SEQ ID NO: 8 and specifically binds to CD20.

[0271] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD3, and the antigen-binding domain is (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 9 (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 10 (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 11 A heavy chain variable region including, (i) HVR-L1 containing the amino acid sequence of SEQ ID NO: 12 (ii) HVR-L2 containing the amino acid sequence of SEQ ID NO: 13 (iii) HVR-L3 containing the amino acid sequence of SEQ ID NO: 14 Includes a light chain variable region.

[0272] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD3, wherein the antigen-binding domain comprises 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 the sequence of SEQ ID NO: 16. In a further embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain that comprises the heavy chain variable region sequence of SEQ ID NO: 15 and the light chain variable region sequence of SEQ ID NO: 16 and specifically binds to CD3.

[0273] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is a) At least one antigen-binding domain that specifically binds to CD20, (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 1 (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 2 (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 3 A heavy chain variable region including; (i) HVR-L1 containing the amino acid sequence of SEQ ID NO: 4 (ii) HVR-L2 containing the amino acid sequence of Sequence ID No. 5 (iii) HVR-L3 containing the amino acid sequence of SEQ ID NO: 6 An antigen-binding domain including a light chain variable region and b) At least one antigen-binding domain that specifically binds to CD3, (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 9 (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 10 (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 11 A heavy chain variable region including; (i) HVR-L1 containing the amino acid sequence of SEQ ID NO: 12 (ii) HVR-L2 containing the amino acid sequence of SEQ ID NO: 13 (iii) HVR-L3 containing the amino acid sequence of SEQ ID NO: 14 It includes an antigen-binding domain that includes a light chain variable region.

[0274] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is (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 (ii) comprising at least one antigen-binding domain that specifically binds to CD3, including the heavy chain variable region sequence of SEQ ID NO: 15 and the light chain variable region sequence of SEQ ID NO: 16.

[0275] In one embodiment, the antigen-binding domain of an anti-CD20 / anti-CD3 bispecific antibody that specifically binds to CD3 is an antibody fragment, particularly a Fab molecule or scFv molecule, especially a Fab molecule. In a particular embodiment, the antigen-binding domain of an anti-CD20 / anti-CD3 bispecific antibody that specifically binds to CD3 is a crossover Fab molecule in which the variable domain or constant domain of the Fab heavy chain and the variable domain or constant domain of the Fab light chain are exchanged (i.e., substituted for each other).

[0276] In one embodiment, the anti-CD20 / anti-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 another embodiment, the anti-CD20 / anti-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 another embodiment, the first antigen-binding domain is a crossover Fab molecule, and the second and third antigen-binding domains are conventional Fab molecules. In another embodiment, the anti-CD20 / anti-CD3 bispecific antibody further comprises an Fc domain. The anti-CD20 / anti-CD3 bispecific antibody may include modifications of the Fc region and / or antigen-binding domain as described herein. In another embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises an IgG1 Fc domain containing one or more amino acid substitutions that reduce binding to and / or effector function of the Fc receptor. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises an IgG1 Fc domain including amino acid substitutions L234A, L235A, and P329G (EU numbering).

[0277] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is (i) An antigen-binding domain that specifically binds to CD3, and is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first subunit of the Fc domain; (ii) A first antigen-binding domain that specifically binds to CD20, wherein the antigen-binding domain is fused to the N-terminus of the Fab heavy chain of an antigen-binding domain that specifically binds to CD3 at the C-terminus of the Fab heavy chain; and (iii) A second antigen-binding domain that specifically binds to CD20, wherein the antigen-binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second subunit of the Fc domain. Includes.

[0278] In certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody is a) A first Fab molecule that specifically binds to CD3, particularly CD3 epsilon, wherein the variable domains VL and VH of the Fab light chain and Fab heavy chain are substituted for each other. b) A second Fab molecule and a third Fab molecule that specifically bind to CD20, wherein in the constant domain CL of the second and third Fab molecules, the amino acid at position 124 is substituted with lysine (K) (Kabat numbering), the amino acid at position 123 is substituted with lysine (K) or arginine (R), particularly arginine (R) (Kabat numbering), in the constant domain CH1 of the second and third Fab molecules, the amino acid at position 147 is substituted with glutamic acid (E) (EU numbering), the amino acid at position 213 is substituted with glutamic acid (E) (EU numbering), and c) Fc domain consisting of a first subunit and a second subunit capable of stable association. Includes.

[0279] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises two antigen-binding domains that specifically bind to CD20 and one antigen-binding domain that specifically binds to CD3.

[0280] In one embodiment, an anti-CD20 / anti-CD3 bispecific antibody is used, which is divalent against CD20 and monovalent against CD3.

[0281] In one embodiment, the first Fab molecule in a) is fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain in c), the second Fab molecule in 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 in a), and the third Fab molecule in b) is fused at the C-terminus of the Fab heavy chain to the N-terminus of the other subunit of the Fc domain in c). In one embodiment, the first Fab molecule in a) includes a heavy chain variable region which is at least 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 15, and a light chain variable region which is at least 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 16.

[0282] In a further embodiment, the first Fab molecule of a) includes the heavy chain variable region sequence of SEQ ID NO: 15 and the light chain variable region sequence of SEQ ID NO: 16.

[0283] In one embodiment, the second Fab molecule and the third Fab molecule of b) each include a heavy chain variable region which is at least 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 7, and a light chain variable region which is at least 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 8.

[0284] In one embodiment, the second Fab molecule and the third Fab molecule of b) each include the heavy chain variable region sequence of SEQ ID NO: 7 and the light chain variable region sequence of SEQ ID NO: 8, respectively.

