Subcutaneous administration of Anti-CD20 / Anti-CD3 bispecific antibodies

Abstract

To provide a method for treating a subject having CD20-positive cell proliferative disorder.SOLUTION: The method comprises subcutaneously administering a bispecific antibody that binds to CD20 and CD3 to the subject in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, where (a) the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2) and a third subcutaneous dose (C1D3) of the bispecific antibody, where: (i) the C1D1 is no greater than the C1D2 and less than the C1D3; (ii) the C1D2 is no greater than the C1D3; and (iii) the C1D1 is from about 0.1 mg to about 10 mg, the C1D2 is from about 5 mg to about 80 mg, and the C1D3 is from about 10 mg to about 300 mg; and (b) the second dosing cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, where the C2D1 is equal to or greater than the C1D3 and is from about 10 mg to about 300 mg.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy created on November 1, 2021, is titled 50474-235WO4_Sequence_Listing_11_1_21_ST25 and is 35,329 bytes in size.

[0002] FIELD OF THE INVENTION The present invention relates to the treatment of CD20-positive cell proliferative disorders. More specifically, the present invention relates to the treatment of subjects with CD20-positive cell proliferative disorders by subcutaneous administration of a bispecific antibody that binds to cluster of differentiation 20 (CD20) and cluster of differentiation 3 (CD3). [Background technology]

[0003] background Cancer is characterized by the uncontrolled proliferation of a cell subpopulation. Cancer is the leading cause of death in developed countries and the second leading cause of death in developing countries, with over 14 million new cancer cases diagnosed and over 8 million cancer deaths occurring each year. Therefore, cancer care represents a significant and growing societal burden.

[0004] CD20-positive cell proliferative disorders, such as B-cell proliferative disorders, are a leading cause of cancer-related deaths. For example, non-Hodgkin's lymphoma (NHL) is rapidly progressive and, if untreated, fatal. In the United States, B-cell lymphomas account for approximately 80% to 85% of all NHL cases. Diffuse large B-cell lymphoma (DLBCL) is the most common type of NHL, accounting for approximately 30% to 40% of all NHL diagnoses, followed by follicular lymphoma (FL; 20% to 25% of all NHL diagnoses) and mantle cell lymphoma (MCL; 6% to 10% of all NHL diagnoses). B-cell chronic lymphocytic leukemia (CLL) is the most common leukemia in adults, with approximately 15,000 new cases each year in the United States (American Cancer Society 2015).

[0005] Bispecific antibodies are capable of simultaneously binding cell surface antigens on cytotoxic cells (e.g., T cells via binding to cluster of differentiation 3 (CD3)) and cancer cells (e.g., B cells via binding to CD20) with the intent that the bound cytotoxic cells will destroy the bound cancer cells. However, antibody-based immunotherapies such as these can be limited by undesirable effects, including cytokine-driven toxicity (e.g., cytokine release syndrome (CRS)), infusion-related reactions (IRR), severe tumor lysis syndrome (TLS), and central nervous system (CNS) toxicity.

[0006] Thus, there is an unmet need in the art to develop effective methods of administering therapeutic bispecific antibodies (e.g., bispecific antibodies that bind to CD20 and CD3) for the treatment of CD20-positive cell proliferative disorders (e.g., B-cell proliferative disorders) that achieve a more favorable benefit-risk profile. Summary of the Invention

[0007] The present invention relates to a method of treating a subject with a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder) by subcutaneously administering a bispecific antibody that binds to cluster of differentiation 20 (CD20) and cluster of differentiation 3 (CD3).

[0008] In one aspect, the invention provides a method of treating a subject with a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first administration cycle and a second administration cycle, wherein: (a) the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein: (i) C1D1 is equal to or less than C1D2 and less than C1D3; (ii) C1D2 is equal to or less than C1D3; and (iii) C1D1 is between about 0.1 mg and about 10 mg (e.g., between about 0.1 mg and about 7 mg, between about 0.2 mg and about 10 mg, between about 0.5 mg and about 15 mg, or between about 0.6 mg and about 15 mg).5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (for example, about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 (b) the second administration cycle comprises a single subcutaneous dose (2D1) of the bispecific antibody, C2D1 being equal to or greater than C1D3 and is between about 10 mg and about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 25 mg to about 300 mg, about 30 mg to about 300 mg, about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg); The dose is about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).

[0009] In some embodiments, C1D1 is less than C1D2. In some embodiments, C1D2 is equivalent to C1D3. In some embodiments, (a) C1D1 is about 2 mg to about 8 mg, and C1D2 is about 10 mg to about 75 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg). (b) C1D3 is about 20 mg to about 75 mg (e.g., about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 45 mg); and (b) C2D1 is about 20 mg to about 75 mg (e.g., about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 45 mg). In some embodiments, C1D1 is about 5 mg. In some embodiments, C1D3 is about 25 mg to about 75 mg. In some embodiments, C1D3 is about 30 mg, about 45 mg, or about 60 mg. In some embodiments, C2D1 is about 40 mg to about 75 mg. In some embodiments, C2D1 is about 30 mg, about 45 mg, or about 60 mg. In some embodiments, C1D2 is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 45 mg, or about 60 mg.

[0010] In some embodiments, C1D1 is about 5 mg, C1D2 is about 45 mg, C1D3 is about 45 mg, and C2D1 is about 45 mg; C1D1 is about 5 mg, C1D2 is about 10 mg, C1D3 is about 30 mg, and C2D1 is about 30 mg; C1D1 is about 5 mg, C1D2 is about 15 mg, C1D3 is about 45 mg, and C2D1 is about 45 mg; C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 40 mg, and C2D1 is about 40 mg; or C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 60 mg, and C2D1 is about 60 mg. In some embodiments, C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 60 mg, and C2D1 is about 60 mg.

[0011] In some embodiments, C1D1 is equal to C1D2 (e.g., C1D1 is about 5 mg, C1D2 is about 5 mg, C1D3 is about 45 mg, and C2D1 is about 45 mg). In some embodiments, C1D1 is equal to C1D2 (e.g., C1D1 is about 5 mg, C1D2 is about 5 mg, C1D3 is about 60 mg, and C2D1 is about 60 mg). In other embodiments, C1D2 is equal to C1D3 (e.g., C1D1 is about 5 mg, C1D2 is about 60 mg, C1D3 is about 60 mg, and C2D1 is about 60 mg). In other embodiments, C1D2 is equal to C1D3 (e.g., C1D1 is about 5 mg, C1D2 is about 45 mg, C1D3 is about 45 mg, and C2D1 is about 60 mg).

[0012] In some embodiments, the method includes administering C1D2 to the subject about 7 days after C1D1. In some embodiments, the method includes administering C1D3 to the subject about 7 days after C1D2. In some embodiments, the method includes administering C2D1 to the subject about 7 days after C1D3. In some embodiments, the method includes administering C1D1, C1D2, and C1D3 to the subject on days 1, 8, and 15, or about days 1, 8, and 15, respectively, of a first administration cycle. In some embodiments, the method includes administering C2D1 to the subject on day 1 of a second administration cycle.

[0013] In some embodiments, the first and second dosing cycles are 21 day dosing cycles.

[0014] In some embodiments, the first and second administration cycles are 28-day administration cycles, hi some embodiments, the first administration cycle is a 21-day administration cycle and the second administration cycle is a 28-day administration cycle.

[0015] In another aspect, the invention provides a method of treating a subject with a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first and a second dosing cycle, wherein (a) the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, where (i) C1D1 is about 5 mg, (ii) C1D2 is equal to or greater than C1D1 and equal to or less than C1D3, and (iii) C1D3 is about 60 mg; and (b) the second dosing cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), where C2D1 is about 60 mg. In some embodiments, C1D3 is about 45 mg.

[0016] In some embodiments, C1D2 is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 45 mg, or about 60 mg. In some embodiments, C1D2 is about 15 mg. In some embodiments, C1D2 is about 45 mg. In some embodiments, the first administration cycle and the second administration cycle are 21-day administration cycles. In some embodiments, the method comprises administering C1D1, C1D2, and C1D3 to a subject on days 1, 8, and 15, or about days 1, about 8, and about 15, of the first administration cycle, respectively. In some embodiments, the method comprises administering C2D1 to a subject on day 1 of the second administration cycle. In some embodiments, the first administration cycle and the second administration cycle are 21-day administration cycles. In some embodiments, the first administration cycle and the second administration cycle are 28-day administration cycles. In some embodiments, the first administration cycle is a 21 day administration cycle and the second administration cycle is a 28 day administration cycle.

[0017] In another aspect, the invention features a method of treating a subject with a CD20-positive proliferative disorder (e.g., a B-cell proliferative disorder), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first subcutaneous dose of the bispecific antibody (C1D1) on day 1 of the first dosing cycle, a second subcutaneous dose of the bispecific antibody (C1D2) on day 8 of the first dosing cycle, and a third subcutaneous dose of the bispecific antibody (C1D3) on day 15 of the first dosing cycle, wherein (i) C1D1 is about 5 mg, (ii) C1D2 is greater than or equal to C1D1 and less than or equal to C1D3, and (iii) C1D3 is about 45 mg; and (b) the second dosing cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1) on day 1 of the second dosing cycle, wherein C2D1 is about 45 mg.

[0018] In another aspect, the invention features a method of treating a subject with a CD20-positive proliferative disorder (e.g., a B-cell proliferative disorder), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first subcutaneous dose of the bispecific antibody (C1D1) on day 1 of the first dosing cycle, a second subcutaneous dose of the bispecific antibody (C1D2) on day 8 of the first dosing cycle, and a third subcutaneous dose of the bispecific antibody (C1D3) on day 15 of the first dosing cycle, wherein (i) C1D1 is about 5 mg, (ii) C1D2 is greater than or equal to C1D1 and less than or equal to C1D3, and (iii) C1D3 is about 60 mg; and (b) the second dosing cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1) on day 1 of the second dosing cycle, wherein C2D1 is about 60 mg. In some embodiments, the C1D3 is about 45 mg.

[0019] In some embodiments, C1D2 is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 45 mg, or about 60 mg. In some embodiments, C1D2 is about 15 mg. In some embodiments, C1D2 is about 45 mg. In some embodiments, each additional administration cycle is a 21-day administration cycle. In some embodiments, each additional administration cycle is a 28-day administration cycle.

[0020] In some embodiments, each of the one or more additional administration cycles comprises a single subcutaneous dose of the bispecific antibody. In some embodiments, the method comprises administering to the subject a single subcutaneous dose on day 1 of each of the one or more additional administration cycles.

[0021] In some embodiments of any of the foregoing methods, the CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder) is non-Hodgkin's lymphoma (NHL) or chronic lymphocytic leukemia (CLL). In some embodiments, the NHL is diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL). In some embodiments, the NHL is previously untreated (1L) NHL. In some embodiments, the NHL is relapsed or refractory NHL (R / R NHL). In some embodiments, the DLBCL is 1L DLBCL. In some embodiments, the DLBCL is relapsed or refractory DLBCL. In some embodiments, the DLBCL is Richter's transformed. In some embodiments, the FL is 1L FL. In some embodiments, the FL is relapsed or refractory FL. In some embodiments, the FL is transformed FL. In some embodiments, the NHL is high-grade B-cell lymphoma. In some embodiments, the NHL is Ann Arbor Stage III or IV NHL. In some embodiments, the subject has previously received at least one (e.g., at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or more) prior line of systemic therapy. In some embodiments, the subject has received between one and nine (e.g., one, two, three, four, five, six, seven, eight, or nine) prior lines of systemic therapy. In some embodiments, the subject has received three prior lines of systemic therapy. In some embodiments, at least one (e.g., one, two, three, four, five, six, seven, eight, or nine) prior line of systemic therapy included an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is rituximab or obinutuzumab. In some embodiments, the prior line of systemic therapy comprising the anti-CD20 antibody additionally comprises an alkylating agent or an anthracycline. In some embodiments, the alkylating agent is cyclophosphamide or bendamustine.In some embodiments, the anthracycline is daunomycin or doxorubicin. In some embodiments, the prior line of systemic therapy comprising an anti-CD20 antibody additionally comprises: (i) cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP); (ii) cyclophosphamide, vincristine, and prednisone (CVP); (iii) fludarabine; or (iv) bendamustine. In some embodiments, at least one (e.g., one, two, three, four, five, six, seven, eight, or nine) prior line of systemic therapy included a Bruton's tyrosine kinase (BTK) inhibitor.

[0022] In another aspect of the invention, there is provided a method of treating a subject with DLBCL, comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first 21-day dosing cycle and a second 21-day dosing cycle, wherein: (a) the first 21-day dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) C1D2, C1D3, and C1D4; D1 is equal to or less than C1D2 and less than C1D3, (ii) C1D2 is equal to or less than C1D3, and (iii) C1D1 is about 0.1 mg to about 10 mg (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; e.g., about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg). g to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg;(b) a second 21-day administration cycle comprising a single subcutaneous dose (2D1) of the bispecific antibody, wherein C2D1 is equal to or greater than C1D3 and is between about 10 mg and about 300 mg (e.g., between about 25 mg and about 300 mg, between about 50 mg and about 300 mg, between about 100 mg and about 300 mg, between about 200 mg and about 300 mg, between about 50 mg and about 250 mg, between about 10 ... In some embodiments, the dose of C1D1 is about 0 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; e.g., about 30 mg, about 45 mg, or about 60 mg). In some embodiments, the DLBCL is 1L DLBCL, or relapsed or refractory DLBCL. In some embodiments, the DLBCL is Richter's transformed. In some embodiments, the method comprises administering C1D2 to the subject about 7 days after C1D1. In some embodiments, the method comprises administering C1D3 to the subject about 7 days after C1D2. In some embodiments, the method includes administering C2D1 to the subject about 7 days after C1D3. In some embodiments, the method includes administering C1D1, C1D2, and C1D3 to the subject on or about days 1, 8, and 15, respectively, of a first administration cycle.

[0023] In another aspect, the invention provides a method of treating a subject with FL, comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first 28-day dosing cycle and a second 28-day dosing cycle, wherein: (a) the first 28-day dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein: (i) C1D1 is equal to or less than C1D2 and less than C1D3; (ii) C1D2 is equal to or less than C1D3; and (iii) C1D1 is between about 0.1 mg and about 10 mg (e.g., between about 0.1 mg and about 7 mg, between about 0.2 mg and about 10 mg, between about 0.5 mg and about 15 mg, or between about 0.5 mg and about 15 mg).5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (for example, about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg , about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg (b) a second 28-day administration cycle comprising a single subcutaneous dose (C2D1) of bispecific antibody, C2D1 being equal to or greater than C1D3 and ranging from about 10 mg to about 300 mg (e.g., about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg); g, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; e.g., about 30 mg, about 45 mg, or about 60 mg). In some embodiments, the FL is previously untreated (1L) FL or relapsed or refractory FL. In some embodiments, the FL is previously untreated (1L) FL. In some embodiments, the FL is transformed FL.

[0024] In another aspect, the invention provides a method of treating a subject with FL, comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first 21-day dosing cycle and a second 28-day dosing cycle, wherein: (a) the first 21-day dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein: (i) C1D1 is equal to or less than C1D2 and less than C1D3; (ii) C1D2 is equal to or less than C1D3; and (iii) C1D1 is between about 0.1 mg and about 10 mg (e.g., between about 0.1 mg and about 7 mg, between about 0.2 mg and about 10 mg, between about 0.5 mg and about 15 mg, or between about 0.5 mg and about 15 mg).5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (for example, about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg , about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg (b) a second 28-day administration cycle comprising a single subcutaneous dose (C2D1) of bispecific antibody, C2D1 being equal to or greater than C1D3 and ranging from about 10 mg to about 300 mg (e.g., about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg); g, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; e.g., about 30 mg, about 45 mg, or about 60 mg). In some embodiments, the FL is previously untreated (1L) FL or relapsed or refractory FL. In some embodiments, the FL is previously untreated (1L) FL. In some embodiments, the FL is transformed FL.

[0025] In some embodiments, the method comprises administering C1D2 to the subject about 7-10 days after C1D1. In some embodiments, the method comprises administering C1D3 to the subject about 7-10 days after C1D2. In some embodiments, the method comprises administering C2D1 to the subject about 7-10 days after C1D3. In some embodiments, C1D1 is less than C1D2. In other embodiments, C1D2 is about equal to or less than C1D3. In some embodiments, (a) C1D1 is about 2 mg to about 8 mg (e.g., about 5 mg), and C1D2 is about 10 mg to about 75 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg). g, about 65 mg, about 70 mg, or about 75 mg), C1D3 is about 20 mg to about 75 mg (e.g., about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 45 mg), and (b) C2D1 is about 20 mg to about 75 mg (e.g., about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 45 mg). In some embodiments, C1D1 is about 5 mg. In some embodiments, C1D3 is about 25 mg to about 75 mg. In some embodiments, C1D3 is about 30 mg, about 45 mg, or about 60 mg. In some embodiments, C2D1 is about 40 mg to about 75 mg (e.g., about 30 mg, about 45 mg, or about 60 mg), and C1D2 is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 45 mg, or about 60 mg.

[0026] In some embodiments, C1D1 is about 5 mg, C1D2 is about 10 mg, C1D3 is about 30 mg, and C2D1 is about 30 mg; C1D1 is about 5 mg, C1D2 is about 15 mg, C1D3 is about 45 mg, and C2D1 is about 45 mg; C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 40 mg, and C2D1 is about 40 mg; C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 45 mg, and C2D1 is about 60 mg; or C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 60 mg, and C2D1 is about 60 mg. In some embodiments, C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 45 mg, and C2D1 is about 60 mg. In some embodiments, C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 60 mg, and C2D1 is about 60 mg. In some embodiments, C1D1 is equal to C1D2. In some embodiments, C1D1 is about 5 mg, C1D2 is about 5 mg, C1D3 is about 60 mg, and C2D1 is about 60 mg. In some embodiments, C1D1 is about 5 mg, C1D2 is about 5 mg, C1D3 is about 45 mg, and C2D1 is about 60 mg. In some embodiments, C1D2 is equal to C1D3 (e.g., C1D1 is about 5 mg, C1D2 is about 60 mg, C1D3 is about 60 mg, and C2D1 is about 60 mg, or e.g., C1D1 is about 5 mg, C1D2 is about 45 mg, C1D3 is about 45 mg, and C2D1 is about 60 mg). In some embodiments, the method comprises administering C2D1 to the subject on day 1 of a second administration cycle.

[0027] In some embodiments, the dosing regimen includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) additional administration cycles (e.g., 1 to 15 additional administration cycles, 8 to 17 additional administration cycles, or 6 to 15 additional administration cycles). In some embodiments, the dosing regimen includes 6 additional administration cycles. In some embodiments, the dosing regimen includes 15 additional administration cycles. In some embodiments, the dosing regimen includes 2 to 17 (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17) total administration cycles. In some embodiments, the dosing regimen includes 8 total administration cycles. In some embodiments, the dosing regimen includes 17 total administration cycles. In some embodiments, each additional administration cycle is a 21-day administration cycle. In some embodiments, each additional administration cycle is a 28-day administration cycle. In some embodiments, each additional administration cycle comprises administering an additional dose of the bispecific antibody. In some embodiments, each additional dose of the bispecific antibody is an amount approximately equal to C2D1. In some embodiments, each additional dose of the bispecific antibody is about 45 mg. In some embodiments, the method comprises administering each additional dose of the bispecific antibody to the subject on day 1 of each respective additional administration cycle.

[0028] In some embodiments of any of the preceding aspects, the bispecific antibody is administered to the subject as monotherapy.

[0029] In other embodiments of any of the preceding aspects, the bispecific antibody is administered to the subject as a combination therapy. In some embodiments, the bispecific antibody is administered to the subject simultaneously with one or more additional therapeutic agents. In some embodiments, the bispecific antibody is administered to the subject prior to administration of the one or more additional therapeutic agents. In some embodiments, the bispecific antibody is administered to the subject after administration of the one or more additional therapeutic agents. In some embodiments, the additional therapeutic agent is a CD79b antibody drug conjugate (ADC), e.g., polatuzumab vedotin or anti-CD79b-MC-vc-PAB-MMAE. In some embodiments, the additional therapeutic agent is a PD-1 axis binding antagonist (e.g., a PD-L1 antagonist antibody). In some embodiments, the additional therapeutic agent is obinutuzumab (GAZYVA®). In some embodiments, the additional therapeutic agent is lenalidomide.

[0030] In some embodiments of any one of the preceding aspects, the subject has a cytokine release syndrome event, and the method further comprises treating symptoms of the cytokine release syndrome event while withholding treatment with the bispecific antibody. In some embodiments, the method further comprises administering to the subject an effective amount of tocilizumab to treat the cytokine release syndrome event. In some embodiments, tocilizumab is administered intravenously to the subject as a single dose of about 8 mg / kg. In some embodiments, the cytokine release syndrome event does not resolve or worsens within 24 hours of treating the symptoms of the cytokine release syndrome event, and the method further comprises administering to the subject one or more additional doses of tocilizumab to manage the cytokine release syndrome event. In some embodiments, the one or more additional doses of tocilizumab are administered intravenously to the subject at a dose of about 8 mg / kg. In some embodiments, each dose of tocilizumab does not exceed 800 mg / dose. In some embodiments, the method further comprises administering to the subject an effective amount of a corticosteroid (e.g., methylprednisolone or dexamethasone). In some embodiments, a corticosteroid (e.g., methylprednisolone or dexamethasone) is administered intravenously to a subject. In some embodiments, methylprednisolone is administered at a single dose of about 2 mg / kg per day. In some embodiments, dexamethasone is administered at a dose of about 10 mg to about 100 mg (e.g., about 10 mg).

[0031] In another aspect, the invention provides a method of treating a population of subjects having a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder), comprising subcutaneously administering to one or more subjects a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein: (i) C1D1 is equal to or less than C1D2 and less than C1D3; (ii) C1D2 is equal to or less than C1D3; and (iii) C1D1 is between about 0.1 mg and about 10 mg (e.g., between about 0.1 mg and about 7 mg, between about 0.2 mg and about 10 mg, between about 0.5 mg and about 15 mg, or between about 0.6 mg and about 15 mg).C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg). g, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 1 (b) the second administration cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, C2D1 being equal to or greater than C1D3 and ranging from about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, The method is characterized in that the dose is about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).

[0032] In another aspect, the invention features a method of treating a population of subjects having a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, where (i) C1D1 is about 5 mg, (ii) C1D2 is greater than or equal to C1D1 and less than or equal to C1D3, and (iii) C1D3 is about 45 mg; and (b) the second dosing cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), where C2D1 is about 45 mg.

[0033] In another aspect, the invention features a method of treating a population of subjects having a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, where (i) C1D1 is about 5 mg, (ii) C1D2 is greater than or equal to C1D1 and less than or equal to C1D3, and (iii) C1D3 is about 45 mg or about 60 mg; and (b) the second dosing cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), where C2D1 is about 45 mg or 60 mg.

[0034] In another aspect, the invention provides a method of treating a population of subjects having a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first subcutaneous dose (C1D1) of the bispecific antibody on day 1 of the first dosing cycle, a second subcutaneous dose (C1D2) of the bispecific antibody on day 8 of the first dosing cycle, and a third subcutaneous dose (C1D3) of the bispecific antibody on day 8 of the first dosing cycle. and a third subcutaneous dose (C1D3) of the bispecific antibody on day 15 of the first administration cycle, wherein (i) C1D1 is about 5 mg, (ii) C1D2 is greater than or equal to C1D1 and less than or equal to C1D3, and (iii) C1D3 is about 45 mg or about 60 mg; and (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1) on day 1 of the second administration cycle, wherein C2D1 is about 45 mg or about 60 mg.

[0035] In some embodiments, the CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder) is non-Hodgkin's lymphoma (NHL) or chronic lymphocytic leukemia (CLL). In some embodiments, the NHL is diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL). In some embodiments, the NHL is previously untreated (1L) NHL. In some embodiments, the NHL is CLL. In some embodiments, the DLBCL is 1L DLBCL. In some embodiments, the DLBCL is relapsed or refractory DLBCL. In some embodiments, the DLBCL is Richter's transformed. In some embodiments, the FL is 1L FL. In some embodiments, the FL is relapsed or refractory FL. In some embodiments, the FL is transformed FL. In some embodiments, the NHL is an aggressive B-cell lymphoma. In some embodiments, the NHL is Ann Arbor Stage III or IV NHL. In some embodiments, the subject has previously received at least one (e.g., at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or more) prior line of systemic therapy. In some embodiments, the subject has received between one and nine (e.g., one, two, three, four, five, six, seven, eight, or nine) prior lines of systemic therapy. In some embodiments, the subject has received three prior lines of systemic therapy. In some embodiments, at least one (e.g., one, two, three, four, five, six, seven, eight, or nine) prior line of systemic therapy included an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is rituximab or obinutuzumab. In some embodiments, the prior line of systemic therapy comprising an anti-CD20 antibody additionally comprises an alkylating agent or an anthracycline. In some embodiments, the alkylating agent is cyclophosphamide or bendamustine. In some embodiments, the anthracycline is daunomycin or doxorubicin.In some embodiments, the prior line of systemic therapy comprising an anti-CD20 antibody additionally comprises: (i) cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP); (ii) cyclophosphamide, vincristine, and prednisone (CVP); (iii) fludarabine; or (iv) bendamustine. In some embodiments, at least one (e.g., one, two, three, four, five, six, seven, eight, or nine) prior line of systemic therapy included a Bruton's tyrosine kinase (BTK) inhibitor.

[0036] In another aspect, the invention provides a method of treating a population of subjects with DLBCL, comprising subcutaneously administering to one or more subjects a bispecific antibody that binds CD20 and CD3 (e.g., mosunetuzumab) 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 subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) C1D1 is equal to or less than C1D2 and less than C1D3, (ii) C1D2 is equal to or less than C1D3, and (iii) C1D1 is about 0.1 mg to about 10 mg (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; e.g., about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg). , about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 1 00mg to about 300mg, about 200mg to about 300mg, about 50mg to about 250mg, about 100mg to about 250mg, about 100mg to about 200mg, about 10mg to about 250mg, about 10mg to about 200mg, about 10mg to about 180mg, about 10mg to about 160mg, about 10mg to about 150mg, about 10mg to about 140mg, about 20mg to about 130mg, about 30mg to about 120mg, about 40mg to about 100mg, or about 25mg to about 75mg;(b) the second administration cycle comprises a single subcutaneous dose (2D1) of the bispecific antibody, wherein C2D1 is equal to or greater than C1D3 and is between about 10 mg and about 300 mg (e.g., between about 25 mg and about 300 mg, between about 50 mg and about 300 mg, between about 100 mg and about 300 mg, between about 200 mg and about 300 mg, between about 50 mg and about 250 mg, between about 100 mg and about 250 mg, between about 100 mg and about 250 mg, between about 100 mg and about 100 mg), and In some embodiments, the dose is about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; e.g., about 30 mg, about 45 mg, or about 60 mg). In some embodiments, the DLBCL is relapsed or refractory DLBCL. In some embodiments, the DLBCL is previously untreated (1L) DLBCL. In some embodiments, the DLBCL is Richter's transformation. In some embodiments, the complete response rate is at least about 10% (e.g., at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, or at least about 65%; e.g., about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 10% to about 30%, about 10% to about 20%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 20% to about 30%, about 30% to about 50%, about 30% to about 60%, or about 40% to about 60%; e.g., about 15%, about 20%, about 25%, about 30%, about 35%, or about 40%). In some embodiments, the complete response rate is about 10 to about 90% (e.g., about 10 to about 80%, about 10 to about 70%, about 10 to about 60%, about 10 to about 50%, about 10 to about 40%, about 10 to about 30%, about 20 to about 80%, about 30 to about 80%, about 40 to about 80%, about 50 to about 80%, about 30 to about 70%, about 30 to about 60%, about 40 to about 60%, about 30 to about 50%, about 15 to about 40%, about 20 to about 40%, about 60 to about 90%, about 45 to about 55%, or about 45 to about 50%;For example, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. In a specific embodiment, the complete response rate for a population of subjects with 1L DLBCL is about 40%. In another specific embodiment, the complete response rate for a population of subjects with R / R DLBCL is about 20%.

