Dosing for combination treatment with Anti-cd20 / Anti-cd3 bispecific antibody and Anti-cd79b antibody drug conjugate

Abstract

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

Description

technical field

[0001] The present invention relates to methods for treating diseases, particularly B cell proliferative disorders, by administering anti-CD20 / anti-CD3 bispecific antibodies and anti-CD79b antibody-drug conjugates, and methods for reducing reactive anti-CD20 / anti-CD3 bispecific antibodies Methods of Adverse Effects of Administration of Antibody and Anti-CD79b Antibody Drug Conjugates. prior art

[0002] B-cell proliferative disorders reveal a heterogeneous population of malignancies, including leukemias and lymphomas. Lymphoma develops from lymphocytes and includes two types: Hodgkin's lymphoma (HL) and non-Hodgkin's lymphoma (NHL). Lymphomas of B-cell origin account for approximately 80-85% of all non-Hodgkin's lymphoma cases in the United States, and there is considerable variation within B-cell subsets based on the genotypic and phenotypic expression patterns of the B-cell of origin qualitative. For example, the subgroup of B-cell lymphomas includes slow-growing indolent and incurable diseases, such as follicular lymphoma (FL) or chronic lymphocytic leukemia (CLL), as well as the more aggressive subtype, mantle cell lymphoma. lymphoma (MCL) and diffuse large B-cell lymphoma (DLBCL). Diffuse large B-cell lymphoma (DLBCL) is the most common type of NHL, accounting for approximately 30%-40% of all NHL diagnoses, followed by follicular lymphoma (FL; accounting for 20%-25% of all NHL diagnoses) and mantle cell lymphoma (MCL; 6%-10% of all NHL diagnoses). B-cell chronic lymphocytic leukemia (CLL) is the most common leukemia in adults, with approximately 15,000 new cases each year in the United States (American Cancer Society 2015).

[0003] Bispecific antibodies can simultaneously bind cell surface antigens on cytotoxic cells (e.g., T cells, by binding to cluster of differentiation 3 (CD3)) and cancer cells (e.g., B cells, by binding to CD20), with the purpose of The bound cytotoxic cells destroy the bound cancer cells. Glofitamab is a T cell engaging bispecific (TCB) antibody that targets CD20 expressed on B cells and the CD3 epsilon chain (CD3e) present on T cells.

[0004] However, immunotherapy with anti-CD20 / anti-CD3 bispecific antibodies such as grifidumab may be limited by adverse effects, including cytokine-driven toxicity (e.g., cytokine release syndrome (CRS)), transfusion Injection-related reactions (IRR), severe tumor lysis syndrome (TLS), and central nervous system (CNS) toxicity.

[0005] Therefore, development of anti-CD20 / anti-CD3 bispecific antibodies (e.g., Griffin) for the treatment of CD20-positive B-cell proliferative disorders (e.g., non-Hodgkin's lymphoma, NHL) that achieve a more favorable benefit-risk profile There is an unmet need in this field for an effective delivery method for the drug. Contents of the invention

[0006] In one aspect, the invention features a method of treating an individual having a CD20-positive cell proliferative disorder comprising administering to the individual an anti-CD79b on a dosing regimen comprising at least a first dosing cycle and a second dosing cycle Antibody-drug conjugates and bispecific antibodies that bind to CD20 and CD3, wherein: (a) the first dosing cycle includes a first dose of the bispecific antibody (C1D1) and a second dose of the bispecific antibody (C1D2) , wherein the C1D1 of the bispecific antibody is about 2.5 mg and the C1D2 of the bispecific antibody is about 10 mg; and (b) the second dosing cycle comprises a single dose of the bispecific antibody (C2D1), wherein the bispecific antibody The C2D1 is about 10 mg, about 16 mg or about 30 mg.

[0007] In another aspect, the invention features an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 for use in a method of treating an individual with a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody The drug conjugate and the bispecific antibody binding to CD20 and CD3 are administered with a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle comprises the bispecific antibody A first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody, wherein the C1D1 of the bispecific antibody is about 2.5 mg, and the C1D2 of the bispecific antibody is about 10 mg; and (b) the second dose of The drug cycle comprises a single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is about 10 mg, about 16 mg or about 30 mg.

[0008] In another aspect, the present invention is characterized by the use of anti-CD79b antibody-drug conjugates and bispecific antibodies that bind to CD20 and CD3 in the treatment of individuals with CD20-positive cell proliferative disorders, wherein the anti-CD79b antibody-drug conjugates and The bispecific antibody that binds to CD20 and CD3 is administered with a dosing regimen that includes at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle includes a first dose of the bispecific antibody (C1D1) and a second dose (C1D2) of the bispecific antibody, wherein C1D1 of the bispecific antibody is about 2.5 mg and C1D2 of the bispecific antibody is about 10 mg; and (b) the second dosing cycle includes bispecific A single dose (C2D1) of the specific antibody, wherein the C2D1 of the bispecific antibody is about 10 mg, about 16 mg or about 30 mg.

[0009] In another aspect, the invention features anti-CD79b antibody-drug conjugates and the use of bispecific antibodies that bind to CD20 and CD3 in the manufacture of a medicament for treating individuals with CD20-positive cell proliferative disorders, wherein the anti-CD79b Antibody-drug conjugates and bispecific antibodies that bind to CD20 and CD3 are administered with a dosing regimen that includes at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle includes bispecific A first dose of the antibody (C1D1) and a second dose of the bispecific antibody (C1D2), wherein the C1D1 of the bispecific antibody is about 2.5 mg and the C1D2 of the bispecific antibody is about 10 mg; and (b) the second dose of The dosing cycle comprises a single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is about 10 mg, about 16 mg or about 30 mg.

[0010] In some embodiments, the C2D1 of the bispecific antibody is about 10 mg. In some embodiments, the C2D1 of the bispecific antibody is about 16 mg. In some embodiments, the C2D1 of the bispecific antibody is about 30 mg.

[0011] In some embodiments, the first dosing cycle comprises a single dose of C1D1 of the anti-CD79b antibody drug conjugate. In some embodiments, a single dose of anti-CD79b antibody drug conjugate C1D1 is about 0.1 mg / kg to about 2.4 mg / kg (e.g., about 0.1 mg / kg to about 2.2 mg / kg, about 0.1 mg / kg to about 2.0 mg / kg, about 0.5 mg / kg to about 2.2 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.2 mg / kg, about 1.2 mg / kg to about 2.2 mg / kg, about 1.4 mg / kg to about 2.2 mg / kg, about 1.6 mg / kg to about 2.2 mg / kg, about 1.8 mg / kg to about 2.0 mg / kg, about 0.1 mg / kg to about 1.6 mg / kg , about 0.5 mg / kg to about 1.6 mg / kg, or about 1 mg / kg to about 1.8 mg / kg; for example, about 1 mg / kg, about 1.2 mg / kg, about 1.6 mg / kg, or about 1.8 mg / kg). In some embodiments, a single dose of anti-CD79b antibody drug conjugate C1D1 is about 1.8 mg / kg. In some embodiments, the C1D1 of the anti-CD79b antibody drug conjugate is or will be administered on or about day 2 (± day 1) of the dosing cycle. In some embodiments, the second dosing cycle comprises a single dose of C2D1 of the anti-CD79b antibody drug conjugate. In some embodiments, a single dose of anti-CD79b antibody drug conjugate C2D1 is about 0.1 mg / kg to about 2.4 mg / kg (e.g., about 0.1 mg / kg to about 2.2 mg / kg, about 0.1 mg / kg to about 2.0 mg / kg, about 0.5 mg / kg to about 2.2 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.2 mg / kg, about 1.2 mg / kg to about 2.2 mg / kg, about 1.4 mg / kg to about 2.2 mg / kg, about 1.6 mg / kg to about 2.2 mg / kg, about 1.8 mg / kg to about 2.0 mg / kg, about 0.1 mg / kg to about 1.6 mg / kg , about 0.5 mg / kg to about 1.6 mg / kg, or about 1 mg / kg to about 1.8 mg / kg; for example, about 1 mg / kg, about 1.2 mg / kg, about 1.6 mg / kg, or about 1.8 mg / kg). In some embodiments, a single dose of anti-CD79b antibody drug conjugate C2D1 is about 1.8 mg / kg.

[0012] In some embodiments, C1D1 of the bispecific antibody and C1D2 of the bispecific antibody are or will be administered on or around day 8 (± 1 day) and day 15 (± 1 day) of the first administration cycle, respectively. 1 day) administered to individuals. In some embodiments, C2D1 of the bispecific antibody is or will be administered to the individual on or about day 1 (± 1 day) of the second dosing cycle. In some embodiments, the C2D1 of the bispecific antibody is or will be administered after the administration of the C2D1 of the anti-CD79b antibody drug conjugate is complete. In some embodiments, the C2D1 of the bispecific antibody is or will be between about 60 and 120 minutes (e.g., between about 60 and 100 minutes, about 60 to 120 minutes) after administration of the C2D1 of the anti-CD79b antibody drug conjugate is complete. Between 90 minutes, between about 60 and 80 minutes, between about 90 and 120 minutes, between about 80 and 100 minutes, between about 80 and 120 minutes, between about 75 and 105 minutes, or between about 85 and 95 minutes Between minutes; for example, about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes, about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes ) cast. In some embodiments, C2D1 of the bispecific antibody is or will be administered about 90 minutes after administration of C2D1 of the anti-CD79b antibody drug conjugate is complete. In some embodiments, the C1D1 of the anti-CD79b antibody drug conjugate is or will be administered to the individual on or about day 2 (± 1 day) of the first dosing cycle, and the C2D1 of the anti-CD79b antibody drug conjugate is or will be Administer to individuals on or around Day 1 (± 1 day) of the second dosing cycle. In some embodiments, the first and second dosing cycles are 14-day (eg, 14±3-day) dosing cycles. In some embodiments, the first and second dosing cycles are 21 day (eg, 21 ± 3 day) dosing cycles.

[0013] In some embodiments, the dosing regimen comprises one or more additional dosing cycles. In some embodiments, the dosing regimen comprises six to ten additional dosing cycles (e.g., six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles). In some embodiments, the dosing regimen comprises ten additional dosing cycles. In some embodiments, the additional dosing cycle is a 14 day (eg, 14 ± 3 day) dosing cycle. In some embodiments, the additional dosing cycle is a 21 day (eg, 21 ± 3 day) dosing cycle.

[0014] In some embodiments, one or more of the additional dosing cycles comprises an additional single dose of the bispecific antibody and an additional single dose of the anti-CD79b antibody drug conjugate. In some embodiments, the additional single dose of the anti-CD79b antibody drug conjugate is about the same amount of C2D1 of the anti-CD79b antibody drug conjugate. In some embodiments, the additional single dose of the anti-CD79b antibody drug conjugate is about 1.8 mg / kg. In some embodiments, the additional single dose of the anti-CD79b antibody drug conjugate is or will be administered on or around (± 1 day) day 1 of each additional dosing cycle comprising the additional dose of the anti-CD79b antibody drug conjugate individual.

[0015] In some embodiments, the additional single dose of the bispecific antibody for each additional dosing cycle comprising the additional dose of the anti-CD79b antibody drug conjugate is or will be after completion of the administration of the additional single dose of the anti-CD79b antibody drug conjugate cast. In some embodiments, the additional single dose of the bispecific antibody for each additional dosing cycle comprising the additional dose of the anti-CD79b antibody drug conjugate is or will be after completion of the administration of the additional single dose of the anti-CD79b antibody drug conjugate Between about 60 and 120 minutes (e.g., between about 60 and 100 minutes, between about 60 and 90 minutes, between about 60 and 80 minutes, between about 90 and 120 minutes, between about 80 and 100 minutes, Between about 80 and 120 minutes, between about 75 and 105 minutes, or between about 85 and 95 minutes; e.g., about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes , about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes) administration. In some embodiments, the additional single dose of the bispecific antibody for each additional dosing cycle comprising the additional dose of the anti-CD79b antibody drug conjugate is or will be after completion of the administration of the additional single dose of the anti-CD79b antibody drug conjugate Administer for approximately 90 minutes.

[0016] In some embodiments, the dosing regimen comprises at least four additional dosing cycles comprising additional single doses of the anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen comprises four to ten additional dosing cycles (e.g., four additional dosing cycles, five additional dosing cycles, six additional dosing cycles, seven additional dosing cycles, Eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles) comprising an additional single dose of the anti-CD79b antibody drug conjugate.

[0017] In some embodiments, one or more of the additional dosing cycles comprises an additional single dose of the bispecific antibody and does not comprise the administration of an anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen comprises at least two additional dosing cycles comprising additional single doses of the bispecific antibody and excluding administration of the anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen comprises two to ten additional dosing cycles (e.g., two additional dosing cycles, three additional dosing cycles, four additional dosing cycles, five additional dosing cycles, Six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles), the additional dosing cycles comprising additional doses of the bispecific antibody Single dose and does not include administration of anti-CD79b antibody drug conjugate.

[0018] In some embodiments, the additional single dose of the bispecific antibody is about equivalent to the amount of C2D1 of the bispecific antibody. In some embodiments, the additional single dose of the bispecific antibody is about 30 mg. In some embodiments, the additional single dose of the bispecific antibody is or will be administered to the subject on or around (± 1 day) day 1 of each additional dosing cycle comprising the additional dose of the bispecific antibody.

[0019] In some embodiments, the dosing regimen comprises six or more additional dosing cycles (e.g., six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles or ten additional dosing cycles), each of the six or more additional dosing cycles comprises a single dose of the bispecific antibody, and no more than four of the six or more additional dosing cycles Administration of one (e.g., none, not more than one, not more than two, not more than three, or not more than four; e.g., none, one, two, three, or four) comprising an anti-CD79b antibody drug conjugate give. In some embodiments, the dosing regimen comprises six additional dosing cycles, wherein each of the six or more additional dosing cycles comprises a single dose of the bispecific antibody, and wherein in the six additional dosing cycles Not more than four (e.g., none, not more than one, not more than two, not more than three, or not more than four; e.g., none, one, two, three, or four) pharmaceuticals containing an anti-CD79b antibody Administration of Conjugates.

[0020] In one aspect, the invention features a method of treating an individual having a CD20-positive cell proliferative disorder comprising administering to the individual an anti-CD79b on a dosing regimen comprising at least a first dosing cycle and a second dosing cycle Antibody-drug conjugates and bispecific antibodies that bind to CD20 and CD3, wherein: (a) the first dosing cycle comprises: (i) a single dose of an anti-CD79b antibody-drug conjugate (C1D1); and (ii) bispecific A first dose (C1D1) of the anti-CD79b antibody and a second dose (C1D2) of the bispecific antibody, wherein C1D1 and C1D2 of the bispecific antibody are each administered to the individual after C1D1 of the anti-CD79b antibody drug conjugate, wherein the bispecific The C1D1 of the anti-CD79b antibody is about 2.5 mg, and the C1D2 of the bispecific antibody is about 10 mg; and (b) the second dosing cycle comprises: (i) a single dose (C2D1) of the anti-CD79b antibody drug conjugate; and (ii ) A single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is about 10 mg, about 16 mg or about 30 mg.

[0021] In another aspect, the invention features an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 for use in a method of treating an individual with a CD20-positive cell proliferative disorder, comprising: wherein the anti- The CD79b antibody-drug conjugate and the bispecific antibody that binds to CD20 and CD3 are administered to the individual with a dosing regimen that includes at least the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes: (i) a single dose of the anti-CD79b antibody drug conjugate (C1D1); and (ii) the first dose of the bispecific antibody (C1D1) and the second dose of the bispecific antibody (C1D2), wherein the bispecific antibody C1D1 and C1D2 are each to be administered to the individual after C1D1 of the anti-CD79b antibody drug conjugate, wherein C1D1 of the bispecific antibody is about 2.5 mg and C1D2 of the bispecific antibody is about 10 mg; and (b) a second dose The drug cycle includes: (i) a single dose (C2D1) of the anti-CD79b antibody drug conjugate; and (ii) a single dose (C2D1) of the bispecific antibody, wherein the C2D1 of the bispecific antibody is about 10 mg, about 16 mg or about 30 mg.

[0022] In another aspect, the present invention is characterized in that the anti-CD79b antibody-drug conjugate and the bispecific antibody that binds to CD20 and CD3 are used for treating individuals suffering from CD20-positive cell proliferative disorders, wherein the anti-CD79b antibody-drug conjugate and The bispecific antibody that binds to CD20 and CD3 is administered to the individual with a dosing regimen that includes at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle includes: (i) anti-CD79b antibody A single dose of the drug conjugate (C1D1); and (ii) a first dose of the bispecific antibody (C1D1) and a second dose of the bispecific antibody (C1D2), wherein C1D1 and C1D2 of the bispecific antibody are each Administering to the individual after C1D1 of the anti-CD79b antibody drug conjugate, wherein C1D1 of the bispecific antibody is about 2.5 mg and C1D2 of the bispecific antibody is about 10 mg; and (b) a second dosing cycle comprising: (i ) a single dose (C2D1) of an anti-CD79b antibody drug conjugate; and (ii) a single dose (C2D1) of a bispecific antibody, wherein the C2D1 of the bispecific antibody is about 10 mg, about 16 mg or about 30 mg.

[0023] In another aspect, the invention features anti-CD79b antibody-drug conjugates and the use of bispecific antibodies that bind to CD20 and CD3 in the manufacture of a medicament for treating individuals with CD20-positive cell proliferative disorders, wherein the anti-CD79b Antibody-drug conjugates and bispecific antibodies that bind to CD20 and CD3 are administered to individuals with a dosing regimen that includes at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle includes: ( i) a single dose of an anti-CD79b antibody drug conjugate (C1D1); and (ii) a first dose of a bispecific antibody (C1D1) and a second dose of a bispecific antibody (C1D2), wherein C1D1 of the bispecific antibody and C1D2 are each to be administered to the individual after C1D1 of the anti-CD79b antibody drug conjugate, wherein C1D1 of the bispecific antibody is about 2.5 mg and C1D2 of the bispecific antibody is about 10 mg; and (b) a second dose Cycles consist of: (i) a single dose (C2D1) of an anti-CD79b antibody drug conjugate; and (ii) a single dose (C2D1) of a bispecific antibody, wherein the C2D1 of the bispecific antibody is about 10 mg, about 16 mg, or About 30 mg.

[0024] In some embodiments, a single dose of anti-CD79b antibody drug conjugate C1D1 is about 0.1 mg / kg to about 2.4 mg / kg (e.g., about 0.1 mg / kg to about 2.2 mg / kg, about 0.1 mg / kg to about 2.0 mg / kg, about 0.5 mg / kg to about 2.2 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.2 mg / kg, about 1.2 mg / kg to about 2.2 mg / kg, about 1.4 mg / kg to about 2.2 mg / kg, about 1.6 mg / kg to about 2.2 mg / kg, about 1.8 mg / kg to about 2.0 mg / kg, about 0.1 mg / kg to about 1.6 mg / kg , about 0.5 mg / kg to about 1.6 mg / kg, or about 1 mg / kg to about 1.8 mg / kg; for example, about 1 mg / kg, about 1.2 mg / kg, about 1.6 mg / kg, or about 1.8 mg / kg), and the single dose C2D1 of the anti-CD79b antibody drug conjugate is about 0.1 mg / kg to about 2.4 mg / kg (for example, about 0.1 mg / kg to about 2.2 mg / kg, about 0.1 mg / kg to about 2.0 mg / kg, about 0.5 mg / kg to about 2.2 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.2 mg / kg, about 1.2 mg / kg to about 2.2 mg / kg kg, about 1.4 mg / kg to about 2.2 mg / kg, about 1.6 mg / kg to about 2.2 mg / kg, about 1.8 mg / kg to about 2.0 mg / kg, about 0.1 mg / kg to about 1.6 mg / kg, About 0.5 mg / kg to about 1.6 mg / kg, or about 1 mg / kg to about 1.8 mg / kg; for example, about 1 mg / kg, about 1.2 mg / kg, about 1.6 mg / kg, or about 1.8 mg / kg kg). In some embodiments, the single dose C1D1 of the anti-CD79b antibody drug conjugate is about 1.8 mg / kg, and the single dose C2D1 of the anti-CD79b antibody drug conjugate is about 1.8 mg / kg.