[0285] In certain embodiments, the anti-CD20 / anti-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 further certain embodiments, 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 further certain embodiments, 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.

[0286] Specific anti-CD20 / anti-CD3 bispecific antibodies are described in International Publication No. 2016 / 020309 and European Patent Nos. 15188093 and 16169160, respectively (each incorporated herein by reference in its entirety).

[0287] Grofitamab In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody useful in the method provided herein is grofitamab. Glofitamab (proposed international generic name: List 121 WHO Drug Information, Vol. 33, No. 2, 2019, p. 276; alternative names: CD20-TCB, RO7082859 or RG6026; CAS number: 2229047-91-8) is a novel T cell-inducing bispecific (TCB) full-length antibody having a 2:1 molecular configuration, binding divalently to CD20 on B cells and monovalently to CD3, particularly the CD3 epsilon chain (CD3e), on T cells. Its CD3-binding domain is fused head-to-tail with one of the CD20-binding domains via a flexible linker. This structure gives grofitamab superior in vitro efficacy compared to other 1:1 CD20-CD3 bispecific antibodies, resulting in strong antitumor efficacy in preclinical DLBCL models. The bivalent nature of CD20 maintains this efficacy even in the presence of competing anti-CD20 antibodies, providing opportunities for prior or combination therapy with these agents. Glofitamab contains an engineered heterodimeric Fc region in which binding to FcgR and C1q is completely abolished. Glofitamab induces tumor cell lysis, as well as T cell activation, proliferation, and cytokine release, by co-binding to CD3e on the T cell receptor (TCR) complex on human CD20-expressing tumor cells and T cells. B cell-mediated lysis by grofitamab is CD20-specific and does not occur in the absence of CD20 expression or co-binding (crosslinking) of T cells to CD20-expressing cells. In addition to death, T cells are activated by CD3 crosslinking, which can be detected by increased T cell activation markers (CD25 and CD69), cytokine release (IFNγ, TNFα, IL-2, IL-6, IL-10), cytotoxic granule release (granzyme B), and T cell proliferation. A schematic diagram of the molecular structure of grofitamab is shown in Figure 2. The sequence of grofitamab is summarized in Table 2. Table 2. Sequence ID of grofitamab TIFF0007836317000002.tif76170

[0288] B. Anti-CD79b antibody drug conjugateExamples of anti-CD79b antibody drug conjugates useful in the methods described herein (for example, for treating CD20-positive cell proliferative disorders, such as B-cell proliferative disorders (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 and / or refractory MCL)), or CNSL) include any of the anti-CD79b antibody drug conjugates described in U.S. Patent No. 8,088,378 (in whole, incorporated herein by reference). In some examples, the anti-CD79b antibody drug conjugate includes an anti-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 (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26. In some examples, the anti-CD79b antibody drug conjugate contains an anti-CD79b binding domain that includes all of the following six HVRs: (a) HVR-H1 containing the amino acid sequence of GYTFSSYWIE (SEQ ID NO: 21); (b) HVR-H2 containing the amino acid sequence of GEILPGGGDTNYNEIFKG (SEQ ID NO: 22); (c) HVR-H3 containing the amino acid sequence of TRRVPIRLDY (SEQ ID NO: 23); (d) HVR-L1 containing the amino acid sequence of KASQSVDYEGDSFLN (SEQ ID NO: 24); (e) HVR-L2 containing the amino acid sequence of AASNLES (SEQ ID NO: 25); and (f) HVR-L3 containing the amino acid sequence of QQSNEDPLT (SEQ ID NO: 26).In some examples, the anti-CD79b antibody drug conjugate includes at least one (e.g., one, two, three, or four) heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4, each containing the sequences of SEQ ID NOs. 29–32, and / or at least one (e.g., one, two, three, or four) light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4, each containing the sequences of SEQ ID NOs. 33–36. In some examples, the anti-CD79b antibody drug conjugate includes (a) a heavy chain variable (VH) domain containing an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 27 (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) or the amino acid sequence of SEQ ID NO: 27; (b) a light chain variable (VL) domain containing an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 28 (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) or the amino acid sequence of SEQ ID NO: 28; or (c) a VH domain as in (a) and a VL domain as in (b). Therefore, in some examples, the first binding domain includes a VH domain containing the amino acid sequence of SEQ ID NO: 27 and a VL domain containing the amino acid sequence of SEQ ID NO: 28.

[0289] In some examples, the anti-CD79b antibody drug conjugate includes (a) a heavy chain containing an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 37 (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) or the amino acid sequence of SEQ ID NO: 37; (b) a light chain containing an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 38 (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) or the amino acid sequence of SEQ ID NO: 38; or (c) a VH domain as in (a) and a VL domain as in (b). Therefore, in some examples, the first binding domain includes a VH domain containing the amino acid sequence of SEQ ID NO: 37 and a VL domain containing the amino acid sequence of SEQ ID NO: 38.