[0037] In some embodiments, the median progression-free survival is greater than about 4 months (e.g., at least about 4.5 months, at least about 5 months, at least about 5.5 months, at least about 6 months, at least about 6.5 months, at least about 7 months, at least about 7.5 months, at least about 8 months, at least about 8.5 months, at least about 9 months, at least about 9.5 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 20 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, at least about 44 months, at least about 48 months, at least about 50 months, at least about 52 months, at least about 54 months, at least about 56 months, at least about 58 months, at least about 59 months, at least about 60 months, at least about 61 months, at least about 62 months, at least about 63 months, at least about 64 months, at least about 65 months, at least about 66 months, at least about 67 months, at least about 68 months, at least about 69 months, at least about 70 months, at least about 71 months, at least about 72 months, at least about 73 months, at least about 74 months, at least about 75 months, at least about 76 months, at least about 77 months, at least about 78 months, at least about 79 months, at least about 80 months, at least about 81 months, at least about 82 months, at least about 83 months, at least about 84 months, at least about 85 months, at least about 86 months, at least about 87 at least about 48 months, at least about 54 months, or more; for example, between about 4 months and about 48 months, between about 4 months and about 36 months, between about 4 months and about 24 months, between about 4 months and about 12 months, between about 4 months and about 10 months; between about 4 months and about 8 months, between about 8 months and about 24 months, between about 12 months and about 24 months, or between about 8 months and about 16 months; for example, between about 4.5 months, about 5 months, about 5.5 months, approximately 6 months, approximately 6.5 months, approximately 7 months, approximately 7.5 months, approximately 8 months, approximately 8.5 months, approximately 9.0 months, approximately 9.5 months, approximately 10 months, approximately 11 months, approximately 12 months, approximately 13 months, approximately 14 months, approximately 15 months, approximately 16 months, approximately 17 months, approximately 18 months, approximately 20 months, approximately 24 months, approximately 30 months, approximately 36 months, approximately 42 months, approximately 48 months, approximately 54 months, or more). In some embodiments, the median progression-free survival in the population of subjects with R / R FL is greater than about 4 months (e.g., at least about 4.5 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, or at least about 12 months; e.g., between about 4 and about 12 months, between about 4 and about 10 months, between about 4 and about 8 months, between about 4 and about 6 months, between about 8 and about 12 months, between about 6 and about 10 months, between about 6 and about 12 months, or between about 5 and about 9 months; e.g., about 4 months, about 4.5 months, about 5 months, about 5.5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 12 months).In certain embodiments, the median progression-free survival in a population of subjects with R / R FL is greater than about 4 months.

[0038] In some embodiments, the median progression-free survival is greater than about 1 month (e.g., at least about 1.5 months, at least about 2 months, at least about 2.5 months, at least about 3 months, at least about 3.5 months, at least about 4 months, at least about 4.5 months, at least about 5 months, or at least 56 months; e.g., between about 1 month and about 6 months, between about 1 month and about 5 months, between about 1 month and about 4 months, between about 1 month and about 3 months, between about 1 month and about 2 months, between about 2 months and about 4 months, between about 3 months and about 5 months, between about 4 months and about 6 months, or between about 3 months and about 6 months; e.g., about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, or about 6 months). In some embodiments, the median progression-free survival in a population of subjects with DLBCL is greater than about 2 months (e.g., greater than about 2.5 months, greater than about 3 months, greater than about 3.5 months, greater than about 4 months, greater than about 4.5 months, greater than about 5 months, or greater than about 6 months; e.g., between about 2 months and about 12 months, between about 2 months and about 6 months, between about 2 months and about 5 months, between about 2 months and about 4 months, between about 2 months and about 3 months, between about 3 months and about 5 months, or between about 4 months and about 6 months; e.g., about 2.1 months, about 2.5 months, about 3 months, about 3.5 months, about 4 months, about 4.5 months, about 5 months, about 5.5 months, or about 6 months). In one embodiment, the median progression-free survival in a population of subjects with R / R DLBCL is greater than 2 months. In a particular embodiment, the median progression-free survival in a population of subjects with R / R DLBCL is about 2.5 months.

[0039] In some embodiments, the median overall survival is greater than 9.5 months (e.g., at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 20 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, at least about 48 months, at least about 54 months, or more; e.g., at least about 9 months). In some embodiments, the first administration cycle is a 21-day administration cycle, and the second administration cycle is a 21-day administration cycle.

[0040] In another aspect, the invention provides a method of treating a population of subjects with FL, comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, and (i) C1D1 is less than or equal to C1D2; (ii) C1D2 is equal to or less than C1D3, and (iii) C1D1 is about 0.1 mg to about 10 mg (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; e.g., about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg). , about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 3 00 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg;(b) the second administration cycle comprises a single subcutaneous dose (2D1) of the bispecific antibody, wherein C2D1 is equal to or greater than C1D3 and is between about 10 mg and about 300 mg (e.g., between about 25 mg and about 300 mg, between about 50 mg and about 300 mg, between about 100 mg and about 300 mg, between about 200 mg and about 300 mg, between about 50 mg and about 250 mg, between about 100 mg and about 250 mg, between about 100 mg and about 250 mg, between about 100 mg and about 100 mg), and The method is characterized in that the FL is administered at a dose of about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg). In some embodiments, the FL is relapsed or refractory FL. In some embodiments, the FL is transformed FL. In some embodiments, the complete response rate is at least about 40% (e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, or at least about 75%; e.g., about 40% to about 80%, about 40% to about 70%, about 40% to about 60%, about 40% to about 50%, about 50% to about 70%, about 60% to about 80%, or about 50% to about 80%; e.g., about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, or about 80%). In certain embodiments, the complete response rate in the population of subjects with R / R FL is about 45% to about 50%.

[0041] In some embodiments, the objective response rate at about 20 months after initiating treatment is at least about 70% (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95%; e.g., about 70% to about 80%, about 70% to about 90%, about 70% to about 95%, or about 70% to about 100%; e.g., about 70%, about 75%, about 80%, about 85%, or about 90%). In some embodiments, the objective response rate at about 24 months after initiating treatment is at least about 75% (e.g., at least about 80%, at least about 85%, at least about 90%, or at least about 95%; e.g., about 75% to about 80%, about 75% to about 90%, about 75% to about 95%, about 75% to about 100%, about 80% to about 100%, or about 90% to about 100%; e.g., about 75%, about 80%, about 85%, or about 90%). In some embodiments, the objective response rate at about 12 months after initiating treatment is at least about 60% (e.g., at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95%; e.g., about 60% to about 70%, about 60% to about 80%, about 60% to about 90%, or about 60% to about 100%; e.g., about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%).

[0042] In some embodiments, the subject population has relapsed or refractory NHL and the objective response rate is at least 34% (e.g., at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more; e.g., between 34% and 95%, between 34% and 85%, between 34% and 75%, between 34% and 65%, between 34% and 55%, between 35% and 65%). The objective response rate may be between 0%, 35% and 75%, 55% and 95%, 75% and 95%, 40% and 50%, 45% and 64%, 34% and 45%, or 34% and 40%, e.g., about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. In some embodiments, the objective response rate is at least 44%. In some embodiments, the objective response rate is between 35% and 55%. In some embodiments, the objective response rate is about 45%.

[0043] In some embodiments, the subject population has relapsed or refractory FL and the objective response rate is at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%; e.g., 70%-80%, 70%-90%, 70%-95%, or 70%-100%; e.g., about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, or about 95%). In some embodiments, the objective response rate is at least 80%. In some embodiments, the subject population has relapsed or refractory FL and the objective response rate is between 70% and 90%. In some embodiments, the objective response rate is about 80%.

[0044] In some embodiments, the subject population has relapsed or refractory DLBCL or transformed FL and the objective response rate is at least 25% (e.g., at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more; e.g., between 25% and 95%, between 25% and 75%, between 25% and 55%, between 25% and 50%, between 25% and 45%, between 25% and 40%). between 25% and 35%, between 25% and 30%, between 30% and 75%, between 35% and 75%, between 40% and 75%, between 30% and 40%, between 30% and 45%, between 30% and 50%, or between 50% and 70%; for example, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. In some embodiments, the objective response rate is at least 35%. In some embodiments, the subject population has relapsed or refractory DLBCL and the objective response rate is between 25% and 45%. In some embodiments, the objective response rate is about 35%.

[0045] In some embodiments, a population of subjects exhibits cytokine release syndrome after administration of the bispecific antibody, and the rate of cytokine release syndrome in the population of subjects is about 40% or less (e.g., about 35% or less, about 30% or less, about 25% or less, about 20% or less, about 15% or less, about 10% or less, about 5% or less, or about 3% or less; e.g., between about 0% and about 40%, between about 0% and about 30%, about Between 0% and about 20%, between about 0% and about 10%, between about 0% and about 5%, between about 10% and about 20%, between about 10% and about 30%, between about 20% and about 40%, between about 15% and about 35%, or between about 5% and about 15%; for example, about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, about 10%, about 7%, about 5%, about 4%, about 3%, about 2%, about 1%, or about 0%). In some embodiments, the rate of cytokine release syndrome of Grade 2 or higher (as defined by the American Society for Transplantation and Cellular Therapy, 2018; ASTCT) is about 10% or less (e.g., about 7% or less, about 5% or less, about 3% or less, or about 1% or less; e.g., about 0% to about 10%, about 0% to about 7%, about 0% to about 5%, about 0% to about 3%, about 1% to about 3%, about 3% to about 5%, about 5% to about 7%, about 5% to about 10%, about 3% to about 7%; e.g., about 10%, about 7%, about 5%, about 4%, about 3%, about 2%, about 1%, or about 0%).

[0046]

[0010] Another aspect of the invention provides methods for reducing the rate of a particular adverse event in a population of subjects with a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder) to which a bispecific antibody that binds CD20 and CD3 is administered, the method comprising administering the bispecific antibody subcutaneously using a stepwise dosing regimen, wherein the rate of adverse events is reduced in the population of subjects compared to a reference population of subjects to which the bispecific antibody is administered intravenously. In some embodiments, the stepwise dosing regimen is selected from the group consisting of: (I) at least a first dosing cycle and a second dosing cycle, wherein (a) the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein (i) C1D1 is equal to or less than C1D2 and less than C1D3, and (ii) C1D2 is equal to or less than C1D3. (iii) C1D1 is about 0.1 mg to about 10 mg (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg). g to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg). mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg;(b) the second administration cycle comprises a single subcutaneous dose (2D1) of the bispecific antibody, wherein C2D1 is equal to or greater than C1D3 and is between about 10 mg and about 300 mg (e.g., between about 25 mg and about 300 mg, between about 50 mg and about 300 mg, between about 100 mg and about 300 mg, between about 200 mg and about 300 mg, between about 50 mg and about 250 mg, between about 100 mg and about 250 mg, between about 100 mg and about 200 mg, between about 10 mg and about 250 mg, between about 10 mg and about 200 mg, between about 10 mg and about 180 mg, mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg); (II) at least a first administration cycle and a second administration cycle, wherein (a) the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) C1D1 is about 5 mg, (ii) C1D2 is equal to or greater than C1D1 and equal to or less than C1D3; (iii) C1D3 is about 45 mg or about 60 mg, (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), and C2D1 is 45 mg or about 60 mg; and (III) at least a first administration cycle and a second administration cycle, wherein (a) the first administration cycle comprises a first subcutaneous dose of the bispecific antibody (C1D1) on day 1 of the first administration cycle, a second subcutaneous dose of the bispecific antibody (C1D2) on day 8 of one administration cycle, and a third subcutaneous dose of the bispecific antibody (C1D3) on day 15 of the first administration cycle, where (i) C1D1 is about 5 mg, (ii) C1D2 is greater than or equal to C1D1 and less than or equal to C1D3, and (iii) C1D3 is about 45 mg or about 60 mg; and (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1) on day 1 of the second administration cycle, where C2D1 is about 45 mg or about 60 mg;

[0047] In some embodiments, the CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder) is non-Hodgkin's lymphoma (NHL) or chronic lymphocytic leukemia (CLL). In some embodiments, the NHL is diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL). In some embodiments, the NHL is previously untreated (1L) NHL. In some embodiments, the NHL is CLL. In some embodiments, the DLBCL is 1L DLBCL. In some embodiments, the DLBCL is relapsed or refractory DLBCL. In some embodiments, the DLBCL is Richter's transformed. In some embodiments, the FL is 1L FL. In some embodiments, the FL is relapsed or refractory FL. In some embodiments, the FL is transformed FL. In some embodiments, the NHL is an aggressive B-cell lymphoma. In some embodiments, the NHL is Ann Arbor Stage III or IV NHL. In some embodiments, the subject has previously received at least one (e.g., at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or more) prior line of systemic therapy. In some embodiments, the subject has received between one and nine (e.g., one, two, three, four, five, six, seven, eight, or nine) prior lines of systemic therapy. In some embodiments, the subject has received three prior lines of systemic therapy. In some embodiments, at least one (e.g., one, two, three, four, five, six, seven, eight, or nine) prior line of systemic therapy included an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is rituximab or obinutuzumab. In some embodiments, the prior line of systemic therapy comprising an anti-CD20 antibody additionally comprises an alkylating agent or an anthracycline. In some embodiments, the alkylating agent is cyclophosphamide or bendamustine. In some embodiments, the anthracycline is daunomycin or doxorubicin.In some embodiments, the prior line of systemic therapy comprising an anti-CD20 antibody additionally comprises: (i) cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP); (ii) cyclophosphamide, vincristine, and prednisone (CVP); (iii) fludarabine; or (iv) bendamustine. In some embodiments, at least one (e.g., one, two, three, four, five, six, seven, eight, or nine) prior line of systemic therapy included a Bruton's tyrosine kinase (BTK) inhibitor.

[0048] In some embodiments, a population of subjects exhibits cytokine release syndrome after administration of the bispecific antibody, and the rate of cytokine release syndrome in the population of subjects is about 40% or less (e.g., about 35% or less, about 30% or less, about 25% or less, about 20% or less, about 15% or less, about 10% or less, about 5% or less, or about 3% or less; e.g., between about 0% and about 40%, between about 0% and about 30%). , between about 0% and about 20%, between about 0% and about 10%, between about 0% and about 5%, between about 10% and about 20%, between about 10% and about 30%, between about 20% and about 40%, between about 15% and about 35%, or between about 5% and about 15%; for example, about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, about 10%, about 7%, about 5%, about 4%, about 3%, about 2%, about 1%, or about 0%). In some embodiments, the rate of cytokine release syndrome of Grade 2 or higher (as defined by the American Society for Transplantation and Cellular Therapy, 2018; ASTCT) is about 10% or less (e.g., about 7% or less, about 5% or less, about 3% or less, about 1% or less; e.g., about 0% to about 10%, about 0% to about 7%, about 0% to about 5%, about 0% to about 3%, about 1% to about 3%, about 3% to about 5%, about 5% to about 7%, about 5% to about 10%, about 3% to about 7%; e.g., about 10%, about 7%, about 5%, about 4%, about 3%, about 2%, about 1% or about 0%).

[0049] In some embodiments, the complete response rate is at least about 10% (e.g., at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, or more; For example, about 10% to about 40%, about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 10% to about 30%, about 15% to about 30%, about 20% to about 40%, or more; for example, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 25%, about 30%, about 35%, about 40%, or more). In some embodiments, the complete response rate is at least about 42% (e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or more; e.g., 42% to 45%, 45% to 50%, about 50% to about 55%, about 55% to about 60%, about 60% to about 65%, about 65% to about 70%, about 70% to about 75%, or more; e.g., about 42%, about 45%, about 50%, about 55%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more). In some embodiments, the complete response rate is at least about 20%. In some embodiments, the complete response rate is at least about 40%. In some embodiments, the complete response rate is at least about 55%. In some embodiments, the objective response rate at about 24 months after initiation of treatment is at least about 75% (e.g., at least about 80%, at least about 85%, at least about 90%, or at least about 95%; e.g., about 75% to about 80%, about 75% to about 90%, about 75% to about 95%, about 75% to about 100%, about 80% to about 100%, or about 90% to about 100%; e.g., about 75%, about 80%, about 85%, or about 90%).In some embodiments, the objective response rate at about 20 months after initiation of treatment is at least about 70% (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95%; e.g., 70% to 80%, 70% to 90%, 70% to 95%, or 70% to 100%; e.g., about 70%, about 75%, about 80%, about 85%, or about 90%). In some embodiments, the objective response rate at about 12 months after initiation of treatment is at least about 60% (e.g., at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95%; e.g., about 60% to about 70%, about 60% to about 80%, about 60% to about 90%, or about 60% to about 100%; e.g., about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%).

[0050] In some embodiments, the subject population has relapsed or refractory NHL and the objective response rate is at least 34% (e.g., at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more; e.g., between 34% and 95%, between 34% and 85%, between 34% and 75%, between 34% and 65%, between 34% and 55%, between 35% and 65%). The objective response rate may be between 0%, 35% and 75%, 55% and 95%, 75% and 95%, 40% and 50%, 45% and 64%, 34% and 45%, or 34% and 40%, e.g., about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. In some embodiments, the objective response rate is at least 44%. In some embodiments, the objective response rate is between 35% and 55%. In some embodiments, the objective response rate is about 45%.

[0051] In some embodiments, the subject population has relapsed or refractory FL and the objective response rate is at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%; e.g., 70%-80%, 70%-90%, 70%-95%, or 70%-100%; e.g., about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, or about 95%). In some embodiments, the objective response rate is at least 80%. In some embodiments, the subject population has relapsed or refractory FL and the objective response rate is between 70% and 90%. In some embodiments, the objective response rate is about 80%.

[0052] In some embodiments, the subject population has relapsed or refractory DLBCL or transformed FL and the objective response rate is at least 25% (e.g., at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more; e.g., between 25% and 95%, between 25% and 75%, between 25% and 55%, between 25% and 50%, between 25% and 45%, between 25% and 40%). between 25% and 35%, between 25% and 30%, between 30% and 75%, between 35% and 75%, between 40% and 75%, between 30% and 40%, between 30% and 45%, between 30% and 50%, or between 50% and 70%; for example, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. In some embodiments, the objective response rate is at least 35%. In some embodiments, the subject population has relapsed or refractory DLBCL and the objective response rate is between 25% and 45%. In some embodiments, the objective response rate is about 35%.

[0053] In some embodiments of any of the preceding aspects, the bispecific antibody comprises an anti-CD20 arm comprising a first binding domain comprising the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence of GYTFTSYNMH (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of AIYPGNGDTSYNQKFKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of VVYYSNSYWYFDV (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of RASSSVSYMH (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of APSNLAS (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QQWSFNPPT (SEQ ID NO: 6). In some embodiments, the bispecific antibody comprises an anti-CD20 arm comprising: (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 7; (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 8; or (c) a first binding domain comprising a VH domain as in (a) and a VL domain as in (b). In some embodiments, the first binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 7 and a VL domain comprising the amino acid sequence of SEQ ID NO: 8. In some embodiments, the bispecific antibody comprises an anti-CD3 arm with a second binding domain comprising the following six HVRs: (a) HVR-H1 comprising the amino acid sequence of NYYIH (SEQ ID NO: 9); (b) HVR-H2 comprising the amino acid sequence of WIYPGDGNTKYNEKFKG (SEQ ID NO: 10); (c) HVR-H3 comprising the amino acid sequence of DSYSNYYFDY (SEQ ID NO: 11); (d) HVR-L1 comprising the amino acid sequence of KSSQSLLNSRTRKNYLA (SEQ ID NO: 12); (e) HVR-L2 comprising the amino acid sequence of WASTRES (SEQ ID NO: 13); and (f) HVR-L3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 14).In some embodiments, the bispecific antibody comprises an anti-CD3 arm comprising: (a) a VH domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 15; (b) a VL domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16; or (c) a second binding domain comprising a VH domain as in (a) and a VL domain as in (b). In some embodiments, the second binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 15 and a VL domain comprising the amino acid sequence of SEQ ID NO: 16. In some embodiments, the bispecific antibody comprises (a) an anti-CD20 arm comprising (i) a heavy chain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 51 and (ii) a light chain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 52, and (b) an anti-CD3 arm comprising (i) a heavy chain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 53 and (ii) a light chain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 54. In some antibodies, the (a) anti-CD20 arm comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51 and a light chain comprising the amino acid sequence of SEQ ID NO: 52, and the (b) anti-CD3 arm comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 53 and a light chain comprising the amino acid sequence of SEQ ID NO: 54.

[0054] In some embodiments of any of the preceding aspects, the bispecific antibody is a humanized antibody. In some embodiments, the bispecific antibody is a chimeric antibody. In some embodiments, the bispecific antibody is an antibody fragment that binds to CD20 and CD3. In some embodiments, the antibody fragment is selected from the group consisting of a Fab fragment, a Fab'-SH fragment, an Fv fragment, an scFv fragment, and a (Fab')2 fragment.

[0055] In some embodiments, the bispecific antibody is a full-length antibody. In some embodiments, the bispecific antibody is an IgG antibody. In some embodiments, the IgG antibody is an IgG1 antibody. In some embodiments, the IgG antibody comprises a mutation at amino acid residue N297 (EU numbering) that results in the absence of glycosylation. In some embodiments, the mutation at amino acid residue N297 is a substitution mutation. In some embodiments, the mutation at amino acid residue N297 reduces effector function of the Fc region. In some embodiments, the mutation is an N297G or N297A mutation. In some embodiments, the bispecific antibody comprises a mutation in the Fc region that reduces effector function. In some embodiments, the mutation is a substitution mutation. In some embodiments, the substitution mutation is at amino acid residues L234, L235, D265, and / or P329 (EU numbering). In some embodiments, the substitution mutation is selected from the group consisting of L234A, L235A, D265A, and P329G. In some embodiments, the bispecific antibody comprises one or more heavy chain constant domains, wherein the one or more heavy chain constant domains are selected from a first CH1 (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 form a protrusion or a cavity, and the protrusion or cavity in the CH31 domain is positionable in the cavity or protrusion in the CH32 domain, respectively. In some embodiments, the CH31 domain and the CH32 domain contact at the interface between the protrusion and the cavity. In some embodiments, the CH21 domain and the CH22 domain each comprise a protrusion or a cavity, and the protrusion or cavity in the CH21 domain is positionable in the cavity or protrusion in the CH22 domain, respectively. In some embodiments, the CH21 and CH22 domains associate at the interface between the protrusion and the cavity.

[0056] In some embodiments, the anti-CD20 arm of the bispecific antibody further comprises the following substitution mutations (EU numbering): T366W and N297G. In some embodiments, the anti-CD3 arm of the bispecific antibody further comprises the following substitution mutations (EU numbering): T366S, L368A, Y407V, and N297G. In some embodiments, (a) the anti-CD20 arm further comprises the following substitution mutations (EU numbering): T366W and N297G, and (b) the anti-CD3 arm further comprises the following substitution mutations (EU numbering): T366S, L368A, Y407V, and N297G.

[0057] In some embodiments, the subjects are humans. In some embodiments, the population of subjects is a population of human subjects. [Brief explanation of the drawings]

[0058] The content of this application file contains at least one drawing executed in color. Copies of this patent or this patent application with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. [Figure 1] Figure 1 is a schematic diagram illustrating the design of the dose escalation portion of the GO29781 study. Initially, mosunetuzumab is given as a single, unfractionated intravenous (IV) dose on day 1 of each cycle (Group A). ​​Cycle 1 dosing is then altered to stop the dose escalation for Group A and perform mosunetuzumab dose escalation as follows: Group B: mosunetuzumab dose escalation utilizing a cycle 1 step-up IV administration scheme; Group D: mosunetuzumab dose escalation utilizing a cycle 1 unfractionated subcutaneous (SC) dosing scheme; Group F: mosunetuzumab dose escalation utilizing a cycle 1 step-up SC dosing scheme. C = cycle; D = day; DL = dose level; MAD = maximum assessed dose. [Figure 2]Figure 2 is a schematic diagram showing the design of the non-Hodgkin's lymphoma (NHL) expansion cohort and chronic lymphocytic leukemia (CLL) dose escalation / expansion cohort for the GO29781 study. DLBCL = diffuse large B-cell lymphoma; FL = follicular lymphoma; MCL = mantle cell lymphoma; NHL = non-Hodgkin's lymphoma; RP2D = recommended Phase II dose; R / R = relapsed / refractory; trFL = transformed follicular lymphoma. a Multiple expansion cohorts based on dose escalation of arms A, B, D, and F may be tested. b Expansion cohorts in R / R DLBCL / trFL will enroll a maximum of approximately 80 patients, except for expansion cohorts based on the arm B RP2D, which will enroll a maximum of approximately 20 patients. c Expansion cohorts in R / R FL will enroll a maximum of approximately 80 patients, except for expansion cohorts based on the arm B RP2D, which will enroll a maximum of approximately 20 patients. Expansion cohorts based on dB group dose escalation only will be studied. e Dose escalation was performed similarly to NHL (see Figure 1). f Multiple expansion cohorts based on dose escalation of groups B, D, and F may be studied. [Figure 3] FIG. 3 is a schematic diagram showing the evaluation window in Arm B of the GO29781 study. [Figure 4] Figure 4 is a set of schematic diagrams showing three exemplary scenarios for the observation of DLTs in Cycle 1 dose escalation (Arm F) in the GO29781 study. The diagrams represent examples showing the timing of two DLTs in a dose escalation cohort of six patients and do not represent all possible scenarios. [Figure 5] Figure 5 is a schematic diagram showing the duration of initial study treatment and options for retreatment or continuation study treatment in the GO29781 study. CR = complete response; PD = progressive disease; PR = partial response; SD = stable disease. a Additional rounds of retreatment are possible according to the treatment trajectory for the initial treatment. b Scans should be scheduled to avoid / minimize any dose delays between cycles 8 and 9 whenever possible. [Figure 6] Figure 6 is a schematic diagram showing the doses of SC mosunetuzumab tested in Arm D of the GO29781 study. D = dose. N = number of patients. [Figure 7]7 is a pair of graphs showing the concentration of mosunetuzumab (μg / mL) in patient serum samples at the indicated doses and time points. The left panel shows samples from Group B of the GO29781 study (IV step-up administration). The right panel shows samples from Group D of the GO29781 study (SC dosing). The dotted line indicates the Cmax of a 1 mg dose delivered by IV administration. [Figure 8] Figure 8 is a set of graphs showing the concentration of IL-6 (pg / mL) in patient peripheral blood samples at the indicated doses and time points. The left panel shows samples from Group A of the GO29781 study (IV dosing). The right panel shows samples from Group D of the GO29781 study (SC dosing). PD: Pre-dose. EoI: End of infusion. [Figure 9] Figure 9 is a set of graphs showing the concentration of IL-6 (pg / mL) in patient peripheral blood samples at the indicated time points. The left and center panels show samples from Group B of the GO29781 study (step-up IV dosing), which received a 1 mg dose of mosunetuzumab on Day 1 of Cycle 1 (C1D1) (left panel: data from the dose escalation phase; center panel: data from the dose expansion phase). The right panel shows samples from Group D of the GO29781 study (SC dosing), which received doses of 1.6 mg, 2.4 mg, 3.6 mg, or 7.2 mg. [Figure 10] 10 is a set of graphs showing the concentration of IL-6 (pg / mL) in peripheral blood samples of subcutaneously administered patients at the indicated doses and time points. Arrows indicate patient 1. PRE: Pre-dose. [Figure 11] Figure 11 is a set of graphs showing the concentrations (in pg / mL) of IL-2 (top left), IL-6 (top right), IFNγ (bottom left), and TNFα (bottom right) in the blood of cynomolgus monkeys that received vehicle intravenously, mosunetuzumab intravenously at doses ranging from 0.01 mg / kg to 1 mg / kg, or mosunetuzumab subcutaneously at a dose of 1 mg / kg. [Figure 12] Figure 12 is a set of graphs showing T cell activation after intravenous or subcutaneous administration of mosunetuzumab. Top panel: quantification of CD4+ / CD69+ / CD25+ T cells; bottom panel: quantification of CD8+ / CD69+ / CD25+ T cells. [Figure 13] Figure 13 is a set of graphs showing B cell depletion in cynomolgus monkeys after a single 1 mg / mL intravenous dose of mosunetuzumab. Top panel: circulating B cells (CD40+); bottom panel: splenic B cells. [Figure 14] Figure 14 is a graph showing the kinetics of circulating B-cell activating factor (BAFF) in serum. BAFF was assessed up to day 8 in all dose groups and up to day 57 in the control and 1 mg / kg intravenous groups. DETAILED DESCRIPTION OF THE INVENTION

[0059] Detailed Description The present invention relates to the treatment of CD20-positive cell proliferative disorders, such as B-cell proliferative disorders (e.g., non-Hodgkin's lymphoma (NHL) (e.g., previously untreated (1L) NHL), diffuse large B-cell lymphoma (DLBCL) (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), follicular lymphoma (FL) (e.g., 1L The present invention includes a method of treating a subject (or population of subjects) with FL, relapsed and / or refractory FL, or transformed FL), mantle cell lymphoma (MCL), high-grade B-cell lymphoma, or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL) or chronic lymphocytic leukemia (CLL) by subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 (e.g., mosunetuzumab) in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle. The first dosing cycle comprises three subcutaneous administrations, wherein the first subcutaneous dose (C1D1) is equal to or less than the second subcutaneous dose (C1D2) and less than the third subcutaneous dose (C1D3), and C1D2 is equal to or less than C1D3.In some examples, C1D1 is about 0.1 mg to about 10 mg (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg). mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg). The second administration cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, where C2D1 is C1D3 or greater and is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).

[0060] The present invention is based, in part, on the discovery that a dosing regimen comprising subcutaneous administration of a bispecific antibody that binds to CD20 and CD3 (e.g., mosunetuzumab) over multiple dosing cycles (e.g., the first dosing cycle is a step-up split dosing cycle) can effectively treat a subject with a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder) while reducing toxicity (e.g., cytokine release syndrome or CNS toxicity).