[0025] In some embodiments, C1D1 of the bispecific antibody and C1D2 of the bispecific antibody are or will be administered on or around day 8 (± 1 day) and day 15 (± 1 day) of the first administration cycle, respectively. 1 day) administered to individuals.

[0026] In some embodiments, C2D1 of the bispecific antibody is or will be administered to the individual on or about day 1 (± 1 day) of the second dosing cycle. In some embodiments, the C1D1 of the anti-CD79b antibody drug conjugate is or will be administered to the individual on or about day 2 (± 1 day) of the first dosing cycle, and the C2D1 of the anti-CD79b antibody drug conjugate is or will be Administer to individuals on or around Day 1 (± 1 day) of the second dosing cycle.

[0027] In some embodiments, the first and second dosing cycles are 14-day (eg, 14±3-day) dosing cycles. In some embodiments, the first and second dosing cycles are 21 day (eg, 21 ± 3 day) dosing cycles.

[0028] In some embodiments, the dosing regimen comprises one or more additional dosing cycles. In some embodiments, the dosing regimen comprises six to ten additional dosing cycles (e.g., six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles).

[0029] In some embodiments, the dosing regimen comprises ten additional dosing cycles. In some embodiments, the additional dosing cycle is a 21 day (eg, 21 ± 3 day) dosing cycle. In some embodiments, the dosing regimen comprises one or more additional dosing cycles. In some embodiments, the dosing regimen comprises six to ten additional dosing cycles. In some embodiments, the dosing regimen comprises ten additional dosing cycles.

[0030] In some embodiments, the additional dosing cycle is a 14 day (eg, 14 ± 3 day) dosing cycle.

[0031] In some embodiments, one or more of the additional dosing cycles comprises an additional single dose of the bispecific antibody and an additional single dose of the anti-CD79b antibody drug conjugate. In some embodiments, the additional single dose of the anti-CD79b antibody drug conjugate is about the same amount of C2D1 of the anti-CD79b antibody drug conjugate. In some embodiments, the additional single dose of the anti-CD79b antibody drug conjugate is or will be administered on or around (± 1 day) day 1 of each additional dosing cycle comprising the additional dose of the anti-CD79b antibody drug conjugate individual.

[0032] In some embodiments, the additional single dose of the bispecific antibody for each additional dosing cycle comprising the additional dose of the anti-CD79b antibody drug conjugate is or will be after completion of the administration of the additional single dose of the anti-CD79b antibody drug conjugate cast. In some embodiments, the additional single dose of the bispecific antibody for each additional dosing cycle comprising the additional dose of the anti-CD79b antibody drug conjugate is or will be after completion of the administration of the additional single dose of the anti-CD79b antibody drug conjugate Between about 60 and 120 minutes (e.g., between about 60 and 100 minutes, between about 60 and 90 minutes, between about 60 and 80 minutes, between about 90 and 120 minutes, between about 80 and 100 minutes, Between about 80 and 120 minutes, between about 75 and 105 minutes, or between about 85 and 95 minutes; e.g., about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes , about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes) administration. In some embodiments, the additional single dose of the bispecific antibody for each additional dosing cycle comprising the additional dose of the anti-CD79b antibody drug conjugate is or will be after completion of the administration of the additional single dose of the anti-CD79b antibody drug conjugate Administer for approximately 90 minutes.

[0033] In some embodiments, the dosing regimen comprises at least four additional dosing cycles comprising an additional single dose of the bispecific antibody and an additional single dose of the anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen comprises four to ten additional dosing cycles (e.g., four additional dosing cycles, five additional dosing cycles, six additional dosing cycles, seven additional dosing cycles, Eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles) comprising an additional single dose of the bispecific antibody and an additional single dose of the anti-CD79b antibody drug conjugate. In some embodiments, one or more of the additional dosing cycles comprises an additional single dose of the bispecific antibody and does not comprise the administration of an anti-CD79b antibody drug conjugate.

[0034] In some embodiments, the dosing regimen comprises at least two additional dosing cycles comprising additional single doses of the bispecific antibody and excluding administration of the anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen comprises between two and ten additional dosing cycles (e.g., two additional dosing cycles, three additional dosing cycles, four additional dosing cycles, five additional dosing cycles, six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles) that include both An additional single dose of specific antibody does not include administration of anti-CD79b antibody drug conjugate.

[0035] In some embodiments, the additional single dose of the bispecific antibody is about equivalent to the amount of C2D1 of the bispecific antibody. In some embodiments, the additional single dose of the bispecific antibody is about 30 mg. In some embodiments, the additional single dose of the bispecific antibody is or will be administered to the subject on or around (± 1 day) day 1 of each additional dosing cycle comprising the additional dose of the bispecific antibody.

[0036] In some embodiments, the dosing regimen comprises six or more additional dosing cycles (e.g., six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles or ten additional dosing cycles), each of the six or more additional dosing cycles comprises a single dose of the bispecific antibody, and no more than four of the six or more additional dosing cycles Administration of one (e.g., none, not more than one, not more than two, not more than three, or not more than four; e.g., none, one, two, three, or four) comprising an anti-CD79b antibody drug conjugate give. In some embodiments, the dosing regimen comprises six to ten additional dosing cycles (e.g., six additional dosing cycles, seven additional dosing cycles, eight additional dosing cycles, nine additional dosing cycles, or ten additional dosing cycles), each of six to ten additional dosing cycles comprises a single dose of the bispecific antibody, and no more than four of the six additional dosing cycles (e.g., none, No more than one, no more than two, no more than three, or no more than four; eg, none, one, two, three, or four) comprising administration of an anti-CD79b antibody drug conjugate. In some embodiments, the dosing regimen comprises ten additional dosing cycles, wherein each of the ten additional dosing cycles comprises a single dose of the bispecific antibody, and wherein no more than four of the ten additional dosing cycles Administration of one (e.g., none, not more than one, not more than two, not more than three, or not more than four; e.g., none, one, two, three, or four) comprising an anti-CD79b antibody drug conjugate give. In some embodiments, the dosing regimen comprises ten additional dosing cycles, wherein each of the ten additional dosing cycles comprises a single dose of the bispecific antibody, and wherein each of the ten additional dosing cycles comprises Administration of Anti-CD79b Antibody Drug Conjugate.

[0037] In one aspect, the invention features a method of treating an individual having a CD20-positive cell proliferative disorder comprising administering to the individual an anti-CD79b antibody drug conjugate and a CD20 antibody drug conjugate on a dosing regimen comprising 12 dosing cycles. and CD3-binding bispecific antibody, wherein: (a) the first administration cycle includes: (i) the first dose of the bispecific antibody (C1D1) and the second dose of the bispecific antibody (C1D2), wherein the bispecific antibody The C1D1 of the specific antibody is about 2.5 mg, and the C1D2 of the bispecific antibody is about 10 mg; and (ii) a single dose (C1D1) of the anti-CD79b antibody drug conjugate; (b) the second to sixth dosing cycles , each comprising a single dose of a bispecific antibody (C2D1 to C6D1) and a single dose of an anti-CD79b antibody drug conjugate (C2D1 to C6D1); and (c) seventh to twelfth dosing cycles, each comprising a bispecific A single dose of the bispecific antibody (C7D1 to C12D1) does not include the administration of the anti-CD79b antibody drug conjugate, and wherein each single dose of the bispecific antibody C2D1 to C12D1 is about 10 mg, about 16 mg or about 30 mg.

[0038] In another aspect, the invention features an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 for use in a method of treating an individual with a CD20-positive cell proliferative disorder, wherein the anti-CD79b antibody The drug conjugates and bispecific antibodies that bind to CD20 and CD3 are administered in a dosing regimen that includes 12 dosing cycles, wherein: (a) the first dosing cycle includes: (i) the first dose of the bispecific antibody dose (C1D1) and a second dose (C1D2) of the bispecific antibody, wherein the C1D1 of the bispecific antibody is about 2.5 mg and the C1D2 of the bispecific antibody is about 10 mg; and (ii) the anti-CD79b antibody drug-conjugated (C1D1); (b) second to sixth dosing cycles each comprising a single dose of the bispecific antibody (C2D1 to C6D1) and a single dose of the anti-CD79b antibody drug conjugate (C2D1 to C6D1) and (c) seventh through twelfth dosing cycles, each comprising a single dose of the bispecific antibody (C7D1 through C12D1) and excluding administration of the anti-CD79b antibody drug conjugate, and wherein the bispecific antibody Each single dose of C2D1 to C12D1 is about 10 mg, about 16 mg or about 30 mg.

[0039] In another aspect, the present invention is characterized by the use of anti-CD79b antibody-drug conjugates and bispecific antibodies that bind to CD20 and CD3 in the treatment of individuals with CD20-positive cell proliferative disorders, wherein the anti-CD79b antibody-drug conjugates and Bispecific antibodies that bind to CD20 and CD3 are administered in a dosing regimen that includes 12 dosing cycles, where: (a) the first dosing cycle includes: (i) the first dose of the bispecific antibody (C1D1) and a second dose (C1D2) of the bispecific antibody, wherein the C1D1 of the bispecific antibody is about 2.5 mg and the C1D2 of the bispecific antibody is about 10 mg; and (ii) a single dose of the anti-CD79b antibody drug conjugate (C1D1); (b) second to sixth dosing cycles, each comprising a single dose of the bispecific antibody (C2D1 to C6D1) and a single dose of the anti-CD79b antibody drug conjugate (C2D1 to C6D1); and (c ) Seventh to twelfth dosing cycles, each comprising a single dose of the bispecific antibody (C7D1 to C12D1) and excluding administration of the anti-CD79b antibody drug conjugate, and wherein each single dose of the bispecific antibody C2D1 To C12D1 is about 10 mg, about 16 mg or about 30 mg.

[0040] In another aspect, the invention features anti-CD79b antibody-drug conjugates and the use of bispecific antibodies that bind to CD20 and CD3 in the manufacture of a medicament for treating individuals with CD20-positive cell proliferative disorders, wherein the anti-CD79b Antibody drug conjugates and bispecific antibodies that bind to CD20 and CD3 are administered in a dosing regimen that includes 12 dosing cycles, wherein: (a) the first dosing cycle includes: (i) the second dose of the bispecific antibody A dose (C1D1) and a second dose (C1D2) of the bispecific antibody, wherein C1D1 of the bispecific antibody is about 2.5 mg and C1D2 of the bispecific antibody is about 10 mg; and (ii) an anti-CD79b antibody drug A single dose of the conjugate (C1D1); (b) second to sixth dosing cycles each comprising a single dose of the bispecific antibody (C2D1 to C6D1) and a single dose of the anti-CD79b antibody drug conjugate (C2D1 to C6D1 ); and (c) seventh through twelfth dosing cycles, each comprising a single dose of the bispecific antibody (C7D1 through C12D1) and excluding administration of the anti-CD79b antibody drug conjugate, and, wherein the bispecific antibody Each single dose of C2D1 to C12D1 is about 10 mg, about 16 mg or about 30 mg.

[0041] In some embodiments, the bispecific antibody has approximately equal amounts of C2D1 to C12D1. In some embodiments, the C2D1 of the bispecific antibody is about 30 mg.

[0042] In some embodiments, the anti-CD79b antibody drug conjugate has approximately equal amounts of C1D1 to C6D1. In some embodiments, each of C1D1 to C6D1 of the anti-CD79b antibody drug conjugate is about 0.1 mg / kg to about 2.4 mg / kg (e.g., about 0.1 mg / kg to about 2.2 mg / kg, about 0.1 mg / kg to about 2.0 mg / kg, about 0.5 mg / kg to about 2.2 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.2 mg / kg, about 1.2 mg / kg to about 2.2 mg / kg, about 1.4 mg / kg to about 2.2 mg / kg, about 1.6 mg / kg to about 2.2 mg / kg, about 1.8 mg / kg to about 2.0 mg / kg, about 0.1 mg / kg to about 1.6 mg / kg, about 0.5 mg / kg to about 1.6 mg / kg, or about 1 mg / kg to about 1.8 mg / kg; for example, about 1 mg / kg, about 1.2 mg / kg, about 1.6 mg / kg, or about 1.8 mg / kg). In some embodiments, each of C1D1 to C6D1 of the anti-CD79b antibody drug conjugate is about 1.8 mg / kg.

[0043] In some embodiments, C1D1 of the bispecific antibody and C1D2 of the bispecific antibody are or will be administered on or around day 8 (± 1 day) and day 15 (± 1 day) of the first administration cycle, respectively. 1 day) administered to individuals. In some embodiments, C2D1 to C12D1 of the bispecific antibody are or will be administered to the individual on or about Day 1 of each dosing cycle. In some embodiments, C1D1 of the anti-CD79b antibody drug conjugate is or will be administered to the individual on or about day 2 (± 1 day) of the first dosing cycle, and C2D1 to C6D1 of the anti-CD79b antibody drug conjugate Administration to the subject is or will be on or about Day 1 of each dosing cycle comprising administration of the anti-CD79b antibody drug conjugate.

[0044] In some embodiments, C2D1 to C6D1 of the bispecific antibody is or will be administered after administration of C2D1 to C6D1 of the anti-CD79b antibody drug conjugate is complete. In some embodiments, C2D1-C6D1 of the bispecific antibody is or will be between about 60 and 120 minutes (e.g., between about 60 and 100 minutes) after administration of C2D1-C6D1 of the anti-CD79b antibody drug conjugate is complete , between about 60 and 90 minutes, between about 60 and 80 minutes, between about 90 and 120 minutes, between about 80 and 100 minutes, between about 80 and 120 minutes, between about 75 and 105 minutes, or Between about 85 and 95 minutes; for example, about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes, about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or approximately 120 minutes) administered. In some embodiments, C2D1 to C6D1 of the bispecific antibody is or will be administered about 90 minutes after administration of C2D1 to C6D1 of the anti-CD79b antibody drug conjugate is complete.

[0045] In some embodiments, each dosing cycle is a 14 day (eg, 14 ± 3 day) dosing cycle. In some embodiments, each dosing cycle is a 21 day (eg, 21 ± 3 day) dosing cycle.

[0046] In some embodiments, the dosing regimen comprises an additional retreatment regimen after completion of 12 dosing cycles of the dosing regimen. In some embodiments, the additional retreatment regimen comprises 12 additional dosing cycles, wherein: (a) the first additional dosing cycle comprises: (i) the first dose of the bispecific antibody (C13D1 ) and the bispecific antibody The second dose (C13D2) of the bispecific antibody, wherein the C13D1 of the bispecific antibody is about 2.5 mg, and the C13D2 of the bispecific antibody is about 10 mg; and (ii) a single dose of the anti-CD79b antibody drug conjugate (C13D1); ( b) the second to sixth additional dosing cycles each comprising a single dose of the bispecific antibody (C14D1 to C18D1) and a single dose of the anti-CD79b antibody drug conjugate (C14D1 to C18D1); and (c) seventh to 12th The additional dosing cycles each comprise a single dose of the bispecific antibody (C19D1 to C24D1) and do not comprise administration of the anti-CD79b antibody drug conjugate, and wherein each single dose of the bispecific antibody C14D1 to C24D1 is about 10 mg, About 16 mg or about 30 mg. In some embodiments, (a) the first additional dosing cycle comprises: (i) the first dose of the bispecific antibody administered on or to be administered on day 8 (± 1 day) of the first additional dosing cycle (C13D1) and a second dose (C13D2) of the bispecific antibody administered or to be administered on day 15 (± 1 day) of the first additional dosing cycle, wherein C13D1 of the bispecific antibody is about 2.5 mg, and C13D2 of the bispecific antibody is about 10 mg; and (ii) a single dose of the anti-CD79b antibody drug conjugate administered on or to be administered on day 2 (± 1 day) of the first additional dosing cycle (C13D1) ; (b) the second to sixth additional dosing cycles each comprising a single dose of the bispecific antibody (C14D1 to C18D1) and a single dose of the anti-CD79b antibody drug conjugate (C14D1 to C18D1); and (c) seventh to The 12th additional dosing cycles each included a single dose of the bispecific antibody (C19D1 to C24D1) and did not include administration of the anti-CD79b antibody drug conjugate, and wherein C14D1 to C24D1 of the bispecific antibody was or would be at each is administered on Day 1 (± 1 day) of the additional dosing cycle, and C14D1 through C18D1 of the anti-CD79b antibody drug conjugate is or will be administered on Day 1 (± 1 day) of each additional dosing cycle, and , wherein each single dose of bispecific antibody C14D1 to C24D1 is about 30 mg, and each single dose of anti-CD79b antibody-drug conjugate C13D1 to C18D1 is about 1.8 mg / kg. In some embodiments, there is a waiting period between the completion of the 12 dosing cycles of the dosing regimen and the start of the 12 additional dosing cycles of the additional re-treatment regimen. In some embodiments, the waiting period is between about one week and about eight weeks. In some embodiments, each additional dosing cycle of the additional re-treatment regimen is a 14-day (eg, 14±3-day) dosing cycle. In some embodiments, each additional dosing cycle of the additional re-treatment regimen is a 21-day (eg, 21 ± 3-day) dosing cycle.

[0047] In some embodiments, the method further comprises administering to the individual one or more additional therapeutic agents. In some embodiments, anti-CD79b antibody drug conjugates and bispecific antibodies are administered with one or more additional therapeutic agents.

[0048] In some embodiments, the one or more additional therapeutic agents comprise one or more chemotherapeutic agents. In some embodiments, the one or more chemotherapeutic agents comprise cyclophosphamide, doxorubicin, and rituximab.

[0049] In some embodiments, the one or more additional therapeutic agents is tocilizumab. In some embodiments, the one or more additional therapeutic agents are corticosteroids. In some embodiments, the corticosteroid comprises prednisolone, prednisolone, methylprednisolone, and dexamethasone.

[0050] In some embodiments, the method further comprises administering to the individual rituximab, cyclophosphamide, doxorubicin, and prednisone (R-CHP). In some embodiments, anti-CD79b antibody drug conjugates and bispecific antibodies are administered with rituximab, cyclophosphamide, doxorubicin, and prednisone (R-CHP).

[0051] In some embodiments, the one or more additional therapeutic agents are antihistamines. In some embodiments, the antihistamine is diphenhydramine. In some embodiments, the one or more additional therapeutic agents comprise isopurinol and rasburicase. In some embodiments, the one or more additional therapeutic agents are antipyretics.

[0052] In some embodiments, the one or more additional therapeutic agents is obinutuzumab. In some embodiments, obinutuzumab is or will be administered prior to administration of the bispecific antibody. In some embodiments, obinutuzumab is or will be administered about seven days (± 1 day) prior to administration of the bispecific antibody. In some embodiments, obinutuzumab is or will be administered in a single dose of about 1000 mg. In some embodiments, obinutuzumab is administered at a first dose of about 1000 mg and a second dose of about 1000 mg. In some embodiments, the first dose of obinutuzumab is administered about seven days (± 1 day) prior to the C1D1 administration of the bispecific antibody. In some embodiments, the second dose of obinutuzumab is administered about one day (± 1 day) prior to the C1D1 administration of the bispecific antibody.