[0290] The sequence of polatuzumab vedotin, an anti-CD79b antibody, is summarized in Table 3 below. Table 3. Sequence IDs of the anti-CD79b antibody polatuzumab vedotin TIFF0007836317000003.tif36170

[0291] In some cases, anti-CD79b antibodies bind to toxins such as monomethyl auristatin E (MMAE, i.e., vedotin). In some cases, anti-CD79b antibody drug conjugates are defined in the International Name of Drugs (INN) List 110 (WHO Drug Information, Vol. 27, No. 4, 2016, p. 443) as follows: polatuzumab vedotin (an immunoglobulin G1-kappa auristatin E conjugate, conjugated to monomethyl auristatin E (MMAE) with an average of 3-4 cysteinyl molecules via a cleavable maleimidocaproyl-valyl-citrullinyl-p-aminobenzyloxycarbonyl (mc-val-cit-PABC) type linker, anti-[Homo sapiens CD79b (immunoglobulin-associated CD79 beta)], a humanized monoclonal antibody conjugated to auristatin E; gamma single-chain (1-447) [humanized VH (Homo sapiens IGHV3-23 * 04(76.50%)-(IGHD)-IGHJ4 * 01)[8.8.10](1-117)-Homo sapiens IGGH1 * 03(CH1 R120>K(214)(118-215), hinge(216-230), CH2(231-340), CH3(341-445), CHS(446-447))(118-447)], (220-218')-disulfide having kappa light chain(1'-218') [humanized V-KAPPA (Homo sapiens IGKV1-39 * 01(85.90%)-IGKJ1 * 01)[10.3.9](1'-111')-Homo sapiens IGKC *01(112'-218'); dimer (226-226”:229-229”)-bis-disulfide; also known as RG-7596 or RO5541077-000). Polatuzumab vedotin is also known as IUPHAR / BPS number 8404, KEGG number D10761 or CAS registry number 1313206-42-6. Polatuzumab vedotin is also interchangeably referred to as "polatuzumab vedotin-piiq", "huMA79bv28-MC-vc-PAB-MMAE" or "DCDS4501A". In some embodiments, the anti-CD79b antibody (e.g., anti-CD79b ADC) contains the heavy chain sequence of SEQ ID NO: 37 and the light chain sequence of SEQ ID NO: 38.

[0292] In some examples, anti-CD79b antibody drug conjugates include the following formula: In formula TIFF0007836317000004.tif26170, Ab is an anti-CD79b antibody containing (i) the hypervariable region H1 (HVR-H1) containing the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 22; (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 24; (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 25; and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 26, and p is between 1 and 8.

[0293] 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 the 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 conjugate comprises: (i) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23 and / or (ii) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24. In some embodiments, the antibody-drug conjugate comprises an anti-CD79b antibody comprising (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 (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26.

[0294] In some embodiments, the antibody-drug conjugate comprises HVR-H3 containing the amino acid sequence of SEQ ID NO: 23 and / or HVR-L1 containing the amino acid sequence of SEQ ID NO: 24. In some embodiments, the antibody-drug conjugate comprises an anti-CD79b antibody comprising (a) HVR-H1 containing the amino acid sequence of SEQ ID NO: 21, (b) HVR-H2 containing the amino acid sequence of SEQ ID NO: 22, (c) HVR-H3 containing the amino acid sequence of SEQ ID NO: 23, (d) HVR-L1 containing the amino acid sequence of SEQ ID NO: 24, (e) HVR-L2 containing the amino acid sequence of SEQ ID NO: 25, and (f) HVR-L3 containing the amino acid sequence of SEQ ID NO: 26.

[0295] 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 as in 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 human VL kappa 1 (VLK1) framework and / or a VH framework VHIII. In some practical embodiments, the 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 (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, the humanized anti-CD79b antibody comprises (a) HVR-H1 containing the amino acid sequence of SEQ ID NO: 21, (b) HVR-H2 containing the amino acid sequence of SEQ ID NO: 22, (c) HVR-H3 containing the amino acid sequence of SEQ ID NO: 23, (d) HVR-L1 containing the amino acid sequence of SEQ ID NO: 24, (e) HVR-L2 containing the amino acid sequence of SEQ ID NO: 25, and (f) HVR-L3 containing the amino acid sequence of SEQ ID NO: 26.

[0296] In some embodiments, the antibody-drug conjugate (e.g., anti-CD79b antibody-drug conjugate) comprises an anti-CD79 antibody containing a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 27. In some embodiments, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence of SEQ ID NO: 27 contains substitutions (e.g., conservative substitutions), insertions, or deletions compared to the reference sequence, but the anti-CD79b antibody-drug conjugate containing that sequence retains the ability to bind to CD79b. In some embodiments, a total of 1 to 10 amino acids are substituted, inserted, and / or deleted in SEQ ID NO: 27. In some embodiments, a total of 1 to 5 amino acids are substituted, inserted, and / or deleted in SEQ ID NO: 27. In some embodiments, the substitution, insertion, or deletion occurs in the region outside the HVR (i.e., within the FR, e.g., SEQ ID NOs: 29-32). In some embodiments, the antibody-drug conjugate (e.g., The anti-CD79b antibody drug conjugate comprises an anti-CD79b antibody containing 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 containing the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 containing the amino acid sequence of SEQ ID NO: 22; and (c) HVR-H3 containing the amino acid sequence of SEQ ID NO: 23.

[0297] In some embodiments, the antibody-drug conjugate (e.g., anti-CD79b antibody-drug conjugate) comprises an anti-CD79b antibody containing a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 28. In certain embodiments, the VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence of SEQ ID NO: 28 contains substitutions (e.g., conservative substitutions), insertions, or deletions compared to the reference sequence, but the anti-CD79b antibody-drug conjugate containing that sequence retains its ability to bind to CD79b. In certain embodiments, a total of 1 to 10 amino acids are substituted, inserted, and / or deleted in SEQ ID NO: 28. In certain embodiments, a total of 1 to 5 amino acids are substituted, inserted, and / or deleted in SEQ ID NO: 28. In certain embodiments, the substitutions, insertions, or deletions occur in the outer region of the HVR (i.e., within the FR, e.g., SEQ ID NOs: 33-36). In some embodiments, the anti-CD79b antibody drug conjugate comprises an anti-CD79b antibody containing 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 containing the amino acid sequence of SEQ ID NO: 24; (b) HVR-L2 containing the amino acid sequence of SEQ ID NO: 25; and (c) HVR-L3 containing 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 having the amino acid sequence of SEQ ID NO: 24; (b) HVR-L2 having the amino acid sequence of SEQ ID NO: 25; and (c) HVR-L3 having the amino acid sequence of SEQ ID NO: 26.