[0061] I. General techniques The techniques and procedures described or referenced herein are generally well understood and can be found, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual 3rd edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology (F.M.A.usubel, et al. eds., (2003)); the series Methods in Enzymology (Academic Press, Inc.): PCR 2: A Practical Approach (M.J. MacPherson, B.D. Hames and G.R. Taylor eds. (1995)), Harlow and Lane, eds. (1988); Antibodies, A Laboratory Manual, and Animal Cell Culture (R.I. Freshney, ed. (1987)); Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook(JECellis,ed.,1998)Academic Press;Animal Cell Culture(RIFreshney),ed.,1987);Introduction to Cell and Tissue Culture(JPMather and PERoberts,1998)Plenum Press;Cell and Tissue Culture:Laboratory Procedures(A.Doyle,JBGriffiths,and DGNewell,eds.,1993-8)J.Wiley and Sons; Handbook of Experimental Immunology (DM Weir and CC Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (JMMiller and MPCalos, eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994); Current Protocols in Immunology (JEColigan et al., eds., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (CA Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (D. Catty., ed., IRL Press, 1988-1989); Monoclonal Antibodies: A Practical Approach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow and D.Lane(Cold Spring Harbor Laboratory Press,1999);The Antibodies(M.Zanetti and These methods are commonly employed by those skilled in the art using conventional methodologies, such as the widely used methodologies described in J.D. Capra, eds., Harwood Academic Publishers, 1995; and Cancer: Principles and Practice of Oncology (VT DeVita et al., eds., J.B. Lippincott Company, 1993).

[0062] II. Definition It will be understood that aspects and embodiments of the invention described herein include "comprising," "consisting of," and "consisting essentially of" aspects and embodiments.

[0063] As used herein, the singular forms "a," "an," and "the" include plural referents unless otherwise indicated.

[0064] As used herein, the term "about" refers to a normal error range for the respective value, which would be readily understood by one of ordinary skill in the art. Reference herein to a value or parameter preceded by "about" includes (and describes) embodiments that are directed to that value or parameter itself.

[0065] The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, but are not limited to, hematological cancers such as mature B-cell cancers, e.g., Hodgkin's lymphoma, but not limited to non-Hodgkin's lymphoma (NHL), e.g., diffuse large B-cell lymphoma (DLBCL), which may be relapsed or refractory DLBCL or Richter's transformation. Other specific examples of cancer include germinal center B-cell-like (GCB) diffuse large B-cell lymphoma (DLBCL), activated B-cell-like (ABC) DLBCL, follicular lymphoma (FL), transformed FL, mantle cell lymphoma (MCL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), marginal zone lymphoma (MZL), transformed MZL, high-grade B-cell lymphoma, primary mediastinal (thymic) large B-cell lymphoma (PMLBCL), small lymphocytic leukemia (SLL), lymphoplasmacytic lymphoma (LL), transformed LL, Waldenstrom's hypergammaglobulinemia (WM), and thyroid cancer. Central nervous system lymphoma (CNSL), Burkitt lymphoma (BL), B-cell prolymphocytic leukemia, splenic marginal zone lymphoma, hairy cell leukemia, splenic lymphoma / leukemia, unclassifiable, diffuse red pulp small B-cell lymphoma, hairy cell leukemia variant, heavy chain disease, alpha heavy chain disease, gamma heavy chain disease, mu heavy chain disease, plasma cell myeloma, isolated bone plasmacytoma, extraskeletal plasmacytoma, extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma), nodal marginal zone lymphoma, pediatric nodal marginal zone lymphoma, pediatric follicular lymphoma, primary cutaneous follicle center lymphoma, T-cell / histiocyte-rich large B-cell lymphoma, primary CNS Includes DLBCL, primary cutaneous DLBCL, leg type, EBV-positive DLBCL of the elderly, chronic inflammation-associated DLBCL, lymphomatoid granulomatosis, intravascular large B-cell lymphoma, ALK-positive large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma due to HHV8-associated multicentric Castleman disease, primary effusion lymphoma; unclassifiable B-cell lymphoma with features intermediate between DLBCL and Burkitt lymphoma; and unclassifiable B-cell lymphoma with features intermediate between DLBCL and classical Hodgkin lymphoma.Further examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and lymphoid malignancies, including leukemia or B-cell lymphoma. More specific examples of such cancers include, but are not limited to, multiple myeloma (MM); low-grade / follicular NHL; small lymphocytic (SL) NHL; intermediate-grade / follicular NHL; intermediate-grade diffuse NHL; high-grade immunoblastic NHL; high-grade lymphocytic NHL; high-grade small non-dividing cell NHL; bulky mass disease NHL; AIDS-related lymphoma; and acute lymphocytic leukemia (ALL); chronic myeloblastic leukemia; and post-transplant lymphoproliferative disorder (PTLD).

[0066] "Tumor," as used herein, refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor" are not mutually exclusive when referred to herein.

[0067] A "disorder" is any condition that would benefit from treatment, including, but not limited to, chronic and acute disorders or diseases, including conditions that predispose a mammal to the disorder in question.

[0068] The terms "cell proliferative disorder" and "proliferative disorder" refer to disorders associated with some degree of abnormal cell proliferation. In one embodiment, the cell proliferative disorder is cancer. In another embodiment, the cell proliferative disorder is a tumor.

[0069] The terms "B-cell proliferative disorder" or "B-cell malignancy" refer to diseases associated with some degree of abnormal B-cell proliferation and include, for example, lymphoma, leukemia, myeloma, and myelodysplastic syndrome. In one embodiment, the B-cell proliferative disorder is a lymphoma, such as, for example, non-Hodgkin's lymphoma (NHL), including diffuse large B-cell lymphoma (DLBCL) (e.g., relapsed or refractory DLBCL or Richter's transformation), FL (e.g., relapsed and / or refractory FL or transformed FL), MCL, high-grade B-cell lymphoma, or PMLBCL. In another embodiment, the B-cell proliferative disorder is a leukemia, such as chronic lymphocytic leukemia (CLL).

[0070] As used herein, "treatment" (and grammatical variations thereof, such as "treat" or "treating") refers to clinical intervention in an attempt to alter the natural course of the treated subject and can be performed prophylactically or during the course of clinical pathology. Desired effects of treatment include, but are not limited to, preventing the onset or recurrence of disease, alleviating symptoms, attenuating any direct or indirect pathological consequences of the disease, preventing metastasis, slowing the rate of disease progression, remission or palliation of disease symptoms, and recovery or improved prognosis. In some embodiments, the antibodies of the invention are used to delay the onset of disease or to slow the progression of disease.

[0071] As used herein, "delaying the progression" of a disorder or disease means postponing, preventing, slowing, retarding, stabilizing, and / or delaying the development of the disease or disorder (e.g., a CD20-positive cell proliferative disorder, e.g., a B-cell proliferative disorder, e.g., NHL, e.g., DLBCL or FL). This delay can be of varying duration depending on the history of the disease and / or the individual being treated. As will be apparent to one of skill in the art, a sufficient or significant delay can, in effect, encompass prevention, in that the individual does not develop the disease. For example, late-stage cancer, such as the development of metastases, can be delayed.

[0072] "Reducing" or "inhibiting" refers to the ability to produce an overall decrease, e.g., of 20% or more, 50% or more, or 75%, 85%, 90%, 95%, or more. In certain embodiments, reducing or inhibiting refers to a reduction or inhibition of undesirable events such as cytokine-driven 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 following treatment with an anti-CD20 / anti-CD3 bispecific antibody using a split-escalating dose regimen of the invention compared to intravenous administration with the bispecific antibody. In other embodiments, reducing or inhibiting can refer to antibody effector functions mediated by the antibody Fc region, including complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC), and antibody-dependent cellular phagocytosis (ADCP).

[0073] As used herein, "administering" refers to a method of providing a dosage of a compound (e.g., a bispecific antibody) or composition (e.g., a pharmaceutical composition, e.g., a pharmaceutical composition comprising a bispecific antibody) to a subject. The compounds and / or compositions utilized in the methods described herein can be administered subcutaneously (e.g., by subcutaneous injection).

[0074] A "fixed" or "constant" dose of a therapeutic agent (e.g., a bispecific antibody) herein refers to a dose administered to a patient without regard to the patient's weight or body surface area (BSA). Thus, a fixed or flat dose can be expressed as a mg / kg dose or a mg / m 2 It is not provided as a dose, but as an absolute amount (eg, mg) of therapeutic agent.

[0075] A "subject" or "individual" is a mammal. Mammals include, but are not limited to, primates (e.g., humans and non-human primates, such as monkeys), livestock (e.g., cows, sheep, cats, dogs, and horses), rabbits, and rodents (e.g., mice and rats). In certain embodiments, the subject or individual is a human.

[0076] An "individual response" or "response" can include, but is not limited to, (1) disease progression (e.g., a CD20-positive cell proliferative disorder, e.g., a B-cell proliferative disorder (e.g., non-Hodgkin's lymphoma (NHL) (e.g., previously untreated (1L) NHL), diffuse large B-cell lymphoma (DLBCL) (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), follicular lymphoma (FL) (e.g., 1L (1) inhibition to some extent (including slowing and complete halting) of the progression of FL, relapsed and / or refractory FL, or transformed FL, mantle cell lymphoma (MCL), high-grade B-cell lymphoma, or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL) or chronic lymphocytic leukemia (CLL); (2) reduction in tumor size; (3) suppression (i.e., reduction, slowing, or complete halt) of cancer cell invasion into adjacent peripheral organs and / or tissues; (4) inhibition (i.e., reduction, slowing, or complete halt) of metastasis; (5) suppression of a CD20-positive cell proliferative disorder, such as a B-cell proliferative disorder (e.g., previously untreated (1L) NHL, diffuse large B-cell lymphoma (DLBCL) (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), follicular lymphoma (FL) (e.g., 1L Any endpoint that indicates benefit to the subject can be used to assess treatment, including (1) some alleviation of one or more symptoms associated with FL, relapsed and / or refractory FL, or transformed FL), mantle cell lymphoma (MCL), high-grade B-cell lymphoma, or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL)) or chronic lymphocytic leukemia (CLL); (2) an increase or prolongation of survival, including overall survival and progression-free survival; and / or (3) a reduction in mortality at a given time point after treatment.

[0077] As used herein, "complete response" or "CR" refers to the disappearance of all target lesions (i.e., all evidence of disease).

[0078] As used herein, "partial response" or "PR" refers to at least a 30% reduction in the sum of the longest diameters (SLD) of target lesions referenced to a baseline SLD, or at least a 50% reduction in the product of diameters (SPD) of target lesions referenced to a baseline SPD.

[0079] As used herein, "objective response rate" (ORR) means the sum of the complete response (CR) rate and the partial response (PR) rate.

[0080] As used herein, "duration of objective response" (DOR) is defined as the time from the first occurrence of a documented objective response to disease progression or death from any cause within 30 days of the last dose of treatment, whichever occurs first.

[0081] A "durable response" refers to a sustained effect on reducing tumor growth after cessation of treatment. For example, tumor size may remain the same or may be smaller compared to the size at the beginning of the administration phase. In some embodiments, the durable response has a duration at least equal to the treatment period, at least 1.5, 2.0, 2.5, or 3.0 times the treatment period.

[0082] A subject's "effective response" or subject "responsiveness," and similar words, to treatment with a pharmaceutical agent refers to a clinical or therapeutic benefit conferred on a subject at risk for or suffering from a disease or disorder, such as cancer. In one embodiment, such benefit includes any one or more of extending survival (including overall survival and progression-free survival), producing an objective response (including a complete or partial response), or ameliorating the signs or symptoms of cancer.

[0083] A subject who "does not respond effectively" to treatment refers to a subject who does not have any of the following: an extension of survival (including overall survival and progression-free survival), an objective response (including a complete or partial response), or an improvement in the signs or symptoms of cancer.

[0084] As used herein, the term "survival" refers to the patient being alive, and includes overall survival and progression-free survival.

[0085] As used herein, "overall survival" (OS) refers to the proportion of subjects in a group who are alive after a particular period of time, such as 1 year or 5 years from the time of diagnosis or treatment.

[0086] As used herein, "progression-free survival" (PFS) refers to the length of time during and after treatment that the disease being treated (e.g., a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., an NHL (e.g., previously untreated (1L) NHL, DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), FL (e.g., 1L FL, relapsed and / or refractory FL, or transformed FL), MCL, high-grade B-cell lymphoma or PMLBCL) or CLL)) does not worsen. Progression-free survival can include the amount of time a patient experiences a complete or partial remission, as well as the amount of time a patient experiences stable disease.

[0087] As used herein, "stable disease" or "SD" refers to neither sufficient shrinkage of target lesions to qualify as PR nor sufficient increase to qualify as PD, based on the smallest SLD since treatment initiation.

[0088] As used herein, "progressive disease" or "PD" refers to at least a 20% increase in the SLD of a target lesion, referenced to the smallest SLD, or at least a 50% increase in the SPD of a target lesion, referenced to the smallest SPD recorded after initiation of treatment or the presence of one or more new lesions.

[0089] As used herein, "delaying progression" of a disorder or disease means postponing, preventing, delaying, stabilizing, and / or postponing the onset of a disease or disorder (e.g., a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., NHL (e.g., previously untreated (1L) NHL, DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), FL (e.g., 1L FL, relapsed and / or refractory FL, or transformed FL), MCL, high-grade B-cell lymphoma or PMLBCL) or CLL)). This delay can be of varying duration depending on the disease history and / or the subject being treated. As will be apparent to one of skill in the art, a sufficient or significant delay can, in effect, encompass prevention, in that the subject does not develop the disease. For example, in late-stage cancer, the development of central nervous system (CNS) metastases can be delayed.

[0090] As used herein, the term "reducing or inhibiting cancer recurrence" means reducing or inhibiting the recurrence of a tumor or cancer, or the progression of a tumor or cancer.

[0091] "Reduce or inhibit" refers to the ability to result in an overall decrease of 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or more. Reduction or inhibition can refer to the symptoms of the disorder being treated (e.g., a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., NHL (e.g., previously untreated (1L) NHL, DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), FL (e.g., 1L FL, relapsed and / or refractory FL, or transformed FL), MCL, high-grade B-cell lymphoma or PMLBCL) or CLL)), the presence or size of metastases, or the size of the primary tumor.

[0092] As used herein, the term "Ann Arbor staging" or "Ann Arbor stage" refers to a system for classifying the stages of lymphoma (e.g., non-Hodgkin's lymphoma (NHL); e.g., DLBCL, FL, MCL, high-grade B-cell lymphoma, PMLBCL, or CLL). Lymphoma (e.g., NHL) can be classified as one of four Ann Arbor stages. Stage I refers to lymphoma showing involvement of a single lymph node region or a single extralymphatic organ or site. Stage II refers to lymphoma showing involvement of two or more lymph node regions on the same side of the diaphragm. Stage III refers to lymphoma showing involvement of lymph node regions on both sides of the diaphragm (III) and may be accompanied by focal involvement of an extralymphatic organ or site, or involvement of the spleen, or both. Stage IV refers to lymphomas showing diffuse or disseminated involvement of one or more extralymphatic organs or tissues, with or without associated lymphadenopathy. Liver involvement is always considered diffuse and therefore always considered Ann Arbor Stage IV. Lymphatic structures include lymph nodes, thymus, spleen, appendix, Waldeyer's rings, and Peyer's patches. See Carbone, PP et al., Cancer Res. 1971, 31(11):1860-1861.

[0093] By "prolonged survival" is meant an increase in overall survival or progression-free survival in treated patients relative to untreated patients (e.g., relative to patients not treated with the drug), or relative to patients who do not express the biomarker at the specified level, and / or relative to patients treated with an approved anti-tumor agent. Objective response refers to a measurable response, including a complete response (CR) or partial response (PR).

[0094] The term "antibody" is used herein in the broadest sense and encompasses a variety of antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired antigen-binding activity.

[0095] "Antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments.

[0096] The terms "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein to refer to an antibody having a heavy chain that has a structure substantially similar to a native antibody structure or that contains an Fc region as defined herein.

[0097] "Binding domain" refers to a portion of a compound or molecule that specifically binds to a target epitope, antigen, ligand, or receptor. Binding domains include, but are not limited to, antibodies (e.g., monoclonal, polyclonal, recombinant, humanized, and chimeric antibodies), antibody fragments or portions thereof (e.g., Fab fragments, Fab'2, scFv antibodies, SMIPs, domain antibodies, diabodies, minibodies, scFv-Fc, affibodies, nanobodies, and antibody VH and / or VL domains), receptors, ligands, aptamers, and other molecules with identified binding partners.

[0098] The term "Fc region" is used herein to define a C-terminal region of an immunoglobulin heavy chain containing at least a portion of the constant region. This term includes native sequence Fc regions and variant Fc regions. In one embodiment, a human IgG heavy chain Fc region extends from Cys226, or from Pro230, to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, numbering of amino acid residues in the Fc region or constant region follows the EU numbering system (also referred to as the EU index) as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0099] The "class" of an antibody refers to the type of constant domain or constant region carried by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, several of which can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy-chain constant domains that correspond to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0100] As used herein, the term IgG "isotype" or "subclass" means any of the subclasses of immunoglobulins defined by the chemical and antigenic properties of their constant regions.

[0101] "Framework" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The FR of a variable domain typically consists of four FR domains: FR1, FR2, FR3, and FR4. Thus, the HVR and FR sequences typically appear in VH (or VL) in the following order: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0102] A "human consensus framework" is a framework that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. Generally, the subgroup of sequences is a subgroup as in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda, MD (1991), vols. 1-3. In one embodiment, for VL, the subgroup is subgroup kappa I in Kabat et al. (supra). In one embodiment, for VH, the subgroup is subgroup III in Kabat et al. (supra).

[0103] For purposes herein, an "acceptor human framework" is a framework that comprises 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 comprise the same amino acid sequence, or may contain amino acid sequence changes. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the VL acceptor human framework is identical in sequence to the VL human immunoglobulin framework sequence or the human consensus framework sequence.

[0104] A "humanized" antibody refers to a chimeric antibody comprising amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody comprises substantially all of at least one, and typically two, variable domains, in which all or substantially all of the HVRs (e.g., CDRs) correspond to those of a non-human antibody and all or substantially all of the FRs correspond to those of a human antibody. A humanized antibody may optionally comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody, e.g., a non-human antibody, refers to an antibody that has undergone humanization.

[0105] A "human antibody" is an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human or human cell, or of an antibody derived from a non-human source utilizing the human antibody repertoire, or to the sequence encoding another human antibody. This definition of a human antibody specifically excludes humanized antibodies, which contain non-human antigen-binding residues. Human antibodies can be generated using various techniques known in the art, including phage display libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). The methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(1):86-95 (1991) can also be used to prepare human monoclonal antibodies. See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5:368-74 (2001). Human antibodies can be prepared by administering antigen to transgenic animals, e.g., immunized xenomouse, that have been engineered to produce such antibodies in response to antigen challenge, but whose endogenous gene loci have been disabled (see, e.g., U.S. Patent Nos. 6,075,181 and 6,150,584 regarding XENOMOUSE™ technology). See also, e.g., Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006), regarding human antibodies produced by human B-cell hybridoma technology.

[0106] The term "variable region" or "variable domain" refers to the domain of an antibody heavy chain or light chain that is involved in binding the antibody to an antigen. The variable domains of the heavy and light chains (VH and VL, respectively) of natural antibodies generally have similar structures, and each domain contains four conserved framework regions (FR) and three hypervariable regions (HVR). See, for example, Kindt et al., Kuby Immunology, 6th , W.H. Freeman and Co., page 91 (2007). A single VH or VL domain may be sufficient to confer antigen-binding specificity. Moreover, antibodies that bind to a specific antigen may be isolated by using the VH or VL domain of an antigen-binding antibody to screen a library of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0107] The term "hypervariable region" or "HVR", as used herein, refers to each region of an antibody variable domain that is hypervariable in sequence ("complementarity determining region" or "CDR") and / or structurally forms defined loops ("hypervariable loops") and / or contains residues that contact the antigen ("antigen contacts"). Generally, antibodies contain six HVRs, three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). Exemplary HVRs of the invention include the following: (a) hypervariable loops occurring 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 occurring at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991)); (c) antigenic 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) comprising 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).

[0108] Unless otherwise indicated, HVR residues and other residues in the variable domain (e.g., FR residues) are numbered herein according to Kabat et al., supra.

[0109] An "immunoconjugate" is an antibody conjugated to one or more heterologous molecules, including, but not limited to, cytotoxic agents.

[0110] The term "isolated antibody," as used to describe various antibodies disclosed herein, refers to an antibody that has been identified, separated, and / or recovered from the cell or cell culture in which it is expressed. Contaminant components of its natural environment are materials that would typically interfere with diagnostic or therapeutic uses for the polypeptide, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In some embodiments, the antibody is purified to greater than 95% or greater than 99% purity, as determined, for example, by electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse-phase HPLC). For a review of methods for assessing antibody purity, see, e.g., Flatman et al., J. Chromatogr. B 848:79-87 (2007). In preferred embodiments, antibodies are purified (1) sufficiently to obtain at least 15 residues of N-terminal or internal amino acid sequence using a spinning cup sequenator, or (2) to homogeneity by SDS-PAGE under non-reducing or reducing conditions using Coomassie blue or, preferably, silver stain. Isolated antibodies include antibodies in situ within recombinant cells, since at least one component of the polypeptide's natural environment will not be present. Ordinarily, however, isolated polypeptides will be prepared by at least one purification step.

[0111] The term "monoclonal antibody," as used herein, refers to an antibody obtained from a substantially homogeneous population of antibodies. That is, except for possible minor variant antibodies, including, for example, naturally occurring mutations or mutations that arise during the production of a monoclonal antibody preparation, the individual antibodies comprising the population are identical and / or bind the same epitope. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. Thus, the modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular 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 the human immunoglobulin loci; such methods and other exemplary methods for producing monoclonal antibodies are described herein.

[0112] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by methods common in the art, including those described herein. Specific illustrative explanations and exemplary embodiments for measuring binding affinity are described below.

[0113] An "affinity matured" antibody refers to an antibody that contains one or more alterations in one or more hypervariable regions (HVRs) compared to a parent antibody that does not contain such alterations, which alterations improve the affinity of the antibody for antigen.

[0114] The terms "anti-CD3 antibody" and "antibody that binds to CD3" refer to an antibody that can bind to CD3 with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent targeting CD3. In one embodiment, the binding of an anti-CD3 antibody to an unrelated, non-CD3 protein is less than about 10% of the binding of the antibody to CD3, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, an antibody that binds to CD3 has an affinity 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 ~10 -13 M, e.g. 10 -9 M~10 -13 Dissociation constant (K D In certain embodiments, the anti-CD3 antibody binds to an epitope of CD3 that is conserved among CD3 from different species.

[0115] The term "cluster of differentiation 3" or "CD3," as used herein, unless otherwise indicated, refers to any native CD3 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), including, for example, the CD3ε, CD3γ, CD3α, and CD3β chains. The term encompasses "full-length" unprocessed CD3 (e.g., unprocessed or unmodified CD3ε or CD3γ) as well as any form of CD3 resulting from intracellular processing. The term also encompasses naturally occurring variants of CD3, including, for example, splice variants or allelic variants. CD3 includes, for example, the human CD3ε protein, which is 207 amino acids long (NCBI Reference SEQ ID NO: NP_000724), and the human CD3γ protein, which is 182 amino acids long (NCBI Reference SEQ ID NO: NP_000064).

[0116] The terms "anti-CD20 antibody" and "antibody that binds to CD20" refer to an antibody that is capable of binding to CD20 with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent in targeting CD20. In one embodiment, the binding of an anti-CD20 antibody to an unrelated, non-CD20 protein is less than about 10% of the binding of the antibody to CD20 as measured, for example, by radioimmunoassay (RIA). In certain embodiments, an antibody that binds to CD20 has an affinity 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~10 -13 M, e.g. 10 -9 M~10 -13 M) In certain embodiments, the anti-CD20 antibody binds to an epitope of CD20 that is conserved among CD20 from different species.

[0117] As used herein, the term "cluster of differentiation 20" or "CD20," unless otherwise indicated, refers to any native CD20 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CD20 and any form of CD20 resulting from intracellular processing. The term also encompasses naturally occurring variants of CD20, including, for example, splice variants or allelic variants. CD20 includes, for example, the human CD20 protein (see, e.g., NCBI Reference SEQ ID NOs: NP_068769.2 and NP_690605.1), which is, for example, 297 amino acids in length and can be produced from, for example, a variant mRNA transcript lacking a portion of the 5' UTR (see, e.g., NCBI Reference SEQ ID NO: NM_021950.3), or a longer mutant mRNA transcript (see, e.g., NCBI Reference SEQ ID NO: NM_152866.2).

[0118] The terms "anti-CD20 / anti-CD3 bispecific antibody," "bispecific anti-CD20 / anti-CD3 antibody," and "antibody that binds to CD20 and CD3," or variants thereof, refer to a multispecific antibody (e.g., a bispecific antibody) that is capable of binding to CD20 and CD3 with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent when targeting CD20 and / or CD3. In one embodiment, the extent of binding of a bispecific antibody that binds CD20 and CD3 to unrelated non-CD3 and / or non-CD20 proteins is less than about 10% of the binding of the antibody to CD3 and / or CD20, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, a bispecific antibody that binds CD20 and CD3 has an affinity of 1 μM or less, 100 nM or less, 10 nM or less, 1 nM or less, 0.1 nM or less, 0.01 nM or less, or 0.001 nM or less (e.g., 10 -8 M or less, e.g. 10 -8 M~10 -13 M, e.g. 10 -9 M~10 -13 In certain embodiments, the bispecific antibody that binds to CD20 and CD3 binds to an epitope of CD3 that is conserved among CD3 from different species and / or an epitope of CD20 that is conserved among CD20 from different species. In one embodiment, the bispecific antibody that binds to CD20 and CD3 is mosunetuzumab.

[0119] As used herein, the term "mosunetuzumab" refers to an anti-CD20 / anti-CD3 bispecific antibody having International Nonproprietary Name (INN) List 117 (WHO Drug Information, Vol. 31, No. 2, 2017, p. 303) or CAS Registry Number 1905409-39-3.

[0120] As used herein, the terms "bind," "specifically bind to," or "specific for" refer to a measurable and reproducible interaction, such as binding between a target and an antibody, that is determinative of the presence of the target in the presence of a heterogeneous population of molecules, including biomolecules. For example, an antibody that specifically binds to a target (which may be an epitope) is one that binds to this target with higher affinity, avidity, more readily, and / or with a longer duration than it binds to other targets. In one embodiment, the extent to which an antibody binds to an unrelated target is less than about 10% of the binding of the antibody to the target, as measured by radioimmunoassay (RIA). In certain embodiments, an antibody that specifically binds to a target has a dissociation constant (K) of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, or ≦0.1 nM. D In certain embodiments, the antibody specifically binds to an epitope on a protein that is conserved among proteins from different species. In another embodiment, specific binding can include, but is not required to be, exclusive binding. As used herein, the term refers to, for example, binding to a target. -4 M or less, or 10 -5 M or less, or 10 -6 M or less, or 10 -7 M or less, or 10 -8 M or less, or 10 -9 M or less, or 10 -10 M or less, or 10 -11 M or less, or 10 -12 K below M D , or 10 -4 M~10 -6 M or 10 -6 M~10 -10 M or 10 -7 M~10 -9 M range K D As will be appreciated by those skilled in the art, affinity and K D The values ​​are inversely correlated: a high affinity for the antigen corresponds to a low K DIn one embodiment, the term "specific binding" refers to binding when a molecule binds to a particular polypeptide or epitope on a particular polypeptide without substantially binding to any other polypeptides or polypeptide epitopes.

[0121] "Percent (%) amino acid sequence identity" with respect to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to those in the reference polypeptide, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, without considering any conservative substitutions as part of the sequence identity. Alignment for determining percent amino acid sequence identity can be obtained by a variety of methods within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN® (DNASTAR®) software. Those skilled in the art can determine appropriate parameters for sequence alignment, including any algorithms needed to achieve maximum alignment over the full length of the sequences being compared. However, for purposes herein, percent amino acid sequence identity values ​​are generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was written by Genentech, Inc., and the source code, together 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 publicly available from Genentech, Inc. (South San Francisco, California), or can be compiled from the source code. The ALIGN-2 program should be compiled for use on UNIX operating systems, including Digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary.

[0122] 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 (alternatively, it may be written as a given amino acid sequence A having or comprising a particular % amino acid sequence identity to, with, or relative to a given amino acid sequence B) is calculated as follows: 100 x fraction X / Y

[0123] In this case, X is the number of amino acid residues scored as identical matches by the sequence alignment program ALIGN-2 in that program's alignment of A and B, and Y is the total number of amino acid residues in B. It will be understood that if the length of amino acid sequence A differs from the length of amino acid sequence B, the % amino acid sequence identity of A to B will differ from the % amino acid sequence identity of B to A. Unless otherwise specified, all % amino acid sequence identity values ​​as used herein are obtained as described in the immediately preceding paragraph using the ALIGN-2 computer program.

[0124] The term "pharmaceutical formulation" refers to a preparation in which the biological activity of the active ingredient contained in the preparation is in a form such that it is effective, and which does not contain additional components that are unacceptably toxic to the subject to which the formulation is administered.