[0053] In some embodiments, the anti-CD79b antibody drug conjugate is polatuzumab vedotin or anti-CD79b-MC-vc-PAB-MMAE. In some embodiments, the anti-CD79b antibody drug conjugate is parotuzumab.

[0054] In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds to CD20, the Fab molecule comprising the following six hypervariable regions (HVR): (a) HVR-H1 comprising YSWIN (SEQ ID NO: the amino acid sequence of 1); (b) HVR-H2, it comprises the amino acid sequence of RIFPGDGDTDYNGKFKG (SEQ ID NO: 2); (c) HVR-H3, it comprises NVFDGYWLVY (SEQ ID NO: 3) (d) HVR-L1, which comprises the amino acid sequence of RSSKSLLHSNGITYLY (SEQ ID NO: 4); (e) HVR-L2, which comprises the amino acid sequence of QMSNLVS (SEQ ID NO: 5) sequence; and (f) HVR-L3, which comprises the amino acid sequence of AQNLELPYT (SEQ ID NO: 6). In some embodiments, the bispecific antibody comprises at least one Fab molecule specifically binding to CD20, the Fab molecule comprising (a) a heavy chain variable VH domain comprising the amino acid sequence of SEQ ID NO: 7 having At least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%; e.g., 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity Amino acid sequence; (b) a light chain variable (VL) domain comprising at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98% or at least 99%; for example, 95%, 96%, 97%, 98%, 99% or 100%) sequence identity of amino acid sequences; or (c) the VH domain as in (a) and Such as the VL domain in (b). In some embodiments, the Fab molecule that specifically binds to CD20 comprises: (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 7; and (b) a VL domain comprising the amino acid sequence of SEQ ID NO: 8 amino acid sequence.

[0055] In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds CD3, the Fab molecule comprising the following six HVRs: (a) HVR-H1 comprising the amine of TYAMN (SEQ ID NO: 9) (b) HVR-H2, which comprises the amino acid sequence of RIRSKYNNYATYYADSVKG (SEQ ID NO: 10); (c) HVR-H3, which comprises the amino acid sequence of HGNFGNSYVSWFAY (SEQ ID NO: 11); (d) HVR-L1, it comprises the amino acid sequence of GSSTGAVTTSNYAN (SEQ ID NO: 12); (e) HVR-L2, it comprises the amino acid sequence of GTNKRAP (SEQ ID NO: 13); And (f) HVR-L3, which comprises the amino acid sequence of ALWYSNLWV (SEQ ID NO: 14). In some embodiments, the bispecific antibody comprises at least one Fab molecule specifically binding to CD3, the Fab molecule comprising (a) a heavy chain variable VH domain comprising the amino acid sequence of SEQ ID NO: 15 having At least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%; e.g., 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity Amino acid sequence; (b) a light chain variable (VL) domain comprising at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98% or at least 99%; for example, 95%, 96%, 97%, 98%, 99% or 100%) sequence identity of amino acid sequences; or (c) the VH domain as in (a) and Such as the VL domain in (b). In some embodiments, the Fab molecule that specifically binds to CD3 comprises: (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 15; and (b) a VL domain comprising the amino acid sequence of SEQ ID NO: 16 amino acid sequence.

[0056] In some embodiments, the bispecific antibody comprises a Fab molecule that specifically binds CD3, wherein (a) the variable domains of the Fab heavy and light chains are swapped or (b) the constant domains of the Fab heavy and light chains are swapped exchange. In some embodiments, the bispecific antibody comprises at least one Fab molecule that specifically binds to CD20, wherein in the constant domain CL of the Fab molecule, the amino acid at position 124 is replaced by lysine (K) (according to Kabat numbering), and the amino acid at position 123 is substituted by arginine (R) or lysine (K) (according to Kabat numbering), and, in the constant domain CH1 of the Fab molecule, at position 147 The amino acid at position 213 is substituted by glutamic acid (E) (EU number) and the amino acid at position 213 is substituted by glutamic acid (EU number). In some embodiments, the bispecific antibody is bivalent to CD20 and monovalent to CD3. In some embodiments, the bispecific antibody comprises two Fab molecules that specifically bind CD20 and one Fab molecule that specifically binds CD3.

[0057] In some embodiments, the bispecific antibody comprises (a) a first Fab molecule that specifically binds CD20; (b) a second Fab molecule that specifically binds CD3; (c) a third Fab molecule that specifically binds CD20. Fab molecules; and (d) an Fc domain consisting of first and second units capable of stable association; wherein the third Fab molecule in (c) is the same as the first Fab molecule in (a); wherein in ( In the constant domain CL of the first Fab molecule in a) and the third Fab molecule in (c), the amino acid at position 124 was replaced by lysine (K) (according to Kabat numbering), and at position 123 where the amino acid is substituted by arginine (R) or lysine (K) (according to Kabat numbering); and, wherein the first Fab molecule in (a) and the third Fab molecule in (c) In the constant domain CH1, the amino acid at position 147 is substituted by glutamic acid (E) (EU numbering) and the amino acid at position 213 is substituted by glutamic acid (E) (EU numbering); and, wherein the first Fab molecule in (a) is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab in (b), and the second Fab molecule in (b) and (c) Each of the third Fab molecules is fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain in (d).

[0058] In some embodiments, bispecific antibodies are humanized antibodies. In some embodiments, bispecific antibodies are chimeric antibodies.

[0059] In some embodiments, the bispecific antibody comprises an Fc domain, wherein the Fc domain is an IgG Fc domain. In some embodiments, the IgG Fc domain is an IgG1 Fc domain. In some embodiments, the IgG Fc domain comprises a mutation at amino acid residue N297 (EU numbering) that results in loss of glycosylation. In some embodiments, the mutation at amino acid residue N297 is a substitution mutation. In some embodiments, the mutation at amino acid residue N297 reduces the effector function of the Fc region. In some embodiments, the mutation is a N297G or N297A mutation. In some embodiments, the bispecific antibody comprises substitution mutations in the Fc region that reduce 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 mutation is selected from the group consisting of: L234A, L235A, D265A, and P329G.

[0060] 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 (CH1 1 ) domain, a first CH2 (CH2 1 ) domain, A first CH3 (CH3 1 ) domain, a second CH1 (CH1 2 ) domain, a second CH2 (CH2 2 ) domain, and a second CH3 (CH3 2 ) 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 CH3 1 and CH3 2 domains each comprise a protuberance or cavity, wherein the protuberance or cavity in the CH3 1 domain is located within the cavity or protuberance of the CH3 2 domain, respectively. In some embodiments, the CH3 1 and CH3 2 domains meet at the interface between the protuberance and the lumen. In some embodiments, the CH2 1 and CH2 2 domains each comprise a protuberance or cavity, wherein the protuberance or cavity in the CH2 1 domain is located within the cavity or protuberance of the CH2 2 domain, respectively. In some embodiments, the CH2 1 domain and the CH2 2 domain meet at the interface between the protuberance and the lumen.

[0061] In some embodiments, the bispecific antibody is grafiduzumab.

[0062] In some embodiments, bispecific antibodies are administered or administered intravenously. In some embodiments, an anti-CD79b antibody drug conjugate is administered or administered intravenously. In some embodiments, if the bispecific antibody and the anti-CD79b antibody drug conjugate are or will be administered on the same day, the bispecific antibody is or will be administered after administration of the anti-CD79b antibody drug conjugate is complete . In some embodiments, the bispecific antibody is or will be between about 60 and 120 minutes (e.g., between about 60 and 100 minutes, between about 60 and 90 minutes) after administration of the anti-CD79b antibody drug conjugate is complete , between about 60 and 80 minutes, between about 90 and 120 minutes, between about 80 and 100 minutes, between about 80 and 120 minutes, between about 75 and 105 minutes, or between about 85 and 95 minutes; For example, about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes, about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes) of administration. In some embodiments, the bispecific antibody is or will be administered about 90 minutes after administration of the anti-CD79b antibody drug conjugate is complete.

[0063] In one aspect, the invention features a method of treating a subject with a CD20-positive cell proliferative disorder comprising administering to the subject Pascal with a dosing regimen comprising at least a first dosing cycle and a second dosing cycle Rotuzumab and gefitumumab, wherein: (a) the first dosing cycle includes the first dose of gefitumumab administered on the 8th day (± 1 day) of the first dosing cycle (C1D1) and the second dose (C1D2) of grafiduzumab administered on the 15th day (± 1 day) of the first dosing cycle, wherein the C1D1 of grafiduzumab is about 2.5 mg. The C1D2 of grafitumumab was about 10 mg; (b) the second dosing cycle consisted of a single dose of grafitumumab administered on day 1 (± 1 day) of the second dosing cycle (C2D1) and A single dose (C2D1) of parotuzumab administered on day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of grafituzumab is about 10 mg, about 16 mg or about 30 mg .

[0064] In another aspect, the invention features parotuzumab and grifituzumab for use in a method of treating an individual with a CD20-positive cell proliferative disorder, wherein parotuzumab and grifituzumab It is administered with a dosing regimen that includes at least the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes the dose that will be administered on the 8th day (± 1 day) of the first dosing cycle The first dose of grafitumumab (C1D1) and the second dose of grafitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle, where The C1D1 of Fetuzumab is about 2.5 mg, and the C1D2 of Fetuzumab is about 10 mg; (b) The second dosing cycle includes administration on the first day (± 1 day) of the second dosing cycle A single dose of grafiduzumab (C2D1) and a single dose of parotuzumab (C2D1) to be administered on day 1 (± 1 day) of the second dosing cycle, in which grafiduzumab The anti-C2D1 is about 10 mg, about 16 mg or about 30 mg.

[0065] In another aspect, the invention features the use of parotuzumab and grifituzumab for the treatment of an individual suffering from a proliferative disorder of CD20 positive cells, wherein parotuzumab and grifituzumab are formulated in the form of A dosing regimen that includes at least the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes the drug that will be administered on the 8th day (± 1 day) of the first dosing cycle. The first dose of grafitumumab (C1D1) and the second dose of grafitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle, in which grafitumumab The C1D1 of the monoclonal antibody is about 2.5 mg, and the C1D2 of the grafiduzumab is about 10 mg; (b) The second dosing cycle includes the dose that will be administered on the first day (± 1 day) of the second dosing cycle A single dose of Fetuzumab (C2D1) and a single dose of Parotuzumab (C2D1) to be administered on Day 1 (± 1 day) of the second dosing cycle, of which Fetuzumab C2D1 is about 10 mg, about 16 mg, or about 30 mg.

[0066] In another aspect, the invention features the use of parotuzumab and grafitumumab in the manufacture of a medicament for treating an individual with a CD20-positive cell proliferative disorder, wherein parotuzumab and grafitumumab are Tomumab is administered with a dosing regimen that includes at least the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes the 8th day (± 1 day) of the first dosing cycle. The first dose of grafitumumab (C1D1) to be administered on day 1) and the second dose of grafitumumab to be administered on day 15 (± 1 day) of the first dosing cycle (C1D2) , wherein the C1D1 of grafitumumab is about 2.5 mg, and the C1D2 of grafitumumab is about 10 mg; (b) the second dosing cycle includes the first day (± 1 day) of the second dosing cycle ) single dose of graftuzumab administered (C2D1) and single dose of parotuzumab administered on day 1 (± 1 day) of the second dosing cycle (C2D1), where The C2D1 of fituzumab is about 10 mg, about 16 mg, or about 30 mg.

[0067] In one aspect, the invention features a method of treating a subject with a CD20-positive cell proliferative disorder comprising administering to the subject Pascal with a dosing regimen comprising at least a first dosing cycle and a second dosing cycle Rotuzumab and grafituzumab, wherein: (a) the first dosing cycle includes: (i) Parotuzumab administered on the second day (± 1 day) of the first dosing cycle and (ii) the first dose of grafitumumab administered on day 8 (± 1 day) of the first dosing cycle (C1D1) and A second dose of grafitumumab (C1D2) administered on day 15 (± 1 day), wherein C1D1 of grafitumumab is about 2.5 mg and C1D2 of grafitumumab is about 10 mg; and ( b) The second dosing cycle comprises: (i) a single dose of Parotuzumab administered on Day 1 (± 1 day) of the second dosing cycle (C2D1); A single dose (C2D1) of grafitumumab administered on day 1 (± 1 day) of the dosing cycle, wherein the C2D1 of grafitumumab is about 10 mg, about 16 mg or about 30 mg, and Pa C1D1 and C2D1 of rotuzumab were each about 1.8 mg / kg.

[0068] In another aspect, the invention features parotuzumab and grifituzumab for use in a method of treating an individual with a CD20-positive cell proliferative disorder, wherein parotuzumab and grifituzumab It is administered with a dosing regimen comprising at least the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes: (i) will be administered on the second day of the first dosing cycle (± A single dose of parotuzumab (C1D1) administered on day 1); and (ii) the first dose of grafitumumab to be administered on day 8 (± 1 day) of the first dosing cycle dose (C1D1) and the second dose (C1D2) of grafitumumab administered on the 15th day (± 1 day) of the first dosing cycle, wherein the C1D1 of grafitumumab is about 2.5 mg, The C1D2 of grafitumumab is approximately 10 mg; and (b) the second dosing cycle comprises: (i) Parotuzumab to be administered on Day 1 (± 1 day) of the second dosing cycle (C2D1); and (ii) a single dose of grafitumumab to be administered on day 1 (± 1 day) of the second dosing cycle (C2D1), wherein C2D1 was about 10 mg, about 16 mg, or about 30 mg, and each of Parotuzumab's C1D1 and C2D1 was about 1.8 mg / kg.

[0069] In another aspect, the invention features the use of parotuzumab and grifituzumab in the treatment of an individual with a proliferative disorder of CD20 positive cells, wherein parotuzumab and grifituzumab are formulated with At least include the administration of the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes: (i) will be administered on the second day (± 1 day) of the first dosing cycle ) a single dose of parotuzumab (C1D1); and (ii) the first dose of graftuzumab to be administered on day 8 (± 1 day) of the first dosing cycle ( C1D1) and the second dose of Griffithumab (C1D2) to be administered on the 15th day (± 1 day) of the first dosing cycle, wherein the C1D1 of Griffithumab is about 2.5 mg, Griffithumab The C1D2 of Parotuzumab is approximately 10 mg; and (b) the second dosing cycle comprises: (i) the C1D2 of Parotuzumab to be administered on Day 1 (± 1 Day) of the second dosing cycle A single dose (C2D1); and (ii) a single dose of grafitumumab (C2D1) to be administered on day 1 (± 1 day) of the second dosing cycle, wherein C2D1 of grafitumumab is About 10 mg, about 16 mg, or about 30 mg, and the C1D1 and C2D1 of Parotuzumab are each about 1.8 mg / kg.

[0070] In another aspect, the invention features the use of parotuzumab and grafitumumab in the manufacture of a medicament for treating an individual with a CD20-positive cell proliferative disorder, wherein parotuzumab and grafitumumab are Tomumab is administered with a dosing regimen that includes at least the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes: (i) will be administered on the second day of the first dosing cycle (± 1 day) of the single dose of parotuzumab administered (C1D1); The first dose (C1D1) and the second dose (C1D2) of grafitumumab to be administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of grafitumumab is about 2.5 mg, the C1D2 of grafitumumab is approximately 10 mg; and (b) the second dosing cycle includes: (i) Palotol to be administered on Day 1 (± 1 day) of the second dosing cycle A single dose of zizumab (C2D1); and (ii) a single dose of grafidumab to be administered on day 1 (± 1 day) of the second dosing cycle (C2D1), wherein grafidumab The C2D1 of the antibody was about 10 mg, about 16 mg, or about 30 mg, and the C1D1 and C2D1 of parotuzumab were each about 1.8 mg / kg.

[0071] In one aspect, the invention features a method of treating a subject with a CD20-positive cell proliferative disorder comprising administering to the subject parotuzumab and grafidox on a dosing regimen comprising 12 dosing cycles Monoclonal antibody, wherein: (a) the first dosing cycle includes the first dose (C1D1) of grafitumumab administered on the 8th day (± 1 day) of the first dosing cycle, Second dose of grafiduzumab (C1D2) administered on cycle day 15 (± 1 day) and parotuzumab administered on day 2 (± 1 day) of the first dosing cycle A single dose (C1D1), wherein the C1D1 of grafitumumab is between about 1 mg to about 5 mg, and the C1D2 of grafitumumab is about 10 mg; (b) the second to sixth doses Each drug cycle includes a single dose of grafiduzumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), wherein the C2D1 of grafiduzumab is about 10 mg, about 16 mg or approximately 30 mg; and (c) the seventh to twelfth dosing cycles each included a single dose of grafiduzumab (C7D1 to C12D1) and did not include the administration of parotuzumab, and, wherein grafiduzumab Each single dose of mAb C2D1 to C12D1 was administered on Day 1 (± 1 day) of each dosing cycle, and each single dose of Parotuzumab C2D1 to C6D1 was administered on Day 1 of each dosing cycle. (± 1 day), and wherein each single dose of grafiduzumab C3D1 to C12D1 is approximately equivalent to the amount of C2D1 of grafiduzumab, and each single dose of parotuzumab C1D1 to C6D1 is about 1.8 mg / kg.

[0072] In another aspect, the invention features parotuzumab and grifituzumab for use in a method of treating an individual with a CD20-positive cell proliferative disorder, wherein parotuzumab and grifituzumab Administered with a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle includes the dose of grafitumumab to be administered on day 8 (± 1 day) of the first dosing cycle The first dose (C1D1), the second dose of grafitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle and the second dose (C1D2) to be administered on day 2 of the first dosing cycle A single dose (C1D1) of Parotuzumab administered (± 1 day), wherein the C1D1 of grafiduzumab is about 2.5 mg, and the C1D2 of grafiduzumab is about 10 mg; (b) The second to sixth dosing cycles each comprised a single dose of grafiduzumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), wherein the C2D1 of grafiduzumab was about 10 mg, about 16 mg, or about 30 mg; and (c) the seventh through twelfth dosing cycles each comprised a single dose of grafitumumab (C7D1 through C12D1) and did not include the administration of parotuzumab, And, each single dose of C2D1 to C12D1 of grafiduzumab will be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of C2D1 to C6D1 of parotuzumab will be administered on each The first day (± 1 day) of the dosing cycle is administered, and each single dose of C3D1 to C12D1 of grafiduzumab is approximately equivalent to the C2D1 of grafiduzumab, and the amount of parotuzumab Each single dose of C1D1 to C6D1 was approximately 1.8 mg / kg.

[0073] In another aspect, the invention features the use of parotuzumab and grifituzumab in the treatment of an individual with a proliferative disorder of CD20-positive cells, wherein parotuzumab and grifituzumab are formulated in the form of A dosing regimen of 12 dosing cycles, where: (a) the first dosing cycle included the first dose of grafitumumab to be administered on Day 8 (± 1 day) of the first dosing cycle. Dose (C1D1), a second dose of grafitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle and a second dose (C1D2) to be administered on day 2 (± 1 day) a single dose (C1D1) of parotuzumab administered, wherein the C1D1 of grafiduzumab was about 2.5 mg, and the C1D2 of grafiduzumab was about 10 mg; (b) the second Each of the to sixth dosing cycles comprises a single dose of grafiduzumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), wherein the C2D1 of grafiduzumab is about 10 mg, about 16 mg or about 30 mg; and (c) seventh through twelfth dosing cycles each comprising a single dose of grafitumumab (C7D1 through C12D1) and excluding administration of parotuzumab, and, Each single dose C2D1 to C12D1 of grafiduzumab will be administered on the first day (± 1 day) of each dosing cycle, and each single dose C2D1 to C6D1 of parotuzumab will be administered on each dosing cycle. The first day (± 1 day) of the cycle was administered, and each single dose of C3D1 to C12D1 of grafiduzumab was approximately equivalent to C2D1 of grafiduzumab, and each single dose of parotuzumab The dose for C1D1 to C6D1 is about 1.8 mg / kg.