[0298] In some embodiments, the antibody-drug conjugate (e.g., anti-CD79b antibody-drug conjugate) comprises an anti-CD79b antibody including a VH sequence, as in any of the embodiments provided herein, and a VL sequence, as in any of the embodiments provided herein. In some embodiments, the antibody-drug conjugate comprises an anti-CD79b antibody including a VH sequence and a VL sequence (including post-translational modifications of these sequences) in SEQ ID NO: 27 and SEQ ID NO: 28, respectively.

[0299] In some embodiments, the antibody-drug conjugate (e.g., anti-CD79b antibody-drug conjugate) includes an anti-CD79b antibody that binds to the same epitope as the anti-CD79b antibody described herein. For example, in some embodiments, the antibody-drug conjugate (e.g., anti-CD79b antibody-drug conjugate) includes 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.

[0300] In some embodiments, the antibody-drug conjugate comprises an anti-CD79b antibody, which is a monoclonal antibody, a chimeric antibody, a humanized antibody, or a human antibody. In some embodiments, the antibody-drug conjugate comprises an antigen-binding fragment of the anti-CD79b antibody described herein, e.g., Fv, Fab, Fab', scFv, diabody, or F(ab')2 fragment. In some embodiments, the antibody-drug conjugate comprises a substantially full-length anti-CD79b antibody, e.g., an IgG1 antibody or other antibody classes or isotypes described elsewhere herein. Anti-CD79b antibody-drug conjugates can be produced using recombinant methods and compositions, for example, as described in U.S. Patent No. 4816567.

[0301] In some examples, an anti-CD79b antibody drug conjugate according to any of the embodiments described above may incorporate, individually or in combination, the features described below.

[0302] C. Antibody format 1. Anti-CD20 / anti-CD3 bispecific antibody The components of anti-CD20 / anti-CD3 bispecific antibodies can be fused together in various configurations. An example configuration is shown in Figure 1.

[0303] In certain embodiments, the antigen-binding moiety contained in the anti-CD20 / anti-CD3 bispecific antibody is a Fab molecule. In such embodiments, the antigen-binding moieties, such as the first, second, and third, may be referred to herein as the first, second, and third Fab molecules, respectively. Furthermore, in certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody includes an Fc domain composed of a first subunit and a second subunit capable of stable association.

[0304] In one embodiment, the first Fab molecule is fused to the N-terminus of the first or second subunit of the Fc domain at the C-terminus of the Fab heavy chain.

[0305] In one such embodiment, the second Fab molecule is fused to the N-terminus of the Fab heavy chain of the first Fab molecule at the C-terminus of the Fab heavy chain. In a particular such embodiment, the anti-CD20 / anti-CD3 bispecific antibody essentially consists of a first Fab molecule and a second Fab molecule, where the Fc domain is composed of a first subunit and a second subunit and optionally one or more peptide linkers, the first Fab molecule is fused to the N-terminus of the first or second subunit of the Fc domain at the C-terminus of the Fab heavy chain, and the second Fab molecule is fused to the N-terminus of the Fab heavy chain of the first Fab molecule at the C-terminus of the Fab heavy chain. Such a configuration is schematically shown in Figures 1G and 1K. 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.

[0306] In another embodiment, the second Fab molecule is fused to the N-terminus of the first or second subunit of the Fc domain at the C-terminus of the Fab heavy chain. In a particular such embodiment, the antibody substantially consists of the first and second Fab molecules, an Fc domain composed of the first and second subunits, and one or more optional peptide linkers, with each of the first and second Fab molecules fused to the N-terminus of one of the subunits of the Fc domain at the C-terminus of the Fab heavy chain. Such configurations are schematically shown 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 particular embodiment, the first and second Fab molecules are each fused to the Fc domain via an immunoglobulin hinge region. In a specific embodiment, the immunoglobulin hinge region is the human IgG1 hinge region, in particular when the Fc domain is an IgG1Fc domain.

[0307] 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 subunit of the Fc domain. 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 a specific embodiment, the antibody essentially consists of first and second Fab molecules, an Fc domain composed of first and second subunits, and optionally one or more peptide linkers, where 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 subunit of the Fc domain. Such configurations are schematically shown in Figures 1H and 1L. 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.

[0308] Multiple Fab molecules can be fused to each other or to their Fc domains directly, or via a peptide linker containing one or more amino acids, typically about 2 to 20 amino acids. Peptide linkers are known in the art and are described herein. Suitable non-immunogenic peptide linkers include, for example, (G4S) n (SG4) n (G4S) n Or G4 (SG4) n A peptide linker is included. "n" is usually an integer from 1 to 10, typically from 2 to 4. In some embodiments, the peptide linker has an amino acid length of at least 5, in some embodiments from 5 to 100, and in further embodiments from 10 to 50. In some embodiments, the peptide linker is (GxS) n or (GxS) n G m [wherein G = glycine, S = serine, (x = 3, n = 3, 4, 5 or 6, m = 0, 1, 2 or 3) or (x = 4, n = 2, 3, 4 or 5, m = 0, 1, 2 or 3)], where in one embodiment x = 4, n = 2 or 3, and in a further embodiment x = 4, n = 2. In one embodiment, the peptide linker is (G4S)2. A specific suitable peptide linker for fusing the Fab light chains of the first and second Fab molecules together is (G4S)2. An exemplary peptide linker suitable for linking the Fab heavy chains of the first and second Fab fragments includes sequence (D)-(G4S)2. Another suitable such linker includes sequence (G4S)4. In addition, the linker may include (part of) an immunoglobulin hinge region. In particular, when the Fab molecule is fused to the N-terminus of the Fc domain subunit, it may be fused via the immunoglobulin hinge region or a portion thereof, with or without a further peptide linker.