[0125] A "pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation, other than an active ingredient, that is non-toxic to a subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0126] As used herein, the term "chemotherapeutic agent" refers to a compound useful in the treatment of cancer, such as a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., a relapsed or refractory B-cell proliferative disorder), e.g., non-Hodgkin's lymphoma (NHL; e.g., diffuse large B-cell lymphoma (DLBCL; e.g., Richter's transformed), 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)), or chronic lymphocytic leukemia (CLL), 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)), or relapsed or refractory CLL). Examples of chemotherapeutic agents include EGFR inhibitors (including small molecule inhibitors such as erlotinib (TARCEVA®, Genentech / OSI Pharm.)); PD 183805 (CI 1033, 2-propenamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-dihydrochloride, Pfizer Inc.); ZD1839, gefitinib (IRESSA®) 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca); ZM 105180 ((6-amino-4-(3-methylphenylamino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-(1-methylpiperidin-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]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide);Dual EGFR / HER2 tyrosine kinase inhibitors such as EKB-569 (N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolinyl]-4-(dimethylamino)-2-butenamide) (Wyeth); AG1478 (Pfizer); AG1571 (SU 5271; Pfizer); and lapatinib (TYKERB®, GSK572016, or N-[3-chloro-4-[(3fluorophenyl)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 such as TAK165 (Takeda); CP- 724,714, an oral selective inhibitor of ErbB2 receptor tyrosine kinase (Pfizer and OSI); dual HER inhibitors such as EKB-569 (available from Wyeth), which preferentially bind to EGFR but inhibit both HER2 and EGFR-overexpressing cells; PKI-166 (Novartis); pan-HER inhibitors such as canertinib (CI-1033; Pharmacia); and ISIS-5132, an antisense agent that inhibits Raf-1 signaling (ISIS Raf-1 inhibitors such as Raf-1 inhibitors such as Raf-1 inhibitors (Raf-1 inhibitors) and Raf-2 inhibitors (Raf-2 inhibitors) and Raf-3 inhibitors (Raf-3 inhibitors) and Raf-4 inhibitors (Raf-4 inhibitors) and Raf-5 inhibitors (Raf-5 inhibitors) and Raf-6 inhibitors (Raf-6 inhibitors) and Raf-7 inhibitors (Raf-7 inhibitors) and Raf-8 inhibitors (Raf-8 inhibitors) and Raf-9 inhibitors (Raf-9 inhibitors) and Raf-1 inhibitors (Raf-1 inhibitors) and Raf-2 inhibitors (Raf-1 inhibitors) and Raf-3 inhibitors (Raf-1 inhibitors) and Raf-4 inhibitors (Raf-1 inhibitors) and Raf-5 inhibitors (Raf-1 inhibitors) and Raf-1 inhibitors (Raf-1 inhibitors) and Raf-2 inhibitors (Raf-2 inhibitors) and Raf-3 inhibitors (Raf-2 inhibitors) and Raf-4 inhibitors (Raf-2 inhibitors) and Raf-3 inhibitors (Raf-2 inhibitors) and Raf-4 inhibitors (Raf-2 inhibitors) and Raf-5 ... Pyrrolopyrimidines such as 62706; pyrazolopyrimidine, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine; curcumin (diferuloylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tyrphostins containing a nitrothiophene moiety;PD-0183805 (Warner-Lambert); antisense molecules (e.g., those that bind to a nucleic acid encoding a HER); quinoxalines (U.S. Patent No. 5,804,396); tryphostin (U.S. Patent No. 5,804,396); pan-HER inhibitors such as ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering AG); CI-1033 ((Pfizer); Afinitac (ISIS 3521; Isis / Lilly); PKI 166 (Novartis); GW2016 (Glaxo SmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering AG); INC-1C11 (Imclone); and rapamycin (sirolimus, RAPAMUNE®); proteasome inhibitors such as bortezomib (VELCADE®, Millennium Pharm.); disulfiram; epigallocatechin gallate; salinosporamide A; carfilzomib; 17-AAG (geldanamycin); radicicol; lactate dehydrogenase A (LDH-A); fulvestrant (FASLODEX®, AstraZeneca); letrozole (Femara®, Novartis), finasunate (VATALANIB®, Novartis); oxaliplatin (ELOXATIN®, Sanofi); 5-FU (5-fluorouracil); leucovorin; lonafamib (SCH 66336); sorafenib (NEXAVAR®, Bayer Labs); alkylating agents such as AG1478, thiotepa, and CYTOXAN® cyclophosphamide; alkylsulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, metuledopa, and uredopa; ethylenimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimeromelamine; acetogenins (especially bullatacin and bullatacinone); camptothecins (including topotecan and irinotecan); bryostatin; kallistatin;CC-1065 (including its synthetic analogs, adozelesin, carzelesin, and bizelesin); cryptophycins (especially cryptophycin 1 and cryptophycin 8); corticosteroids (including prednisone and prednisolone); cyproterone acetate; 5α-reductase inhibitors, including finasteride and dutasteride; vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat dolastatins; aldesleukin, talc duocarmycin (including synthetic analogs, KW-2189 and CB1-TM1); elute Robin; pancratistatin; sarcodictin; spongistatin; nitrogen mustards such as chlorambucil, chromafazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobembine, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; enediyne antibiotics (e.g., calicheamicin, especially calicheamicin) antibiotics such as calicheamicin γ1 and calicheamicin ω1; dynemicins, including dynemicin A; bisphosphonates such as cyclosporin; esperamicin; and neocarzinostatin chromophores and related chromoproteins (enediyne antibiotic chromophores), aclacinomycin, actinomycin, autramycin, azaserine, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycin, dactinomycin, detorubicin, 6-diazo-5-oxo-L-norleucine, morpholino-doxorubicin , cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), mitomycins such as epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, chelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU);Folic acid analogs such as denopterin, methotrexate, pteropterin, and trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and floxuridine; androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testolactone; anti-adrenals such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as folic acid; acetonitrile Laton; aldophosphamide glycosides; aminolevulinic acid; eniluracil; amsacrine; bestravcil; bisantrene; edatraxate; defofamine; demecolcine; diaziconazole; elfomitin; elliptinium acetate; epothilone; etoglucide; gallium nitrate; hydroxyurea; lentinan; lonidynin; maytansinoids such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidamol; nitrerin; pentostatin; fenamet; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products); Razoxane; Rhizoxin; Schizofuran; Spirogermanium; Tenuazonic acid; Triazicone; 2,2',2"-Trichlorotriethylamine; Trichothecenes (especially T-2 toxin, veracrine A, roridin A, and anguidine); Urethane; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitolactol; Pipobroman; 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 pharmaceutically acceptable salts, acids, prodrugs, and derivatives of any of the above.

[0127] Chemotherapeutic agents also include (i) antihormonal agents that act to regulate or inhibit hormone action on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including tamoxifen (including NOLVADEX®; tamoxifen citrate), raloxifene, droloxifene, iodoxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and FARESTON® (toremifine citrate); (ii) aromatase inhibitors that inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as 4(5)-imidazole, aminoglutethimide, MEGASE® (megestrol acetate), AROMASIN® (exemestane; Pfizer), formestany, fadrozole, RIVISOR® (vorozole), FEMARA® (letrozole; Novartis), and ARIMIDEX. (registered trademark) (Anastrozole; AstraZeneca); (iii) antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, tripterelin, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, all-trans retinoic acid, fenretinide, and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly those that inhibit the expression of genes in signal transduction 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., ALLOVECTIN®, LEUVECTIN®, and VAXID®;(ix) growth inhibitors, including vincas (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, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C); (x) including pharmaceutically acceptable salts, acids, prodrugs, and derivatives of any of the above;

[0128] The term "cytotoxic agent," as used herein, refers to any agent that is detrimental to cells (e.g., causes cell death, inhibits growth, or otherwise interferes with cell function). Cytotoxic agents include radioisotopes (e.g., At 211 , I 131 , I 125 , Y 90 ,Re 186 ,Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212, and radioactive isotopes of Lu), chemotherapeutic agents, enzymes such as nucleases and fragments thereof, and toxins such as small molecule or enzymatically active toxins of bacterial, fungal, plant, or animal origin, including fragments and / or variants thereof. Exemplary cytotoxic agents may be selected from anti-microtubule agents, platinum coordination complexes, alkylating agents, antibiotic agents, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormone analogs, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, proapoptotic agents, LDH-A inhibitors, fatty acid biosynthesis inhibitors, cell cycle signaling inhibitors, HDAC inhibitors, proteasome inhibitors, and cancer metabolism inhibitors. In one example, the cytotoxic agent is a platinum-based chemotherapeutic agent (e.g., carboplatin or cisplatin). In one example, the cytotoxic agent is an EGFR antagonist, such as N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (e.g., erlotinib). In one example, the cytotoxic agent is a RAF inhibitor, such as a BRAF and / or CRAF inhibitor. In one example, the RAF inhibitor is vemurafenib. In one example, the cytotoxic agent is a PI3K inhibitor.

[0129] The term "PD-1 axis binding antagonist" refers to a molecule that inhibits the interaction of a PD-1 axis binding partner with any one or more of its binding partners to eliminate T cell dysfunction resulting from signaling on the PD-1 signaling axis, thereby restoring or enhancing T cell function (e.g., proliferation, cytokine production, target cell killing). As used herein, PD-1 axis binding antagonists include PD-1 binding antagonists, PD-L1 binding antagonists, and PD-L2 binding antagonists.

[0130] The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, impairs, or prevents signaling that occurs as a result of 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 particular aspects, PD-1 binding antagonists inhibit the binding of PD-1 to PD-L1 and / or PD-L2. For example, PD-1 binding antagonists include anti-PD-1 antibodies, antigen-binding fragments thereof, immunoadhexins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, impair, or prevent signaling that results from the interaction of PD-1 with PD-L1 and / or PD-L2. In one embodiment, the PD-1 binding antagonist reduces negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes that mediate PD-1-mediated signaling, rendering dysfunctional T cells less dysfunctional (e.g., enhancing the effector response to antigen recognition). In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody. In a particular aspect, the PD-1 binding antagonist is MDX-1106 (nivolumab). In another particular aspect, the PD-1 binding antagonist is pembrolizumab (formerly lambrolizumab (MK-3475)). In another particular aspect, the PD-1 binding antagonist is AMP-224. In some embodiments, the PD-1 binding antagonist is MDX-1106 (nivolumab). In some embodiments, the PD-1 binding antagonist is MK-3475 (pembrolizumab). In some embodiments, the PD-1 binding antagonist is MED1-0680. In some examples, the PD-1 binding antagonist is PDR001 (spartalizumab). In some examples, the PD-1 binding antagonist is REGN 2810 (cemiplimab). In some examples, the PD-1 binding antagonist is BGB-108. In other examples, the PD-1 binding antagonist is prorugolimab, camrelizumab, sintilimab, tislelizumab, or toripalimab.

[0131] Further examples of PD-1 axis binding antagonists include cemiplimab, prorugolimab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostarimab, retifanlimab, spartalizumab, southernlimab, penprimimab, CS1003, HLX10, SCT-I10A, SHR-1316, CS1001, embafolimab, TQB2450, ZKAB001, LP-002, zimberelimab, balstilimab, genolimuzumab, BI754091, cetrelimab, YBL-006, and BAT13. 06, HX008, CX-072, IMC-001, KL-A167, budicalimab, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, APL-502, cosibelimab, lodapolimab, GS-4224, INCB086550, FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, MAX-10181, RC98, BION-004, AM0001, CB201, ENUM 244C8, ENUM 388D4, AUNP-012, STI-1110, ADG104, AK-103, LBL-006, hAb21, AVA-004, PDL-GEX, INCB090244, KD036, KY1003, LYN192, MT-6035, VXM10, YBL-007, ABSK041, GB7003, JS-003, and HS-636.

[0132] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, abrogates, or prevents 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 partners. In particular aspects, PD-L1 binding antagonists inhibit the binding of PD-L1 to PD-1 and / or B7-1. In some embodiments, PD-L1 binding antagonists include anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, abrogate, or prevent signaling resulting from the interaction of PD-L1 with one or more of its binding partners, such as PD-1 or B7-1. In one embodiment, the PD-L1 binding antagonist reduces the negative costimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes that mediate PD-L1-mediated signaling, thereby alleviating the dysfunctional state of 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 a specific embodiment, the anti-PD-L1 antibody is atezolizumab (CAS Registry Number: 1422185-06-5), also known as MPDL3280A and described herein. In another specific embodiment, the anti-PD-L1 antibody is MDX-1105, described herein. In yet another specific aspect, the anti-PD-L1 antibody is MEDI4736, described herein.

[0133] As used herein, the term "atezolizumab" refers to the anti-PD-L1 antagonist antibody having International Nonproprietary Name (INN) List 112 (WHO Drug Information, Vol. 28. No. 4, 2014, p. 488) or CAS Registry Number 1380723-44-3.

[0134] The term "PD-L2 binding antagonist" refers to a molecule that reduces, blocks, inhibits, abrogates, or prevents signaling resulting from the interaction of PD-L2 with any one or more of its binding partners, e.g., 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 particular aspects, PD-L2 binding antagonists inhibit the binding of PD-L2 to PD-1. In some embodiments, PD-L2 antagonists include anti-PD-L2 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, abrogate, or prevent signaling resulting from the interaction of PD-L2 with any one or more of its binding partners, e.g., PD-1. In one embodiment, the PD-L2 binding antagonist reduces the negative costimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-L2, reducing the dysfunction of dysfunctional T cells (e.g., enhancing the effector response to antigen recognition). In some embodiments, the PD-L2 binding antagonist is an immunoadhesin.

[0135] The term "package insert" is used to refer to instructions customarily included in commercial packaging of a therapeutic product that contain information regarding the indications, uses, dosage, administration, concomitant therapy, contraindications and / or warnings regarding the use of such therapeutic product.

[0136] III. Treatment method Provided herein are methods of treating a subject with a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder, e.g., NHL (e.g., DLBCL or FL) or CLL), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle. In some examples, the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, and the second dosing cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody. In some examples, C1D1 is less than or equal to C1D2 and less than C1D3, and C1D2 is less than or equal to C1D3. In some examples, C1D1 is about 0.1 mg to about 10 mg (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg). mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).In some examples, C2D1 is greater than or equal to C1D3 and is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; e.g., about 30 mg, about 45 mg, or about 60 mg). In some examples, the methods provided herein include treating a subject with CLL, wherein the treatment comprises subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosage regimen comprising 0.1 mg of C1D1.

[0137] Also provided herein are methods of treating a population of subjects having a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder, e.g., NHL (e.g., DLBCL or FL) or CLL), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle. In some examples, the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, and the second dosing cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody. In some examples, C1D1 is less than or equal to C1D2 and less than C1D3, and C1D2 is less than or equal to C1D3. In some examples, C1D1 is about 0.1 mg to about 10 mg (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, or about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 150 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 100 mg, about 15 ...150 mg, about 150 mg, about 150 mg, about 150 mg, 0 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).In some examples, C2D1 is greater than or equal to C1D3 and is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, about 40 mg to about 100 mg, or about 25 mg to about 75 mg; e.g., about 30 mg, about 45 mg, or about 60 mg). In some examples, the methods provided herein include treating a population of subjects with CLL, wherein the treatment comprises subcutaneously administering to the subjects a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising 0.1 mg of C1D1.

[0138] A. Therapeutic Methods for Administration of Anti-CD20 / Anti-CD3 Bispecific Antibodies The present invention relates to the treatment of CD20-positive cell proliferative disorders, such as B-cell proliferative disorders (e.g., non-Hodgkin's lymphoma (NHL) (e.g., previously untreated (1L) NHL), diffuse large B-cell lymphoma (DLBCL) (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), follicular lymphoma (FL) (e.g., 1L Provided are methods of treating a subject with FL, relapsed and / or refractory FL, or transformed FL), mantle cell lymphoma (MCL), high-grade B-cell lymphoma, or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL) or chronic lymphocytic leukemia (CLL), comprising administering an anti-CD20 / anti-CD3 bispecific antibody (e.g., mosunetuzumab) to the subject, e.g., in a split-dose escalation regimen, or, e.g., in a split-step-up regimen for the first dosing cycle. In some examples, the method is used to treat a subject with relapsed and / or refractory NHL (e.g., aggressive NHL (e.g., relapsed and / or refractory DLBCL, or relapsed and / or refractory FL)). In some examples, the subject has a documented history of at least 6 months of response (e.g., complete or partial response) in the period since completion of treatment, followed by one or more The subject has relapsed to (e.g., one, two, three, or more) prior therapies (e.g., one or more prior systemic therapies, e.g., one or more prior systemic chemotherapies (e.g., one or more prior systemic therapies including administration of an anthracycline), one or more prior stem cell therapies, or one or more prior CAR-T cell therapies). In some examples, the subject is refractory to any prior therapies (e.g., has not had a response to the prior therapies or has progressed within 6 months of completing the last dose of therapy). Thus, in some embodiments, the dosing regimen is a second-line (2L) therapy. In some embodiments, the dosing regimen is a third-line (3L) therapy. In some embodiments, the subject has transformed FL that is refractory to standard therapies for transformed FL. In some embodiments, the FL is graded FL (e.g., grade 1 FL, grade 2 FL, grade 3a FL, or grade 3b FL).In some embodiments, the methods are used to treat subjects with non-relapsed and non-refractory NHL, and the dosing regimen is first-line (1L) therapy.

[0139] In some examples, the present invention provides a method for treating a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., NHL (e.g., previously untreated (1L) NHL), DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), FL (e.g., 1L NHL), or FL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), comprising subcutaneously administering to a subject a bispecific antibody that binds to CD20 and CD3 (e.g., mosunetuzumab) in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle. (ii) treating a subject with FL, relapsed and / or refractory FL, or transformed FL), MCL, high-grade B-cell lymphoma or PMLBCL) or CLL), wherein the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein C1D1 is equal to or less than C1D2 and less than C1D3; (ii) C1D2 is equal to or less than C1D3; and (iii) ) C1D1 is about 0.1 mg to about 10 mg (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg). about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg) , about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).The second administration cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, where C2D1 is C1D3 or greater and is about 10 mg to about 300 mg (e.g., about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg , about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg). In some examples, C1D1 is less than C1D2. In some examples, C1D1 is approximately equivalent to C1D3. In some examples, C1D1 is about 2 mg to about 8 mg (e.g., about 3 mg to about 7 mg, about 4 mg to about 6 mg; e.g., about 5 mg), and C1D2 is about 10 mg to about 75 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg). , about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), C1D3 is about 20 mg to about 75 mg (e.g., about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 45 mg), and C2D1 is about 20 mg to about 75 mg (e.g., about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 45 mg). In a specific embodiment, C1D1 is about 5 mg, C1D2 is about 45 mg, C1D3 is about 45 mg, and C2D1 is about 45 mg. In another embodiment, C1D1 is about 5 mg, C1D2 is about 15 mg, C1D3 is about 45 mg, and C2D1 is about 45 mg. In another embodiment, C1D1 is about 5 mg, C1D2 is about 10 mg, C1D3 is about 30 mg, and C2D1 is about 30 mg.In other embodiments, C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 40 mg, and C2D1 is about 40 mg. In yet other embodiments, C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 60 mg, and C2D1 is about 60 mg. In yet other embodiments, C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 45 mg, and C2D1 is about 60 mg.

[0140] In some examples, C1D1 is equal to C1D2 (e.g., C1D1 is about 5 mg, C1D2 is about 5 mg, C1D3 is about 45 mg or 60 mg, and C2D1 is about 45 mg or 60 mg).

[0141] In other examples, C1D2 is equal to C1D3 (e.g., C1D1 is about 5 mg, C1D2 is about 45 mg or 60 mg, C1D3 is about 45 mg or 60 mg, and C2D1 is about 45 mg or 60 mg). In some examples, the methods include administering C1D2 to the subject about 7 days after C1D1. In some examples, the methods include administering C1D3 to the subject about 7 days after C1D2. In some examples, the methods include administering C2D1 to the subject about 7 days after C1D3. For example, in some embodiments, the methods of the invention include administering C1D1, C1D2, and C1D3 to the subject on days 1, 8, and 15, or about days 1, 8, and 15, respectively, of a first administration cycle. In some examples, the methods include administering C2D1 to the subject on day 1 of a second administration cycle. In some examples, the first and second administration cycles are 21-day administration cycles. In some examples, the first administration cycle is a 21-day administration cycle and the second administration cycle is a 28-day administration cycle. Alternatively, in some examples, the first and second administration cycles are 28-day administration cycles.

[0142] In some examples, the present invention provides a method for treating a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., NHL (e.g., previously untreated (1L) NHL), DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), FL (e.g., 1L NHL), or FL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), comprising subcutaneously administering to a subject a bispecific antibody that binds to CD20 and CD3 (e.g., mosunetuzumab) in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle. The present invention includes treating a subject with FL, relapsed and / or refractory FL, or transformed FL), MCL, high-grade B-cell lymphoma or PMLBCL, or CLL, wherein the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, where (i) C1D1 is about 5 mg, (ii) C1D2 is equal to or greater than C1D1 and equal to or less than C1D3, and (iii) C1D3 is about 45 mg. The second administration cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, where C2D1 is about 45 mg. In some examples, C1D2 is about 5 mg, about 10 mg, about 15 mg, about 25 mg, about 30 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 120 mg, about 140 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 450 mg, about 50 ... In some examples, the first administration cycle and the second administration cycle are 21-day administration cycles (e.g., a 21-day administration cycle in which C1D1, C1D2, and C1D3 are administered on days 1, 8, and 15, or about days 1, 8, and 15, respectively, of the first administration cycle, and C2D1 is administered on day 1 of the second administration cycle). In some examples, the first administration cycle is a 21-day administration cycle and the second administration cycle is a 28-day administration cycle. In some examples, the first administration cycle and the second administration cycle are 28-day administration cycles.

[0143] In some examples, the present invention provides a method for treating a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., NHL (e.g., previously untreated (1L) NHL), DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), FL (e.g., 1L NHL), or FL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), comprising subcutaneously administering to a subject a bispecific antibody that binds to CD20 and CD3 (e.g., mosunetuzumab) in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle. and (c) treating a subject with FL, relapsed and / or refractory FL, or transformed FL), MCL, high-grade B-cell lymphoma or PMLBCL) or CLL), wherein the first administration cycle comprises a first subcutaneous dose of the bispecific antibody (C1D1) on day 1 of the first administration cycle, a second subcutaneous dose of the bispecific antibody (C1D2) on day 8 of the first administration cycle, and a third subcutaneous dose of the bispecific antibody (C1D3) on day 15 of the first administration cycle, wherein (i) C1D1 is about 5 mg, and (i) (i) C1D2 is greater than or equal to C1D1 and less than or equal to C1D3, and (iii) C1D3 is about 45 mg. The second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1) on day 1 of the second administration cycle, where C2D1 is about 45 mg. 31 In some examples, C1D2 is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 45 mg, or about 60 mg. In some embodiments, C1D2 is about 15 mg. In some embodiments, C1D2 is about 45 mg.

[0144] In some embodiments, the dosing regimen includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) additional administration cycles (e.g., 1 to 15 additional administration cycles, 8 to 17 additional administration cycles, or 6 to 15 additional administration cycles). In some embodiments, the dosing regimen includes 6 additional administration cycles. In some embodiments, the dosing regimen includes 15 additional administration cycles. In some embodiments, the dosing regimen includes 2 to 17 (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17) total administration cycles. In some embodiments, the dosing regimen includes 8 total administration cycles. In some embodiments, the dosing regimen includes 17 total administration cycles. In some embodiments, each additional administration cycle is a 21-day administration cycle. In some embodiments, each additional administration cycle is a 28-day administration cycle. In some embodiments, each additional administration cycle comprises administering an additional dose of the bispecific antibody. In some embodiments, each additional dose of the bispecific antibody is an amount approximately equal to C2D1. In some embodiments, each additional dose of the bispecific antibody is about 45 mg. In some embodiments, the method comprises administering each additional dose of the bispecific antibody to the subject on day 1 of each respective additional administration cycle.

[0145] In some examples, each additional administration cycle is a 21-day administration cycle. In some examples, the first administration cycle is a 21-day administration cycle and the second administration cycle is a 28-day administration cycle. Alternatively, each additional administration cycle is a 28-day administration cycle.

[0146] In some examples, each of the one or more additional administration cycles comprises a single subcutaneous dose of the bispecific antibody, e.g., a single subcutaneous dose on day 1 of each of the one or more additional administration cycles.

[0147] In a particular example, provided herein is a method of treating a subject with DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation) comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 (e.g., mosunetuzumab) in a dosing regimen comprising at least a first 21-day dosing cycle and a second 21-day dosing cycle, wherein the first 21-day dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein (i) C1D1 is equal to or less than C1D2 and less than C1D3, (ii) C1D2 is equal to or less than C1D3, and (iii) C1D1 is equal to or less than C1D1. is about 0.1 mg to about 10 mg (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg). mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).The second 21-day administration cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, where C2D1 is C1D3 or greater and is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg , about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg). In some examples, the method includes administering C1D2 to the subject about 7 days after C1D1. In some examples, the method includes administering C1D3 to the subject about 7 days after C1D2. In some examples, the method includes administering C2D1 to the subject about 7 days after C1D3. In some examples, the method includes administering C1D1, C1D2, and C1D3 to a subject on or about days 1, 8, and 15 of a first administration cycle, respectively.

[0148] In a particular example, provided herein is a method of treating a subject with FL (e.g., 1L FL, relapsed and / or refractory FL, or transformed FL), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 (e.g., mosunetuzumab) in a dosing regimen comprising at least a first 28-day dosing cycle and a second 28-day dosing cycle, wherein the first 28-day dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein (i) C1D1 is equal to or less than C1D2 and less than C1D3, and (ii) C1D2 is equal to or less than C1D3. (iii) C1D1 is about 0.1 mg to about 10 mg (e.g., about 0.5 to about 10 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 300 mg). about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).The second 28-day administration cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, where C2D1 is C1D3 or greater and is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg , about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).

[0149] In a particular example, provided herein is a method of treating a subject with FL (e.g., 1L FL, relapsed and / or refractory FL, or transformed FL), comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 (e.g., mosunetuzumab) in a dosing regimen comprising at least a first 21-day dosing cycle and a second 28-day dosing cycle, wherein the first 21-day dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein (i) C1D1 is equal to or less than C1D2 and less than C1D3, and (ii) C1D2 is equal to or less than C1D3. (iii) C1D1 is about 0.1 mg to about 10 mg (e.g., about 0.5 to about 10 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 300 mg). about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).The second 28-day administration cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, where C2D1 is C1D3 or greater and is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg , about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).

[0150] In some examples, C1D1 is less than C1D2. In some examples, C1D2 is approximately equivalent to C1D3. In some examples, C1D1 is about 2 mg to about 8 mg (e.g., about 3 mg to about 7 mg, about 4 mg to about 6 mg; e.g., about 5 mg), and C1D2 is about 10 mg to about 75 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg). , about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), C1D3 is about 20 mg to about 75 mg (e.g., about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 45 mg), and C2D1 is about 20 mg to about 75 mg (e.g., about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 45 mg). In certain embodiments, C1D1 is about 5 mg. In some embodiments, C1D3 is about 25 mg to about 75 mg. In some embodiments, C1D3 is about 30 mg, about 45 mg, or about 60 mg. In some embodiments, C2D1 is about 40 mg to about 75 mg. In some embodiments, C2D1 is about 30 mg, about 45 mg, or about 60 mg. In some embodiments, C1D2 is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, or about 60 mg.In some examples, (a) C1D1 is about 5 mg, C1D2 is about 45 mg, C1D3 is about 45 mg, and C2D1 is about 45 mg; (b) C1D1 is about 5 mg, C1D2 is about 10 mg, C1D3 is about 30 mg, and C2D1 is about 30 mg; (c) C1D1 is about 5 mg, C1D2 is about 15 mg, C1D3 is about 45 mg, and C2D1 is about 45 mg. (d) C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 40 mg, and C2D1 is about 40 mg; (e) C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 45 mg, and C2D1 is about 60 mg; or (f) C1D1 is about 5 mg, C1D2 is about 20 mg, C1D3 is about 60 mg, and C2D1 is about 60 mg.

[0151] In certain embodiments, C1D1 is about 5 mg, C1D2 is about 45 mg, C1D3 is about 45 mg, and C2D1 is about 45 mg. In some embodiments, C1D1 is equal to C1D2, e.g., C1D1 is about 5 mg, C1D2 is about 5 mg, C1D3 is about 45 mg or about 60 mg, and C2D1 is about 45 mg or about 60 mg.

[0152] In other examples, C1D2 is equal to C1D3, for example, C1D1 is about 5 mg, C1D2 is about 45 mg or about 60 mg, C1D3 is about 45 mg or about 60 mg, and C2D1 is about 45 mg or about 60 mg.

[0153] In some examples, the methods include administering C2D1 to the subject on day 1 of a second administration cycle.

[0154] In some examples, the dosing regimen includes one or more additional administration cycles (additional administration cycles beyond the second administration cycle) (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, or 17 or more additional administration cycles, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 additional administration cycles). In particular examples, the dosing regimen includes 8 to 17 additional administration cycles (e.g., 10 to 19 total administration cycles). In particular examples, the dosing regimen includes 6 to 15 additional administration cycles (e.g., 8 to 17 total administration cycles).