[0074] In another aspect, the invention features the use of parotuzumab and grafitumumab in the manufacture of a medicament for treating an individual with a CD20-positive cell proliferative disorder, wherein parotuzumab and grafitumumab are Tomumab was administered in a dosing regimen consisting of 12 dosing cycles, where: (a) the first dosing cycle included grafidox to be administered on day 8 (± 1 day) of the first dosing cycle The first dose of monoclonal antibody (C1D1), the second dose of gfitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle and A single dose (C1D1) of Parotuzumab administered on day 2 (± 1 day), wherein the C1D1 of grafitumumab is about 2.5 mg, and the C1D2 of grafitumumab is about 10 mg; (b) The second to sixth dosing cycles each include a single dose of grafiduzumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), of which C2D1 of grafiduzumab is about 10 mg, about 16 mg, or about 30 mg; and (c) each of the seventh to twelfth dosing cycles includes a single dose of grafiduzumab (C7D1 to C12D1) and does not include parotuzumab administration, and wherein each single dose of grafituzumab C2D1 to C12D1 will be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of parotuzumab C2D1 to C6D1 It will be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of grafitumumab C3D1 to C12D1 is approximately equivalent to the amount of C2D1 of grafitumumab, and Parotrolizumab Each single dose of mAbs C1D1 to C6D1 was approximately 1.8 mg / kg.

[0075] In one aspect, the invention features a method of treating a subject with a CD20-positive cell proliferative disorder comprising administering to the subject parotuzumab and grafidox on a dosing regimen comprising 12 dosing cycles Monoclonal antibody, wherein: (a) the first dosing cycle includes: (i) the first dose of grafitumumab (C1D1) administered on day 8 (± 1 day) of the first dosing cycle and The second dose of grafitumumab (C1D2) administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of grafitumumab is about 2.5 mg and the C1D2 of grafitumumab is about 10 mg; and (ii) a single dose of parotuzumab administered on day 2 (± 1 day) of the first dosing cycle (C1D1); (b) the second to sixth doses Cycles each consisted of a single dose of grafiduzumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), wherein the C2D1 of grafiduzumab was about 10 mg, about 16 mg, or about 30 mg; and (c) Cycles 7 through 12 each included a single dose of grafiduzumab (C7D1 to C12D1) and did not include the administration of parotuzumab, and, where grafiduzumab Each single dose of parotuzumab C2D1 to C12D1 was administered on day 1 (± 1 day) of each dosing cycle, and each single dose of parotuzumab C2D1 to C6D1 was administered on day 1 of each dosing cycle (± 1 day) administration, and wherein each single dose C3D1 to C12D1 of grafiduzumab is approximately equivalent to C2D1 of grafiduzumab, and each single dose C1D1 to C6D1 of parotuzumab is About 1.8 mg / kg.

[0076] In another aspect, the invention features parotuzumab and grifituzumab for use in a method of treating an individual with a CD20-positive cell proliferative disorder, wherein parotuzumab and grifituzumab Administered with a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) Griffin to be administered on day 8 (± 1 day) of the first dosing cycle The first dose of gfitumumab (C1D1) and the second dose of gfitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle, of which gfitumumab C1D1 of approximately 2.5 mg and C1D2 of grafitumumab approximately 10 mg; and (ii) a single dose of parotuzumab to be administered on day 2 (± 1 day) of the first dosing cycle (C1D1); (b) The second to sixth dosing cycles each include a single dose of grafiduzumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), of which The C2D1 of the mAb is about 10 mg, about 16 mg, or about 30 mg; and (c) the seventh to twelfth dosing cycles each include a single dose of grafitumumab (C7D1 to C12D1) and do not include Pareto The administration of zizumab, and each single dose of grafituzumab C2D1 to C12D1 will be administered on the first day (± 1 day) of each dosing cycle, and each single dose of parotuzumab Doses C2D1 through C6D1 will be administered on Day 1 (± 1 day) of each dosing cycle, and wherein each single dose of grafitumumab C3D1 through C12D1 is approximately equivalent to C2D1 of grafitumumab, and Each single dose of Parotuzumab C1D1 to C6D1 was approximately 1.8 mg / kg.

[0077] In another aspect, the invention features the use of parotuzumab and grifituzumab in the treatment of an individual with a proliferative disorder of CD20-positive cells, wherein parotuzumab and grifituzumab are formulated in the form of A dosing regimen of 12 dosing cycles, wherein: (a) the first dosing cycle includes: (i) Griffithram to be administered on day 8 (± 1 day) of the first dosing cycle The first dose (C1D1) of the antibody and the second dose (C1D2) of the grafiduzumab to be administered on the 15th day (± 1 day) of the first dosing cycle, wherein the C1D1 of the grafiduzumab is approximately 2.5 mg and C1D2 of grafiduzumab approximately 10 mg; and (ii) a single dose of parotuzumab (C1D1) to be administered on day 2 (± 1 day) of the first dosing cycle ); (b) The second to sixth dosing cycles each included a single dose of grafitumumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), wherein grafitumumab The C2D1 is about 10 mg, about 16 mg, or about 30 mg; and (c) each of the seventh to twelfth dosing cycles includes a single dose of grafitumumab (C7D1 to C12D1) and does not include parotuzumab Antibody administration, and wherein each single dose of grafituzumab C2D1 to C12D1 will be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of parotuzumab C2D1 C6D1 to C6D1 will be administered on day 1 (± 1 day) of each dosing cycle, and wherein each single dose of grafiduzumab C3D1 to C12D1 is approximately equivalent to C2D1 of grafiduzumab, and Parot Each single dose of tocilizumab C1D1 to C6D1 was approximately 1.8 mg / kg.

[0078] In another aspect, the invention features the use of parotuzumab and grafitumumab in the manufacture of a medicament for treating an individual with a CD20-positive cell proliferative disorder, wherein parotuzumab and grafitumumab are Tomumab was administered in a dosing regimen comprising 12 dosing cycles, where: (a) the first dosing cycle consisted of: (i) will be administered on day 8 (± 1 day) of the first dosing cycle The first dose of grafitumumab (C1D1) and the second dose of grafitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle, wherein The C1D1 of Mab is approximately 2.5 mg and the C1D2 of Griffithumab is approximately 10 mg; and (ii) Parotuzumab to be administered on Day 2 (± 1 Day) of the first dosing cycle (C1D1); (b) The second to sixth dosing cycles each include a single dose of grafitumumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), wherein C2D1 of grafitumumab is about 10 mg, about 16 mg, or about 30 mg; and (c) the seventh to twelfth dosing cycles each include a single dose of grafitumumab (C7D1 to C12D1) and do not include Administration of Parotuzumab, and where each single dose of Griffithumab C2D1 to C12D1 will be administered on Day 1 (± 1 Day) of each dosing cycle, and Parotuzumab Each single dose of C2D1 to C6D1 will be administered on the first day (± 1 day) of each dosing cycle, and wherein each single dose of grafitumumab C3D1 to C12D1 is approximately equivalent to C2D1, and Palotol Each single dose of zizumab C1D1 to C6D1 was about 1.8 mg / kg.

[0079] In one aspect, the invention features a method of treating a subject with a CD20-positive cell proliferative disorder comprising administering to the subject parotuzumab and grafidox on a dosing regimen comprising 12 dosing cycles Monoclonal antibody, wherein: (a) the first dosing cycle includes: (i) the first dose of grafitumumab (C1D1) administered on day 8 (± 1 day) of the first dosing cycle and The second dose of grafitumumab (C1D2) administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of grafitumumab is about 2.5 mg and the C1D2 of grafitumumab is about 10 mg; and (ii) a single dose of parotuzumab administered on day 2 (± 1 day) of the first dosing cycle (C1D1); (b) the second to sixth doses Cycles each consisted of a single dose of grafiduzumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), where C2D1 of grafiduzumab was approximately 30 mg; and (c) Dosing cycles 7 to 12 each included a single dose of grafiduzumab (C7D1 to C12D1) and did not include the administration of parotuzumab, and, where each single dose of grafiduzumab C2D1 to C12D1 was administered on Day 1 (± 1 day) of each dosing cycle, and each single dose of Parotuzumab C2D1 to C6D1 was administered on Day 1 (± 1 day) of each dosing cycle, And, wherein each single dose of C3D1 to C12D1 of grafiduzumab is about the same amount as C2D1 of grafiduzumab, and each single dose of parotuzumab from C1D1 to C6D1 is about 1.8 mg / kg.

[0080] In another aspect, the invention features parotuzumab and grifituzumab for use in a method of treating an individual with a CD20-positive cell proliferative disorder, wherein parotuzumab and grifituzumab Administered with a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) Griffin to be administered on day 8 (± 1 day) of the first dosing cycle The first dose of gfitumumab (C1D1) and the second dose of gfitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle, of which gfitumumab C1D1 of approximately 2.5 mg and C1D2 of grafitumumab approximately 10 mg; and (ii) a single dose of parotuzumab to be administered on day 2 (± 1 day) of the first dosing cycle (C1D1); (b) The second to sixth dosing cycles each include a single dose of grafiduzumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), of which The C2D1 of the mAb was about 30 mg; and (c) the seventh through 12 dosing cycles each included a single dose of grafitumumab (C7D1 through C12D1) and did not include the administration of parotuzumab, and , wherein each single dose of grafitumumab C2D1 to C12D1 will be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of parotuzumab C2D1 to C6D1 will be administered on each given day The first day (± 1 day) of the drug cycle was administered, and each single dose of C3D1 to C12D1 of grafiduzumab was approximately equivalent to C2D1 of grafiduzumab, and each dose of parotuzumab A single dose of C1D1 to C6D1 is approximately 1.8 mg / kg.

[0081] In another aspect, the invention features the use of parotuzumab and grifituzumab in the treatment of an individual with a proliferative disorder of CD20-positive cells, wherein parotuzumab and grifituzumab are formulated in the form of A dosing regimen of 12 dosing cycles, wherein: (a) the first dosing cycle includes: (i) Griffithram to be administered on day 8 (± 1 day) of the first dosing cycle The first dose (C1D1) of the antibody and the second dose (C1D2) of the grafitumumab to be administered on the 15th day (± 1 day) of the first dosing cycle, wherein the C1D1 of the grafitumumab is approximately 2.5 mg and C1D2 of grafiduzumab approximately 10 mg; and (ii) a single dose of parotuzumab (C1D1) to be administered on day 2 (± 1 day) of the first dosing cycle ); (b) The second to sixth dosing cycles each included a single dose of grafitumumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), wherein grafitumumab The C2D1 is about 30 mg; and (c) the seventh to twelfth dosing cycles each include a single dose of grafiduzumab (C7D1 to C12D1) and do not include the administration of parotuzumab, and, wherein Each single dose of grafitumumab C2D1 to C12D1 will be administered on Day 1 (± 1 day) of each dosing cycle, and each single dose of parotuzumab C2D1 to C6D1 will be administered on each dosing cycle administered on the first day (± 1 day), and wherein each single dose of grafiduzumab C3D1 to C12D1 is approximately equivalent to the amount of C2D1 of grafiduzumab, and each single dose of parotuzumab C1D1 to C6D1 is about 1.8 mg / kg.

[0082] In another aspect, the invention features the use of parotuzumab and grafitumumab in the manufacture of a medicament for treating an individual with a CD20-positive cell proliferative disorder, wherein parotuzumab and grafitumumab are Tomumab was administered in a dosing regimen comprising 12 dosing cycles, where: (a) the first dosing cycle consisted of: (i) will be administered on day 8 (± 1 day) of the first dosing cycle The first dose of grafitumumab (C1D1) and the second dose of grafitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle, wherein The C1D1 of Mab is approximately 2.5 mg and the C1D2 of Griffithumab is approximately 10 mg; and (ii) Parotuzumab to be administered on Day 2 (± 1 Day) of the first dosing cycle (C1D1); (b) The second to sixth dosing cycles each include a single dose of grafitumumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1), wherein C2D1 of grafitumumab was approximately 30 mg; and (c) dosing cycles 7 through 12 each included a single dose of grafitumumab (C7D1 to C12D1) and did not include an administration of parotuzumab and, wherein each single dose of grafituzumab C2D1 to C12D1 will be administered on day 1 (± 1 day) of each dosing cycle, and each single dose of parotuzumab C2D1 to C6D1 will be administered Administered on the first day (± 1 day) of each dosing cycle, and each single dose of grifituzumab C3D1 to C12D1 is approximately equivalent to C2D1, and each single dose of parotuzumab C1D1 To C6D1 is about 1.8 mg / kg.

[0083] In some embodiments, the dosing cycle is a 14 day (eg, 14 ± 3 day) dosing cycle. In some embodiments, the dosing cycle is a 21 day (eg, 21 ± 3 day) dosing cycle.

[0084] In some embodiments, grafitumumab is administered intravenously. In some embodiments, parotuzumab is administered intravenously. In some embodiments, if grafituzumab is or will be administered on the same day as parotuzumab, then grafituzumab is or will be administered after completion of parotuzumab administration cast. In some embodiments, grafiduzumab is or will be administered between about 60 and 120 minutes (e.g., between about 60 and 100 minutes, between about 60 and 90 minutes) after administration of parotuzumab is complete between about 60 to 80 minutes, about 90 to 120 minutes, about 80 to 100 minutes, about 80 to 120 minutes, about 75 to 105 minutes, or about 85 to 95 minutes ; for example, about 60 minutes, about 70 minutes, about 80 minutes, about 85 minutes, about 88 minutes, about 90 minutes, about 92 minutes, about 95 minutes, about 100 minutes, about 110 minutes, or about 120 minutes) . In some embodiments, grafitumumab is or will be administered about 90 minutes after administration of parotuzumab is complete.

[0085] In some embodiments, the method further comprises administering obinutuzumab to the subject. In some embodiments, parotuzumab and grafituzumab are administered with obinutuzumab. In some embodiments, obinutuzumab is or will be administered prior to administration of grafiduzumab. In some embodiments, obinutuzumab is or will be administered about seven days (± 1 day) prior to administration of grafiduzumab. In some embodiments, obinutuzumab is or will be administered in a single dose of about 1000 mg.

[0086] In some embodiments, the proliferative disorder of CD20 positive cells is a proliferative disorder of B cells. In some embodiments, the B cell proliferative disorder is non-Hodgkin's lymphoma (NHL) or central nervous system lymphoma (CNSL). In some embodiments, NHL is relapsed and / or refractory. In some embodiments, NHL is diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), high-grade B-cell lymphoma primary mediastinal (thymic) large B-cell lymphoma (PMLBCL), diffuse B-cell lymphoma, or small lymphocytic lymphoma.

[0087] In some embodiments, NHL is DLBCL. In some embodiments, DLBCL is recurrent or metastatic DLBCL.

[0088] In some embodiments, NHL is FL. In some embodiments, FL is recurrent or metastatic FL. In some embodiments, FL is converted FL.

[0089] In some embodiments, the NHL is the MCL. In some embodiments, the MCL is recurrent or metastatic MCL.

[0090] In some embodiments, the CD20-positive cell proliferative disorder is not chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), Richter's transformation, Burch's lymphoma, or lymphoplasmacytic lymphoma.

[0091] In one aspect, the invention features a method of treating a population of individuals with R / R NHL comprising administering to the individual an anti-CD79b antibody on a dosing regimen comprising at least a first dosing cycle and a second dosing cycle Drug conjugates and bispecific antibodies that bind to CD20 and CD3, wherein: (a) the first administration cycle includes the second dose of the bispecific antibody administered on day 8 (± 1 day) of the first administration cycle. One dose (C1D1), a second dose of the bispecific antibody (C1D2) administered on day 15 (± 1 day) of the first dosing cycle and a second dose (C1D2) on day 2 (± 1 day) of the first dosing cycle ) a single dose (C1D1) of Parotuzumab administered, wherein C1D1 of the bispecific antibody is about 2.5 mg and C1D2 of the bispecific antibody is about 10 mg; and (b) a second dosing cycle Consisting of a single dose of bispecific antibody (C2D1) administered on day 1 (± 1 day) of the second dosing cycle and Parol administered on day 1 (± 1 day) of the second dosing cycle A single dose (C2D1) of tocilizumab, wherein C2D1 of the bispecific antibody is about 30 mg, and wherein C1D1 and C1D2 of the anti-CD79b antibody drug conjugate are each about 1.8 mg / kg.

[0092] In another aspect, the invention features an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 for use in a method of treating a population of individuals with R / R NHL, wherein the anti-CD79b antibody drug The conjugate and the bispecific antibody that binds to CD20 and CD3 are administered with a dosing regimen that includes at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle includes The first dose of the bispecific antibody (C1D1) administered on the 8th day (± 1 day) of the drug cycle, the dose of the bispecific antibody administered on the 15th day (± 1 day) of the first drug cycle A second dose (C1D2) and a single dose of Parotuzumab (C1D1) to be administered on day 2 (± 1 day) of the first dosing cycle, wherein C1D1 of the bispecific antibody is approximately 2.5 mg , and the C1D2 of the bispecific antibody is about 10 mg; and (b) the second dosing cycle comprises a single dose of the bispecific antibody to be administered on day 1 (± 1 day) of the second dosing cycle ( C2D1 ) and a single dose of parotuzumab (C2D1) to be administered on day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of the bispecific antibody is about 30 mg, and, wherein C1D1 and C1D2 of the anti-CD79b antibody drug conjugate were each about 1.8 mg / kg.

[0093] In another aspect, the present invention is characterized in the use of anti-CD79b antibody-drug conjugates and bispecific antibodies binding to CD20 and CD3 for treating a population of individuals with R / R NHL, wherein the anti-CD79b antibody-drug conjugates and The CD20- and CD3-binding bispecific antibody is administered with a dosing regimen that includes at least the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes the second dosing cycle to be administered in the first dosing cycle. The first dose of the bispecific antibody (C1D1) administered on day 8 (± 1 day), the second dose of the bispecific antibody to be administered on day 15 (± 1 day) of the first dosing cycle ( C1D2) and a single dose of parotuzumab (C1D1) to be administered on the second day (± 1 day) of the first dosing cycle, wherein the C1D1 of the bispecific antibody is about 2.5 mg, and the bispecific The C1D2 of the specific antibody is about 10 mg; and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody to be administered on day 1 (± 1 day) of the second dosing cycle and the A single dose (C2D1) of Parotuzumab administered on day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of the bispecific antibody is about 30 mg, and, wherein the anti-CD79b antibody drug The C1D1 and C1D2 of the conjugate were each about 1.8 mg / kg.