[0309] Antibodies possessing a single antigen-binding moiety (such as a Fab molecule) that can specifically bind to a target cell antigen (e.g., those shown in Figures 1A, 1D, 1G, 1H, 1K, or 1L) are particularly useful when internal translocation of the target cell antigen is expected after binding of the high-affinity antigen-binding moiety. In such cases, the presence of more than one antigen-binding moiety specific to the target cell antigen enhances internal translocation of the target cell antigen, thereby reducing its availability.

[0310] However, in many other cases, for example, to optimize targeting to a target site or to enable crosslinking of target cell antigens, it would be advantageous to have an antibody containing two or more antigen-binding moieties (e.g., Fab molecules) specific to the target cell antigen (see examples shown in Figures 1B, 1C, 1E, 1F, 1I, 1J, 1M, or 1N).

[0311] Therefore, in certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody comprises two anti-CD20 binding moieties, for example, two Fab molecules targeting CD20. In some embodiments, the two Fab molecules targeting CD20 are conventional Fab molecules. In some embodiments, the two Fab molecules targeting CD20 contain the same heavy-chain and light-chain amino acid sequences and have the same domain configuration (i.e., conventional or crossover).

[0312] In an alternative embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises two anti-CD3 binding moieties, for example, two Fab molecules targeting CD3. In such an embodiment, both CD3-targeting Fab molecules are crossover Fab molecules (the variable domains VH and VL of the Fab heavy and light chains, or the constant regions CL and CH1). These are Fab molecules in which the two Fab molecules are exchanged / substituted for each other. In one such embodiment, the two Fab molecules targeting CD3 contain the same heavy-chain amino acid sequence and light-chain amino acid sequence and have the same domain configuration (i.e., conventional or crossover).

[0313] 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.

[0314] In certain embodiments, the second and third Fab molecules are each fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain, 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 certain such embodiments, the antibody essentially consists of the first, second and third Fab molecules, an Fc domain composed 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 subunit of the Fc domain, and the third Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second subunit of the Fc domain. Such configurations are schematically shown in Figures 1B and 1E (an embodiment in which the third Fab molecule is a conventional Fab molecule and is identical to the second Fab molecule), and in Figures 1I and 1M (an embodiment 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 certain embodiments, the second and third Fab molecules are fused to the Fc domain, respectively, via an immunoglobulin hinge region. In certain embodiments, the immunoglobulin hinge region is a human IgG1 hinge region, particularly when the Fc domain is an IgG1Fc domain. Optionally, the Fab light chains of the first Fab molecule and the second Fab molecule may be further fused to each other.

[0315] In another embodiment, the second and third Fab molecules are each fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain, 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 certain such embodiments, the antibody essentially consists of the first, second and third Fab molecules, an Fc domain composed 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 subunit of the Fc domain, and the third Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second subunit of the Fc domain. Such configurations are schematically shown in Figures 1C and 1F (an embodiment in which the third Fab molecule is a conventional Fab molecule and is identical to the second Fab molecule), and Figures 1J and 1N (an embodiment 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 certain embodiments, the second and third Fab molecules are fused to the Fc domain, respectively, via an immunoglobulin hinge region. In certain embodiments, the immunoglobulin hinge region is a human IgG1 hinge region, particularly when the Fc domain is an IgG1Fc domain. Optionally, the Fab light chains of the first and second Fab molecules may be further fused to each other.

[0316] In an antibody configuration in which a Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of each subunit of the Fc domain via an immunoglobulin hinge region, these two Fab molecules, the hinge region, and the Fc domain essentially form an immunoglobulin molecule. In certain embodiments, the immunoglobulin molecule is an IgG-class immunoglobulin. In even more specific embodiments, the immunoglobulin is an IgG1 subclass immunoglobulin. In another embodiment, the immunoglobulin is an IgG4 subclass immunoglobulin. In yet another embodiment, the immunoglobulin is a human immunoglobulin. In yet another embodiment, the immunoglobulin is a chimeric immunoglobulin or a humanized immunoglobulin.

[0317] In some antibodies, the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule are fused to each other, and sometimes via a peptide linker. Depending on the configuration of the first and second Fab molecules, the Fab light chain of the first Fab molecule can be fused at its C-terminus to the N-terminus of the Fab light chain of the second Fab molecule, or the Fab light chain of the second Fab molecule can be fused at its C-terminus to the N-terminus of the Fab light chain of the first Fab molecule. The fusion of the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule further reduces mispairing of unpaired Fab heavy chains and light chains, and also reduces the number of plasmids required for the expression of some of the antibodies of the present invention.

[0318] In a particular embodiment, the antibody is a polypeptide (VL) in which the Fab light chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule (i.e., the first Fab molecule contains a crossover Fab heavy chain, and the heavy chain variable region is replaced by the light chain variable region), and consequently the Fab heavy chain constant region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fc domain subunit. (1) -CH1 (1) -CH2-CH3(-CH4)), and the Fab heavy chain of the second Fab molecule share a carboxy-terminal peptide bond with the Fc domain subunit, forming a polypeptide (VH (2)-CH1 (2) The antibody contains a polypeptide (VH) in which the Fab heavy chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule. (1) -CL (1) ) and the Fab light chain polypeptide (VL) of the second Fab molecule (2) -CL (2) ) further comprises. In certain embodiments, these polypeptides are covalently bonded, for example, by disulfide bonds.