[0155] The present invention also provides methods of treating a population of subjects having a CD20-positive cell proliferative disorder by administering a bispecific antibody that binds CD20 and CD3 (e.g., mosunetuzumab) to one or more subjects according to any of the dosing regimens described herein. In some examples, the present invention provides a method of treating a population of subjects having a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., NHL (e.g., previously untreated (1L) NHL), DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation)), FL (e.g., 1L NHL), or IL-16 (e.g., IL-16)), comprising subcutaneously administering a bispecific antibody that binds CD20 and CD3 (e.g., mosunetuzumab) to one or more subjects in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle. Provided herein are methods of treating a population of subjects with FL, relapsed and / or refractory FL, or transformed FL), MCL, high-grade B-cell lymphoma or PMLBCL) or CLL), wherein the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of a bispecific antibody, wherein (i) C1D1 is equal to or less than C1D2 and less than C1D3, (ii) C1D2 is equal to or less than C1D3, and (iii) C1D1 is between about 0.1 mg and about 10 mg (e.g., between about 0.1 mg and about 7 mg, between about 0.2 mg and about 10 mg, between about 0.5 mg and about 10 mg, or between about 0.6 mg and about 10 mg, and between about 0.7 mg and about 10 mg, and between about 0.8 mg and about 10 mg, and between about 0.9 mg and about 10 mg, and between about 10 mg and about 10 mg, and between about 0.9 ...C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; e.g., about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (e.g., about 25 mg to about 300 mg). g, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, about 40 mg to about 100 mg, about 20 mg to about 100 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 45 mg). The second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), where C2D1 is C1D3 or greater and is about 10 mg to about 300 mg (e.g., about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg ~200mg, 10mg~180mg, 10mg~160mg, 10mg~150mg, 10mg~140mg, 20mg~130mg, 30mg~120mg, 40m g to about 100 mg, about 20 mg to about 100 mg, about 25 mg to about 75 mg, from about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 45 mg). .

[0156] In some examples, the present invention provides a method for treating a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., NHL (e.g., previously untreated (1L) NHL), DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), FL (e.g., 1L NHL), or FL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), comprising subcutaneously administering to a subject a bispecific antibody that binds to CD20 and CD3 (e.g., mosunetuzumab) in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle. In some examples, the present invention features a method of treating a population of subjects with FL, relapsed and / or refractory FL, or transformed FL), MCL, high-grade B-cell lymphoma or PMLBCL), or CLL), wherein the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of a bispecific antibody, where (i) C1D1 is about 5 mg, (ii) C1D2 is greater than or equal to C1D1 and less than or equal to C1D3, and (iii) C1D3 is about 45 mg. In some examples, the second administration cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, where C2D1 is about 45 mg.

[0157] In some examples, the present invention provides a method for treating a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., NHL (e.g., previously untreated (1L) NHL), DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), FL (e.g., 1L NHL), or FL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), comprising subcutaneously administering to a subject a bispecific antibody that binds to CD20 and CD3 (e.g., mosunetuzumab) in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle. and (iii) C1D3 is about 45 mg. In some examples, the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1) on day 1 of the second administration cycle, wherein C2D1 is about 45 mg.

[0158] In some examples, the invention provides a method of treating a population of subjects with DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation) comprising subcutaneously administering a bispecific antibody that binds CD20 and CD3 (e.g., mosunetuzumab) to one or more subjects in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein (i) C1D1 is equal to or less than C1D2 and less than C1D3, (ii) C1D2 is equal to or less than C1D3, and (iii) C1D1 is equal to or less than about 0.1 m g to about 10 mg (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg); For example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).In some examples, the second administration cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, where C2D1 is C1D3 or greater and is between about 10 mg and about 300 mg (about 25 mg and about 300 mg, about 50 mg and about 300 mg, about 100 mg and about 300 mg, about 200 mg and about 300 mg, about 50 mg and about 250 mg, about 100 mg and about 250 mg, about 100 mg and about 200 mg). mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).

[0159] In another example, the invention provides a method of treating a population of subjects with FL (e.g., 1L FL, relapsed and / or refractory FL, or transformed FL), comprising subcutaneously administering to the subjects a bispecific antibody that binds CD20 and CD3 (e.g., mosunetuzumab) in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody, wherein (i) C1D1 is equal to or less than C1D2 and less than C1D3, (ii) C1D2 is equal to or less than C1D3, and (iii) C1D1 is between about 0.1 mg and about 10 mg. (e.g., about 0.1 mg to about 7 mg, about 0.2 mg to about 10 mg, about 0.5 mg to about 10 mg, about 1 mg to about 9 mg, about 2 mg to about 8 mg, about 3 mg to about 7 mg, about 4 mg to about 6 mg; for example, about 5 mg), and C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).The second administration cycle comprises a single subcutaneous dose (C2D1) of the bispecific antibody, where C2D1 is C1D3 or greater and is about 10 mg to about 300 mg (about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, or about 40 mg to about 100 mg, or about 25 mg to about 75 mg; for example, about 30 mg, about 45 mg, or about 60 mg).

[0160] The dosing regimens provided herein may also reduce the rate of certain adverse events in a population of subjects with a CD20-positive cell proliferative disorder (e.g., a B-cell proliferative disorder (e.g., an NHL (e.g., previously untreated (1L) NHL), DLBCL (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), FL (e.g., 1L FL, relapsed and / or refractory FL, or transformed FL), MCL, high-grade B-cell lymphoma, or PMLBCL) or CLL). For example, in some examples, the present invention provides a method for administering a bispecific antibody that binds CD20 and CD3 (e.g., mosunetuzumab) to a subject ... DLBCL, or and a method of reducing the rate of a particular adverse event in a population of subjects with FL, relapsed and / or refractory FL, or transformed FL), MCL, high-grade B-cell lymphoma or PMLBCL) or CLL, the method comprising administering a bispecific antibody subcutaneously using a step-up dosing regimen, wherein the rate of adverse events is reduced in the population of subjects compared to a reference population of subjects to whom the bispecific antibody is administered intravenously or a reference population of subjects to whom the bispecific antibody is administered subcutaneously at a fixed dose (i.e., not a step-up dose). In some examples, the stepwise dosing regimen includes at least a first administration cycle and a second administration cycle, wherein (a) the first administration cycle includes a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of a bispecific antibody, wherein (i) C1D1 is equal to or less than C1D2 and less than C1D3, (ii) C1D2 is equal to or less than C1D3, and (iii) C1D1 is between about 0.1 mg and about 10 mg (e.g., between about 0.1 mg and about 7 mg, between about 0.2 mg and about 10 mg, between about 0.C1D2 is about 5 mg to about 80 mg (e.g., about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg, e.g., about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 75 mg, 0 mg, or about 75 mg), and C1D3 is about 10 mg to about 300 mg (e.g., about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein C2D1 is C1D3 or greater and is about 10 mg to about 300 mg (e.g., about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 100 mg to about 300 mg, about 200 mg to about 300 mg, about 50 mg to about 300 mg, about 50 mg to about 300 mg, about 40 mg to about 75 mg, about 45 mg to about 450 mg, about 50 mg to about 450 mg, about 45 mg to about 500 mg, about 50 ... about 250 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 180 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 20 mg to about 130 mg, about 30 mg to about 120 mg, about 40 mg to about 100 mg, about 20 mg to about 100 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, or about 40 mg to about 75 mg; for example, about 45 mg).

[0161] In some examples, the stepwise dosing regimen comprises at least a first dosing cycle and a second dosing cycle, wherein (a) the first dosing cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of a bispecific antibody, where (i) C1D1 is about 5 mg, (ii) C1D2 is greater than or equal to C1D1 and less than or equal to C1D3, and (iii) C1D3 is about 45 mg; and (b) the second dosing cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), where C2D1 is about 45 mg.

[0162] In some examples, the stepwise dosing regimen comprises at least a first dosing cycle and a second dosing cycle, wherein (a) the first dosing cycle comprises a first subcutaneous dose of the bispecific antibody (C1D1) on day 1 of the first dosing cycle, a second subcutaneous dose of the bispecific antibody (C1D2) on day 8 of the first dosing cycle, and a third subcutaneous dose of the bispecific antibody (C1D3) on day 15 of the first dosing cycle, wherein (i) C1D1 is about 5 mg, (ii) C1D2 is greater than or equal to C1D1 and less than or equal to C1D3, and (iii) C1D3 is about 45 mg; and (b) the second dosing cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1) on day 1 of the second dosing cycle, wherein C2D1 is about 45 mg.

[0163] Any of the methods described herein may include monitoring the subject for cytokine release syndrome (CRS) (e.g., a CRS event after initiation of any of the above methods). Current clinical responses focus on treating individual signs and symptoms, providing supportive care, and attempting to attenuate the inflammatory response using high doses of corticosteroids. However, this approach is not always successful, especially in cases of late intervention. The CRS grading criteria used by the methods described herein define mild, moderate, severe, or life-threatening CRS and have been published by the American Society for Transplantation and Cellular Therapy (ASTCT) to harmonize reporting across clinical trials and enable rapid recognition and treatment of CRS (Lee et al., Biology of Blood and Marrow Transplantation. 25(4):625-638, 2019). The ASTCT criteria are objective, easy to apply, and intended to more accurately classify the severity of CRS. This revised CRS grading system is shown in Table 1 below. [Table 1] ASTCT = American Society for Transplantation and Cellular Therapy; BiPAP = bilevel positive airway pressure; CPAP = continuous positive airway pressure; CRS = cytokine release syndrome; CTCAE = Common Terminology Criteria for Adverse Events.

[0164] Fever is defined as a temperature of 38°C or higher not attributable to other causes. In subjects with CRS, if they then receive antipyretic or anticytokine therapy, such as tocilizumab or steroids, fever is no longer required to grade the severity of their subsequent CRS. In this case, CRS grading is determined by hypotension and / or hypoxia.

[0165] CRS grade is determined by the more severe events, hypotension, or hypoxia not attributable to other causes. For example, a subject with a temperature of 39.5°C, hypotension requiring one vasopressor, and hypoxia requiring a low-flow nasal cannula is classified as having Grade 3 CRS.

[0166] Low-flow nasal cannulae are defined as oxygen delivered at ≤6 L / min. Low-flow also includes blow-by oxygen delivery, sometimes used in pediatrics. High-flow nasal cannulae are defined as oxygen delivered at >6 L / min.

[0167] CRS is associated with elevations in a variety of cytokines, with marked increases in IFNγ, IL-6, and TNF-α levels. Emerging evidence specifically implicates IL-6 as a central mediator in CRS. IL-6 is a proinflammatory, multifunctional cytokine produced by a variety of cell types, and this cytokine has been shown to be involved in a wide variety of physiological processes, including T cell activation. Regardless of the inducer, CRS is associated with elevated IL-6 levels (Nagorsen et al. Cytokine. 25(1):31-5, 2004; Lee et al. Blood. 124(2):188-95, 2014; Doesegger et al. Clin. Transl. Immunology. 4(7):e39, 2015), and IL-6 correlates with CRS severity, with subjects experiencing grade 4 or 5 CRS events having significantly higher IL-6 levels compared with subjects who do not experience CRS or who experience milder CRS (grades 0-3) (Chen et al. J. Immunol. Methods. 434:1-8, 2016).

[0168] Therefore, blocking the inflammatory effects of IL-6 using an agent that inhibits IL-6-mediated signaling to manage CRS observed in subjects during a two-phase, fractionated, ascending-dose regimen is an alternative to steroid treatment that is not expected to adversely affect T cell function or reduce the efficacy or clinical benefit of anti-CD20 / anti-CD3 bispecific antibody therapy in the treatment of CD20-positive cell proliferative disorders (e.g., B-cell proliferative disorders).

[0169] Tocilizumab (ACTEMRA® / RoACTEMRA®) is a recombinant, humanized, anti-human monoclonal antibody directed against the soluble, membrane-bound IL-6R that inhibits IL-6-mediated signaling (see, e.g., WO1992 / 019579, which is incorporated herein by reference in its entirety).

[0170] If the subject experiences a cytokine release syndrome (CRS) event after administration of the bispecific antibody, the method may further comprise administering to the subject an effective amount of an interleukin-6 receptor (IL-6R) antagonist (e.g., an anti-IL-6R antibody, e.g., tocilizumab (ACTEMRA® / RoACTEMRA®)) to manage the event. In some examples, tocilizumab is administered intravenously to the subject as a single dose of about 8 mg / kg. In some examples, each dose of tocilizumab does not exceed 800 mg / dose. Other anti-IL-6R antibodies that may be used instead of or in combination with tocilizumab include sarilumab, bovalilizumab (ALX-0061), satralizumab (SA-237), and variants thereof.

[0171] If the subject has a CRS event that does not resolve or worsen within 24 hours of administering the IL-6R antagonist to treat the symptoms of the CRS event, the method may further comprise administering one or more additional doses of an IL-6R antagonist (e.g., an anti-IL-6R antibody, e.g., tocilizumab) to the subject to manage the CRS event. If the CRS event is not managed through administration of the IL-6R antagonist, the subject may be administered a corticosteroid, such as methylprednisolone or dexamethasone.

[0172] Management of the CRS event can be adjusted based on the stage of CRS and the presence of comorbidities. For example, if, after administration of the bispecific antibody, the subject experiences a Grade 2 cytokine release syndrome (CRS) event in the absence of comorbidities or in the presence of minimal comorbidities, the method can further comprise treating the symptoms of the Grade 2 CRS event while withholding treatment with the bispecific antibody. If the Grade 2 CRS event resolves to a Grade ≦1 CRS event for at least three consecutive days, the method can further comprise resuming treatment with the bispecific antibody at the same dose. On the other hand, if the Grade 2 CRS event does not resolve or worsen to a Grade ≧3 event within 24 hours of treating the symptoms of the Grade 2 CRS event, the method can further comprise administering to the subject an effective amount of an interleukin-6 receptor (IL-6R) antagonist (e.g., an anti-IL-6R antibody, e.g., tocilizumab (ACTEMRA® / RoACTEMRA®)) to manage the Grade 2 or Grade ≧3 CRS event. In some examples, tocilizumab is administered intravenously to a subject as a single dose of about 8 mg / kg. In some examples, each dose of tocilizumab does not exceed 800 mg / dose. Other anti-IL-6R antibodies that can be used instead of or in combination with tocilizumab include sarilumab, bovalilizumab (ALX-0061), satralizumab (SA-237), and variants thereof.

[0173] If the subject experiences a Grade 2, 3, or 4 CRS event in the presence of extensive comorbidities after administration of the bispecific antibody, the method may further include art-recognized methods for alleviating the CRS event, such as administering to the subject an initial dose of an IL-6R antagonist (e.g., an anti-IL-6R antibody, e.g., tocilizumab (ACTEMRA® / RoACTEMRA®)) to manage the CRS event while discontinuing treatment with the bispecific antibody. Other anti-IL-6R antibodies that may be used in place of or in combination with tocilizumab include sarilumab, bovalilizumab (ALX-0061), satralizumab (SA-237), and variants thereof. In some examples, the method further includes administering to the subject an effective amount of a corticosteroid, such as methylprednisolone or dexamethasone.

[0174] In some examples, the dosing regimens of the invention result in a median progression-free survival (PFS) in a population of subjects of greater than about 1 month (e.g., at least about 1.5 months, at least about 2 months, at least about 2.5 months, at least about 3 months, at least about 3.5 months or more; e.g., from about 1 month to about 5 months, from about 1 month to about 4 months, from about 1 month to about 3 months, from about 1 month to about 2 months, from about 3 months to about 5 months, from about 2 months to about 4 months, from about 2 months to about 5 months, or from about 2 months to about 3 months; e.g., about 1 month, about 1.5 months, about 2 months, about 2.5 months, about 3 months, about 3.5 months, about 4 months, or more). In some examples, the dosing regimens of the invention provide a therapeutic effect for more than about 4 months (e.g., at least about 4.5 months, at least about 5 months, at least about 5.5 months, at least about 6 months, at least about 6.5 months, at least about 7 months, at least about 7.5 months, at least about 8 months, at least about 8.5 months, at least about 9.0 months, at least about 9.5 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 20 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, at least about 48 months, at least about 54 months, or more; e.g., from about 4 to about 60 months). months, about 8 to about 60 months, about 12 to about 60 months, about 24 to about 60 months, about 48 to about 60 months, about 4 to about 48 months, about 4 to about 24 months, about 4 to about 18 months, about 4 to about 12 months, about 4 to about 8 months, about 8 to about 24 months, about 8 to about 18 months, about 8 to about 12 months, about 4 to about 6 months, about 6 to about 8 months, about 6 to about 12 months, or about 6 to about 10 months; for example, about 4.5 months, about 5 months, about 5.5 months, about 6 months , about 6.5 months, about 7 months, about 7.5 months, about 8 months, about 8.5 months, about 9.0 months, about 9.5 months, about 10 months, about 11 months, about 12 months, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 20 months, about 24 months, about 30 months, about 36 months, about 42 months, about 48 months, about 54 months, or more) median progression-free survival (PFS) for a population of subjects.

[0175] In some examples, the dosing regimens of the invention provide a therapeutic effect of more than about 4 months (e.g., at least about 4.5 months, at least about 5 months, at least about 5.5 months, at least about 6 months, at least about 6.5 months, at least about 7 months, at least about 7.5 months, at least about 8 months, at least about 8.5 months, at least about 9.0 months, at least about 9.5 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 20 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, at least about 48 months, at least about 54 months, or more; e.g., from about 4 to about 60 months, from about 8 to about 9 months, 60 months, about 12 to about 60 months, about 24 to about 60 months, about 48 to about 60 months, about 4 to about 48 months, about 4 to about 24 months, about 4 to about 18 months, about 4 to about 12 months, about 4 to about 8 months, about 8 to about 24 months, about 8 to about 18 months, about 8 to about 12 months, about 4 to about 6 months, about 6 to about 8 months, about 6 to about 12 months, or about 6 to about 10 months; for example, about 4.5 months, about 5 months, about 5.5 months, about 6 months, about 6.5 months, about 7 months months, about 7.5 months, about 8 months, about 8.5 months, about 9.0 months, about 9.5 months, about 10 months, about 11 months, about 12 months, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 20 months, about 24 months, about 30 months, about 36 months, about 42 months, about 48 months, about 54 months, or more) in a population of subjects with FL (e.g., relapsed and / or refractory FL).

[0176] In some examples, the dosing regimens of the invention result in a median PFS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of greater than about 1 month (e.g., at least about 1.5 months, at least about 2 months, at least about 2.5 months, at least about 3 months, at least about 3.5 months or more; e.g., from about 1 month to about 5 months, from about 1 month to about 4 months, from about 1 month to about 3 months, from about 1 month to about 2 months, from about 3 months to about 5 months, from about 2 months to about 4 months, from about 2 months to about 5 months, or from about 2 months to about 3 months; e.g., about 1 month, about 1.5 months, about 2 months, about 2.5 months, about 3 months, about 3.5 months, about 4 months, or more).In some examples, the dosing regimens of the present invention provide a therapeutic effect of more than about 4 months (e.g., at least about 4.5 months, at least about 5 months, at least about 5.5 months, at least about 6 months, at least about 6.5 months, at least about 7 months, at least about 7.5 months, at least about 8 months, at least about 8.5 months, at least about 9.0 months, at least about 9.5 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 20 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, at least about 48 months, at least about 54 months, or more; e.g., from about 4 to about 60 months, from about 8 to about 60 months, months, about 12 to about 60 months, about 24 to about 60 months, about 48 to about 60 months, about 4 to about 48 months, about 4 to about 24 months, about 4 to about 18 months, about 4 to about 12 months, about 4 to about 8 months, about 8 to about 24 months, about 8 to about 18 months, about 8 to about 12 months, about 4 to about 6 months, about 6 to about 8 months, about 6 to about 12 months, or about 6 to about 10 months; for example, about 4.5 months, about 5 months, about 5.5 months, about 6 months, about 6.5 months, about 7 months, about The present invention provides a PFS of about 7.5 months, about 8 months, about 8.5 months, about 9.0 months, about 9.5 months, about 10 months, about 11 months, about 12 months, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 20 months, about 24 months, about 30 months, about 36 months, about 42 months, about 48 months, about 54 months, or more) in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL).

[0177] In some examples, the dosing regimens of the invention result in a median PFS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of greater than about 1 month (e.g., at least about 1.5 months, at least about 2 months, at least about 2.5 months, at least about 3 months, at least about 3.5 months or more; e.g., from about 1 month to about 5 months, from about 1 month to about 4 months, from about 1 month to about 3 months, from about 1 month to about 2 months, from about 3 months to about 5 months, from about 2 months to about 4 months, from about 2 months to about 5 months, or from about 2 months to about 3 months; e.g., about 1 month, about 1.5 months, about 2 months, about 2.5 months, about 3 months, about 3.5 months, about 4 months, or more). In some examples, the dosing regimens of the invention result in a median PFS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of at least 1 month. In some examples, the dosing regimens of the invention result in a median PFS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of at least 1.5 months. In some examples, the dosing regimens of the invention result in a median PFS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of at least 2 months. In some examples, the dosing regimens of the invention result in a median PFS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of at least 2.5 months. In some examples, the dosing regimens of the invention result in a median PFS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of at least 3 months.In some examples, the dosing regimens of the invention provide a sustained response of more than about 6.3 months (e.g., at least about 6.5 months, at least about 6.7 months, at least about 7 months, at least about 7.3 months, at least about 7.5 months, at least about 8 months, at least about 8.5 months, at least about 9.0 months, at least about 9.5 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 20 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, at least about 48 months, at least about 54 months, or more; e.g., Between about 6 months and about 48 months, between about 6 months and about 36 months, between about 6 months and about 24 months, between about 6 months and about 12 months, between about 6 months and about 10 months; between about 6 months and about 8 months, between about 8 months and about 24 months, between about 12 months and about 24 months, or between about 8 months and about 16 months; for example, about 6.3 months, about 6.5 months, about 7 months, about 7.5 months, about 8 months, about 8.5 months, about 9 months. In some examples, the dosing regimens of the invention result in a median PFS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of at least 6.7 months. In some examples, the dosing regimens of the invention result in a median PFS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of at least 7.3 months. In some examples, the dosing regimens of the present invention result in a median PFS in a population of subjects with DLBCL (eg, relapsed and / or refractory DLBCL) of at least 8.0 months.

[0178] In some examples, the dosing regimens of the present invention are for more than 9.5 months (e.g., at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 20 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, at least about 48 months, at least about 54 months, or more; e.g., between about 9 months and about 48 months, at least about 54 months, or more). between about 36 months, between about 9 months and about 24 months, between about 9 months and about 12 months, between about 10 months and about 18 months; between about 12 months and about 24 months, between about 18 months and about 36 months, between about 12 months and about 36 months, or between about 24 months and about 48 months; e.g., about 10 months, about 11 months, about 12 months, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 20 months, about 24 months, about 30 months, about 36 months, about 42 months, about 48 months, about 54 months, or more).

[0179] In some examples, the dosing regimens of the present invention are for a duration of more than 9.5 months (e.g., at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 20 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, at least about 48 months, at least about 54 months, or more; e.g., between about 9 months and about 48 months, between about 9 months and about 36 months, at least about 54 months, or more). between about 9 months and about 24 months, between about 9 months and about 12 months, between about 10 months and about 18 months; between about 12 months and about 24 months, between about 18 months and about 36 months, between about 12 months and about 36 months, or between about 24 months and about 48 months; e.g., about 10 months, about 11 months, about 12 months, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 20 months, about 24 months, about 30 months, about 36 months, about 42 months, about 48 months, about 54 months, or more) in a population of subjects with FL (e.g., recurrent and / or refractory FL).

[0180] In some examples, the dosing regimens of the present invention are for more than 9.5 months (e.g., at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 20 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, at least about 48 months, at least about 54 months, or more; e.g., between about 9 months and about 48 months, between about 9 months and about 36 months, between about 9 months and about 54 months, or more). between about 12 months and about 24 months, between about 9 months and about 12 months, between about 10 months and about 18 months; between about 12 months and about 24 months, between about 18 months and about 36 months, between about 12 months and about 36 months, or between about 24 months and about 48 months; e.g., about 10 months, about 11 months, about 12 months, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 20 months, about 24 months, about 30 months, about 36 months, about 42 months, about 48 months, about 54 months, or more) in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL).

[0181] In some examples, the dosing regimens of the present invention are for more than 12.5 months (e.g., at least about 13 months, at least about 14 months, at least about 14.6 months, at least about 15 months, at least about 15.8 months, at least about 16 months, at least about 17 months, at least about 17.3 months, at least about 18 months, at least about 20 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, at least about 48 months, at least about 54 months, or more; e.g., between about 13 months and about 48 months, at least about 13 months, The present invention provides a median OS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of about 36 months, about 13 months to about 24 months, about 16 months to about 60 months, about 24 months to about 36 months, about 12 months to about 24 months, about 18 months to about 36 months, about 24 months to about 36 months, or about 24 months to about 48 months; e.g., about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 20 months, about 24 months, about 30 months, about 36 months, about 42 months, about 48 months, about 54 months, or more. In some examples, the present dosing regimens provide a median OS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of greater than about 14.6 months. In some examples, the dosing regimens of the invention result in a median OS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of greater than about 15.8 months. In some examples, the dosing regimens of the invention result in a median OS in a population of subjects with DLBCL (e.g., relapsed and / or refractory DLBCL) of greater than about 17.3 months.

[0182] In some examples, the dosing regimens of the present invention provide a reduction in the incidence of HIV-1 infection in a population of subjects of at least about 10% (e.g., at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, or more; e.g., , about 10% to about 40%, about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 10% to about 30%, about 15% to about 30%, about 20% to about 40%, or more; for example, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 25%, about 30%, about 35%, about 40%, or more) of a complete response (CR). In some examples, the dosing regimens of the present invention result in a CR rate of at least about 42% (e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or more, e.g., about 42% to about 45%, about 45% to about 50%, about 50% to about 55%, about 55% to about 60%, about 60% to about 65%, about 65% to about 70%, about 70% to about 75%, or more, e.g., about 42%, about 45%, about 50%, about 55%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more) in a population of subjects.

[0183] In some examples, the dosing regimens of the invention result in a CR in a rate of at least about 20% (e.g., at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, or more; e.g., about 20% to about 50%, about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, about 20% to about 40%, about 30% to about 50%, or more; e.g., about 20%, about 25%, about 30%, about 35%, about 45%, about 50%, or more) in a population of subjects with FL (e.g., 1L FL, or relapsed and / or refractory FL). In some examples, the dosing regimens of the present invention result in a CR rate of at least about 55% (e.g., at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or more, e.g., about 55% to about 60%, about 60% to about 65%, about 65% to about 70%, about 70% to about 75%, or more, e.g., about 42%, about 45%, about 50%, about 55%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more) in a population of subjects with FL (e.g., 1L FL, or relapsed and / or refractory FL). In certain embodiments, the complete response rate in a population of subjects with R / R FL is about 45% to about 50%.

[0184] In some examples, the dosing regimens of the invention achieve a reduction in at least about 10% (e.g., at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, or more) of DLBCL in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). For example, about 10% to about 40%, about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 10% to about 30%, about 15% to about 30%, about 20% to about 40%, or more; for example, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 25%, about 30%, about 35%, about 40%, or more. In some examples, the dosing regimens of the present invention are used to treat patients with DLBCL (e.g., 1L In a population of DLBCL, or relapsed and / or refractory DLBCL, the compound results in a CR rate of at least about 42% (e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or more, e.g., about 42% to about 45%, about 45% to about 50%, about 50% to about 55%, about 55% to about 60%, about 60% to about 65%, about 65% to about 70%, about 70% to about 75%, or more, e.g., about 42%, about 45%, about 50%, about 55%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more).

[0185] In some embodiments, the subject population has relapsed or refractory FL and the objective response rate is at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%; e.g., 70%-80%, 70%-90%, 70%-95%, or 70%-100%; e.g., about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, or about 95%). In some embodiments, the objective response rate is at least 80%. In some embodiments, the subject population has relapsed or refractory FL and the objective response rate is between 70% and 90%. In some embodiments, the objective response rate is about 80%.

[0186] In some examples, the dosing regimens of the invention result in a CR rate of at least about 50% (e.g., at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or more, e.g., 50% to 55%, 55% to 60%, 60% to 65%, 65% to 70%, 70% to 75%, or more, e.g., about 42%, about 45%, about 50%, about 55%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more) in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). In some examples, the dosage regimens of the present invention result in a CR at a rate of at least about 15% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). In some examples, the dosage regimens of the present invention result in a CR at a rate of at least about 20% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). In some examples, the dosage regimens of the present invention result in a CR at a rate of at least about 25% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). In some examples, the dosage regimens of the present invention result in a CR at a rate of at least about 30% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). In some examples, the dosage regimens of the present invention result in a CR at a rate of at least about 35% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). In some examples, the dosing regimens of the invention result in a CR rate of at least about 40% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). In some examples, the dosing regimens of the invention result in a CR rate of at least about 45% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL).In some examples, the dosing regimens of the invention result in a CR at a rate of at least about 50% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). In some examples, the dosing regimens of the invention result in a CR at a rate of at least about 55% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). In some examples, the dosing regimens of the invention result in a CR at a rate of at least about 60% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL). In some examples, the dosing regimens of the invention result in a CR at a rate of at least about 65% in a population of subjects with DLBCL (e.g., 1L DLBCL, or relapsed and / or refractory DLBCL).

[0187] In some embodiments, the subject population has relapsed or refractory DLBCL or transformed FL and the objective response rate is at least 25% (e.g., at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more; e.g., between 25% and 95%, between 25% and 75%, between 25% and 55%, between 25% and 50%, between 25% and 45%, between 25% and 40%). between 25% and 35%, between 25% and 30%, between 30% and 75%, between 35% and 75%, between 40% and 75%, between 30% and 40%, between 30% and 45%, between 30% and 50%, or between 50% and 70%; for example, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. In some embodiments, the objective response rate is at least 35%. In some embodiments, the subject population has relapsed or refractory DLBCL and the objective response rate is between 25% and 45%. In some embodiments, the objective response rate is about 35%.