[0094] In another aspect, the invention features the use of anti-CD79b antibody-drug conjugates and bispecific antibodies that bind to CD20 and CD3 in the manufacture of a medicament for treating a population of individuals with R / R NHL, wherein the anti-CD79b antibody The drug conjugate and the bispecific antibody binding to CD20 and CD3 are administered with a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) the first dosing cycle includes The first dose of the bispecific antibody (C1D1) administered on day 8 (± 1 day) of the dosing cycle, the bispecific antibody to be administered on day 15 (± 1 day) of the first dosing cycle The second dose (C1D2) of the bispecific antibody and the single dose (C1D1) of parotuzumab to be administered on the second day (± 1 day) of the first dosing cycle, wherein the C1D1 of the bispecific antibody is about 2.5 mg, and the C1D2 of the bispecific antibody is about 10 mg; and (b) the second dosing cycle comprises a single dose of the bispecific antibody to be administered on day 1 (± 1 day) of the second dosing cycle (C2D1) and a single dose of Parotuzumab (C2D1) to be administered on Day 1 (± 1 day) of the second dosing cycle, wherein the C2D1 of the bispecific antibody is approximately 30 mg, and, The C1D1 and C1D2 of the anti-CD79b antibody-drug conjugate are each about 1.8 mg / kg.

[0095] In one aspect, the invention features a method of treating a population of individuals with R / R NHL comprising administering to the individual an anti-CD79b antibody drug conjugate in combination with CD20 and CD3-binding bispecific antibody, wherein: (a) the first administration cycle comprises: (i) the first dose of the bispecific antibody administered on day 8 (± 1 day) of the first administration cycle ( C1D1 ) and the second dose (C1D2) of the bispecific antibody to be administered on the 15th day (± 1 day) of the first administration cycle, wherein the C1D1 of the bispecific antibody is about 2.5 mg, and the bispecific antibody The C1D2 of the antibody is approximately 10 mg; and (ii) a single dose of the anti-CD79b antibody-drug conjugate administered on day 2 (± 1 day) of the first dosing cycle (C1D1); (b) the second to second Each of the six dosing cycles comprises a single dose of the bispecific antibody (C2D1 to C6D1) and a single dose of the anti-CD79b antibody drug conjugate (C2D1 to C6D1); and (c) the seventh through 12 dosing cycles each comprise the bispecific Single doses of anti-CD79b antibodies (C7D1 to C12D1) do not include the administration of anti-CD79b antibody drug conjugates, and wherein each single dose of bispecific antibodies C2D1 to C12D1 is on day 1 of each dosing cycle (± 1 day), and C2D1 to C6D1 of the anti-CD79b antibody drug conjugate is administered on the first day (± 1 day) of each administration cycle, and wherein each single dose of the bispecific antibody C2D1 to C12D1 is about 30 mg, and each single dose of anti-CD79b antibody-drug conjugate C1D1 to C6D1 is about 1.8 mg / kg.

[0096] In another aspect, the invention features an anti-CD79b antibody drug conjugate and a bispecific antibody that binds to CD20 and CD3 for use in a method of treating a population of individuals with R / R NHL, wherein the anti-CD79b antibody drug Conjugates and bispecific antibodies that bind to CD20 and CD3 are administered in a dosing regimen that includes 12 dosing cycles, wherein: (a) the first dosing cycle includes: (i) will be administered after the first dosing cycle The first dose of bispecific antibody (C1D1) administered on day 8 (± 1 day) and the second dose of bispecific antibody to be administered on day 15 (± 1 day) of the first dosing cycle (C1D2), wherein the C1D1 of the bispecific antibody is about 2.5 mg, and the C1D2 of the bispecific antibody is about 10 mg; and (ii) will be administered on day 2 (± 1 day) of the first dosing cycle A single dose of the anti-CD79b antibody drug conjugate (C1D1); (b) the second to sixth dosing cycles each comprising a single dose of the bispecific antibody (C2D1 to C6D1) and a single dose of the anti-CD79b antibody drug conjugate ( C2D1 to C6D1); and (c) seventh to twelfth dosing cycles each comprising a single dose of the bispecific antibody (C7D1 to C12D1) and excluding administration of an anti-CD79b antibody drug conjugate, and, wherein the bispecific Each single dose of C2D1 through C12D1 of the antibody will be administered on Day 1 (± 1 day) of each dosing cycle, and C2D1 through C6D1 of the anti-CD79b antibody drug conjugate will be administered on Day 1 (± 1 day) of each dosing cycle. day) administration, and wherein each single dose of bispecific antibody C2D1 to C12D1 is about 30 mg, and each single dose of anti-CD79b antibody drug conjugate C1D1 to C6D1 is about 1.8 mg / kg.

[0097] In another aspect, the present invention is characterized in the use of anti-CD79b antibody-drug conjugates and bispecific antibodies binding to CD20 and CD3 for treating a population of individuals with R / R NHL, wherein the anti-CD79b antibody-drug conjugates and The CD20 and CD3 binding bispecific antibody is administered with a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) will be administered on day 8 of the first dosing cycle ( ± 1 day) of the first dose of the bispecific antibody administered (C1D1) and the second dose of the bispecific antibody administered on day 15 (± 1 day) of the first dosing cycle (C1D2), wherein the C1D1 of the bispecific antibody is about 2.5 mg, and the C1D2 of the bispecific antibody is about 10 mg; and (ii) the anti-CD79b antibody to be administered on day 2 (± 1 day) of the first dosing cycle Single dose of drug conjugate (C1D1); (b) second to sixth dosing cycles each comprising a single dose of bispecific antibody (C2D1 to C6D1) and a single dose of anti-CD79b antibody drug conjugate (C2D1 to C6D1) and (c) the seventh through twelfth dosing cycles each comprised a single dose of the bispecific antibody (C7D1 through C12D1) and did not include administration of the anti-CD79b antibody drug conjugate, and, wherein each single dose of the bispecific antibody Doses C2D1 through C12D1 will be administered on Day 1 (± 1 day) of each dosing cycle and C2D1 through C6D1 of the anti-CD79b antibody drug conjugate will be administered on Day 1 (± 1 day) of each dosing cycle , and wherein each single dose of the bispecific antibody C2D1 to C12D1 is about 30 mg, and each single dose of the anti-CD79b antibody drug conjugate C1D1 to C6D1 is about 1.8 mg / kg.

[0098] In another aspect, the invention features the use of anti-CD79b antibody-drug conjugates and bispecific antibodies that bind to CD20 and CD3 in the manufacture of a medicament for treating a population of individuals with R / R NHL, wherein the anti-CD79b antibody The drug conjugates and bispecific antibodies that bind to CD20 and CD3 are administered in a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle comprises: (i) will be administered in the first dosing cycle The first dose of bispecific antibody (C1D1) administered on day 8 (± 1 day) of the first dosing cycle and the second dose of bispecific antibody administered on day 15 (± 1 day) of the first dosing cycle Dose (C1D2), wherein the C1D1 of the bispecific antibody is about 2.5 mg, and the C1D2 of the bispecific antibody is about 10 mg; and (ii) will be administered on day 2 (± 1 day) of the first dosing cycle A single dose of anti-CD79b antibody-drug conjugate (C1D1); (b) second to sixth dosing cycles each comprising a single dose of bispecific antibody (C2D1 to C6D1) and a single dose of anti-CD79b antibody-drug conjugate (C2D1 to C6D1); and (c) seventh to twelfth dosing cycles each comprising a single dose of the bispecific antibody (C7D1 to C12D1) and excluding administration of the anti-CD79b antibody drug conjugate, and, wherein the bispecific Each single dose of C2D1 to C12D1 of the anti-CD79b antibody drug conjugate will be administered on day 1 (± 1 day) of each dosing cycle, and C2D1 to C6D1 of the anti-CD79b antibody drug conjugate will be administered on day 1 (± 1 day) of each dosing cycle 1 day), and wherein each single dose of the bispecific antibody C2D1 to C12D1 is about 30 mg, and each single dose of the anti-CD79b antibody drug conjugate C1D1 to C6D1 is about 1.8 mg / kg.

[0099] In one aspect, the invention features a method of treating a population of individuals with R / R NHL, the method comprising administering to the individual Pareto with a dosing regimen comprising at least a first dosing cycle and a second dosing cycle. zizumab and grafitumumab, wherein: (a) the first dosing cycle includes the first dose of gefitumumab administered on day 8 (± 1 day) of the first dosing cycle (C1D1 ), the second dose of grafitumumab administered on day 15 (± 1 day) of the first dosing cycle (C1D2) and administered on day 2 (± 1 day) of the first dosing cycle A single dose of Parotuzumab (C1D1), wherein the C1D1 of grafiduzumab is about 2.5 mg, and the C1D2 of grafiduzumab is about 10 mg; and (b) the second dosing cycle comprises A single dose of grafitumumab (C2D1) administered on day 1 (± 1 day) of the second dosing cycle and Parol administered on day 1 (± 1 day) of the second dosing cycle A single dose (C2D1) of tocilizumab, wherein the C2D1 of grafiduzumab is about 30 mg, and, wherein the C1D1 and C2D1 of parotuzumab are each about 1.8 mg / kg.

[0100] In another aspect, the invention features parotuzumab and grifituzumab for use in a method of treating a population of individuals with R / R NHL, wherein parotuzumab and grifituzumab are A dosing regimen that includes at least the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes the drug that will be administered on the 8th day (± 1 day) of the first dosing cycle. The first dose of grafitumumab (C1D1), the second dose of grafitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle and A single dose (C1D1) of parotuzumab administered on the second day (± 1 day) of the drug cycle, wherein the C1D1 of grafiduzumab was about 2.5 mg, and the C1D2 of grafiduzumab was about 2.5 mg. 10 mg; and (b) a second dosing cycle consisting of a single dose of grafitumumab (C2D1) to be administered on day 1 (± 1 day) of the second dosing cycle and to be administered on the second dosing cycle A single dose (C2D1) of Parotuzumab administered on Day 1 (± 1 day) of the cycle, wherein the C2D1 of grafitumumab is approximately 30 mg, and wherein the C1D1 of Parotuzumab and C2D1 are each about 1.8 mg / kg.

[0101] In another aspect, the invention features the use of parotuzumab and grifituzumab in the treatment of a population of individuals with R / R NHL, wherein parotuzumab and grifituzumab are formulated with at least The administration of the dosing regimen of the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes Griffin that will be administered on the 8th day (± 1 day) of the first dosing cycle The first dose of gfitumumab (C1D1), the second dose of gfitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle and A single dose (C1D1) of parotuzumab administered on the second day (± 1 day), wherein the C1D1 of grafiduzumab was about 2.5 mg, and the C1D2 of grafiduzumab was about 10 mg and (b) a second dosing cycle comprising a single dose of grafitumumab (C2D1) to be administered on Day 1 (± 1 day) of the second dosing cycle and A single dose (C2D1) of Parotuzumab administered on day 1 (± 1 day), wherein C2D1 of grafitumumab is approximately 30 mg, and wherein C1D1 and C2D1 of Parotuzumab Each is about 1.8 mg / kg.

[0102] In another aspect, the invention features the use of parotuzumab and grifituzumab in the manufacture of a medicament for the treatment of a population of individuals with R / R NHL, wherein parotuzumab and grafituzumab The monoclonal antibody is administered with a dosing regimen that includes at least the first dosing cycle and the second dosing cycle, wherein: (a) the first dosing cycle includes the day 8 (± 1 day) of the first dosing cycle The first dose of grafitumumab administered (C1D1), the second dose of grafitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle, and A single dose (C1D1) of parotuzumab administered on the second day (± 1 day) of the first dosing cycle, wherein the C1D1 of grafiduzumab is about 2.5 mg, and the C1D2 is approximately 10 mg; and (b) the second dosing cycle consists of a single dose of grafitumumab to be administered on Day 1 (± 1 day) of the second dosing cycle (C2D1) and will be administered on Day 1 of the second dosing cycle A single dose (C2D1) of parotuzumab administered on day 1 (± 1 day) of the two dosing cycles, wherein the C2D1 of grafituzumab is about 30 mg, and wherein parotuzumab Anti-C1D1 and C2D1 were each about 1.8 mg / kg.

[0103] In one aspect, the invention features a method of treating a population of individuals with R / R NHL, the method comprising administering to the individual parotuzumab and grafidox on a dosing regimen comprising 12 dosing cycles. Monoclonal antibody, wherein: (a) the first dosing cycle includes: (i) the first dose of grafitumumab (C1D1) administered on day 8 (± 1 day) of the first dosing cycle and The second dose of grafitumumab (C1D2) administered on day 15 (± 1 day) of the first dosing cycle, wherein the C1D1 of grafitumumab is about 2.5 mg and the C1D2 of grafitumumab is about 10 mg; and (ii) a single dose of parotuzumab administered on day 2 (± 1 day) of the first dosing cycle (C1D1); (b) the second to sixth doses Cycles each consisted of a single dose of grafiduzumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1); and (c) cycles 7 to 12 each contained grafiduzumab Single doses of grafitumumab (C7D1 to C12D1) that did not include the administration of parotuzumab, and where each single dose of grafitumumab from C2D1 to C12D1 was administered on day 1 of each dosing cycle (± 1 day), and each single dose of Parotuzumab C2D1 to C6D1 is administered on Day 1 (± 1 day) of each dosing cycle, and each single dose of Griffithumab C2D1 C12D1 to C12D1 were approximately 30 mg, and each single dose of parotuzumab C1D1 to C6D1 was approximately 1.8 mg / kg.

[0104] In another aspect, the invention features parotuzumab and grifituzumab for use in a method of treating a population of individuals with R / R NHL, wherein parotuzumab and grifituzumab are Administration of a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle includes: (i) Griffith to be administered on day 8 (± 1 day) of the first dosing cycle The first dose (C1D1) of monoclonal antibody and the second dose (C1D2) of grafitumumab to be administered on the 15th day (± 1 day) of the first dosing cycle, wherein the C1D1 of grafitumumab and (ii) a single dose of parotuzumab to be administered on day 2 (± 1 day) of the first dosing cycle ( C1D1); (b) the second to sixth dosing cycles each consisted of a single dose of grafitumumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1); and (c) Dosing cycles 7 through 12 each included a single dose of grafiduzumab (C7D1 to C12D1) and did not include the administration of parotuzumab, and, where each single dose of grafiduzumab C2D1 to C12D1 will be administered on Day 1 (± 1 Day) of each dosing cycle, and each single dose of Parotuzumab C2D1 to C6D1 will be administered on Day 1 (± 1 Day) of each dosing cycle, And, wherein each single dose of grafitumumab C2D1 to C12D1 is about 30 mg, and each single dose of parotuzumab C1D1 to C6D1 is about 1.8 mg / kg.

[0105] In another aspect, the invention features the use of parotuzumab and grifituzumab in the treatment of a population of individuals with R / R NHL, wherein parotuzumab and grifituzumab are comprised of 12 Administration of a dosing regimen for two dosing cycles, wherein: (a) the first dosing cycle includes: (i) grafitumumab to be administered on day 8 (± 1 day) of the first dosing cycle The first dose (C1D1) of the first dosing cycle and the second dose (C1D2) of grafiduzumab administered on the 15th day (± 1 day) of the first dosing cycle, wherein the C1D1 of grafiduzumab is about 2.5 mg and the C1D2 of grafitumumab is approximately 10 mg; and (ii) a single dose of parotuzumab (C1D1) to be administered on day 2 (± 1 day) of the first dosing cycle ; (b) the second to sixth dosing cycles each comprised a single dose of grafitumumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1); and (c) seventh to The 12th dosing cycle will each consist of a single dose of grafitumumab (C7D1) and will not include the administration of parotuzumab, and where each single dose of grafitumumab, C2D1 through C12D1, will be given at each dose Day 1 (± 1 day) of the drug cycle, and each single dose of Parotuzumab C2D1 to C6D1 will be administered on the first day (± 1 day) of each drug cycle, and, where Each single dose of Fertuzumab C2D1 to C12D1 was about 30 mg, and each single dose of Parotuzumab C1D1 to C6D1 was about 1.8 mg / kg.

[0106] In another aspect, the invention features the use of parotuzumab and grifituzumab in the manufacture of a medicament for the treatment of a population of individuals with R / R NHL, wherein parotuzumab and grafituzumab The monoclonal antibody is administered with a dosing regimen comprising 12 dosing cycles, wherein: (a) the first dosing cycle includes: (i) will be administered on day 8 (± 1 day) of the first dosing cycle The first dose of grafitumumab (C1D1) and the second dose of grafitumumab (C1D2) to be administered on day 15 (± 1 day) of the first dosing cycle, wherein grafitumumab The C1D1 of the antibody is about 2.5 mg and the C1D2 of grafitumumab is about 10 mg; and (ii) the C1D2 of Parotuzumab to be administered on day 2 (± 1 day) of the first dosing cycle Single dose (C1D1); (b) Second to sixth dosing cycles each comprising a single dose of grafitumumab (C2D1 to C6D1) and a single dose of parotuzumab (C2D1 to C6D1); and ( c) The seventh to twelfth dosing cycles each included a single dose of grafiduzumab (C7D1) and did not include the administration of parotuzumab, and where each single dose of grafiduzumab C2D1 to C12D1 will be administered on Day 1 (± 1 day) of each dosing cycle and each single dose of parotuzumab C2D1 through C6D1 will be administered on Day 1 (± 1 day) of each dosing cycle , and wherein each single dose of C2D1 to C12D1 of grafiduzumab is about 30 mg, and each single dose of C1D1 to C6D1 of parotuzumab is about 1.8 mg / kg.

[0107] In some embodiments, the complete response rate is at least 20% (e.g., at least 25%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%; e.g., 20-100% Between %, Between 40-100%, Between 60-100%, Between 80-100%, Between 20-80%, Between 20-60%, Between 20-40%, Between 40-80% Between, between 40-60%, between 30-50%, or between 35-45%; for example about 20%, about 25%, about 30%, about 35%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 50%, about 60% or higher). In some embodiments, the complete response rate is at least 40%. In a particular embodiment, the complete response rate in the population of individuals with R / R NHL is about 42%. In some embodiments, the overall response rate is at least 30% (e.g., at least 35%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%; e.g., 30-100% Between %, Between 50-100%, Between 70-100%, Between 30-90%, Between 30-70%, Between 30-50%, Between 40-80%, Between 40-60% Between, between 45-55%, or between 35-45%; for example about 30%, about 35%, about 40%, about 45%, about 48%, about 49%, about 50%, about 51% , about 52%, about 55%, about 60%, about 70% or higher). In some embodiments, the overall response rate is at least 50%. In certain embodiments, the overall response rate in the population of individuals with R / R NHL is about 50%.

[0108] In some embodiments, the B cell proliferative disorder is R / R MCL. In some embodiments, R / R NHL is R / R MCL. In some embodiments, the complete response rate is at least 60% (e.g., at least 60%, at least 70%, at least 80%, or at least 90%; e.g., between 60-100%, between 70-100%, 80% Between -100%, Between 90-100%, Between 60-90%, Between 60-80%, Between 60-70%, Between 70-90%, Between 80-90%, Between 80- Between 100%, or between 90-100%; for example about 60%, about 70%, about 75%, about 80%, about 83%, about 85%, about 87%, about 90%, about 95%, about 97%, about 98%, about 99% or higher). In some embodiments, the complete response rate is at least 80%. In specific embodiments, the complete response rate in the population of individuals with R / R NHL is at least about 85%. In specific embodiments, the complete response rate in the population of individuals with R / R MCL is at least about 85%. In certain embodiments, the complete response rate in the population of individuals with R / R MCL is about 100%. In some embodiments, the overall response rate is at least 60% (e.g., at least 60%, at least 70%, at least 80%, or at least 90%; e.g., between 60-100%, between 70-100%, 80% Between -100%, Between 90-100%, Between 60-90%, Between 60-80%, Between 60-70%, Between 70-90%, Between 80-90%, Between 80- Between 100%, or between 90-100%; for example about 60%, about 70%, about 75%, about 80%, about 83%, about 85%, about 87%, about 90%, about 95%, about 97%, about 98%, about 99% or higher). In specific embodiments, the overall response rate in the population of individuals with R / R NHL is at least about 85%. In specific embodiments, the overall response rate in the population of individuals with R / R MCL is at least about 85%. In certain embodiments, the overall response rate in the population of individuals with R / R MCL is about 100%. In some embodiments, the overall response rate is at least 80%.