[0319] In a particular embodiment, the antibody is a polypeptide in which the Fab heavy chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule (i.e., the first Fab molecule contains a crossover Fab heavy chain, and the heavy chain constant region is replaced by the light chain constant region), and consequently, the Fab light chain constant region of the first Fab molecule shares a carboxy-terminal peptide bond with an Fc domain subunit (VH (1) -CL (1) -CH2-CH3(-CH4)), and the Fab heavy chain of the second Fab molecule share a carboxy-terminal peptide bond with the Fc domain subunit, forming a polypeptide (VH (2) -CH1 (2) The antibody contains a polypeptide (VL) in which the Fab light chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule. (1) -CH1 (1) ) and the Fab light chain polypeptide (VL) of the second Fab molecule (2) -CL (2) ) further includes. In certain embodiments, polypeptides are covalently bonded, for example, by disulfide bonds.

[0320] In some embodiments, the antibody has a Fab light chain variable region of a first Fab molecule sharing a carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule (i.e., the first Fab molecule includes a crossover Fab heavy chain with the heavy chain variable region replaced by the light chain variable region), and thus the Fab heavy chain constant region of the first Fab molecule sharing a carboxy-terminal peptide bond with the Fab heavy chain of a second Fab molecule, and thus the Fab heavy chain of the second Fab molecule sharing a carboxy-terminal peptide bond with an Fc domain subunit (VL (1) -CH1 (1) -VH (2) -CH1 (2) -CH2-CH3(-CH4)) polypeptide. In other embodiments, the antibody has a Fab heavy chain of a second Fab molecule sharing a carboxy-terminal peptide bond with the Fab light chain variable region of a first Fab molecule, and thus the Fab light chain variable region of the first Fab molecule sharing a carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule (i.e., the first Fab molecule includes a crossover Fab heavy chain with the heavy chain variable region replaced by the light chain variable region), and thus the Fab heavy chain constant region of the first Fab molecule sharing a carboxy-terminal peptide bond with an Fc domain subunit polypeptide (VH (2) -CH1 (2) -VL (1) -CH1 (1) -CH2-CH3(-CH4)).

[0321] In some of these embodiments, the antibody has a crossover Fab light chain polypeptide of the first Fab molecule (VH (1) -CL (1) ) in which the Fab heavy chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule and a Fab light chain polypeptide of the second Fab molecule (VL (2) -CL (2)) further comprises. In other embodiments of these embodiments, the antibody optionally has a Fab heavy chain variable region of the first Fab molecule sharing a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule, and thereby the Fab light chain constant region of the first Fab molecule sharing a carboxy-terminal peptide bond with the Fab light chain polypeptide of the second Fab molecule (VH (1) -CL (1) -VL (2) -CL (2) The polypeptide, or 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, and consequently the Fab heavy chain variable region of the first Fab molecule 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) ) Further contains polypeptides.

[0322] The antibodies according to these embodiments are (i) Fc domain subunit polypeptide (CH2-CH3(-CH4)) or (ii) polypeptide in which the Fab heavy chain of the third Fab molecule shares a carboxy-terminal peptide bond with the Fc domain subunit (VH (3) -CH1 (3) -CH2-CH3(-CH4)) and the Fab light chain polypeptide (VL) of the third Fab molecule (3) -CL (3) ) may further include. In certain embodiments, the polypeptides are covalently bonded, for example, by disulfide bonds.

[0323] In some embodiments, the antibody is a polypeptide (VH) in which the variable region of the Fab heavy chain of the second Fab molecule shares a carboxy-terminal peptide bond with the constant region of the Fab light chain of the first Fab molecule (i.e., the first Fab molecule contains a crossover Fab heavy chain, and the constant region of the heavy chain is replaced by the constant region of the light chain), and consequently the constant region of the Fab light chain of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of the second Fab molecule, and consequently the Fab heavy chain of the second Fab molecule shares a carboxy-terminal peptide bond with an Fc domain subunit. (1) -CL (1) -VH (2) -CH1 (2) In other embodiments, the antibody contains a polypeptide ((VH)) in which the Fab heavy chain of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain variable region of the first Fab molecule, and the Fab heavy chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule (i.e., the first Fab molecule contains a crossover Fab heavy chain, and the heavy chain constant region is replaced by the light chain constant region), and the Fab light chain constant region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fc domain subunit. (2) -CH1 (2) -VH (1) -CL (1) Includes -CH2-CH3(-CH4)).

[0324] In some of these embodiments, the antibody is a crossover Fab light chain polypeptide (VL) of the first Fab molecule, in which the Fab light chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule. (1) -CH1 (1) ) and the Fab light chain polypeptide (VL) of the second Fab molecule (2) -CL (2)) further comprises. In other embodiments of these embodiments, the antibody optionally comprises a polypeptide (VL) in which the Fab light chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule, and thereby the Fab heavy chain constant region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain polypeptide of the second Fab molecule. (1) -CH1 (1) -VL (2) -CL (2) ) or 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, and consequently the Fab heavy chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule v(VL (2) -CL (2) -VH (1) -CL (1) ) further includes.