[0188] In some embodiments, the subject population has relapsed or refractory NHL and the objective response rate is at least 34% (e.g., at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more; e.g., between 34% and 95%, between 34% and 85%, between 34% and 75%, between 34% and 65%, between 34% and 55%, between 35% and 65%). The objective response rate may be between 0%, 35% and 75%, 55% and 95%, 75% and 95%, 40% and 50%, 45% and 64%, 34% and 45%, or 34% and 40%, e.g., about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. In some embodiments, the objective response rate is at least 44%. In some embodiments, the objective response rate is between 35% and 55%. In some embodiments, the objective response rate is about 45%.

[0189] B. Bispecific antibody that binds to CD20 and CD3 The present invention provides bispecific antibodies that bind CD20 and CD3 (i.e., anti-CD20 / anti-CD3 antibodies) that are useful for the treatment of CD20-positive cell proliferative disorders, such as B-cell proliferative disorders (e.g., non-Hodgkin's lymphoma (NHL) (e.g., previously untreated (1L) NHL, diffuse large B-cell lymphoma (DLBCL) (e.g., 1L DLBCL, relapsed and / or refractory DLBCL, or Richter's transformation), follicular lymphoma (FL) (e.g., 1L FL, relapsed and / or refractory FL, or transformed FL), mantle cell lymphoma (MCL), high-grade B-cell lymphoma, or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL)) or chronic lymphocytic leukemia (CLL).

[0190] In some examples, the present invention provides a bispecific antibody comprising an anti-CD20 arm having a first 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 GYTFTSYNMH (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of AIYPGNGDTSYNQKFKG (SEQ ID NO: 2); (c) HVR-H3 (SEQ ID NO: 3) comprising the amino acid sequence of VVYYSNSYWYFDV; (d) HVR-L1 comprising the amino acid sequence of RASSSVSYMH (SEQ ID NO: 4); (e) HVR-L2 (SEQ ID NO: 5) comprising the amino acid sequence of APSNLAS; and (f) HVR-L3 (SEQ ID NO: 6) comprising the amino acid sequence of QQWSFNPPT. In some examples, the anti-CD20 / anti-CD3 antibody comprises at least one (e.g., one, two, three, or four) of heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 17-20, respectively, and / or at least one (e.g., one, two, three, or four) of light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 21-24, respectively. In some examples, the bispecific antibody comprises: (a) a heavy chain variable (VH) domain having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 7, or the amino acid sequence of SEQ ID NO: 7; (b) a light chain variable (VL) domain having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 8, or the amino acid sequence of SEQ ID NO: 8; or (c) an anti-CD20 arm comprising a first binding domain comprising a VH domain as in (a) and a VL domain as in (b). Thus, in some examples, the first binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 7 and a VL domain comprising the amino acid sequence of SEQ ID NO: 8.

[0191] In some examples, the present invention provides a bispecific antibody comprising an anti-CD3 arm having a second binding domain comprising at least one, two, three, four, five, or six HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of NYYIH (SEQ ID NO: 9); (b) HVR-H2 comprising the amino acid sequence of WIYPGDGNTKYNEKFKG (SEQ ID NO: 10); (c) HVR-H3 comprising the amino acid sequence of DSYSNYYFDY (SEQ ID NO: 11); (d) HVR-L1 comprising the amino acid sequence of KSSQSLLNSRTRKNYLA (SEQ ID NO: 12); (e) HVR-L2 comprising the amino acid sequence of WASTRES (SEQ ID NO: 13); or (f) HVR-L3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 14). In some examples, the anti-CD20 / anti-CD3 antibody comprises at least one (e.g., one, two, three, or four) of heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 25-28, respectively, and / or at least one (e.g., one, two, three, or four) of light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 29-32, respectively. In some examples, the bispecific antibody comprises: (a) a VH domain having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 15, or the amino acid sequence of SEQ ID NO: 15; (b) a VL domain having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 16, or the amino acid sequence of SEQ ID NO: 16; or (c) an anti-CD3 arm comprising a second binding domain comprising a VH domain as in (a) and a VL domain as in (b). Thus, in some examples, the second binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 15 and a VL domain comprising the amino acid sequence of SEQ ID NO: 16.

[0192] In some examples, the present invention provides an anti-CD20 arm having a first binding domain comprising at least one, two, three, four, five, or six HVRs selected from: (1) HVR-H1 comprising the amino acid sequence of GYTFTSYNMH (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of AIYPGNGDTSYNQKFKG (SEQ ID NO: 2); (c) HVR-H3 (SEQ ID NO: 3) comprising the amino acid sequence of VVYYSNSYWYFDV; (d) HVR-L1 comprising the amino acid sequence of RASSSVSYMH (SEQ ID NO: 4); (e) HVR-L2 (SEQ ID NO: 5) comprising the amino acid sequence of APSNLAS; and (f) HVR-L3 (SEQ ID NO: 6) comprising the amino acid sequence of QQWSFNPPT; and (2) and an anti-CD3 arm having a second binding domain comprising at least one, two, three, four, five, or six HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of NYYIH (SEQ ID NO: 9); (b) HVR-H2 comprising the amino acid sequence of WIYPGDGNTKYNEKFKG (SEQ ID NO: 10); (c) HVR-H3 comprising the amino acid sequence of DSYSNYYFDY (SEQ ID NO: 11); (d) HVR-L1 comprising the amino acid sequence of KSSQSLLNSRTRKNYLA (SEQ ID NO: 12); (e) HVR-L2 comprising the amino acid sequence of WASTRES (SEQ ID NO: 13); and (f) HVR-L3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 14). In some examples, an anti-CD20 / anti-CD3 bispecific antibody comprises (1) at least one (e.g., 1, 2, 3, or 4) of heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 17-20, respectively, and / or at least one (e.g., 1, 2, 3, or 4) of light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 21-24, respectively, and (2) at least one (e.g., 1, 2, 3, or 4) of heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 25-28, respectively, and / or at least one (e.g., 1, 2, 3, or 4) of light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 29-32, respectively.In some examples, the anti-CD20 / anti-CD3 bispecific antibody comprises: (1) (a) a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 7, or the amino acid sequence of SEQ ID NO: 7; (b) a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 8, or the amino acid sequence of SEQ ID NO: 8; and (c) a first binding domain comprising a VH as in (a) and a VL as in (b). an anti-CD20 arm and (2) (a) a VH domain having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 15, or the amino acid sequence of SEQ ID NO: 15; (b) a VL domain having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 16, or the amino acid sequence of SEQ ID NO: 16; or (c) an anti-CD3 arm comprising a second binding domain comprising a VH as in (a) and a VL as in (b). In some examples, the anti-CD20 / anti-CD3 bispecific antibody comprises (1) a first binding domain comprising a VH domain comprising the amino acid sequence of SEQ ID NO:7 and a VL domain comprising the amino acid sequence of SEQ ID NO:8, and (2) a second binding domain comprising a VH domain comprising the amino acid sequence of SEQ ID NO:15 and a VL domain comprising the amino acid sequence of SEQ ID NO:16.

[0193] In some examples, the anti-CD20 / anti-CD3 bispecific antibody is mosunetuzumab, which has International Nonproprietary Name (INN) List 117 (WHO Drug Information, Vol. 31, No. 2, 2017, p. 303) or CAS Registry Number 1905409-39-3, and has (1) an anti-CD20 arm comprising the heavy chain and light chain sequences of SEQ ID NOs: 51 and 52, respectively, and (2) an anti-CD3 arm comprising the heavy chain and light chain sequences of SEQ ID NOs: 53 and 54, respectively. In some examples, an anti-CD20 / anti-CD3 bispecific antibody comprises: (1) (a) a heavy chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 51, or the amino acid sequence of SEQ ID NO: 51; (b) a light chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 52, or the amino acid sequence of SEQ ID NO: 52; (c) a first binding domain comprising a heavy chain as in (a) and a light chain as in (b). and (2) an anti-CD20 arm comprising: (a) a heavy chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 53, or the amino acid sequence of SEQ ID NO: 53; (b) a light chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 54, or the amino acid sequence of SEQ ID NO: 54; or (c) an anti-CD3 arm comprising a second binding domain comprising a heavy chain as in (a) and a light chain as in (b). In some examples, the anti-CD20 / anti-CD3 bispecific antibody comprises (1) an anti-CD20 arm comprising a first binding domain comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 51 and a light chain comprising the amino acid sequence of SEQ ID NO: 52, and (2) an anti-CD3 arm comprising a second binding domain comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 53 and a light chain comprising the amino acid sequence of SEQ ID NO: 54.

[0194] The amino acid sequence of mosunetuzumab is summarized in Table 2 below. [Table 2]

[0195] Anti-CD20 / anti-CD3 bispecific antibodies can be produced using recombinant methods and compositions, for example, as described in US Pat. No. 4,816,567.

[0196] In some examples, an anti-CD20 / anti-CD3 bispecific antibody according to any of the above embodiments may incorporate any of the features, alone or in combination, as described in Section C below.

[0197] C. Antibody Formats and Characteristics The methods described herein may further include any of the antibodies described above, where the antibody comprises any of the characteristics as described below, either alone or in combination.

[0198] 1. Antibody affinity In particular examples, the anti-CD20 / anti-CD3 bispecific antibody has a concentration 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~10 -13 M, e.g. 10 -9 M~10 -13 Dissociation constant (K D )

[0199] In one example, K D is measured by a radiolabeled antigen binding assay (RIA). In one example, an RIA is performed using an antibody of interest and a Fab version of its antigen. For example, the solution binding affinity of the Fab for the antigen is determined by measuring the binding affinity of the Fab to the antigen at the lowest concentration ( 125I) Fab is equilibrated with labeled antigen, followed by capturing the bound antigen on a plate coated with an anti-Fab antibody (see, e.g., Chen et al., J. Mol. Biol. 293:865-881 (1999)). To establish assay conditions, MICROTITER® multiwell plates (Thermo Scientific) are coated overnight with 5 μg / mL of capture anti-Fab antibody (Cappel Labs) in 50 mM sodium carbonate (pH 9.6), followed by blocking with 2% (w / v) bovine serum albumin in PBS for 2-5 hours at room temperature (approximately 23°C). In non-adsorbent plates (Nunc #269620), 100 pM or 26 pM [ 125 [I]-antigen is mixed with serial dilutions of the Fab of interest (e.g., consistent with the evaluation of the anti-VEGF antibody Fab-12 in Presta et al., Cancer Res. 57:4593-4599 (1997)). The Fab of interest is then incubated overnight; however, incubation may be continued for a longer period (e.g., approximately 65 hours) to reach equilibrium. The mixture is then transferred to a capture plate for incubation at room temperature (e.g., 1 hour). The solution is then removed, and the plate is washed eight times with 0.1% polysorbate 20 (TWEEN-20®) in PBS. When the plate has dried, 150 μL / well of scintillant (MICROSCINT-20™; Packard) is added, and the plate is counted for 10 minutes in a TOPCOUNT™ gamma counter (Packard). The concentration of each Fab that results in 20% or less of maximum binding is selected for use in the competitive binding assay.

[0200] According to another example, K Dis measured using a BIACORE® surface plasmon resonance assay. For example, assays using a BIACORE®-2000 or BIACORE®-3000 (BIACORE®, Inc., Piscataway, NJ) are performed at 25°C using an immobilized antigen CM5 chip at approximately 10 response units (RU). In one example, a carboxymethylated dextran biosensor chip (CM5, BIACORE®, Inc.) is activated with N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the supplier's instructions. The antigen is diluted to 5 μg / ml (approximately 0.2 μM) with 10 mM sodium acetate, pH 4.8, and then injected at a flow rate of 5 μl / min, achieving approximately 10 response units (RU) of coupled protein. After antigen injection, 1 M ethanolamine is injected to block unreacted groups. For kinetic measurements, two-fold serial dilutions of Fab (0.78 nM to 500 nM) are injected in PBS containing 0.05% polysorbate 20 (TWEEN-20®) surfactant (PBST) at 25°C and a flow rate of approximately 25 μl / min. The association rate (k on ) and dissociation rate (k off ) is calculated by simultaneously fitting the association and dissociation sensorgrams using a simple one-to-one Langmuir binding model (BIACORE® Evaluation Software version 3.2). D ) is k off / k on See, for example, Chen et al., J. Mol. Biol. 293:865-881 (1999). When the on-rate by the surface plasmon resonance assay is 10 6 M- 1 s- 1If the on rate exceeds , the on rate can be determined using a fluorescence quenching technique that measures the increase or decrease in fluorescence emission intensity (excitation = 295 nm, emission = 340 nm, 16 nm bandpass) of 20 nM anti-antigen antibody (Fab form) in PBS (pH 7.2) at 25°C in the presence of increasing antigen concentrations as measured in a spectrometer such as a spectrophotometer equipped with stopped flow (Aviv Instruments) or an 8000 Series SLM-AMINCO™ spectrophotometer (ThermoSpectronic) equipped with a stirred cuvette.

[0201] 2. Antibody fragment In certain examples, the anti-CD20 / anti-CD3 bispecific antibodies provided herein are antibody fragments. Antibody fragments include, but are not limited to, Fab fragments, Fab' fragments, Fab'-SH fragments, F(ab')2 fragments, Fv fragments, and scFv fragments, 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 reference to scFv fragments, see, e.g., Pluckthuen, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York), pp. 269-315 (1994); see also WO 93 / 16185; and U.S. Patent Nos. 5,571,894 and 5,587,458. See US Pat. No. 5,869,046 for a discussion of Fab and F(ab')2 fragments that comprise salvage receptor binding epitope residues and have increased in vivo half-lives.

[0202] Diabodies are antibody fragments with two antigen-binding sites that can be bivalent or bispecific. See, e.g., EP 404,097, WO 1993 / 01161, Hudson et al. Nat. Med. 9:129-134 (2003); and Hollinger et al. Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al. Nat. Med. 9:129-134 (2003).

[0203] Single domain antibodies are antibody fragments that contain all or part of the heavy chain variable domain or all or part of the light chain variable domain of an antibody. In particular examples, single domain antibodies are human single domain antibodies (Domantis, Inc., Waltham, Massachusetts; see, e.g., U.S. Patent No. 6,248,516 B1).

[0204] Antibody fragments can be produced by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies and production by recombinant host cells (e.g., E. coli or phage), as described herein.

[0205] 3. Chimeric and humanized antibodies In certain examples, the anti-CD20 / anti-CD3 bispecific antibodies provided herein are chimeric antibodies. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567; and Morrison et al. Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984). In one example, a chimeric antibody comprises 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 a further example, a chimeric antibody is a "class-switched" antibody in which the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0206] In particular examples, a chimeric antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. Generally, a humanized antibody comprises one or more variable domains in which the HVRs, e.g., CDRs (or portions thereof), are derived from a non-human antibody and the FRs (or portions thereof) are derived from a human antibody sequence. Optionally, the humanized antibody also comprises at least a portion of a human constant region. In some examples, some FR residues in a humanized antibody are substituted with corresponding residues from a non-human antibody (e.g., the antibody from which the HVR residues are derived), e.g., to restore or improve antibody specificity or affinity.

[0207] Humanized antibodies and methods for their production are reviewed by Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and further described in Riechmann et al., Nature 332:323-329 (1988); Queen et al., Proc. Natl. Acad. Sci. USA 86:10029-10033 (1989); U.S. Patent Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005)) (describing specificity-determining region (SDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (describing resurfacing); Dall'Acqua et al., Methods 36:43-60 (2005) (describing "FR shuffling"); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing a "guided selection" approach to FR shuffling).

[0208] Human framework regions that can be used for humanization include, but are not limited to, framework regions selected using the "best-fit" method (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from consensus sequences of human antibodies of particular subgroups of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol., 151:2623 (1993)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and framework regions derived from screening of FR libraries (see, e.g., Baca et al. al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996).

[0209] 4. Human antibodies In a particular example, the anti-CD20 / anti-CD3 bispecific antibody is a human antibody. Human antibodies can be produced using various techniques known in the art. Human antibodies are generally described in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5:368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20:450-459 (2008).

[0210] Human antibodies may be prepared by administering an immunogen to transgenic animals that have been engineered 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 human immunoglobulin loci that replace endogenous immunoglobulin loci, or that are present extrachromosomally or randomly integrated into the animal's chromosomes. In such transgenic mice, the endogenous immunoglobulin loci are generally inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125 (2005). See also, e.g., U.S. Patent Nos. 6,075,181 and 6,150,584, which describe XENOMOUSE™ technology; U.S. Patent No. 5,770,429, which describes HuMab® technology; U.S. Patent No. 7,041,870, which describes KM MOUSE® technology; and U.S. Patent Application Publication No. 2007 / 0061900, which describes VelociMouse® technology. The human variable regions from intact antibodies produced by such animals may be further modified, for example, by combining with different human constant regions.

[0211] Human antibodies can also be produced by hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for producing human monoclonal antibodies have been described. (See, e.g., Kozbor J. Immunol., 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147:86 (1991)). Human antibodies generated via human B cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006). Additional methods include, for example, U.S. Patent No. 7,189,826 (which describes the production of monoclonal human IgM antibodies from hybridoma cell lines), and Ni, Xiandai Mianyixue, 26(4):265-268 (2006), which describes human-human hybridomas. Human hybridoma technology (trioma technology) is also described in 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).

[0212] Human antibodies can also be generated by isolating Fv clone variable domain sequences selected from human-derived phage display libraries. These variable domain sequences can then be combined with desired human constant domains. Techniques for selecting human antibodies from antibody libraries are described below.

[0213] 5. Library-derived antibodies Anti-CD20 / anti-CD3 bispecific antibodies of the invention can be isolated by screening combinatorial libraries for antibodies with the desired activity(ies). For example, various methods are known in the art for generating phage display libraries and screening such libraries for antibodies with the desired binding characteristics. Such methods are reviewed, for example, by Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, 2001), and by 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. 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).

[0214] In a specific phage display method, repertoires of VH and VL genes are cloned separately by polymerase chain reaction (PCR) and randomly recombined in phage libraries, which can then be screened for antigen-binding phage as described by Winter et al., Ann. Rev. Immunol., 12:433-455 (1994). Phage typically display antibody fragments as either single-chain Fv (scFv) fragments or Fab fragments. Libraries from immune sources provide high-affinity antibodies against immunogens without the need for hybridoma construction. 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 also self antigens, as described by Griffiths et al., EMBO J., 12:725-734 (1993). Finally, naive libraries can also be generated synthetically by cloning unrearranged V gene segments from stem cells and using PCR primers containing random sequences to encode highly variable CDR3 regions to achieve rearrangement in vitro, as described in Hoogenboom and Winter, J. Mol. Biol., 227:381-388 (1992). Patent publications describing human antibody phage libraries include, for example: U.S. Pat. No. 5,750,373, and U.S. Patent Application Publication Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360.

[0215] Anti-CD20 / anti-CD3 bispecific antibodies or antibody fragments isolated from a human antibody library are considered human antibodies or human antibody fragments herein.

[0216] 6. Antibody Variants In certain examples, amino acid sequence variants of the anti-CD20 / anti-CD3 bispecific antibodies of the present invention are contemplated. As described in detail herein, anti-TIGIT antagonist antibodies, PD-1 axis-binding antagonist antibodies (e.g., anti-PD-L1 antagonist antibodies), and / or anti-VEGF antibodies can be optimized based on desired structural and functional properties. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody. Amino acid sequence variants of antibodies can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions from, and / or insertions into, and / or substitutions of, residues within the amino acid sequence of the antibody. Any combination of deletions, insertions, and substitutions can be made to arrive at the final construct, provided that the final construct possesses the desired characteristics, e.g., antigen binding.

[0217] Substitution, insertion, and deletion variants In particular examples, anti-CD20 / anti-CD3 bispecific antibody variants with one or more amino acid substitutions are provided. Sites of interest for substitutional mutagenesis include HVRs and FRs. Conservative substitutions are shown in Table 3 under the heading of "Preferred Substitutions." More substantial changes are provided in Table 3 under the heading of "Exemplary Substitutions," and are as further described below with reference to amino acid side chain classes. Amino acid substitutions can be introduced into the antibody of interest, and the products can be screened for a desired activity, such as retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC. [Table 3]

[0218] Amino acids can be classified according to general side chain properties. (1) Hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile; (2) Neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) Acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) Residues affecting chain orientation: Gly, Pro; (6) Aromatic: Trp, Tyr, Phe.

[0219] Non-conservative substitutions involve exchanging a member of one of these classes for another class.

[0220] Certain substitutional variants involve substituting one or more hypervariable region residues of a parent antibody (e.g., a humanized or human antibody). Generally, the resulting variant(s) selected for further study will have a modification (e.g., an improvement) in a particular biological property (e.g., increased affinity, decreased immunogenicity) compared to the parent antibody and / or will substantially retain a particular biological property of the parent antibody. An exemplary substitutional variant is an affinity-matured antibody, which may be conveniently generated using, for example, phage-display-based affinity maturation techniques as described herein. Briefly, one or more HVR residues are mutated, and the variant antibodies are displayed on phage and screened for a particular biological activity (e.g., binding affinity).

[0221] Alterations (e.g., substitutions) may be made, for example, in HVRs to improve antibody affinity. Such alterations may be made within HVR "hot spots," i.e., residues encoded by codons that undergo frequent mutation during the somatic maturation process (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and / or residues that contact the antigen, and the resulting variant VH or VL are tested for binding affinity. Affinity maturation by construction of and reselection from secondary libraries is described, for example, in Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, (2001)). In some examples of affinity maturation, diversity is introduced into the variable genes selected for maturation by any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide-directed mutagenesis). A secondary library is then generated. This library is then screened to identify any antibody variants with the desired affinity. Another method for introducing diversity involves an HVR-directed approach, in which several HVR residues (e.g., 4-6 residues at a time) are randomized. HVR residues involved in antigen binding can be specifically identified, for example, using alanine scanning mutagenesis or modeling. CDR-H3 and CDR-L3 are often targeted in particular.

[0222] In certain examples, substitutions, insertions, or deletions may be made within one or more HVRs, so long as such changes do not substantially reduce the ability of the antibody to bind to the antigen. For example, conservative changes (e.g., conservative substitutions provided herein) that do not substantially reduce binding affinity may be made in the HVRs. Such changes may, for example, be outside of the antigen-contact residues within the HVRs. In certain examples of the variant VH and VL sequences described above, each HVR is either unaltered or has no more than one, two, or three amino acid substitutions.

[0223] A useful method for identifying antibody residues or regions that can be targeted for mutagenesis is called "alanine scanning mutagenesis," as described by Cunningham and Wells (1989) Science, 244:1081-1085. In this method, a residue or group of target residues (e.g., charged residues, e.g., Arg, Asp, His, Lys, and Glu) is identified and replaced with neutral or negatively charged amino acids (e.g., alanine or polyalanine) to determine whether the interaction of the antibody with the antigen is affected. Further substitutions may be introduced at amino acid positions that demonstrate functional sensitivity to the initial substitution. Alternatively, or in addition, a crystal structure of the antigen-antibody complex may be used to identify contact points between the antibody and antigen. Such contact and adjacent residues may be targeted or eliminated as candidates for substitution. Variants may be screened to determine whether they possess desired properties.

[0224] Amino acid sequence insertions include amino- and / or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing 100 or more residues, as well as intrasequence insertions of single or multiple amino acid residues. An example of a terminal insertion includes an antibody with an N-terminal methionyl residue. Other insertional variants of antibody molecules include the fusion to the N- or C-terminus of the antibody to an enzyme (e.g., for ADEPT) or a polypeptide which increases the serum half-life of the antibody.

[0225] B glycosylation variants In certain instances, anti-CD20 / anti-CD3 bispecific antibodies of the invention can be altered to increase or decrease the extent to which the antibody is glycosylated. Addition or deletion of glycosylation sites to an anti-CD20 / anti-CD3 bispecific antibody of the invention can be conveniently accomplished by altering the amino acid sequence to create or remove one or more glycosylation sites.

[0226] If the antibody contains an Fc region, the carbohydrate attached thereto can be altered. Natural antibodies produced by mammalian cells typically contain branched, biantennary oligosaccharides that are commonly attached to Asn297 in the CH2 domain of the Fc region via an N-linkage. See, e.g., Wright et al., TIBTECH 15:26-32 (1997). The oligosaccharides can include various carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose attached to the GlcNAc in the "stem" of the biantennary oligosaccharide structure. In some instances, oligosaccharide modifications in the antibodies of the present invention are performed to generate antibody variants with improved specific properties.

[0227] In one example, an anti-CD20 / anti-CD3 bispecific antibody variant is provided having a carbohydrate structure attached (directly or indirectly) to the Fc region that lacks fucose. For example, the amount of fucose in such an antibody can be 1% to 80%, 1% to 65%, 5% to 65%, or 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose in the glycan at Asn297 relative to the sum of all glycan structures attached to Asn297 (e.g., complex, hybrid, and high-mannose structures) as measured by MALDI-TOF mass spectrometry, for example, as described in WO 2008 / 077546. Asn297 refers to the asparagine residue located at approximately position 297 of the Fc region (EU numbering of Fc region residues); however, Asn297 may also be located upstream or downstream of position 297, i.e., approximately ±3 amino acids between positions 294 and 300, due to minor sequence variations in the antibody. Such fucosylation variants may have improved ADCC function. See, for example, U.S. Patent Application Publication Nos. 2003 / 0157108 (Presta, L.); 2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd.). Examples of publications relating to "defucosylated" or "fucose-deficient" antibody variants include: U.S. Patent Application Publication Nos. 2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; U.S. Patent Application Publication Nos. 2003 / 0115614; 2002 / 0164328; 2004 / 0093621; and 2004 / 0093622. 132140; 2004 / 0110704; 2004 / 0110282; 2004 / 0109865; WO 2003 / 085119; 2003 / 084570; 2005 / 035586; 2005 / 035778; 2005 / 053742; 2002 / 031140; Okazaki et al. J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004).Examples of cell lines capable of producing defucosylated antibodies include Lec13 CHO cells, which are deficient in protein fucosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); U.S. Patent Application Publication No. 2003 / 0157108 A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., especially Example 11), and knockout cell lines, such as α-1,6-fucosyltransferase gene, FUT8, knockout CHO cells (see, e.g., Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO 2003 / 085107).

[0228] In view of the above, in some examples, the methods of the invention comprise administering to a subject a variant of an anti-CD20 / anti-CD3 bispecific antibody having a mutation in an aglycosylation site in accordance with a split-dose escalation regimen. In some examples, the aglycosylation site mutation reduces the effector function of the antibody. In some examples, the aglycosylation site mutation is a substitution mutation. In some examples, the antibody comprises a substitution mutation in the Fc region that reduces effector function. In some examples, the substitution mutation is at amino acid residues N297, L234, L235, and / or D265 (EU numbering). In some examples, the substitution mutation is selected from the group consisting of N297G, N297A, L234A, L235A, D265A, and P329G (EU numbering). In some examples, the substitution mutation is at amino acid residue N297 (EU numbering). In a preferred example, the substitution mutation is N297A (EU numbering). In some embodiments, the anti-CD20 arm of the anti-CD20 / anti-CD3 bispecific antibody further comprises substitution mutations of T366W and N297G (EU numbering). In some embodiments, the anti-CD3 arm of the anti-CD20 / anti-CD3 bispecific antibody further comprises substitution mutations of T366S, L368A, Y407V, and N297G (EU numbering). In some embodiments, (a) the anti-CD20 arm further comprises substitution mutations of T366W and N297G, and (b) the anti-CD3 arm further comprises substitution mutations of T366S, L368A, Y407V, and N297G (EU numbering).

[0229] Further provided are anti-CD20 / anti-CD3 bispecific antibody variants having bisected oligosaccharides, e.g., biantennary oligosaccharides attached to the Fc region of the antibody are bisected by GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described, for example, in WO 2003 / 011878 (Jean-Mairet et al.); U.S. Pat. No. 6,602,684 (Umana et al.); and U.S. Patent Application Publication No. 2005 / 0123546 (Umana et al.). Antibody variants having at least one galactose residue in the oligosaccharide attached to the Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described, for example, in WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.).

[0230] c. Fc region variants In a particular example, one or more amino acid modifications are introduced into the Fc region of an anti-CD20 / anti-CD3 bispecific antibody of the invention, thereby creating an Fc region variant (see, e.g., U.S. Patent Application Publication No. 2012 / 0251531). The Fc region variant may comprise a human Fc region sequence (e.g., a human IgG1, IgG2, IgG3, or IgG4 Fc region) comprising an amino acid modification (e.g., a substitution) at one or more amino acid positions.