[0109] In some embodiments, the B cell proliferative disorder is R / R DLBCL. In some embodiments, R / R NHL is R / R MCL. In some embodiments, the R / R NHL is R / R DLBCL. In some embodiments, the complete response rate is at least 60% (e.g., at least 60%, at least 70%, at least 80%, or at least 90%; e.g., between 60-100%, between 70-100%, 80% Between -100%, between 90-100%, between 60-90%, between 60-80%, between 60-70%, between 60-65%, between 65-75%, or between 75 Between -85%; for example about 60%, about 61%, about 62%, about 65%, about 68%, about 69%, about 70%, about 71%, about 72%, about 73%, about 74% , about 75%, about 76%, about 77%, about 80%, about 85%, about 90% or higher). In some embodiments, the complete response rate is at least 65%. In some embodiments, the complete response rate is at least 70%. In certain embodiments, the complete response rate in the population of individuals with R / R DLBCL is about 60%. In a particular embodiment, the complete response rate in the population of individuals with R / R DLBCL is about 65%. In certain embodiments, the complete response rate in the population of individuals with R / R DLBCL is about 70%. In certain embodiments, the complete response rate in the population of individuals with R / R DLBCL is about 75%. In some embodiments, the overall response rate is at least 60% (e.g., at least 60%, at least 70%, at least 80%, or at least 90%; e.g., between 60-100%, between 70-100%, 80% Between -100%, Between 90-100%, Between 60-90%, Between 60-80%, Between 60-70%, Between 60-65%, Between 65-75%, Between 70- Between 90%, or between 75-85%; for example about 60%, about 63%, about 64%, about 65%, about 66%, about 67%, about 70%, about 73%, about 74%, about 75%, about 76%, about 77%, about 80%, about 83%, about 84%, about 85%, about 86%, about 87%, about 90% or more). In some embodiments, the overall response rate is at least 70%. In some embodiments, the overall response rate is at least 80%. In certain embodiments, the overall response rate in the population of individuals with R / R DLBCL is about 65%. In a particular embodiment, the overall response rate in the population of individuals with R / R DLBCL is about 73%. In certain embodiments, the overall response rate in the population of individuals with R / R DLBCL is about 75%. In certain embodiments, the overall response rate in the population of individuals with R / R DLBCL is about 85%.

[0110] In some embodiments, the B cell proliferative disorder is R / R DLBCL. In some embodiments, the R / R NHL is R / R DLBCL. In some embodiments, the complete response rate is at least 35% (e.g., at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or higher; e.g., between 30-100%) Between, Between 50-100%, Between 70-100%, Between 35-90%, Between 45-90%, Between 35-70%, Between 35-50%, Between 40-80% , between 40-60%, between 45-55%, or between 35-45%; for example, about 35%, about 40%, about 45%, about 48%, about 49%, about 50%, about 51%, about 52%, about 55%, about 60%, about 70%, about 75%, about 80%, about 85% or higher). In some embodiments, the complete response rate is at least 45%. In some embodiments, the complete response rate is at least 55%. In some embodiments, the complete response rate is at least 75%. In some embodiments, the complete response rate is at least 85%. In some embodiments, the complete response rate is at least 90%. In a particular embodiment, the complete response rate in the population of individuals with R / R DLBCL is about 46%. In a particular embodiment, the complete response rate in the population of individuals with R / R DLBCL is about 52%. In a particular embodiment, the complete response rate in the population of individuals with R / R DLBCL is about 86%. In some embodiments, the overall response rate is at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%) %, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% or higher; for example, between 85-100%, between 87-100%, between 90-100%, 95% Between -100%, between 85-97%, between 85-95%, between 85-90%, between 85-87%, between 90-95%, or between 93-97%; e.g. , about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or higher). In some embodiments, the overall response rate is at least 85%. In some embodiments, the overall response rate is at least 90%. In a particular embodiment, the overall response rate in the population of individuals with R / R DLBCL is about 86%.

[0111] In some embodiments, the complete response rate is higher than the reference complete response rate for a reference population of individuals treated with a combination therapy comprising an anti-CD20 / anti-CD3 bispecific antibody and an anti-PD-L1 antagonist antibody and not comprising an anti-CD79b antibody drug conjugate. response rate. In some embodiments, the objective response rate is higher than the reference objective response rate for a reference population of individuals treated with a combination therapy comprising an anti-CD20 / anti-CD3 bispecific antibody and an anti-PD-L1 antagonist antibody and not comprising an anti-CD79b antibody drug conjugate response rate. In some embodiments, the complete response rate is higher than the reference complete response rate for a reference population of individuals treated with a combination therapy comprising grafiduzumab and atezolizumab without parotuzumab . In some embodiments, the objective response rate is higher than the reference objective response rate for a reference population of individuals treated with a combination therapy comprising grifituzumab and atezolizumab without parotuzumab.

[0112] In some embodiments, the individual is a human. In some embodiments, each individual in the population of individuals is a human. In some embodiments, each individual in the reference population of individuals is a human. In some embodiments, the individual or group of individuals has received at least two prior systemic therapies (eg, two, three, four, five, six or more prior systemic therapies). In some embodiments, the individual or group of individuals is not eligible for autologous stem cell transplant (SCT). Brief description of the diagram

[0113] This application file contains at least one drawing in color. Copies of this patent or patent application with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. [picture] [1A-1N] are schematic diagrams showing the configuration of exemplary anti-CD20 / anti-CD3 bispecific antibodies. [picture] [2] is a schematic diagram showing the structure of grafitumumab. [picture] [3] is a graph providing an overview of the study as revealed in Example 1. Atezo = atezolizumab; CRM = continuous reassessment approach; DLBCL = diffuse large B-cell lymphoma; ECOG = Eastern Cooperative Oncology Group; Parotuzumab; R / R = relapsed and / or refractory; SCT = Society for Clinical Trials; TCB = grafitumumab. [picture] [4A-4C] are diagrams showing the atezolizumab group (Fig. 4A), the dose-escalation phase of the parotuzumab group (Fig. 4B) and the expansion phase of the parotuzumab group (Fig. 4C). Schematic representation of the study design, as revealed in Example 1. Atezo = atezolizumab; CR = complete response; DE = dose escalation; DLBCL = diffuse large B-cell lymphoma; EoS = end of study; F / U = follow-up; Non-Hodgkin's lymphoma; Pola = parotuzumab; PD = progressive disease; PR = partial response; Pts = patients; RP2D = recommended phase II dose; R / R = relapsed and / or refractory Sex; SD = stable disease. [picture] [5A] [and picture] [5B] is a schematic diagram showing the timing of dosing for the atezolizumab group (Fig. 5A) and the parotuzumab group (Fig. 5B) in the study disclosed in Example 1. Atezo = atezolizumab; Glofit = grafituzumab; Gpt = GAZVAYA® pretreatment (obinutuzumab pretreatment); Pola = parotuzumab. [picture] [6] It is an adverse event reported in cohort 1 and cohort 2 in safety-evaluable patients who have received grafiduzumab + parotuzumab treatment, with an incidence rate ≥ 10% or NCI-CTCAE grade 5 adverse events A graph of the frequency of (AE). The color indicates the grade of AE. All AEs in the study are reported on the left. Only AEs considered to be related to study treatment (e.g., grafiduzumab or parotuzumab) are reported on the right. Figure 7 provides updated efficacy data as of the clinical cutoff date for dose escalation cohorts 1 and 2 and the RP2D expansion cohort. ; Glofit = grafituzumab. Pola = Parotuzumab. Implementation

[0114] sequence listing

[0115] This application contains a Sequence Listing, which has been filed electronically in ASCII format and is hereby incorporated by reference in its entirety. Created on April 26, 2021, this ASCII copy is named 51177-033001_Sequence_Listing_4.26.21_ST25 and is 32,706 bytes in size.

[0116] The present invention provides for the treatment of patients with CD20 positive cell proliferative disorders (e.g., B cell proliferative disorders (e.g., non-Hodgkin's lymphoma (NHL) (e.g., relapsed and / or refractory NHL, diffuse large B-cell lymphoma (DLBCL) (eg, relapsed and / or refractory DLBCL), follicular lymphoma (FL) (eg, relapsed and / or refractory FL or transformed FL), or mantle cell lymphoma (MCL) (e.g., relapsed or refractory MCL)), or central nervous system lymphoma (CNSL))), comprising administering to the individual an anti-CD79b antibody drug conjugate and / or anti-CD20 / anti-CD3 The bispecific antibody is administered, for example, in a divided escalating dose regimen. The method comprises at least a first administration cycle and a second administration cycle, wherein: (a) the first administration cycle comprises a first dose (C1D1) of the bispecific antibody and a second dose (C1D2) of the bispecific antibody , wherein the C1D1 of the bispecific antibody is about 2.5 mg and the C1D2 of the bispecific antibody is about 10 mg; and (b) the second dosing cycle comprises a single dose of the bispecific antibody (C2D1), wherein the bispecific antibody The C2D1 is about 10 mg, about 16 mg or about 30 mg.

[0117] The present invention is based, in part, on the discovery that it involves administering a bispecific antibody that binds to CD20 and CD3 (e.g., Griffin®) over multiple dosing cycles (e.g., where the first dosing cycle is an ascending, split dosing cycle). A divided dose escalation regimen of B-cell monoclonal antibody) is very effective in treating individuals with CD20-positive cell proliferative disorders (e.g., B-cell proliferative disorders) and has an acceptable safety profile (e.g., with respect to cytokine release syndrome). [I.] [General Technology] []

[0118] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are within the skill of the art. Such techniques are fully described in, for example, "Molecular Cloning: Laboratory Manual (Second Edition)" ("Molecular Cloning: Laboratory Manual", Sambrook et al., 1989); "Oligonucleotide Synthesis" ( "Oligonucleotide Synthesis", edited by M.J.Gait, 1984); "Animal Cell Culture", edited by R. I. Freshney, 1987); "Methods in Enzymology", American Academy Publishing Company ( Academic Press, Inc.)); "Current Protocols in Molecular Biology", edited by F. M. Ausubel et al., 1987, and regularly updated); PCR: Polymerase Chain Reaction ("PCR: The Polymerase Chain Reaction", Mullis et al., eds., 1994); "A Practical Guide to Molecular Cloning", Perbal Bernard V., 1988); Phage Display: A Laboratory Manual (" Phage Display: A Laboratory Manual", Barbas et al., 2001). [II.] [definition] []

[0119] Definitions Unless otherwise defined below, the terms used herein are those commonly used in the technical field.

[0120] Unless otherwise stated, the term "cluster of differentiation 20" or "CD20" as used herein refers to any native CD20 from any vertebrate source, including mammals, such as primates (e.g., humans) and rodents (eg, mice and rats). CD20 (also known as B-lymphocyte antigen CD20, B-lymphocyte surface antigen B1, Leu-16, Bp35, BM5, and LF5; human protein characterized under UniProt database entry P11836) is a hydrophobic protein with a molecular weight of approximately 35 kD Transmembrane protein expressed on pre- and mature B-lymphocytes (Valentine, M.A. et al., J. Biol. Chem. 264 (1989) 11282-11287; Tedder, T.F., et al., Proc. Natl. Acad . Sci. U.S.A. 85(1988) 208-212; Stamenkovic, I., et al., J. Exp. Med. 167 (1988) 1975-1980; Einfeld, D.A., et al., EMBO J. 7 (1988) 711- 717; Tedder, T.F., et al., J. Immunol. 142 (1989) 2560-2568). The corresponding human gene is transmembrane 4 domain, subfamily A, member 1, also known as MS4A1. This gene encodes a member of the transmembrane 4A gene family. Members of this nascent protein family are characterized by shared structural features and similar intron / exon splice boundaries, and display distinct patterns of expression in hematopoietic cells and nonlymphoid tissues. This gene encodes a B lymphocyte surface molecule that plays a role in B cell development and differentiation into plasma cells. This family member maps to 11q12 in the family member cluster. The term encompasses "full-length", unprocessed CD20 as well as any form of CD20 that has been processed in the cell. The term also covers naturally occurring CD20 variants, such as splice variants or allelic variants. Alternative splicing of this gene results in two transcript variants encoding the same protein. In one embodiment, the CD20 is human CD20.

[0121] The terms "anti-CD20 antibody" and "antibody that binds to CD20" refer to an antibody that is capable of binding CD20 with sufficient affinity such that the antibody can be used as a diagnostic and / or therapeutic agent targeting CD20. In one embodiment, an anti-CD20 antagonist antibody binds to an irrelevant, non-CD20 protein to a degree that is about 10% less than the antibody binds to CD20, as measured by, for example, a radioimmunoassay (RIA). In certain embodiments, the dissociation constant (KD) of the antibody that binds to CD20 is ≤ 1 μM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM (e.g., 10 -8M or less, eg, 10 -8M to 10 -13M, eg, 10 -9M to 10 -13M). In certain embodiments, the anti-CD20 antagonist antibody binds to an epitope of CD20 that is conserved in CD20 across species.

[0122] "Type II anti-CD20 antibody" refers to an anti-CD20 antibody having the binding properties and biological activity of a type II anti-CD20 antibody, such as Cragg et al., Blood103 (2004) 2738-2743; Cragg et al., Blood101 (2003) 1045-1052; disclosed in Klein et al., mAbs 5 (2013), 22-33, and summarized in Table 1 below. [surface] [1.] [Article] [I] [Type and No.] [II] [type resistance] [CD20] [Characteristics of Antibodies] [] [Article] [I] [type resistance] [CD20] [Antibody] [Article] [II] [type resistance] [CD20] [Antibody] Binds class I CD20 epitopes Binds class II CD20 epitope Localization of CD20 to lipid rafts Does not localize CD20 to lipid rafts High CDC* Low CDC* ADCC activity* ADCC activity* Full binding ability to B cells About half of the binding capacity for B cells weak homozygous aggregation Homotype aggregation low cell death induction Strong cell death induction * if IgG 1 isotype

[0123] Examples of type II anti-CD20 antibodies include e.g. obinutuzumab (GA101), tositumumab (B1), humanized B-Ly1 antibody IgG1 (chided as disclosed in WO 2005 / 044859 Humanized IgG1 antibody), 11B8 IgG1 (as disclosed in WO 2004 / 035607) and AT80 IgG1.

[0124] Examples of Type I anti-CD20 antibodies include, e.g., rituximab, ofatumumab, veltuzumab, ocaratuzumab, ocrelizumab ( ocrelizumab), PRO131921, ublituximab, HI47 IgG3 (ECACC, fusionoma), 2C6 IgG1 (as disclosed in WO 2005 / 103081), 2F2 IgG1 (as disclosed in WO 2004 / 035607 and WO 2005 / 103081 disclosed) and 2H7 IgG1 (as disclosed in WO 2004 / 056312).

[0125] Unless otherwise stated, "CD3" refers to any native CD3 derived from any vertebrate, including mammals such as primates (e.g. humans), non-human primates (e.g. cynomolgus monkeys) and rodents (such as mice and rats). The term encompasses "full-length", unprocessed CD3 as well as any form of CD3 that has been obtained by processing the cells. The term also covers naturally occurring CD3 variants, for example, splice variants or allele variants. In one embodiment, the CD3 is human CD3, particularly the epsilon subunit of human CD3 (CD3ε). The amino acid sequence of human CD3ε is shown in UniProt (www.uniprot.org) accession number P07766 (version 144) or NCBI (www.ncbi.nlm.nih.gov / ) RefSeq NP_000724.1. The amino acid sequence of cynomolgus monkey [Macaca fascicularis] CD3ε is shown in NCBI GenBank Accession BAB71849.1.

[0126] The terms "anti-CD20 / anti-CD3 bispecific antibody" and "bispecific antibody that binds to CD20 and CD3" are capable of binding both CD20 and CD3 with sufficient affinity such that the antibody can be used as a target for CD20 and / or Or a bispecific antibody of a CD3 diagnostic agent and / or therapeutic agent. In one embodiment, the bispecific antibody that binds CD20 and CD3 binds to an unrelated non-CD3 protein and / or non-CD20 protein to a lower extent than does the antibody binds to Approximately 10% of CD3 and / or CD20 binding. In certain embodiments, the bispecific antibody that binds CD20 and CD3 has a dissociation constant (KD) of ≤ 1 μM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM (eg 10 -8M or lower, eg 10 -8M to 10 -13M, eg 10 -9 to 10 -13M ). In certain embodiments, the bispecific antibody that binds CD20 and CD3 binds to a CD3 epitope that is conserved between CD3 from different species and / or a CD20 epitope that is conserved between CD20 from different species . An example of an anti-CD20 / anti-CD3 bispecific antibody is Griffithumab (Proposed INN: List 121 WHO Drug Information, Vol. 33, No. 2, 2019, p. 276, also known as CD20-TCB, RO7082859 or RG6026; CAS #: 2229047-91-8).

[0127] Unless otherwise stated, the term "cluster of differentiation 79b" or "CD79b" as used herein refers to any native CD79b from any vertebrate source, including mammals, such as primates (e.g., humans) and rodents (eg, mice and rats). The term encompasses "full-length" unprocessed CD79b as well as any form of CD79b produced by processing in the cell. The term also encompasses naturally occurring CD79b variants, for example, splice variants or allele variants. CD79b includes, for example, the human CD79b protein (NCBI RefSeq No. NP_000617), which is 229 amino acids in length.

[0128] The terms "anti-CD79b antibody" and "antibody that binds to CD79b" refer to an antibody that is capable of binding CD79b with sufficient affinity such that the antibody can be used as a diagnostic and / or therapeutic agent targeting CD79b. In one embodiment, an anti-CD79b antagonist antibody binds to an irrelevant, non-CD79b protein to a degree that is about 10% less than the antibody binds to CD79b, as measured by, for example, a radioimmunoassay (RIA). In certain embodiments, the dissociation constant (KD) of an antibody that binds to CD79b is ≤ 1 μM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM (eg, 10 -8M or lower, such as 10 -8M to 10 -13M, or such as 10 -9 to 10 -13M). In certain embodiments, the anti-CD79b antagonist antibody binds to an epitope of CD79b that is conserved in CD79b across species.

[0129] As used herein, the term "cytokine release" or "cytokine release" is synonymous with "cytokine storm" or "cytokine release syndrome" (abbreviated "CRS"), and refers to symptoms that occur during or after the administration of a therapeutic agent. Soon (e.g., within 1 day), cytokines, especially tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), interleukin-6 (IL-6), Increased levels of interleukin-10 (IL-10), interleukin-2 (IL-2) and / or interleukin-8 (IL-8), leading to adverse symptoms. Cytokine release is defined as the supraphysiological response resulting in the activation or engagement of endogenous or infused T cells and / or other immune effector cells following the administration of any immunotherapy. Symptoms may be progressive, always beginning with fever, and may include hypotension, capillary leak (hypoxia), and end-organ dysfunction (Lee et al., 2019). In some cases, such as after administration of CAR-T cells, CRS may also occur when the CAR-T cells expand several days after administration. Incidence and severity usually decrease with subsequent infusions. Symptoms may range from symptomatic discomfort to fatal events and may include fever, chills, dizziness, hypertension, hypotension, dyspnea, restlessness, sweating, flushing, rash, tachycardia, shortness of breath, headache, tumor pain, nausea , vomiting and / or organ failure.