[0325] The antibodies according to these embodiments are (i) Fc domain subunit polypeptide (CH2-CH3(-CH4)) or (ii) polypeptide in which the Fab heavy chain of the third Fab molecule shares a carboxy-terminal peptide bond with the Fc domain subunit (VH (3) -CH1 (3) -CH2-CH3(-CH4)) and the Fab light chain polypeptide (VL) of the third Fab molecule (3) -CL (3) ) may further contain. In certain embodiments, these polypeptides are covalently bonded, for example, by disulfide bonds.

[0326] In a particular embodiment, the antibody is a polypeptide (VH) in which the Fab heavy chain of the first Fab molecule shares a carboxy-terminal peptide bond with the variable region of the Fab light chain of the second Fab molecule, and consequently the variable region of the Fab light chain of the second Fab molecule 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 contains a crossover Fab heavy chain, and the heavy chain variable region is replaced by the light chain variable region). (1) -CH1 (1) -VL (2)-CH1 (2) ) includes. In some embodiments, the antibody is a polypeptide (VH) in which the Fab heavy chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the second Fab molecule. (2) -CL (2) ) and the Fab light chain polypeptide (VL) of the first Fab molecule (1) -CL (1) ) further includes.

[0327] In a particular embodiment, the antibody is a polypeptide (VL) in which the Fab light chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the second Fab molecule (i.e., the second Fab molecule contains a crossover Fab heavy chain, and the heavy chain variable region is replaced by the light chain variable region), and consequently the Fab heavy chain constant region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule. (2) -CH1 (2) -VH (1) -CH1 (1) ) includes. In some embodiments, the antibody is a polypeptide (VH) in which the Fab heavy chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the second Fab molecule. (2) -CL (2) ) and the Fab light chain polypeptide (VL) of the first Fab molecule (1) -CL (1) ) further includes.

[0328] In a particular embodiment, the antibody is a polypeptide (VH) in which the variable region of the Fab heavy chain of the second Fab molecule 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 contains a crossover Fab heavy chain, and the constant region of the heavy chain is replaced by the constant region of the light chain), and consequently the constant region of the Fab light chain of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule. (2) -CL (2) -VH (1) -CH1 (1)) includes. In some embodiments, the antibody is a polypeptide (VL) in which the Fab light chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the second Fab molecule. (2) -CH1 (2) ) and the Fab light chain polypeptide (VL) of the first Fab molecule (1) -CL (1) ) further includes.

[0329] In certain embodiments, the antibody is a polypeptide (VH) in which the Fab heavy chain of the third Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule, and consequently the Fab heavy chain of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain variable region of the second Fab molecule, and consequently the Fab light chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the second Fab molecule (i.e., the second Fab molecule contains a crossover Fab heavy chain, and the heavy chain variable region is replaced by the light chain variable region). (3) -CH1 (3) -VH (1) -CH1 (1) -VL (2) -CH1 (2) ) includes. In some embodiments, the antibody is a polypeptide (VH) in which the Fab heavy chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the second Fab molecule. (2) -CL (2) ) and the Fab light chain polypeptide (VL) of the first Fab molecule (1) -CL (1) ) further comprises. In some embodiments, the antibody is a third Fab molecule Fab light chain polypeptide (VL (3) -CL (3) ) further includes.

[0330] In certain embodiments, the antibody is a polypeptide (VH) in which the Fab heavy chain of the third Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule, and consequently the Fab heavy chain of the first Fab molecule shares a carboxy-terminal peptide bond with the variable region of the Fab heavy chain of the second Fab molecule, and consequently the variable region of the Fab heavy chain of the second Fab molecule 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 contains a crossover Fab heavy chain, and the constant region of the heavy chain is replaced by the constant region of the light chain). (3) -CH1 (3) -VH (1) -CH1 (1) -VH (2) -CL (2) ) includes. In some embodiments, the antibody is a polypeptide (VL) in which the Fab light chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the second Fab molecule. (2) -CH1 (2) ) and the Fab light chain polypeptide (VL) of the first Fab molecule (1) -CL (1) ) further comprises. In some embodiments, the antibody is a third Fab molecule Fab light chain polypeptide (VL (3) -CL (3) ) further includes.

[0331] In certain embodiments, the antibody is a polypeptide (VL) in which the Fab light chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the second Fab molecule (i.e., the second Fab molecule contains a crossover Fab heavy chain, and the heavy chain variable region is replaced by the light chain variable region), and consequently the Fab heavy chain constant region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule, and consequently the Fab heavy chain of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of the third Fab molecule. (2) -CH1 (2) -VH (1) -CH1 (1) -VH (3) -CH1 (3)) includes. In some embodiments, the antibody is a polypeptide (VH) in which the Fab heavy chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the second Fab molecule. (2) -CL (2) ) and the Fab light chain polypeptide (VL) of the first Fab molecule (1) -CL (1) ) further comprises. In some embodiments, the antibody is a third Fab molecule Fab light chain polypeptide (VL (3) -CL (3) ) further includes.

[0332] In certain embodiments, the antibody is a polypeptide (VH) in which the variable region of the Fab heavy chain of the second Fab molecule 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 contains a crossover Fab heavy chain, and the constant region of the heavy chain is replaced by the constant region of the light chain), and consequently the constant region of the Fab light chain of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule, and consequently the Fab heavy chain of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy ch...

Claims

1. A pharmaceutical agent comprising polatuzumab vedotin and grofitamab for the treatment of a subject or population of subjects having diffuse large B-cell lymphoma (DLBCL), wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) The first administration cycle comprises a first dose of 2.5 mg of grofitamab (C1D1) and a second dose of 10 mg of grofitamab (C1D2), and a single dose of 1.8 mg / kg of polatuzumab vedotin; and (b) The second administration cycle includes a single dose of 10 mg or 30 mg of grofitamab (C2D1) and a single dose of 1.8 mg / kg of polatuzumab vedotin, Pharmaceuticals.