[0231] In certain instances, the present invention contemplates anti-CD20 / anti-CD3 bispecific antibody variants that retain some, but not all, effector functions, making them desirable candidates for applications in which in vivo antibody half-life is important, but certain effector functions (such as complement and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity assays can be performed to confirm reduced / absent CDC and / or ADCC activity. For example, Fc receptor (FcR) binding assays can be performed to confirm that the antibody lacks FcγR binding (and thus likely lacks ADCC activity) but retains FcRn binding ability. NK cells, the primary cells for mediating ADCC, express FcRIII only, whereas monocytes express FcR1, FcR1I, and FcR1I. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991). Non-limiting examples of in vitro assays to assess ADCC activity of a molecule of interest include those described in U.S. Pat. No. 5,500,362 (see, e.g., Hellstrom, I. et al., Proc. Natl. Acad. Sci. USA 83:7059-7063 (1986)) and Hellstrom, I. et al., Proc. Natl. Acad. Sci. USA 82:1499-1502 (1985); 5,821,337 (see Bruggemann, M. et al., Proc. Natl. Acad. Sci. USA 82:1499-1502 (1985)). al., J. Exp. Med. 166:1351-1361 (1987). Alternatively, non-radioactive assay methods may be used (see, e.g., ACTI™ Non-Radioactive Cytotoxicity Assay for Flow Cytometry (CellTechnology, Inc., Mountain View, CA; and CYTOTOX 96® Non-Radioactive Cytotoxicity Assay (PROMEGA®, Madison, WI)). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells.Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al. Proc. Natl Acad. Sci. USA 95:652-656 (1998). C1q binding assays may also be performed to confirm that the antibody is unable to bind C1q and lacks CDC activity. See, e.g., C1q and C3c binding ELISAs in WO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, a CDC assay may be performed (see, e.g., Gazzano-Santoro et al. J. Immunol. Methods 202:163 (1996); Cragg, MS et al. Blood. 101:1045-1052 (2003); and Cragg, MS and MJ Glennie Blood. 103:2738-2743 (2004)). Determination of FcRn binding and in vivo clearance / half-life can also be performed using methods known in the art (see, e.g., Petkova, SB et al. Int'l. Immunol. 18(12):1759-1769 (2006)).

[0232] Antibodies with reduced effector function include antibodies with substitutions at one or more of Fc region residues 238, 265, 269, 270, 297, 327, and 329 (U.S. Patent Nos. 6,737,056 and 8,219,149). Such Fc mutants include Fc mutants with substitutions at two or more of amino acid positions 265, 269, 270, 297, and 327, including the so-called "DANA" Fc mutant, in which residues 265 and 297 are substituted with alanine (U.S. Patent Nos. 7,332,581 and 8,219,149).

[0233] In certain examples, the proline at position 329 of the wild-type human Fc region in the antibody is substituted with an amino acid residue large enough to disrupt the proline sandwich within the Fc / Fcγ receptor interface formed between proline 329 of the Fc and tryptophan residues Trp87 and Trp110 of FcγRIII (Sondermann et al.: Nature 406, 267-273 (20 Jul. 2000)), or with glycine or arginine. In certain examples, the antibody further comprises at least one amino acid substitution. In one example, the additional amino acid substitution is S228P, E233P, L234A, L235A, L235E, N297A, N297D, or P331S; in yet another example, the at least one additional amino acid substitution is L234A and L235A in a human IgG1 Fc region, or S228P and L235E in a human IgG4 Fc region (see, e.g., U.S. Patent Application Publication No. 2012 / 0251531); and in yet another example, the at least one additional amino acid substitution is L234A, L235A, and P329G in a human IgG1 Fc region.

[0234] Certain antibody variants have been described with improved or diminished binding to FcRs (see, e.g., U.S. Pat. No. 6,737,056; WO 2004 / 056312; and Shields et al., J. Biol. Chem. 9(2):6591-6604 (2001)).

[0235] In particular examples, the antibody variant comprises an Fc region with one or more amino acid substitutions that improve ADCC, for example, substitutions at positions 298, 333, and / or 334 (EU numbering of residues) of the Fc region.

[0236] In some examples, alterations are made in the Fc region that result in altered (i.e., either improved or decreased) C1q binding and / or complement dependent cytotoxicity (CDC), e.g., as described in U.S. Pat. No. 6,194,551, WO 99 / 51642, and Idusogie et al. J. Immunol. 164:4178-4184 (2000).

[0237] Antibodies with increased half-lives and improved binding to the neonatal Fc receptor (FcRn), which is responsible for transferring maternal IgG to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)), are described in U.S. Patent Application Publication No. 2005 / 0014934 A1 (Hinton et al.). These antibodies comprise an Fc region with one or more substitutions therein that improve binding of the Fc region to FcRn. Such Fc variants include variants having substitutions at one or more of the following Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, ​​413, 424, or 434, e.g., a substitution at Fc region residue 434 (U.S. Patent No. 7,371,826).

[0238] For other examples of Fc region variants, see also Duncan & Winter, Nature 322:738-40 (1988); U.S. Patent No. 5,648,260; U.S. Patent No. 5,624,821; and WO 94 / 29351.

[0239] In some embodiments, the anti-CD20 / anti-CD3 bispecific antibody comprises an Fc region comprising an N297G mutation (EU numbering).

[0240] In some examples, the anti-CD20 / anti-CD3 bispecific antibody comprises one or more heavy chain constant domains, wherein the one or more heavy chain constant domains are selected from a first CH1 (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 examples, at least one of the one or more heavy chain constant domains is paired with another heavy chain constant domain. In some examples, the CH31 domain and the CH32 domain each comprise a protrusion or cavity, and the protrusion or cavity in the CH31 domain can be positioned within the cavity or protrusion in the CH32 domain, respectively. In some examples, the CH31 domain and the CH32 domain associate at the interface between the protrusion and the cavity. In some examples, the CH21 domain and the CH22 domain form a protrusion or a cavity, respectively, and the protrusion or cavity in the CH21 domain is positionable in the cavity or protrusion in the CH22 domain, respectively. In other examples, the CH21 domain and the CH22 domain contact at the interface between the protrusion and the cavity. In some examples, the anti-CD20 / anti-CD3 bispecific antibody is an IgG1 antibody.

[0241] d. Cysteine ​​Engineered Antibody Variants In certain instances, it may be desirable to generate cysteine-engineered anti-CD20 / anti-CD3 bispecific antibodies, e.g., "thioMAbs," in which one or more residues of an antibody are replaced with cysteine ​​residues. In certain instances, the substituted residues occur at accessible sites on the antibody. By replacing these residues with cysteine, reactive thiol groups are thereby placed at accessible sites on the antibody, which may be used to conjugate the antibody to other moieties, such as drug moieties or linker-drug moieties, to generate immunoconjugates, as further described herein. In certain instances, any one or more of the following residues are substituted with cysteine: V205 (Kabat numbering) of the light chain, A118 (EU numbering) of the heavy chain, and S400 (EU numbering) of the heavy chain Fc region. Cysteine ​​engineered antibodies can be generated, for example, as described in U.S. Pat. No. 7,521,541.

[0242] e. Antibody derivative In certain examples, the anti-CD20 / anti-CD3 bispecific antibodies provided herein are further modified to contain additional nonproteinaceous moieties that are known in the art and readily available. Suitable sites for derivatization of antibodies include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include 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, propropylene glycol homopolymer, polypropylene oxide / ethylene oxide copolymer, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may be advantageous during manufacturing due to its stability in water. The polymers may be of any molecular weight and may be branched or unbranched. The number of polymers attached to the antibody may vary, and when multiple polymers are attached, they may be the same or different molecules. In general, the number and / or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular property or function of the antibody to be improved, whether the antibody derivative will be used therapeutically under defined conditions, etc.

[0243] In another example, a conjugate of an antibody and a non-proteinaceous moiety is provided that can be selectively heated by exposure to radiation. In one example, the non-proteinaceous moiety is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102:11600-11605 (2005)). The radiation can be of any wavelength, including, but not limited to, wavelengths that are not harmful to normal cells but heat the antibody non-protective moiety to a temperature that kills cells proximal to the non-protective moiety.

[0244] 7. Recombinant Production Methods The anti-CD20 / anti-CD3 bispecific antibodies of the invention can be produced using recombinant methods and compositions such as those described in, for example, US Pat. No. 4,816,567, which is incorporated herein by reference in its entirety.

[0245] For recombinant production of an anti-CD20 / anti-CD3 bispecific antibody, nucleic acid encoding the anti-antibody is isolated and inserted into one or more vectors for further cloning and / or expression in host cells. Such nucleic acid can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes capable of binding specifically to genes encoding the antibody heavy and light chains).

[0246] Suitable host cells for cloning or expressing antibody-encoding vectors include prokaryotic or eukaryotic cells as described herein. For example, antibodies may be produced in bacteria, particularly if glycosylation and Fc effector functions are not required. For expression of antibody fragments and polypeptides in bacteria, see, e.g., U.S. Patent Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 245-254, which describes the expression of antibody fragments in E. coli.) Following expression, antibodies of the invention may be isolated in a soluble fraction from the bacterial cell paste or further purified.

[0247] In addition to prokaryotes, eukaryotic organisms such as filamentous fungi and yeast are suitable cloning or expression hosts for antibody-encoding vectors, including bacterial and yeast strains that have been "humanized" in their glycosylation pathways to produce antibodies with partially or fully human glycosylation patterns. See Gerngross, Nat. Biotech. 22:1409-1414 (2004) and Li et al., Nat. Biotech. 24:210-215 (2006).

[0248] Suitable host cells for expressing glycosylated antibodies are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant cells and insect cells. Numerous baculovirus strains have been identified that can be used in conjunction with insect cells, particularly for transfection of Spodoptera frugiperda cells.

[0249] Plant cell cultures can also be used as hosts. See, e.g., U.S. Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (PLANTIBODIES™ technology for producing antibodies in transgenic plants).

[0250] Vertebrate cells may also be used as hosts. For example, mammalian cell lines that are adapted to grow in suspension may be useful. Other examples of useful mammalian host cell lines include SV40 (COS-7) transformed monkey kidney CV1 cells, human embryonic kidney cells (e.g., 293 cells or 293 cells as described in Graham et al., J. Gen Virol. 36:59 (1977)), baby hamster kidney cells (BHK), mouse Sertoli cells (e.g., TM4 cells as described in Mather, Biol. Reprod. 23:243-251 (1980)), monkey kidney cells (CV1), African green monkey kidney cells (VERO-76), human cervical carcinoma cells (HELA), canine kidney cells (MDCK), buffalo rat liver cells (BRL 3A), human lung cells (W138), human liver cells (Hep G2), mouse mammary tumor cells (MMT 060562), TRI cells (e.g., Mather et al., Annals NY Acad. Sci. 383:44-68 (1982); MRC5 cells; and FS4 cells. Other useful mammalian host cell lines include DHFR - Examples of suitable host cell lines for antibody production include Chinese hamster ovary (CHO) cells, including CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)), and myeloma cell lines such as Y0, NS0, and Sp2 / 0. For reviews of specific mammalian host cell lines suitable for antibody production, see, for example, Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ), pp. 255-268 (2003);

[0251] 8. Immunoconjugates The present invention also provides immunoconjugates comprising an anti-CD20 / anti-CD3 bispecific antibody of the invention conjugated to one or more cytotoxic agents, e.g., chemotherapeutic agents or drugs, growth inhibitory agents, toxins (e.g., protein toxins, enzymatically active toxins of bacterial, fungal, plant, or animal origin, or fragments thereof), or radioactive isotopes.

[0252] In some examples, the immunoconjugate is an antibody drug conjugate (ADC), in which the antibody is coupled to an auristatin, such as, but not limited to, maytansinoids (see U.S. Pat. Nos. 5,208,020, 5,416,064, and EP 0425235 B1); monomethyl auristatin drug moieties DE and DF (MMAE and MMAF) (see U.S. Pat. Nos. 5,635,645, ... 483, 5,780,588, and 7,498,298); dolastatins; calicheamicin or its derivatives (see U.S. Patent Nos. 5,712,374, 5,714,586, 5,739,116, 5,767,285, 5,770,701, 5,770,710, 5,773,001, and 5,877,296; Hinman et al., Cancer Res. 53:3336-3342 (1993); and Lode et al., Cancer Res. 58:2925-2928 (1998)); anthracyclines such as daunomycin and doxorubicin (see, e.g., Kratz et al., Current Med. Chem. 13:477-523 (2006); Jeffrey et al., Bioorganic & Med. Chem. Letters 16:358-362 (2006); Torgov et al., Bioconj. Chem. 16:717-721 (2005); Nagy et al., Proc. Natl. Acad. Sci. USA 97:829-834 (2000); Dubowchik et al., Bioorg. & Med. Chem. Letters 12:1529-1532 (2002); King et al., J. Med. Chem. 45:4336-4343 (2002); and U.S. Patent No. 6,630,579); methotrexate; vindesine; taxanes such as docetaxel, paclitaxel, larotaxel, tesetaxel, and ortataxel; trichothecenes; and CC1065.

[0253] In another example, the immunoconjugate comprises an anti-CD20 / anti-CD3 bispecific antibody conjugated to an enzymatically active toxin or fragment thereof, including, but not limited to, diphtheria A chain, a nonbinding active fragment of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii protein, diansin protein, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), Momordica charantia inhibitor, curcin, crotin, Sapaonaria officinalis inhibitor, gelonin, mitogenin, restrictocin, phenomycin, enomycin, and a trichothecene.

[0254] In another example, the immunoconjugate comprises an anti-CD20 / anti-CD3 bispecific antibody conjugated to a radioactive atom to form a radioconjugate. A variety of radioisotopes are available for the production of radioconjugates. Examples include: 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 are included. When radioactive substances are used for detection, it is possible to use radioactive atoms for scintigraphy studies, e.g. 99m Tc or 123 I, or spin labels for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, MRI), such as again iodine-123, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese, or iron.

[0255] Conjugates of antibodies and cytotoxic agents can be prepared using, for example, various bifunctional protein coupling agents, such as N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCl), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bisazide compounds (such as bis(p-azidobenzoyl)hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and biactive fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, ricin 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 conjugating radionucleotides to antibodies. See International Publication No. WO 94 / 11026. The linker may also be a "cleavable linker" that facilitates release of the cytotoxic drug within the cell. For example, an acid-labile linker, a peptidase-sensitive linker, a photolabile linker, a dimethyl linker, or a disulfide-containing linker (Chari et al., Cancer Res. 52:127-131 (1992); U.S. Patent No. 5,208,020) can be used.

[0256] The immunoconjugates or ADCs herein expressly contemplate conjugates prepared using cross-linking reagents including, but not limited to, BMPS, EMCS, GMBS, HBVS, LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB, SMPH, sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC and sulfo-SMPB, and SVSB (succinimidyl-(4-vinylsulfone)benzoate), which are commercially available (e.g., from Pierce Biotechnology, Rockford, IL, USA).

[0257] D. Additional Therapeutic Agents In some examples, the methods described herein include administering a bispecific anti-CD20 / anti-CD3 antibody in combination with one or more additional therapeutic agents (e.g., an antibody-drug conjugate (ADC) and / or an additional chemotherapeutic agent). In some examples, the bispecific anti-CD20 / anti-CD3 antibody is co-administered with one or more additional chemotherapeutic agents selected from cyclophosphamide, doxorubicin, rituximab, and prednisolone. In some examples, the bispecific anti-CD20 / anti-CD3 antibody is co-administered with CHOP, where vincristine is replaced with an ADC. In some examples, the bispecific anti-CD20 / anti-CD3 antibody is co-administered with an anti-CD19 antibody, an anti-CD19 antibody-drug conjugate, an anti-CD22 antibody-drug conjugate, an anti-CD45 antibody-drug conjugate, and an anti-CD32 antibody-drug conjugate.

[0258] In some examples, the additional therapeutic agent is an anti-CD79b ADC, such as any of the anti-CD79b antibody-drug conjugates described in U.S. Patent No. 8,088,378, incorporated herein by reference in its entirety. In some examples, the anti-CD79b antibody-drug conjugate comprises 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: 33; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 34; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 35; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 36; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 37; or (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 38. In some examples, the anti-CD79b antibody drug conjugate comprises an anti-79b binding domain that comprises all six of the following HVRs: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 33, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 34, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 35, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 36, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 37, and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 38. In some examples, the anti-CD79b antibody drug conjugate comprises at least one (e.g., one, two, three, or four) of heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 39-42, respectively, and / or at least one (e.g., one, two, three, or four) of light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 43-46, respectively.In some examples, the anti-CD79b antibody drug conjugate comprises: (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 47, or the sequence of SEQ ID NO: 47; (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 48, or the sequence of SEQ ID NO: 48; or (c) a VH domain as in (a) and a VL domain as in (b). Thus, in some examples, the first binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 47 and a VL domain comprising the amino acid sequence of SEQ ID NO: 48.

[0259] In some examples, the anti-CD79b antibody is linked to a toxin, such as monomethyl auristatin E (MMAE, i.e., vedotin). In some examples, the anti-CD79b antibody-drug conjugate is polatuzumab vedotin (immunoglobulin G1-kappa auristatin E conjugate, anti-[Homo sapiens CD79b (immunoglobulin-related CD79 beta)], a humanized monoclonal antibody conjugated to auristatin E; gamma 1 heavy chain (1-447) [humanized VH (Homo sapiens IGHV3-23)], as defined by International Nonproprietary Names (INN) List 110 (WHO Drug Information, Vol. 27, No. 4, 2016, p. 443). * 04(76.50%)-(IGHD)-IGHJ4 * 01)[8.8.10](1-117)-Homo sapiens IGHG1 *03 (CH1 R120>K(214)(118-215), hinge(216-230), CH2(231-340), CH3(341-445), CHS(446-447))(118-447)], kappa light chain (1'-218') with (220-218')-disulfide [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")-bisdisulfide; an average of 3-4 cysteinyls conjugated to monomethyl auristatin E (MMAE) via a cleavable maleimidocaproyl-valyl-citrullinyl-p-aminobenzyloxycarbonyl (mc-val-cit-PABC) linker; 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 referred to interchangeably as "polatuzumab vedotin-piiq," "huMA79bv28-MC-vc-PAB-MMAE," or "DCDS4501A." In some examples, the anti-CD79b antibody or anti-CD79b ADC comprises the heavy chain sequence of SEQ ID NO:49 and the light chain sequence of SEQ ID NO:50.

[0260] In some examples, the additional therapeutic agent is a biological modifier. In one example, the bispecific anti-CD20 / anti-CD3 antibody is a cytotoxic agent, such as a BCL-2 inhibitor (e.g., GDC-0199 / ABT-199), lenalidomide (REVLIMID®), pomalidomide, thalidomide, a PI3K delta inhibitor (e.g., idelalisib (ZYDELIG®)), a PD-1 axis binding antagonist, tremelimumab (also known as ticilimumab or CP-675,206), urelumab (also known as BMS-663513), or a cytotoxic agent against TGF-beta. The antagonist MGA271 is co-administered with one or more biological modifiers selected from, for example, metelimu- mab (also known as CAT-192), fresolimumab (also known as GC1008), LY2157299k, and adoptive transfer of T cells (e.g., cytotoxic T cells or CTLs) expressing a chimeric antigen receptor (CAR) (e.g., adoptive transfer of T cells comprising a dominant-negative TGF beta receptor, e.g., a domina...

Claims

1. 1. A method of treating a subject having a CD20-positive cell proliferative disorder, comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) the C1D1 is equal to or less than the C1D2 and less than the C1D3; (ii) the C1D2 is equal to or less than the C1D3; (iii) the C1D1 is about 0.1 mg to about 10 mg, the C1D2 is about 5 mg to about 80 mg, and the C1D3 is about 10 mg to about 300 mg; (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein the C2D1 is equal to or greater than the C1D3 and is from about 10 mg to about 300 mg.

2. The method of claim 1, wherein said C1D1 is less than said C1D2.

3. 3. The method of claim 1 or 2, wherein the C1D2 is in an equivalent amount to the C1D3.

4. (a) the C1D1 is about 2 mg to about 8 mg, the C1D2 is about 10 mg to about 75 mg, and the C1D3 is about 20 mg to about 75 mg; (b) the C2D1 is about 20 mg to about 75 mg,

5. The method of any one of claims 1 to 4, wherein the C1D1 is about 5 mg.

6. The method of any one of claims 1 to 5, wherein the C1D3 is about 25 mg to about 75 mg.

7. 7. The method of claim 6, wherein the C1D3 is about 30 mg, about 45 mg, or about 60 mg.

8. The method of any one of claims 1 to 7, wherein the C2D1 is from about 40 mg to about 75 mg.

9. 9. The method of claim 8, wherein the C2D1 is about 30 mg, about 45 mg, or about 60 mg.

10. 10. The method of any one of claims 5 to 9, wherein the C1D2 is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 45 mg, or about 60 mg.

11. (a) the C1D1 is about 5 mg, the C1D2 is about 45 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg; (b) the C1D1 is about 5 mg, the C1D2 is about 15 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg; (c) the C1D1 is about 5 mg, the C1D2 is about 10 mg, the C1D3 is about 30 mg, and the C2D1 is about 30 mg; (d) the C1D1 is about 5 mg, the C1D2 is about 20 mg, the C1D3 is about 40 mg, and the C2D1 is about 40 mg; or (e) The method of any one of claims 1 to 10, wherein the C1D1 is about 5 mg, the C1D2 is about 20 mg, the C1D3 is about 45 mg, and the C2D1 is about 60 mg.

12. The method of any one of claims 1 to 11, wherein the C1D1 is about 5 mg, the C1D2 is about 45 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg.

13. The method of claim 1, wherein the C1D1 is in an equivalent amount to the C1D2.

14. 14. The method of claim 13, wherein the C1D1 is about 5 mg, the C1D2 is about 5 mg, the C1D3 is about 45 mg, and the C2D1 is about 60 mg.

15. 15. The method of claim 13 or 14, wherein the C1D2 is in an equivalent amount to the C1D3.

16. 16. The method of claim 15, wherein the C1D1 is about 5 mg, the C1D2 is about 45 mg, the C1D3 is about 45 mg, and the C2D1 is about 60 mg.

17. The method of any one of claims 1 to 16, wherein the method comprises administering the C1D2 to the subject about 7 days after the C1D1.

18. The method of any one of claims 1 to 17, wherein the method comprises administering the C1D3 to the subject about 7 days after the C1D2.

19. 19. The method of any one of claims 1 to 18, wherein the method comprises administering the C2D1 to the subject about 7 days after the C1D3.

20. 20. The method of any one of claims 1 to 19, wherein the method comprises administering the C1D1, the C1D2, and the C1D3 to the subject on, or about, days 1, 8, and 15, respectively, of the first administration cycle.

21. The method of any one of claims 1 to 20, wherein the method comprises administering the C2D1 to the subject on day 1 of the second administration cycle.

22. 22. The method of any one of claims 1 to 21, wherein the first and second administration cycles are 21 day administration cycles.

23. 22. The method of any one of claims 1-21, wherein the first administration cycle is a 21 day administration cycle and the second administration cycle is a 28 day administration cycle.

24. 1. A method of treating a subject having a CD20-positive cell proliferative disorder, comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) the C1D1 is about 5 mg; (ii) C1D2 is equal to or greater than C1D1 and equal to or less than C1D3; (iii) the C1D3 is about 45 mg; (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein the C2D1 is about 45 mg.

25. 25. The method of claim 24, wherein the C1D2 is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 45 mg, or about 60 mg.

26. 26. The method of claim 24 or 25, wherein the first and second dosing cycles are 21 day dosing cycles.

27. 26. The method of claim 24 or 25, wherein the first administration cycle is a 21 day administration cycle and the second administration cycle is a 28 day administration cycle.

28. 28. The method of any one of claims 24 to 27, wherein the method comprises administering the C1D1, the C1D2, and the C1D3 to the subject on, or about, days 1, 8, and 15, respectively, of the first administration cycle.

29. The method of any one of claims 24 to 28, wherein the method comprises administering the C2D1 to the subject on day 1 of the second administration cycle.

30. 1. A method of treating a subject having a CD20-positive cell proliferative disorder, comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) the first administration cycle comprises a first subcutaneous dose of the bispecific antibody (C1D1) on day 1 of the first administration cycle, a second subcutaneous dose of the bispecific antibody (C1D2) on day 8 of the first administration cycle, and a third subcutaneous dose of the bispecific antibody (C1D3) on day 15 of the first administration cycle; (i) the C1D1 is about 5 mg; (ii) C1D2 is equal to or greater than C1D1 and equal to or less than C1D3; (iii) the C1D3 is about 45 mg; (b) the second dosing cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1) on day 1 of the second dosing cycle, wherein the C2D1 is about 45 mg.

31. 31. The method of claim 30, wherein the C1D2 is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 45 mg, or about 60 mg.

32. 32. The method of claim 30 or 31, wherein the C1D2 is about 15 mg.

33. 32. The method of claim 30 or 31, wherein the C1D2 is about 45 mg.

34. 34. The method of any one of claims 30-33, wherein each of the administration cycles is a 21 day administration cycle.

35. 34. The method of any one of claims 30-33, wherein the first administration cycle is a 21 day administration cycle and the second administration cycle is a 28 day administration cycle.

36. The method of any one of claims 1 to 35, wherein the CD20-positive cell proliferative disorder is a B-cell proliferative disorder.

37. 37. The method of any one of claims 1 to 36, wherein the B-cell proliferative disorder is non-Hodgkin's lymphoma (NHL) or chronic lymphocytic leukemia (CLL).

38. 38. The method of claim 37, wherein the NHL is previously untreated (1L) NHL, relapsed or refractory NHL, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL).

39. 39. The method of claim 38, wherein the DLBCL is 1L DLBCL, or relapsed or refractory DLBCL.

40. 40. The method of claim 38 or 39, wherein the DLBCL is Richter's transformed.

41. 39. The method of claim 38, wherein the FL is 1L FL, or relapsed or refractory FL.

42. The method of claim 38 or 41, wherein the FL is a transformed FL.

43. 39. The method of claim 38, wherein the NHL is high-grade B-cell lymphoma.

44. 39. The method of claim 38, wherein the NHL is Ann Arbor stage III or IV NHL.

45. 45. The method of any one of claims 1-44, wherein the subject has previously received at least one prior line of systemic therapy.

46. 46. ​​The method of claim 45, wherein the subject has received between 1 and 9 prior lines of systemic therapy.

47. 47. The method of claim 46, wherein the subject has received three prior lines of systemic therapy.

48. 48. The method of any one of claims 45 to 47, wherein at least one prior line of systemic therapy included an anti-CD20 antibody.

49. 49. The method of claim 48, wherein the anti-CD20 antibody is rituximab or obinutuzumab.

50. 50. The method of claim 48 or 49, wherein the prior line of systemic therapy comprising the anti-CD20 antibody additionally comprises an alkylating agent or an anthracycline.

51. 51. The method of claim 50, wherein the alkylating agent is cyclophosphamide or bendamustine.

52. 51. The method of claim 50, wherein the anthracycline is daunomycin or doxorubicin.

53. the prior line of systemic therapy comprising the anti-CD20 antibody (i) cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP); (ii) cyclophosphamide, vincristine and prednisone (CVP); (iii) fludarabine, or (iv) bendamustine 49. The method of claim 48, further comprising:

54. 48. The method of any one of claims 45-47, wherein at least one prior line of systemic therapy included a Bruton's tyrosine kinase (BTK) inhibitor.

55. 1. A method of treating a subject with DLBCL, comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first 21-day dosing cycle and a second 21-day dosing cycle; (a) the first 21-day administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) the C1D1 is equal to or less than the C1D2 and less than the C1D3; (ii) the C1D2 is equal to or less than the C1D3; (iii) the C1D1 is about 0.1 mg to about 10 mg, the C1D2 is about 5 mg to about 80 mg, and the C1D3 is about 10 mg to about 300 mg; (b) the second 21-day administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein the C2D1 is equal to or greater than the C1D3 and is from about 10 mg to about 300 mg.

56. 56. The method of claim 55, wherein the DLBCL is 1L DLBCL, or relapsed or refractory DLBCL.

57. 57. The method of claim 55 or 56, wherein the DLBCL is Richter's transformed.

58. 58. The method of any one of claims 55 to 57, wherein the method comprises administering the C1D2 to the subject about 7 days after the C1D1.

59. 59. The method of any one of claims 55 to 58, wherein the method comprises administering the C1D3 to the subject about 7 days after the C1D2.

60. 60. The method of any one of claims 55-59, wherein the method comprises administering the C2D1 to the subject about 7 days after the C1D3.

61. 61. The method of any one of claims 55 to 60, wherein the method comprises administering the C1D1, the C1D2, and the C1D3 to the subject on, or about, days 1, 8, and 15, respectively, of the first administration cycle.

62. 1. A method of treating a subject with FL, comprising subcutaneously administering to the subject a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first 21-day dosing cycle and a second 28-day dosing cycle; (a) the first 21-day administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) the C1D1 is equal to or less than the C1D2 and less than the C1D3; (ii) the C1D2 is equal to or less than the C1D3; (iii) the C1D1 is about 0.1 mg to about 10 mg, the C1D2 is about 5 mg to about 80 mg, and the C1D3 is about 10 mg to about 300 mg; (b) the second 28-day administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein the C2D1 is equal to or greater than the C1D3 and is from about 10 mg to about 300 mg.

63. 63. The method of claim 62, wherein the FL is 1L FL, or relapsed or refractory FL.

64. The method of claim 62 or 63, wherein the FL is a transformed FL.

65. 65. The method of any one of claims 55 to 64, wherein said C1D1 is less than said C1D2.

66. 65. The method of any one of claims 55 to 64, wherein the C1D2 is in an equivalent amount to the C1D3.

67. (a) the C1D1 is about 2 mg to about 8 mg, the C1D2 is about 10 mg to about 75 mg, and the C1D3 is about 20 mg to about 75 mg; (b) the C2D1 is about 20 mg to about 75 mg.