[0130] The term "amino acid mutation" as used herein is meant to cover amino acid substitutions, deletions, insertions and modifications. Any combination of substitutions, deletions, insertions, and modifications can be performed to arrive at the final construct, provided that the final construct possesses the desired characteristics, such as reduced binding to Fc receptors. Amino acid sequence deletions and insertions include deletions and insertions at the amino and / or carboxy termini of amino acids. Specific amino acid mutations are amino acid substitutions. To alter e.g. the binding characteristics of the Fc region, particularly preferred are non-conservative amino acid substitutions, i.e. the substitution of one amino acid for another amino acid having different structural and / or chemical properties. Amino acid substitutions include non-naturally occurring amino acids or naturally occurring amino acid derivatives with twenty standard amino acids (e.g., 4-hydroxyproline, 3-methylhistidine, ornithine acid, homoserine, 5-hydroxylysine) replacement. Amino acid mutations can be generated using genetic or chemical methods well known in the art. Genetic methods can include site-directed mutagenesis, PCR, gene synthesis, and the like. It is contemplated that methods other than genetic engineering, such as chemical modification, to alter the side chain groups of amino acids may also be useful. Various names may be used herein to refer to mutations of the same amino acid. For example, a substitution of proline to glycine at position 329 of the Fc region may be represented as 329G, G329, G329, P329G or Pro329Gly.

[0131] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (eg, a receptor) and its binding partner (eg, a ligand). Unless otherwise stated, "binding affinity" as used herein refers to intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., a receptor and a ligand). The affinity of a molecule X for its partner Y can often be expressed in terms of a dissociation constant (KD), which is the ratio of the dissociation rate constant to the association rate constant (koff and kon, respectively). Thus, equivalent affinities can include different rate constants as long as the ratio of rate constants remains the same. Affinity can be measured by established methods known in the art. A particular method used to determine affinity is surface plasmon resonance (SPR).

[0132] The term "affinity matured" antibody refers to an antibody that has one or more changes in one or more complementarity determining regions (HVRs) that cause the antibody to respond to a parental antibody that does not have such changes. Improvement of antigen affinity.

[0133] The term "antigen binding moiety" as used herein refers to a polypeptide molecule that specifically binds to an antigenic determinant. In one embodiment, the antigen binding moiety is capable of directing the entity to which it is attached (e.g., a cytokine or a second antigen binding moiety) to a target site, e.g., to a particular type of tumor cell bearing an epitope or tumor stroma. Antigen binding portions include antibodies and fragments thereof as further defined herein. Preferred antigen binding portions include the antigen binding domain of an antibody comprising an antibody heavy chain variable region and an antibody light chain variable region. In certain embodiments, an antigen binding portion may comprise an antibody constant region as further defined herein and known in the art. Useful heavy chain constant regions include any of five isotypes: alpha, delta, epsilon, gamma, or mu. Useful light chain constant regions include either of two isotypes: kappa and lambda.

[0134] "Binding", "specifically binding" or "specific for" means that the binding is selective for the antigen and can discriminate against unwanted or non-specific interactions. The ability of an antigen-binding moiety to bind to a specific epitope can be determined by enzyme-linked immunosorbent assay (ELISA) or other techniques familiar to those skilled in the art, such as surface plasmon resonance (e.g., on a BIAcore instrument) (Liljeblad et al., Glyco J. 17, 323-329 (2000)) and traditional binding assays (Heeley, Endocr Res. 28, 217-229 (2002)) to measure. In one embodiment, the antigen-binding portion binds an unrelated protein to an extent that is less than about 10% of the antigen-binding portion binds the antigen, e.g., as determined by SPR. In certain embodiments, the dissociation constant (KD) of the antigen-binding portion or the antigen-binding molecule comprising the antigen-binding portion that binds to the antigen is ≤ 1 μM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM (eg, 10 -8M or less, eg, 10 -8M to 10 -13M, eg, 10 -9M to 10 -13M).

[0135] "Reduced binding", eg reduced binding to an Fc receptor, refers to a decrease in the affinity of the respective interaction as measured eg by SPR. For clarity, the term also includes the reduction of the affinity to zero (or below the detection limit of the analytical method), ie, the complete abolition of the interaction. In contrast, "increased binding" refers to an increase in the binding affinity of the respective interaction.

[0136] As used herein, the term "antigen binding molecule" in its broadest sense refers to a molecule that specifically binds an antigenic epitope. Exemplary of antigen binding molecules are immunoglobulins and derivatives (eg, fragments) thereof.

[0137] As used herein, the term "epitope" is synonymous with "antigen" and "epitope" and refers to a site on a polypeptide macromolecule that binds to an antigen-binding moiety to form an antigen-binding moiety-antigen complex (e.g. , an adjacent stretch of amino acids or a configuration consisting of different regions of non-adjacent amino acids). For example, available epitopes may be present on the surface of tumor cells, on the surface of virus-infected cells, on the surface of other diseased cells, absent in serum, and / or present in the extracellular matrix (ECM) middle. Unless otherwise stated, proteins referred to herein as antigens (e.g., CD3) can be proteins in any native form from any vertebrate source, including mammals, such as primates (e.g., humans) and rodents. animals (such as mice and rats). In specific embodiments, the antigen is a human protein. Where a specific protein is referred to herein, the term encompasses "full length", unprocessed protein and any form of protein resulting from processing in cells. The term also encompasses naturally occurring variants of the protein, eg, splice variants or allele variants. An exemplary human protein that can be used as an antigen is CD3, particularly the epsilon subunit of CD3 (see UniProt Accession P07766 (version 130), NCBI RefSeq Accession NP_000724.1, for human sequences; or UniProt Accession Q95LI5 ( 49th edition), NCBI GenBank accession BAB71849.1, for cynomolgus monkey sequences). In certain embodiments, a T cell activating bispecific antigen binding molecule disclosed herein binds to an epitope of a CD3 or target cell antigen that is conserved among CD3 or target cell antigens from different species.

[0138] As used herein, the term "polypeptide" refers to a molecule composed of monomers (amino acids) linked linearly by amide bonds (also known as peptide bonds). The term "polypeptide" refers to any chain of two or more amino acids and does not imply a specific length of the product. Thus, included in the definition of "polypeptide" are peptides, dipeptides, tripeptides, oligopeptides, "proteins", "amino acid chains" or any other term used to refer to a chain of two or more amino acids , and "polypeptide" may be used instead of or interchangeably with any of these terms. The term "polypeptide" also refers to the products of post-expression modifications of polypeptides, including but not limited to glycosylation, acetylation, phosphorylation, amidation, derivatization through known protecting / blocking groups, proteolysis or Non-naturally occurring amino acid modifications. Polypeptides may be derived from natural biological sources or produced by recombinant techniques, but are not necessarily translated from a specified nucleic acid sequence. It can be produced in any way, including through chemical synthesis. The size of the polypeptides of the invention can be about 3 or more, 5 or more, 10 or more, 20 or more, 25 or more, 50 or more , 75 or more, 100 or more, 200 or more, 500 or more, 1,000 or more, or 2,000 or more amino acids. Polypeptides can have defined three-dimensional structures, although they do not necessarily have such structures. Polypeptides that have a defined three-dimensional structure are called folded, whereas polypeptides that do not have a defined three-dimensional structure but can adopt a number of different configurations are called unfolded.

[0139] An "isolated" polypeptide or variant or derivative thereof refers to a polypeptide in a non-native setting. No specific level of purification is required. For example, an isolated polypeptide can be removed from its native or natural environment. For the purposes of the present invention, recombinantly produced antibodies and proteins expressed in host cells are considered isolated, as native or recombinant polypeptides that have been isolated, fractionated, or partially or substantially purified by any suitable technique.

[0140] The "percentage (%) amino acid sequence identity" described relative to the reference polypeptide sequence refers to the percentage of amino acid residues in the candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence. And after introducing differences (if necessary), the maximum percent sequence identity can be achieved, and any conservative substitutions are not considered as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be accomplished by various means that are within the skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR ) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. However, for purposes herein, % amino acid sequence identity values ​​were 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 has been filed with the user documentation in the United States Copyright Office, Washington, DC, 20559, and is registered under US Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California, and can also be compiled from source code. ALIGN-2 programs should be compiled for use on UNIX operating systems (including digital UNIX V4.0D). All sequence comparison parameters were set by the ALIGN-2 program and were unchanged. In the case of amino acid sequence comparisons using ALIGN-2, the % amino acid sequence identity of a given amino acid sequence A to, with, or relative to a given amino acid sequence B (which is optionally expressed as a given amino acid sequence An amino acid sequence A that has or contains a certain % amino acid sequence identity to, with, or with respect to a given amino acid sequence B) is calculated as follows: 100 times the fraction X / Y where X is the number of amino acid residues that the sequence alignment program ALIGN-2 scores as identical matches in the alignment of programs A and B, and Y is the total number of amino acid residues in B. It should be understood that where the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity of A to B will not be equal to the % amino acid sequence identity of B to A sex. Unless specifically stated otherwise, all % amino acid sequence identity values ​​used herein were obtained using the ALIGN-2 computer program as described in the preceding paragraph.

[0141] The term "antibody" herein is used in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (such as bispecific antibodies), and antibody fragments, so long as they exhibit Antigen-binding activity is expected to suffice.

[0142] The terms "full-length antibody", "intact antibody" and "whole antibody" are used interchangeably herein and refer to an antibody having a structure substantially similar to that of a native antibody or having a heavy chain comprising an Fc region as defined herein. Antibody.

[0143] "Antibody fragment" refers to a molecule other than an intact antibody that comprises a portion of an intact antibody that binds the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2, diabodies, linear antibodies, single chain antibody molecules (such as scFv), and multispecific antibodies formed from antigen fragments. Antibody. As used herein, the term "antibody fragment" also encompasses single domain antibodies.

[0144] The term "immunoglobulin molecule" refers to a protein having the structure of a naturally occurring antibody. For example, immunoglobulins of the IgG class are heterotetrameric glycoproteins of approximately 150,000 daltons consisting of two light chains and two heavy chains bonded by disulfide bonds. From N-terminus to C-terminus, each heavy chain has a variable region (VH), also called variable heavy domain or heavy chain variable domain, followed by three constant domains (CH1, CH2 and CH3), also called Heavy chain constant region. Similarly, from the N-terminus to the C-terminus, each light chain has a variable region (VL), also known as a light chain variable domain or a light chain variable domain, followed by a light chain constant (CL) domain, also known as Light chain constant region. The heavy chains of immunoglobulins can be classified into one of five classes, called alpha (IgA), delta (IgD), epsilon (IgE), gamma (IgG), or mu (IgM), some of which can be further classified into Divided into subclasses such as γ 1 (IgG 1), γ 2 (IgG 2), γ 3 (IgG 3), γ 4 (IgG 4), α 1 (IgA 1) and α 2 (IgA 2). Based on the amino acid sequence of their constant domains, the light chains of immunoglobulins can be classified into one of two types, called kappa (κ) and lambda (λ). Immunoglobulins essentially consist of two Fab molecules and an Fc domain linked by the immunoglobulin hinge region.

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

[0146] The term "variable region" or "variable domain" refers to the domain of an antibody's heavy 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 usually have similar structures, and each domain contains four conserved framework regions (FR) and three hypervariable regions (HVR) . See, e.g., Kindt et al., Kuby Immunology, 6th ed., W.H. Freeman and Co., p. 91 (2007). A single VH or VL domain may be sufficient to confer antigen-binding specificity.

[0147] A "human antibody" is an antibody having an amino acid sequence corresponding to that produced by a human or human cell or from a non-human source utilizing the human antibody repertoire or other human antibody coding sequences Amino acid sequence of the derived antibody. This definition of a human antibody specifically excludes humanized antibodies comprising non-human antigen-binding residues.

[0148] A "humanized" antibody is a chimeric antibody comprising amino acid residues from a non-human HVR and amino acid residues from a human FR. In certain embodiments, a humanized antibody will comprise substantially all of at least one (and usually two) variable domains, wherein all or substantially all of the HVRs (e.g., CDRs) correspond to those of a non-human antibody, and all Or substantially all FRs correspond to those of human antibodies. A humanized antibody optionally can comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody (eg, a non-human antibody) refers to an antibody that has been humanized.

[0149] As used herein, the term "hypervariable region" or "HVR" refers to each region of an antibody variable domain that is highly variable in sequence (a "complementarity determining region" or "CDR") and / or or form structurally defined loops ("hypervariable loops") and / or contain antigen contact residues ("antigen contacts"). Typically, antibodies contain six HVRs; three in the VH (H1, H2, H3), and three in the VL (L1, L2, L3). Herein, exemplary HVRs include: (a) Hypervariable loops are present 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 present at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition Public Health Service, National Institutes of Health, Bethesda, MD (1991)); (c) Antigen contacts are present 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) Combinations of (a), (b) and / or (c), including HVR amino acid residues 46-56 (L2), 47-56 (L2), 48-56 (L2), 49-56 (L2), 26-35 (H1), 26-35b (H1), 49-65 (H2), 93-102 (H3), and 94-102 (H3).

[0150] HVR residues and other residues (e.g., FR residues) in variable domains are numbered herein according to Kabat et al. (supra), unless otherwise stated.

[0151] "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. Therefore, HVR and FR sequences usually appear in VH (or VL) in the following order: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0152] The "human consensus framework" is a framework representing the most common amino acid residues in a series of human immunoglobulin VL or VH framework sequences. Typically, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. Typically, a subgroup of sequences is such as in Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, NIH Publication 91-3242, Bethesda MD (1991), vol. 1-3. In one embodiment, for VL, the subgroup is subgroup κI as described by Kabat et al., supra. In one embodiment, for VH, the subgroup is subgroup III as described by Kabat et al., supra.

[0153] For the purposes herein, an "acceptor human framework" is one derived from a human immunoglobulin framework or a human common framework, comprising a light chain variable domain (VL) framework or a heavy chain variable domain, as defined below (VH) The backbone of the amino acid sequence of the backbone. An acceptor human framework "derived from" a human immunoglobulin framework or a human consensus framework may comprise the same amino acid sequence as these, or it may contain changes in the amino acid sequence. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or more less, or 2 or less. In some embodiments, the VL receptor human framework is identical in sequence to a VL human immunoglobulin framework sequence or a human consensus framework sequence.

[0154] The "class" of an antibody refers to the type of constant domain or region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into subclasses (isotypes), such as IgG 1, IgG 2, IgG 3, IgG 4, IgA 1 and IgA2. The heavy-chain constant domains that correspond to the different classes of immunoglobulins are called α, β, ε, γ, and μ, respectively.

[0155] As used herein, the term IgG "isotype" or "subtype" refers to any subtype of an immunoglobulin defined by the chemical and antigenic properties of its constant regions.

[0156] The terms "Fc domain" or "Fc region" are used herein to define the C-terminal region of an immunoglobulin heavy chain comprising at least a portion of the constant region. The term includes native sequence Fc regions and variant Fc regions. The Fc region of a human IgG heavy chain is generally defined as extending from Cys226 or Pro230 to the carboxy-terminus of the heavy chain, although the boundaries of the Fc region of an IgG heavy chain may vary slightly. However, antibodies produced by host cells may undergo post-translational cleavage at one or more, particularly one or two, amino acids at the C-terminus of the heavy chain. Thus, an antibody produced by a host cell expressing a particular nucleic acid molecule encoding a full-length heavy chain may include a full-length heavy chain, or may include a cleavage variant of a full-length heavy chain (also referred to herein as a "cleavage variant heavy chain"). chain"). This is likely the case when the last two C-terminal amino acids of the heavy chain are glycine (G446) and lysine (K447, EU numbering). Thus, the C-terminal lysine (Lys447) or the C-terminal glycine (Gly446) and lysine (K447) of the Fc region may or may not be present. Unless otherwise stated herein, the numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system (also known as the EU index) as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition Public Health Service, National Institutes of Health, Bethesda, MD, 1991 (see also above). A "subunit" of an Fc domain, as used herein, refers to one of the two polypeptides forming a dimeric Fc domain, i.e., a polypeptide comprising the C-terminal constant region capable of stabilizing self-associated heavy chain immunoglobulins. For example, the subunit of the IgG Fc domain contains IgG CH2 and IgG CH3 constant domains.

[0157] "Modifications that promote the association of the first and second units of the Fc domain" are manipulations of the peptide backbone or post-translational modifications of the Fc domain subunit that reduce or prevent the polypeptide comprising the Fc domain subunit Association with the same polypeptide forms a homodimer. As used herein, association-promoting modifications specifically include individual modifications to each of the two Fc domain subunits (i.e., the first and second subunits of the Fc domain) for which association is desired, wherein the modification are complementary to each other in order to facilitate the association of the two Fc domain subunits. For example, association-promoting modifications can alter the structure or charge of one or both Fc domain subunits to make them sterically or electrostatically favorable, respectively. Thus, (hetero)dimerization occurs between a polypeptide comprising a first Fc domain subunit and a polypeptide comprising a second Fc domain subunit, which in turn is further fused to the assembly of each subunit (e.g., an antigen-binding moiety). may vary. In some embodiments, association-promoting modifications include amino acid mutations, particularly amino acid substitutions, in the Fc domain. In a particular embodiment, the association-promoting modification comprises individual amino acid mutations, in particular amino acid substitutions, in each of the two subunits of the Fc domain.

[0158] "Activated Fc receptors" are Fc receptors that, after engagement with the Fc region of an antibody, initiate signaling events that stimulate receptor-bearing cells to perform effector functions. Activating Fc receptors include FcγRIIIa (CD16a), FcγRI (CD64), FcγRIIa (CD32), and FcαRI (CD89).

[0159] When applied to antibodies, the term "effector functions" refers to those biological activities attributable to the Fc region of an antibody, which vary with antibody isotype. Examples of antibody utility functions include: C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), cytokine secretion, Uptake of immune complexes by antigen-presenting cells mediates antigen, downregulation of cell surface receptors (eg, B-cell receptors), and B-cell activation.

[0160] As used herein, the term "effector cell" refers to a population of lymphocytes displaying on their surface effector moiety receptors (e.g., cytokine receptors) and / or Fc receptors through which they bind effector moieties ( For example, cytokines) and / or the Fc region of an antibody and contribute to the destruction of target cells, such as tumor cells. Effector cells may, for example, mediate cytotoxicity or phagocytosis. Effector cells include, but are not limited to, effector T cells, such as CD8+ cytotoxic T cells, CD4+ helper T cells, γδ T cells, NK cells, lymphokine-activated killer (LAK) cells, and macrophages / monocytes.

[0161] The terms "engineer, engineered, engineering" as used herein are taken to include any manipulation of the peptide backbone or post-translational modification of a naturally occurring or recombinant polypeptide or fragment thereof. Engineering includes modifying the amino acid sequence, glycosylation pattern, or side chain groups of individual amino acids, as well as combinations of these methods. "Engineering" (especially with the prefix "glycosyl") and the term "glycosylation engineering" includes metabolic engineering of cellular glycosylation machinery, including genetic manipulation of oligosaccharide synthesis pathways, to achieve Expression of altered glycosylation of glycoproteins. In addition, glycosylation engineering includes mutations and the influence of the cellular environment on glycosylation. In one embodiment, glycosylation engineering is a change in glycosyltransferase activity. In a specific embodiment, the engineering results in altered glucosyltransferase activity and / or fucosyltransferase activity. Glycosylation engineering can be used to obtain "host cells with increased GnTIII activity" (e.g., one or more cells with β(1,4)-N-acetylglucosamine groups that have been manipulated to exhibit increased levels GnTIII (GnTIII) active polypeptide), "host cell with increased ManII activity" (e.g., one or more cells having α-mannosidase II (ManII) that has been manipulated to express increased levels active polypeptide), or "host cells with reduced α(1,6)-fucosyltransferase activity" (e.g., host cells that have been manipulated to express reduced levels of α(1,6)-fucosyltransferase Algal transferase host cells).