2. A pharmaceutical agent comprising grofitamab for the treatment of a subject or population of subjects having DLBCL in combination with polatuzumab vedotin, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) The first administration cycle comprises a first dose of 2.5 mg of grofitamab (C1D1) and a second dose of 10 mg of grofitamab (C1D2), and a single dose of 1.8 mg / kg of polatuzumab vedotin; and (b) The second administration cycle includes a single dose of 10 mg or 30 mg of grofitamab (C2D1) and a single dose of 1.8 mg / kg of polatuzumab vedotin, Pharmaceuticals.

3. A pharmaceutical agent comprising polatuzumab vedotin for the treatment of a subject or population of subjects having DLBCL in combination with grofitamab, wherein polatuzumab vedotin and grofitamab are administered in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) The first administration cycle comprises a first dose of 2.5 mg of grofitamab (C1D1) and a second dose of 10 mg of grofitamab (C1D2), and a single dose of 1.8 mg / kg of polatuzumab vedotin; and (b) The second administration cycle includes a single dose of 10 mg or 30 mg of grofitamab (C2D1) and a single dose of 1.8 mg / kg of polatuzumab vedotin, Pharmaceuticals.

4. The pharmaceutical product according to any one of claims 1 to 3, wherein the single dose C1D1 or C2D1 of polatuzumab vedotin is about 1.8 mg / kg.

5. The pharmaceutical product according to any one of claims 1 to 3, wherein polatuzumab vedotin C1D1 is administered on the second day of the first administration cycle.

6. The pharmaceutical product according to any one of claims 1 to 5, wherein grofitamab C2D1 is administered to the subject on day 1 of the second administration cycle.

7. The pharmaceutical product according to any one of claims 1 to 6, wherein grofitamab C1D1 and grofitamab C1D2 are administered to the subject on the 8th and 15th days, respectively, of the first administration cycle.

8. The pharmaceutical product according to any one of claims 1 to 7, wherein grofitamab C2D1 is administered after the completion of administration of polatuzumab vedotin C2D1.

9. The pharmaceutical product according to any one of claims 1 to 8, wherein the administration regimen includes one or more additional administration cycles.

10. The pharmaceutical product according to claim 9, wherein the administration regimen includes 6 to 10 additional administration cycles.

11. The pharmacopoeia according to claim 9 or 10, wherein one or more of the additional administration cycles comprise an additional single dose of grofitamab and an additional single dose of polatuzumab vedotin.

12. The pharmaceutical product according to claim 11, wherein the additional single dose of polatuzumab vedotin is about 1.8 mg / kg.

13. The pharmacopoeci of any one of claims 9 to 12, wherein the additional single dose of polatuzumab vedotin is administered to the subject on day 1 of each additional administration cycle comprising the additional dose of polatuzumab vedotin.

14. The pharmacopoeia according to any one of claims 9 to 13, wherein the additional single dose of grofitamab in each additional dosing cycle, including an additional dose of polatuzumab vedotin, is administered after the completion of administration of the additional single dose of polatuzumab vedotin.

15. The pharmacopoeia according to any one of claims 9 to 13, wherein one or more of the additional administration cycles comprise an additional single dose of grofitamab and not administration of polatuzumab vedotin.

16. The pharmaceutical product according to claim 15, wherein the additional single dose of grofitamab is about 30 mg.

17. The pharmacopoeia according to claim 9, wherein the administration regimen comprises 10 additional administration cycles, each of which comprises a single dose of grofitamab, and four of the 10 additional administration cycles comprises the administration of polatuzumab vedotin.

18. The pharmaceutical product according to any one of claims 1 to 17, wherein the administration cycle is a 21-day administration cycle.

19. The pharmaceutical product according to any one of claims 1 to 18, wherein polatuzumab vedotin and grofitamab are administered together with one or more additional therapeutic agents selected from one or more chemotherapeutic agents, tocilizumab, corticosteroids, antihistamines, allopurinol, rasburicase, antipyretics, and obinutuzumab.

20. The pharmaceutical product according to claim 19, wherein the one or more chemotherapeutic agents include cyclophosphamide, doxorubicin, and rituximab.

21. The pharmaceutical product according to claim 19, wherein the corticosteroid comprises prednisone, prednisolone, methylprednisolone, and dexamethasone.

22. The pharmaceutical product according to claim 19, wherein polatuzumab vedotin and grofitamab are administered together with rituximab, cyclophosphamide, doxorubicin and prednisone (R-CHP).

23. The pharmaceutical product according to claim 19, wherein obinutuzumab is to be administered before the administration of grofitamab.

24. The pharmaceutical product according to claim 19, wherein obinutuzumab is administered approximately seven days before the administration of grofitamab.

25. The pharmaceutical product according to claim 24, wherein obinutuzumab is administered as a single dose of approximately 1000 mg.

26. The pharmaceutical product according to any one of claims 1 to 25, wherein grofitamab and polatuzumab vedotin are to be administered intravenously.

27. The pharmaceutical product according to any one of claims 1 to 26, wherein the subject or group of subjects has received at least two prior systemic therapies.

28. The pharmaceutical product according to any one of claims 1 to 27, wherein the subject or population of subjects is unsuitable for autologous stem cell transplantation (SCT).

29. The pharmaceutical product according to any one of claims 1 to 28, wherein the complete response rate of the target population is at least 20%, or the overall response rate of the target population is at least 30%.

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