68. 68. The method of any one of claims 55 to 67, wherein the C1D1 is about 5 mg.

69. 69. The method of any one of claims 55-68, wherein the C1D3 is from about 25 mg to about 75 mg.

70. 70. The method of claim 69, wherein the C1D3 is about 30 mg, about 45 mg, or about 60 mg.

71. 71. The method of any one of claims 55-70, wherein the C2D1 is from about 40 mg to about 75 mg.

72. 72. The method of claim 71, wherein the C2D1 is about 30 mg, about 45 mg, or about 60 mg.

73. 73. The method of any one of claims 68-72, wherein the C1D2 is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 45 mg, or about 60 mg.

74. (a) the C1D1 is about 5 mg, the C1D2 is about 45 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg; (b) the C1D1 is about 5 mg, the C1D2 is about 10 mg, the C1D3 is about 30 mg, and the C2D1 is about 30 mg; (c) the C1D1 is about 5 mg, the C1D2 is about 15 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg; (d) the C1D1 is about 5 mg, the C1D2 is about 20 mg, the C1D3 is about 40 mg, and the C2D1 is about 40 mg; or (e) The method of any one of claims 55 to 73, wherein the C1D1 is about 5 mg, the C1D2 is about 20 mg, the C1D3 is about 45 mg, and the C2D1 is about 60 mg.

75. The method of any one of claims 55 to 73, wherein the C1D1 is about 5 mg, the C1D2 is about 45 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg.

76. 65. The method of any one of claims 55 to 64, wherein the C1D1 is in an equivalent amount to the C1D2.

77. 77. The method of claim 76, wherein the C1D1 is about 5 mg, the C1D2 is about 5 mg, the C1D3 is about 45 mg, and the C2D1 is about 60 mg.

78. 65. The method of any one of claims 55 to 64, wherein the C1D2 is in an equivalent amount to the C1D3.

79. 79. The method of claim 78, wherein the C1D1 is about 5 mg, the C1D2 is about 45 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg.

80. 80. The method of any one of claims 55-79, wherein the method comprises administering the C2D1 to the subject on day 1 of the second administration cycle.

81. 81. The method of any one of claims 1-80, wherein the dosing regimen comprises one or more additional dosing cycles.

82. 82. The method of claim 81, wherein the dosing regimen comprises 1 to 15 additional dosing cycles.

83. 83. The method of claim 81 or 82, wherein the dosing regimen comprises six additional dosing cycles.

84. 83. The method of claim 81 or 82, wherein the dosing regimen comprises 15 additional dosing cycles.

85. 85. The method of any one of claims 81-84, wherein each additional administration cycle is a 21 day administration cycle.

86. 85. The method of any one of claims 81-84, wherein each additional administration cycle is a 28 day administration cycle.

87. 87. The method of any one of claims 81-86, wherein each additional administration cycle comprises administration of an additional dose of the bispecific antibody.

88. 88. The method of claim 87, wherein each additional dose of the bispecific antibody is an approximately equal amount of the C2D1.

89. 89. The method of claim 87 or 88, wherein each additional dose of the bispecific antibody is about 45 mg.

90. 90. The method of any one of claims 87-89, wherein the method comprises administering to the subject each additional dose of the bispecific antibody on day 1 of each respective additional administration cycle.

91. 91. The method of any one of claims 1 to 90, wherein the bispecific antibody is administered to the subject as monotherapy.

92. 91. The method of any one of claims 1 to 90, wherein the bispecific antibody is administered to the subject as a combination therapy.

93. 93. The method of claim 92, wherein the bispecific antibody is administered to the subject simultaneously with an additional therapeutic agent.

94. 93. The method of claim 92, wherein the bispecific antibody is administered to the subject prior to the administration of an additional therapeutic agent.

95. 93. The method of claim 92, wherein the bispecific antibody is administered to the subject after the administration of one or more additional therapeutic agents.

96. 96. The method of claim 95, wherein the additional therapeutic agent is obinutuzumab (GAZYVA®).

97. 96. The method of claim 95, wherein the additional therapeutic agent is tocilizumab.

98. 98. The method of any one of claims 1-97, wherein the subject has a cytokine release syndrome event, and the method further comprises treating the symptoms of the cytokine release syndrome event while withholding treatment with the bispecific antibody.

99. 99. The method of claim 98, wherein the method further comprises administering to the subject an effective amount of tocilizumab to treat the cytokine release syndrome event.

100. 100. The method of claim 99, wherein tocilizumab is administered intravenously to the subject as a single dose of about 8 mg / kg, wherein said single dose does not exceed 800 mg.

101. 101. The method of claim 100, wherein the cytokine release syndrome event does not resolve or worsens within 24 hours of treating the symptoms of the cytokine release syndrome event, and the method further comprises administering to the subject one or more additional doses of tocilizumab to manage the cytokine release syndrome event.

102. 102. The method of claim 101, wherein the one or more additional doses of tocilizumab are administered intravenously to the subject at a dose of about 8 mg / kg, wherein the dose does not exceed 800 mg.

103. 103. The method of claim 101 or 102, further comprising administering to the subject an effective amount of a corticosteroid.

104. 104. The method of claim 103, wherein the corticosteroid is administered to the subject intravenously.

105. 105. The method of claim 103 or 104, wherein the corticosteroid is methylprednisolone.

106. 106. The method of claim 105, wherein methylprednisolone is administered at a dose of about 2 mg / kg per day.

107. 105. The method of claim 103 or 104, wherein the corticosteroid is dexamethasone.

108. 108. The method of claim 107, wherein dexamethasone is administered at a dose of about 10 mg to about 100 mg.

109. 109. The method of claim 108, wherein dexamethasone is administered at a dose of about 10 mg.

110. 109. The method of claim 108, wherein the dexamethasone is administered at a dose of about 20 mg.

111. 1. A method of treating a population of subjects having a CD20-positive cell proliferative disorder, comprising subcutaneously administering to said subjects a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) the C1D1 is equal to or less than the C1D2 and less than the C1D3; (ii) the C1D2 is equal to or less than the C1D3; (iii) the C1D1 is about 0.1 mg to about 10 mg, the C1D2 is about 5 mg to about 80 mg, and the C1D3 is about 10 mg to about 300 mg; (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein the C2D1 is equal to or greater than the C1D3 and is from about 10 mg to about 300 mg.

112. 1. A method of treating a population of subjects having a CD20-positive cell proliferative disorder, comprising subcutaneously administering to said subjects a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) the C1D1 is about 5 mg; (ii) C1D2 is equal to or greater than C1D1 and equal to or less than C1D3; (iii) the C1D3 is about 45 mg; (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein the C2D1 is about 45 mg.

113. 1. A method of treating a population of subjects having a CD20-positive cell proliferative disorder, comprising subcutaneously administering to said subjects a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) the first administration cycle comprises a first subcutaneous dose of the bispecific antibody (C1D1) on day 1 of the first administration cycle, a second subcutaneous dose of the bispecific antibody (C1D2) on day 8 of the first administration cycle, and a third subcutaneous dose of the bispecific antibody (C1D3) on day 15 of the first administration cycle; (i) the C1D1 is about 5 mg; (ii) C1D2 is equal to or greater than C1D1 and equal to or less than C1D3; (iii) the C1D3 is about 45 mg; (b) the second dosing cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1) on day 1 of the second dosing cycle, wherein the C2D1 is about 45 mg.

114. The method of any one of claims 111 to 113, wherein the CD20-positive cell proliferative disorder is a B-cell proliferative disorder.

115. 114. The method of any one of claims 111 to 113, wherein the B-cell proliferative disorder is non-Hodgkin's lymphoma (NHL) or chronic lymphocytic leukemia (CLL).

116. 116. The method of claim 115, wherein the NHL is previously untreated (1L) NHL, relapsed or refractory NHL, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), high-grade B-cell lymphoma, or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL).

117. 117. The method of claim 116, wherein the DLBCL is 1L DLBCL, or relapsed or refractory DLBCL.

118. 118. The method of claim 116 or 117, wherein the DLBCL is Richter's transformed.

119. 117. The method of claim 116, wherein the FL is 1L FL, or relapsed or refractory FL.

120. The method of claim 116 or 119, wherein the FL is a transformed FL.

121. 117. The method of claim 116, wherein the NHL is a high-grade B-cell lymphoma.

122. 1. A method of treating a population of subjects with DLBCL, comprising subcutaneously administering to said subjects a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) the C1D1 is equal to or less than the C1D2 and less than the C1D3; (ii) the C1D2 is equal to or less than the C1D3; (iii) the C1D1 is about 0.1 mg to about 10 mg, the C1D2 is about 5 mg to about 80 mg, and the C1D3 is about 10 mg to about 300 mg; (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein the C2D1 is equal to or greater than the C1D3 and is from about 10 mg to about 300 mg.

123. 123. The method of claim 122, wherein the DLBCL is 1L DLBCL, or relapsed or refractory DLBCL.

124. 124. The method of claim 122 or 123, wherein the DLBCL is Richter's transformed.

125. 1. A method of treating a population of subjects with FL, comprising subcutaneously administering to said subjects a bispecific antibody that binds CD20 and CD3 in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle; (a) the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of the bispecific antibody; (i) the C1D1 is equal to or less than the C1D2 and less than the C1D3; (ii) the C1D2 is equal to or less than the C1D3; (iii) the C1D1 is about 0.1 mg to about 10 mg, the C1D2 is about 5 mg to about 80 mg, and the C1D3 is about 10 mg to about 300 mg; (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein the C2D1 is equal to or greater than the C1D3 and is from about 10 mg to about 300 mg.

126. 126. The method of claim 125, wherein the FL is 1L FL, or relapsed or refractory FL.

127. The method of claim 125 or 126, wherein the FL is a transformed FL.

128. (i) the C1D1 is about 5 mg, the C1D2 is about 15 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg; or (ii) The method of any one of claims 111 to 127, wherein the C1D1 is about 5 mg, the C1D2 is about 45 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg.

129. 129. The method of any one of claims 111-128, wherein the first and second administration cycles are 21 day administration cycles.

130. 129. The method of any one of claims 111-128, wherein the first administration cycle is a 21 day administration cycle and the second administration cycle is a 28 day administration cycle.

131. 131. The method of any one of claims 111-130, wherein the dosing regimen comprises one or more additional dosing cycles.

132. 132. The method of claim 131, wherein the dosing regimen comprises 1 to 15 additional dosing cycles.

133. 133. The method of claim 131 or 132, wherein the dosing regimen comprises six additional dosing cycles.

134. 133. The method of claim 131 or 132, wherein the dosing regimen comprises 15 additional dosing cycles.

135. 135. The method of any one of claims 131-134, wherein each additional administration cycle is a 21 day administration cycle.

136. 135. The method of any one of claims 131-134, wherein each additional administration cycle is a 28 day administration cycle.

137. 137. The method of any one of claims 131 to 136, wherein each additional administration cycle comprises administration of an additional dose of the bispecific antibody.

138. 138. The method of claim 137, wherein each additional dose of the bispecific antibody is an approximately equal amount of the C2D1.

139. 139. The method of claim 137 or 138, wherein each additional dose of the bispecific antibody is about 45 mg.

140. 140. The method of any one of claims 137-139, wherein the method comprises administering each additional dose of the bispecific antibody to the population of subjects on day 1 of each respective additional administration cycle.

141. 141. The method of any one of claims 111 to 140, wherein the complete response rate is at least about 20%.

142. 142. The method of any one of claims 111-141, wherein the complete response rate is greater than about 40%.

143. 143. The method of any one of claims 111-142, wherein the complete response rate is greater than about 55%.

144. 144. The method of any one of claims 111-143, wherein the median progression-free survival is greater than about 2 months.

145. 144. The method of any one of claims 111-143, wherein the median overall survival is greater than about 9.5 months.

146. 144. The method of any one of claims 111-143, wherein the objective response rate is at least about 70% at about 20 months after initiation of treatment.

147. 144. The method of any one of claims 111-143, wherein the objective response rate is at least about 60% at about 12 months after initiation of treatment.

148. 141. The method of any one of claims 111-140, wherein the population of subjects has relapsed or refractory NHL and has an objective response rate of at least 34%.

149. 149. The method of claim 148, wherein the objective response rate is at least 44%.

150. 141. The method of any one of claims 111-140, wherein the population of subjects has relapsed or refractory NHL and the objective response rate is between 35% and 55%.

151. 151. The method of claim 150, wherein the objective response rate is about 45%.

152. 141. The method of any one of claims 111-116 and 125-140, wherein the population of subjects has relapsed or refractory FL and has an objective response rate of at least 70%.

153. 153. The method of claim 152, wherein the objective response rate is at least 80%.

154. 141. The method of any one of claims 111-116 and 125-140, wherein the population of subjects has relapsed or refractory FL and the objective response rate is between 70% and 90%.

155. 155. The method of claim 154, wherein the objective response rate is about 80%.

156. 141. The method of any one of claims 111-116, 122-124, and 128-140, wherein the population of subjects has relapsed or refractory DLBCL, or transformed FL, and the objective response rate is at least 25%.

157. 157. The method of claim 156, wherein the objective response rate is at least 35%.

158. 141. The method of any one of claims 111-116, 122-124, and 128-140, wherein the population of subjects has relapsed or refractory DLBCL and the objective response rate is between 25% and 45%.

159. 159. The method of claim 158, wherein the objective response rate is about 35%.

160. 160. The method of any one of claims 111 to 159, wherein the population of subjects exhibits cytokine release syndrome following administration of the bispecific antibody, and the rate of cytokine release syndrome in the population of subjects is about 30% or less.

161. 161. The method of claim 160, wherein the rate of cytokine release syndrome in the population of subjects is about 25% or less.

162. The method of claim 161, wherein the rate of cytokine release syndrome in the subject population is less than or equal to about 10%.

163. The method of claim 162, wherein the rate of cytokine release syndrome in the population of subjects is about 5% or less.

164. The method of claim 163, wherein the rate of cytokine release syndrome in the subject population is about 3% or less.

165. 165. The method of any one of claims 111 to 164, wherein the rate of cytokine release syndrome having Grade 2 or higher (as defined by the American Society for Transplantation and Cellular Therapy, 2018; ASTCT) is about 10% or less.

166. 166. The method of claim 165, wherein the rate of cytokine release syndrome having grade 2 or higher (as defined by the ASTCT) is about 5% or less.

167. 167. The method of claim 166, wherein the rate of cytokine release syndrome having grade 2 or higher (as defined by the ASTCT) is about 3% or less.

168. 168. The method of any one of claims 111-167, wherein the rate of cytokine release syndrome having grade 3 or higher (as defined by the ASTCT) is about 1% or less.

169. The method of claim 168, wherein the rate of cytokine release syndrome having grade 3 or higher (as defined by the ASTCT) is about 0%.

170. 1. A method of reducing the rate of a specific adverse event in a population of subjects with a CD20-positive cell proliferative disorder administered a bispecific antibody that binds to CD20 and CD3, the method comprising administering the bispecific antibody subcutaneously using a phased dosing regimen, wherein the rate of a specific adverse event is reduced in the population of subjects compared to a reference population of subjects to whom the bispecific antibody is administered intravenously.

171. The step-wise administration regimen comprises: (I) at least a first administration cycle and a second administration cycle, (a) the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of an anti-CD20 antibody; (i) the C1D1 is equal to or less than the C1D2 and less than the C1D3; (ii) the C1D2 is equal to or less than the C1D3; (iii) the C1D1 is about 0.1 mg to about 10 mg, the C1D2 is about 5 mg to about 80 mg, and the C1D3 is about 10 mg to about 300 mg; (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein the C2D1 is equal to or greater than the C1D3 and is from about 10 mg to about 300 mg; at least a first administration cycle and a second administration cycle; (II) at least a first administration cycle and a second administration cycle, (a) the first administration cycle comprises a first subcutaneous dose (C1D1), a second subcutaneous dose (C1D2), and a third subcutaneous dose (C1D3) of an anti-CD20 antibody; (i) the C1D1 is about 5 mg; (ii) C1D2 is equal to or greater than C1D1 and equal to or less than C1D3; (iii) the C1D3 is about 45 mg; (b) the second administration cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1), wherein the C2D1 is about 45 mg; at least a first administration cycle and a second administration cycle; and (III) at least a first administration cycle and a second administration cycle, (a) the first administration cycle comprises a first subcutaneous dose of the bispecific antibody (C1D1) on day 1 of the first administration cycle, a second subcutaneous dose of the bispecific antibody (C1D2) on day 8 of the first administration cycle, and a third subcutaneous dose of the bispecific antibody (C1D3) on day 15 of the first administration cycle; (i) the C1D1 is about 5 mg; (ii) C1D2 is equal to or greater than C1D1 and equal to or less than C1D3; (iii) the C1D3 is about 45 mg; (b) the second dosing cycle comprises a single subcutaneous dose of the bispecific antibody (C2D1) on day 1 of the second dosing cycle, wherein the C2D1 is about 45 mg; at least a first administration cycle and a second administration cycle; 171. The method of claim 170, wherein the administration regimen is a stepwise administration regimen selected from the group consisting of:

172. (i) the C1D1 is about 5 mg, the C1D2 is about 15 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg; or (ii) The method of claim 170 or 171, wherein the C1D1 is about 5 mg, the C1D2 is about 45 mg, the C1D3 is about 45 mg, and the C2D1 is about 45 mg.

173. 173. The method of any one of claims 170-172, wherein the first and second administration cycles are 21 day administration cycles.

174. 173. The method of any one of claims 170-172, wherein the first administration cycle is a 21 day administration cycle and the second administration cycle is a 28 day administration cycle.

175. 175. The method of any one of claims 170-174, wherein the dosing regimen comprises one or more additional dosing cycles.

176. 176. The method of claim 175, wherein the dosing regimen comprises 1 to 15 additional dosing cycles.

177. 177. The method of claim 175 or 176, wherein the dosing regimen comprises six additional dosing cycles.

178. 177. The method of claim 175 or 176, wherein the dosing regimen comprises 15 additional dosing cycles.

179. 179. The method of any one of claims 175-178, wherein each additional administration cycle is a 21 day administration cycle.

180. 179. The method of any one of claims 175-178, wherein each additional administration cycle is a 28 day administration cycle.

181. 181. The method of any one of claims 175-180, wherein each additional cycle of administration comprises administration of an additional dose of the bispecific antibody.

182. 182. The method of claim 181, wherein each additional dose of the bispecific antibody is an approximately equal amount of the C2D1.

183. 183. The method of claim 181 or 182, wherein each additional dose of the bispecific antibody is about 45 mg.

184. 184. The method of any one of claims 181-183, wherein the method comprises administering each additional dose of the bispecific antibody to the population of subjects on day 1 of each respective additional administration cycle.

185. The method of any one of claims 170 to 184, wherein the CD20-positive cell proliferative disorder is a B-cell proliferative disorder.

186. 186. The method of claim 185, wherein the B-cell proliferative disorder is non-Hodgkin's lymphoma (NHL) or chronic lymphocytic leukemia (CLL).

187. 187. The method of claim 186, wherein the NHL is previously untreated (1L) NHL, relapsed or refractory R / R NHL, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), high-grade B-cell lymphoma, or primary mediastinal (thymic) large B-cell lymphoma (PMLBCL).

188. 188. The method of claim 187, wherein the DLBCL is 1L DLBCL, or relapsed or refractory DLBCL.

189. The method of claim 187 or 188, wherein the DLBCL is Richter's transformed.

190. 188. The method of claim 187, wherein the FL is 1L FL, or relapsed or refractory FL.

191. The method of claim 187 or 190, wherein the FL is a transformed FL.

192. 188. The method of claim 187, wherein the NHL is high-grade B-cell lymphoma.

193. 193. The method of any one of claims 170 to 192, wherein the population of subjects exhibits cytokine release syndrome following administration of the bispecific antibody, and the rate of cytokine release syndrome in the population of subjects is about 25% or less.

194. 193. The method of any one of claims 170 to 192, wherein the population of subjects exhibits cytokine release syndrome following administration of the bispecific antibody, and the rate of cytokine release syndrome in the population of subjects is about 30% or less.

195. The method of claim 193, wherein the rate of cytokine release syndrome in the population of subjects is about 10% or less.

196. The method of claim 195, wherein the rate of cytokine release syndrome in the population of subjects is less than or equal to about 5%.

197. The method of claim 196, wherein the rate of cytokine release syndrome in the population of subjects is less than or equal to about 3%.

198. 193. The method of any one of claims 170 to 192, wherein the rate of cytokine release syndrome having Grade 2 or higher (as defined by the American Society for Transplantation and Cellular Therapy, 2018; ASTCT) is about 10% or less.

199. The method of claim 198, wherein the rate of cytokine release syndrome having grade 2 or higher (as defined by the ASTCT) is about 5% or less.

200. 200. The method of claim 199, wherein the rate of cytokine release syndrome having grade 2 or higher (as defined by the ASTCT) is about 3% or less.

201. 201. The method of any one of claims 170-200, wherein the rate of cytokine release syndrome having grade 3 or higher (as defined by ASTCT) is about 1% or less.

202. The method of claim 201, wherein the rate of cytokine release syndrome having grade 3 or higher (as defined by the ASTCT) is about 0%.

203. 203. The method of any one of claims 170 to 202, wherein the complete response rate is at least about 20%.

204. 204. The method of any one of claims 170 to 203, wherein the complete response rate is at least about 40%.

205. 205. The method of any one of claims 170-204, wherein the median progression-free survival is greater than about 4 months.

206. 205. The method of any one of claims 170-204, wherein the median overall survival is greater than about 9.5 months.

207. 207. The method of any one of claims 170 to 206, wherein the objective response rate is at least about 75% at about 24 months after initiation of treatment.

208. 207. The method of any one of claims 170 to 206, wherein the objective response rate is at least about 70% at about 24 months after initiation of treatment.

209. 207. The method of any one of claims 170 to 206, wherein the objective response rate is at least about 60% at about 12 months after initiation of treatment.

210. 188. The method of any one of claims 170-187, wherein the population of subjects has relapsed or refractory NHL and has an objective response rate of at least 34%.

211. 211. The method of claim 210, wherein the objective response rate is at least 44%.

212. 188. The method of any one of claims 170-187, wherein the population of subjects has relapsed or refractory NHL and the objective response rate is between 35% and 55%.

213. 213. The method of claim 212, wherein the objective response rate is about 45%.

214. 188. The method of any one of claims 170-187, wherein the population of subjects has relapsed or refractory FL and has an objective response rate of at least 70%.

215. 215. The method of claim 214, wherein the objective response rate is at least 80%.

216. 188. The method of any one of claims 170-187, wherein the population of subjects has relapsed or refractory FL and the objective response rate is between 70% and 90%.

217. 217. The method of claim 216, wherein the objective response rate is about 80%.

218. 188. The method of any one of claims 170-187, wherein the population of subjects has relapsed or refractory DLBCL, or transformed FL, and has an objective response rate of at least 25%.

219. The method of claim 218, wherein the objective response rate is at least 35%.

220. The method of any one of claims 170-187, wherein the population of subjects has relapsed or refractory DLBCL and the objective response rate is between 25% and 45%.

221. 221. The method of claim 220, wherein the objective response rate is about 35%.

222. The bispecific antibody comprises the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence GYTFTSYNMH (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence AIYPGNGDTSYNQKFKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence VVYYSNSYWYFDV (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of RASSSVSYMH (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of APSNLAS (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence QQWSFNPPT (SEQ ID NO: 6); 222. The method of any one of claims 1 to 221, comprising an anti-CD20 arm comprising a first binding domain comprising:

223. 223. The method of any one of claims 1 to 222, wherein the bispecific antibody comprises: (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 7; (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 8; or (c) an anti-CD20 arm comprising a first binding domain comprising the VH domain of (a) and the VL domain of (b).

224. 224. The method of claim 223, wherein the first binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 7 and a VL domain comprising the amino acid sequence of SEQ ID NO:

8.

225. the bispecific antibody comprises the following six HVRs: (a) HVR-H1 comprising the amino acid sequence of NYYIH (SEQ ID NO: 9); (b) HVR-H2 comprising the amino acid sequence of WIYPGDGNTKYNEKFKG (SEQ ID NO: 10); (c) HVR-H3 comprising the amino acid sequence DSYSNYYFDY (SEQ ID NO: 11); (d) HVR-L1 comprising the amino acid sequence KSSQSLLNSRTRKNYLA (SEQ ID NO: 12); (e) HVR-L2 comprising the amino acid sequence of WASTRES (SEQ ID NO: 13); and (f) HVR-L3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 14) 225. The method of any one of claims 1 to 224, comprising an anti-CD3 arm comprising a second binding domain comprising:

226. 226. The method of any one of claims 1 to 225, wherein the bispecific antibody comprises an anti-CD3 arm comprising: (a) a VH domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 15; (b) a VL domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16; or (c) a second binding domain comprising the VH domain of (a) and the VL domain of (b).

227. The method of claim 226, wherein the second binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 15 and a VL domain comprising the amino acid sequence of SEQ ID NO:

16.

228. 228. The method of any one of claims 1 to 227, wherein the bispecific antibody comprises: (a) an anti-CD20 arm comprising: (i) a heavy chain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 51; and (ii) a light chain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 52; and (b) an anti-CD3 arm comprising: (i) a heavy chain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 53; and (ii) a light chain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:

54.

229. The method of claim 228, wherein (a) the anti-CD20 arm comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51 and a light chain comprising the amino acid sequence of SEQ ID NO: 52, and (b) the anti-CD3 arm comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 53 and a light chain comprising the amino acid sequence of SEQ ID NO:

54.

230. The method of any one of claims 1 to 229, wherein the bispecific antibody is a humanized antibody.

231. The method of any one of claims 1 to 230, wherein the bispecific antibody is a chimeric antibody.

232. The method of any one of claims 1 to 231, wherein the bispecific antibody is an antibody fragment that binds to CD20 and CD3.

233. The antibody fragment may be a Fab fragment, a Fab'-SH fragment, an Fv fragment, an scFv fragment, or a (Fab') ２ The method of claim 232, wherein the fragment is selected from the group consisting of:

234. The method of any one of claims 1 to 231, wherein the bispecific antibody is a full-length antibody.

235. 235. The method of any one of claims 1 to 231 and 234, wherein the bispecific antibody is an IgG antibody.

236. The IgG antibody is IgG １ The method of claim 235, which is an antibody.

237. The method of claim 235 or 236, wherein the IgG antibody comprises a mutation at amino acid residue N297 (EU numbering) that results in the absence of glycosylation.

238. 238. The method of claim 237, wherein the mutation at amino acid residue N297 is a substitution mutation.

239. The method of claim 237 or 238, wherein the mutation at amino acid residue N297 reduces the effector function of the Fc region.

240. The method of any one of claims 237 to 239, wherein the mutation is an N297G or N297A mutation (EU numbering).

241. 241. The method of any one of claims 236 to 240, wherein the bispecific antibody comprises a mutation in the Fc region that reduces effector function.

242. 242. The method of claim 241, wherein the mutation is a substitution mutation.

243. 243. The method of claim 242, wherein the substitution mutation is at amino acid residues L234, L235, D265, and / or P329 (EU numbering).

244. 244. The method of claim 243, wherein the substitution mutation is selected from the group consisting of L234A, L235A, D265A, and P329G (EU numbering).

245. The bispecific antibody comprises one or more heavy chain constant domains, wherein the one or more heavy chain constant domains comprise a first CH1 (CH1 １ ) domain, the first CH2 (CH2 １ ) domain, the first CH3 (CH3 １ ) domain, a second CH1 (CH1 ２ ) domain, the second CH2 (CH2 ２ ) domain, and a second CH3 (CH3 ２ 245. The method of any one of claims 1 to 231 and 234 to 244, wherein the domain is selected from the group consisting of:

246. 246. The method of claim 245, wherein at least one of the one or more heavy chain constant domains is paired with another heavy chain constant domain.

247. CH3 １ domain and the CH3 ２ Each domain contains a protrusion or a cavity, and the CH3 １ The protrusion or cavity of the domain is ２ 247. The method of claim 245 or 246, wherein the domain is positionable in the cavity or protrusion, respectively.

248. CH3 １ domain and the CH3 ２ 248. The method of claim 247, wherein the domains associate at the interface between the protrusion and the cavity.

249. CH2 １ domain and the CH2 ２ Each domain comprises a protrusion or a cavity, and the CH2 １ The protrusion or cavity of the domain is ２ 249. A method according to any one of claims 245 to 248, wherein the domain is positionable in the cavity or protrusion, respectively.

250. CH2 １ domain and the CH2 ２ 250. The method of claim 249, wherein the domains associate at the interface between the protrusion and the cavity.

251. 225. The method of claim 223 or 224, wherein the anti-CD20 arm further comprises T366W and N297G substitution mutations (EU numbering).

252. 228. The method of claim 226 or 227, wherein the anti-CD3 arm further comprises T366S, L368A, Y407V and N297G substitution mutations (EU numbering).

253. 230. The method of claim 228 or 229, wherein (a) the anti-CD20 arm further comprises T366W and N297G substitution mutations, and (b) the anti-CD3 arm further comprises T366S, L368A, Y407V and N297G substitution mutations (EU numbering).

254. 254. The method of any one of claims 1-110 or 222-253, wherein the subject is a human.

255. 254. The method of any one of claims 111-221 or 222-253, wherein the subject is a human.

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