[0162] The terms "host cell", "host cell strain" and "host cell culture" are used interchangeably and refer to a cell into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells", which include primary transformed cells and progeny cells derived therefrom, regardless of the number of passages. The nucleic acid content of progeny cells may not be exactly the same as that of parental cells, but may contain mutations. Included herein are mutant progeny cells that have the same function or biological activity as screened or selected from the original transformed cells. A host cell is any type of cellular system that can be used to produce the proteins used in the present invention. In one embodiment, host cells are engineered to allow production of antibodies with modified oligosaccharides. In certain embodiments, the host cell has been manipulated to express increased levels of one or more polypeptides having β(1,4)-N-acetylglucosaminyltransferase III (GnTIII) activity. In certain embodiments, the host cell has been further manipulated to express increased levels of one or more polypeptides having alpha-mannosidase II (ManII) activity. Host cells include cultured cells, e.g., mammalian cultured cells such as CHO cells, BHK cells, NSO cells, SP2 / 0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells, or fusion tumors Cells, yeast cells, insect cells, and plant cells, to name a few, but also cells contained within transgenic animals, transgenic plants, or cultured plant or animal tissues.

[0163] As used herein, the term "polypeptide having GnTIII activity" refers to trimannan capable of catalyzing the addition of N-acetylglucosamine (GlcNAc) residues in β-1,4 linkages to N-linked oligosaccharides. β-linked mannosides of the sugar nucleus. This includes exhibiting the interaction with β(1,4)-N-acetylglucosaminyltransferase III (also known as β-1,4-mannosyl-glycoprotein 4-β-N-acetylglucosaminyl Base transferase (EC 2.4.1.144), according to the International Union of Biochemistry and Molecular Biology (NC-IUBMB) Nomenclature Committee) fusion polypeptide with similar but not necessarily identical enzymatic activity, as measured in specific biological assays , with or without dose dependence. Where a dose dependence does exist, it need not be identical to that of GnTIII, but rather substantially similar to that of GnTIII for a given activity (i.e., the candidate polypeptide will exhibit a greater The activity is reduced by no more than about 25-fold, preferably by no more than about ten-fold, most preferably by no more than about three-fold). In certain embodiments, the polypeptide having GnTIII activity is a fusion polypeptide comprising the catalytic domain of GnTIII and the Golgi localization domain of a heterologous Gorgi-resident polypeptide. In particular, the Gorgi body localization domain is the localization domain of mannosidase II or GnTI, most particularly, the localization domain of mannosidase II. Alternatively, the Gaugi body localization domain is selected from the group consisting of the localization domain of mannosidase I, the localization domain of GnTII, and the localization domain of α1,6 core fucosyltransferase. Methods of generating such fusion polypeptides and using them to generate antibodies with increased effector function are disclosed in WO2004 / 065540, U.S. Provisional Patent Application No. 60 / 495,142, and U.S. Patent Application Publication No. 2004 / 0241817, the entire contents of which are reproduced by References are expressly incorporated herein.

[0164] As used herein, the term "Gaugi-localization domain" refers to the amino acid sequence of a Gaugi-resident polypeptide that is responsible for anchoring the polypeptide at a position within the Gaugi complex. Typically, the localization domain comprises the amine-terminal "tail" of the enzyme.

[0165] As used herein, the term "polypeptide having ManII activity" refers to the terminal 1,3- and 1,6-linked α-D-linked GlcNAcMan 5GlcNAc2 mannose intermediates capable of catalyzing N-linked oligosaccharides. Hydrolyzed polypeptides of mannose residues. This includes exhibiting interactions with Gorgiella α-mannosidase II (also known as mannosyloligosaccharide 1,3-1,6-α-mannosidase II (EC 3.2.1.114), according to the International Biochem. Biological Union (NC-IUBMB) Nomenclature Committee) polypeptides with similar but not necessarily identical enzymatic activity.

[0166] Antibody-dependent cell-mediated cytotoxicity (ADCC) is an immune mechanism that causes immune effector cells to lyse antibody-coated target cells. A target cell is a cell to which an antibody or fragment thereof comprising an Fc region specifically binds, usually through a protein portion that is N-terminal to the Fc region. As used herein, the term "increased / decreased ADCC" is defined as the number of target cells lysed by the ADCC mechanism defined above with a given concentration of antibody in the medium surrounding the target cells within a given time Increase / decrease, and / or decrease / increase of the antibody concentration in the culture medium surrounding the target cells required to achieve lysis of a given number of target cells within a given time period by the ADCC mechanism. The increase / decrease in ADCC is mediated relative to the same antibody (but not yet engineered) produced by the same type of host cell using the same standard methods of production, purification, formulation and storage (known to those of ordinary skill in the art) ADCC. For example, by engineering via the methods disclosed herein to have altered glycosylation patterns (e.g., to express glycosyltransferases) relative to ADCC mediated by the same antibody produced by the same type of unengineered host cell , GnTIII or other glycosyltransferase) host cells produced antibody-mediated increases in ADCC.

[0167] "Antibody having increased antibody-dependent cell-mediated cytotoxicity (ADCC)" means an antibody as defined herein having an increased / decreased ADCC as determined by any suitable method known to those of ordinary skill in the art . An acceptable in vitro ADCC assay is as follows: 1) The assay uses target cells known to express a target antigen recognized by the antigen-binding region of an antibody; 2) The assay uses human peripheral blood mononuclear cells (PBMC) isolated from the blood of randomly selected healthy donors as effector cells; 3) Implement the assay according to the following operating scheme: i) Isolate PBMCs using standard density centrifugation protocols and suspend in RPMI cell culture medium at a concentration of 5 x 106 cells / ml; ii) Target cells were grown by standard tissue culture methods, harvested from exponential growth phase with a survival rate higher than 90%, washed in RPMI cell culture medium, labeled with 100 microcuries of 51Cr, and washed twice in cell culture medium times, and resuspended in cell culture medium at a density of 10 5 cells / ml; iii) Transfer 100 µl of the above final target cell suspension to each well of a 96-well microtiter plate; iv) Serially dilute the antibody from 4000 ng / ml to 0.04 ng / ml in cell culture medium and add 50 µl of the resulting antibody solution to the target cells in a 96-well microtiter plate and test coverage in triplicate Various antibody concentrations for the entire concentration range described above; v) For the maximal release (MR) control, add 50 μl of 2% (V / V) non-ionic detergent in water (Nonidet, Sigma, St. Louis), instead of antibody solution (point iv above); vi) For the spontaneous release (SR) control, add 50 µl of RPMI cell culture medium in place of the antibody solution (point iv above) to the other 3 wells of the plate containing the labeled target cells; vii) Then centrifuge the 96-well microtiter plate at 50 x g for 1 min and incubate at 4 °C for 1 h; viii) Add 50 μl of PBMC suspension (point i above) to each well so that the effector:target cell ratio is 25:1 and place the plate in an incubator with a 5% CO2 atmosphere medium at 37°C for 4 hours; ix) Collect the cell-free supernatant from each well and quantify the experimentally released radioactivity (ER) using a gamma counter; x) Calculate the percent specific lysis for each antibody concentration according to the formula (ER-MR) / (MR-SR) x 100, where ER is the mean radioactivity quantified for that antibody concentration (see point ix above) and MR is the mean radioactivity (see point ix above) quantified against the MR control (see point v above) and SR the mean radioactivity (see point ix above) quantified for the SR control (see point vi above); 4) "Increased / decreased ADCC" is defined as the maximum percentage increase / decrease in specific lysis observed over the above tested antibody concentration range, and / or the observed specificity is achieved over the above tested antibody concentration range Decrease / increase in antibody concentration required to lyse one-half of the maximum percentage. The increase / decrease in ADCC is mediated relative to the same antibody (but not yet engineered) produced by the same type of host cell using the same standard methods of production, purification, formulation and storage (known to those of ordinary skill in the art) The ADCC was quantified using the assay described above.

[0168] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the population comprises individual antibodies that share the same and / or bind to the same epitope, but do not contain, for example, those containing naturally occurring antibodies. Mutations generated or possible variant antibodies produced during the production of monoclonal antibody preparations, such variants usually exist in small amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinant sites (epitopes), monoclonal antibody preparations have each monoclonal antibody directed against a single epitope on the antigen. Thus, the modifier "monoclonal" indicates that the characteristics of the antibody were obtained from a substantially homogeneous population of antibodies and should not be construed as requiring that the antibody be produced by any particular method. For example, monoclonal antibodies intended for use in accordance with the present invention can be produced by a variety of techniques including, but not limited to, the fusionoma method, recombinant DNA methods, phage display methods, and the use of transgenic antibodies comprising all or part of the human immunoglobulin loci. Methods of breeding genetically modified animals are described herein, along with other exemplary methods for making monoclonal antibodies.

[0169] "Naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (eg, a cytotoxic moiety) or a radioactive label. Naked antibodies may be present in pharmaceutical formulations.

[0170] "Native antibody" refers to a naturally occurring immunoglobulin molecule of varying structure. For example, an Ig native IgG antibody is a heterotetrameric glycoprotein of approximately 150,000 daltons consisting of two identical light chains and two identical heavy chains bonded by disulfide bonds. From N-terminus to C-terminus, each heavy chain has a variable region (VH), also known as a variable heavy domain or heavy chain variable domain, followed by three constant domains (CH1, CH2 and CH3). Similarly, from N-terminus to C-terminus, each light chain has a variable region (VL), also known as a variable light domain or light chain variable domain, followed by a light chain constant (CL) domain. Based on the amino acid sequence of their constant domains, the light chains of antibodies can be classified into one of two types, called kappa (κ) and lambda (λ).

[0171] As used herein, the terms "first", "second" or "third", etc. with respect to an antigen-binding portion or domain are used to facilitate the distinction of each type of portion or domain when there is more than one portion or domain. Use of these terms is not intended to impart a particular order or direction unless explicitly stated.

[0172] The terms "multispecific" and "bispecific" mean that the antigen binding molecule is capable of specifically binding at least two different epitopes. Typically, a bispecific antigen binding molecule comprises two antigen binding sites, each of which is specific for a different epitope. In certain embodiments, a bispecific antigen binding molecule is capable of simultaneously binding two epitopes, particularly two epitopes expressed on two different cells.

[0173] The term "valency" or "valence" as used herein means the presence of a specified number of antigen binding sites in an antigen binding molecule. Thus, the term "monovalent binding to an antigen" indicates the presence of one (and not more than one) antigen-binding site specific for the antigen in the antigen-binding molecule.

[0174] "Antigen binding site" refers to the site, ie, one or more amino acid residues, of an antigen binding molecule that provides for interaction with an antigen. For example, the antigen binding site of an antibody comprises amino acid residues from the complementarity determining regions (CDRs). Native immunoglobulin molecules usually have two antigen-binding sites, and Fab molecules usually have a single antigen-binding site.

[0175] "Activating T-cell antigen" as used herein refers to an epitope presented by T-lymphocytes, especially cytotoxic T-lymphocytes, which is capable of inducing or enhancing T-cell activation upon interaction with an antigen-binding molecule. Specifically, the interaction of antigen-binding molecules with activating T-cell antigens can induce T-cell activation by triggering the signaling cascade of the T-cell receptor complex. An exemplary activating T cell antigen is CD3. In a particular embodiment, the activating T cell antigen is CD3, particularly the epsilon subunit of CD3 (see UniProt accession P07766 (version 130), NCBI RefSeq accession NP_000724.1 for human sequences; or UniProt accession Q95LI5 ( 49th edition), NCBI GenBank accession BAB71849.1, for the cynomolgus monkey [Macaca fascicularis] sequence).

[0176] "T cell activation" as used herein refers to one or more cellular responses of T lymphocytes, in particular cytotoxic T lymphocytes, selected from: proliferation, differentiation, cytokine secretion, cytotoxic effector molecules Release, cytotoxic activity and expression of activation markers. The T cell activation therapeutic agent used in the present invention is capable of inducing T cell activation. Suitable assays for measuring T cell activation are known in the art as described herein.

[0177] A "target cell antigen" as used herein refers to an antigenic determinant present on the surface of a target cell (eg, a cell in a tumor, such as a cancer cell or a cell of the tumor stroma). In a specific embodiment, the target cell antigen is CD20, in particular human CD20 (see UniProt number P11836).

[0178] As used herein, "B cell antigen" refers to an antigenic determinant presented on the surface of B lymphocytes, particularly malignant B lymphocytes (in which case the antigen is also referred to as "malignant B cell antigen").

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

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

[0181] "Chimeric Antigen Receptor" or "CAR" refers to a genetically engineered receptor protein comprising an antigen-binding moiety, such as a single-chain variable fragment (scFv) of a targeting antibody, a transmembrane domain, intracellular T cell activation signaling Domain (eg, CD3 ζ chain of T cell receptor) and optionally one or more intracellular co-stimulatory domains (eg, CD28, CD27, CD137 (4-1BB), Ox40). CAR mediates antigen recognition, T cell activation, and (in the case of second-generation CARs) co-stimulation to enhance T cell function and persistence. For a review see eg Jackson et al., Nat Rev Clin Oncol. (2016) 13, 370-383.

[0182] "Fusion" means that components such as a Fab molecule and an Fc domain subunit are linked via peptide bonds, either directly or via one or more peptide linkers.

[0183] An "effective amount" of a drug refers to the amount required to cause physiological changes in the cells or tissues to which it is administered.

[0184] A "therapeutically effective amount" of a medicament such as a pharmaceutical composition refers to an amount effective to achieve the desired therapeutic or preventive effect within the required dosage and time period of administration. A therapeutically effective amount of an agent eg eliminates, reduces, delays, minimizes or prevents the adverse effects of a disease.

[0185] "Therapeutic agent" refers to an active ingredient, such as a pharmaceutical composition, which is administered to an individual in an attempt to alter the natural course of the disease in the individual being treated, and may be used prophylactically or administered during the course of clinical pathology. "Immunotherapeutic agent" refers to a therapeutic agent administered to an individual in an attempt to restore or enhance the individual's immune response, such as an immune response to a tumor.

[0186] The term "pharmaceutical composition" refers to a preparation in a form that permits the biological activity of the active ingredients contained therein to be effective and free of other components that would be unacceptably toxic to the individual to whom the composition will be administered.

[0187] "Pharmaceutically acceptable carrier" refers to ingredients in a pharmaceutical composition other than active ingredients that are non-toxic to individuals. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers or preservatives.

[0188] The terms "package insert" or "instructions for use" are used to refer to instructions commonly included in commercial packages of therapeutic products, which include the indications, usage, dosage, route of administration, combination, Remedies, contraindications and / or warnings.

[0189] As used herein, the term "combination therapy" encompasses combined administration (where two or more therapeutic agents are included in the same or separate formulations) as well as separate administration, in which case the administration is as described herein. Reporting of the antibodies can occur prior to, concurrently with and / or after administration of one or more additional therapeutic agents, preferably one or more antibodies.

[0190] A "crossover" Fab molecule (also referred to as "Crossfab") means a Fab molecule in which the variable or constant domains of the Fab heavy chain and the Fab light chain are exchanged (i.e. replace each other), i.e., a crossfab molecule Contains a peptide chain consisting of the light chain variable domain VL and the heavy chain constant domain 1 CH1 (VL-CH1, in the N-terminal to C-terminal direction), and a peptide consisting of the heavy chain variable domain VH and the light chain constant domain CL chain (VH-CL, in the N-terminal to C-terminal direction). For clarity, in a swapped Fab molecule in which the variable domains of the Fab light chain and the Fab heavy chain are swapped, the peptide chain comprising the heavy chain constant domain 1 CH1 is referred to herein as the "heavy chain" of the (swapped) Fab molecule. ". Conversely, in a swapped Fab molecule in which the constant domains of the Fab light chain and the Fab heavy chain are swapped, the peptide chain comprising the variable domain VH of the heavy chain is referred to herein as the "heavy chain" of the (swapped) Fab molecule.

[0191] In contrast, a "conventional" Fab molecule is meant in its natural form (i.e., comprising a heavy chain (VH-CH1, in the N-terminal to C-terminal direction) consisting of a heavy chain variable domain and a constant domain and consisting of a light chain variable domain. Fab molecule of a light chain (VL-CL, in the N-terminal to C-terminal direction) consisting of a V domain and a constant domain.

[0192] The term "polynucleotide" refers to an isolated nucleic acid molecule or construct, such as messenger RNA (mRNA), RNA of viral origin, or plastid DNA (pDNA). A polynucleotide may contain conventional phosphodiester bonds or non-conventional bonds (eg, amide bonds, such as those found in peptide nucleic acids (PNAs)). The term "nucleic acid molecule" refers to any one or more nucleic acid segments, such as DNA or RNA segments, present in a polynucleotide.

[0193] An "isolated" nucleic acid molecule or polynucleotide refers to a nucleic acid molecule (DNA or RNA) that has been isolated from its natural environment. For example, a recombinant polynucleotide encoding a polypeptide contained in a vector is considered isolated for purposes of the present invention. Further examples of isolated polynucleotides include recombinant polynucleotides maintained in heterologous host cells or purified (partially or substantially) polynucleotides in solution. An isolated polynucleotide includes a polynucleotide molecule contained in a cell that ordinarily contains the polynucleotide molecule, but the polynucleotide molecule is present extrachromosomally or at a chromosomal location that differs from the natural chromosomal location. Isolated RNA molecules include in vivo or in vitro RNA transcripts of the invention, as well as positive- and negative-strand forms, and double-stranded forms. Isolated polynucleotides or nucleic acids according to the invention further include such molecules produced synthetically. Furthermore, a polynucleotide or nucleic acid may be or may include a regulatory element, such as a promoter, ribosomal binding site, or transcription terminator.

[0194] By a nucleic acid or polynucleotide having a nucleotide sequence that is at least, e.g., 95% "identical" to a reference nucleotide sequence of the present invention, it is meant that the nucleotide sequence of the polynucleotide is identical to the reference sequence , in addition to every 100 nucleotides of the reference nucleotide sequence, the polynucleotide sequence may contain up to five point mutations. In other words, to obtain a polynucleotide having a nucleotide sequence that is at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or replaced with another nucleotide, Or insert up to 5% of the total number of nucleotides in the reference sequence into the reference sequence. These alterations of the reference sequence may occur at the 5' or 3' positions of the reference nucleotide sequence or anywhere in between, both interspersed between residues in the reference sequence and at positions within the reference sequence. in one or more consecutive groups. In practice, whether any particular polynucleotide sequence is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a nucleotide sequence of the invention can be determined using established Known computer programs are routinely determined, such as the programs discussed above for polypeptides (eg, ALIGN-2).

[0195] The term "expression cassette" refers to a recombinantly or synthetically produced polynucleotide having a series of specific nucleic acid elements that permit transcription of a specific nucleic acid in a target cell. Recombinant expression cassettes can be introduced into plastids, chromosomes, mitochondrial DNA, chromosomal DNA, viruses or nucleic acid fragments. Typically, the recombinant expression cassette portion of an expression vector includes, among other sequences, a nucleic acid sequence to be transcribed and a promoter. In certain embodiments, an expression cassette of the invention comprises a polynucleotide sequence encoding a bispecific antigen binding molecule of the invention or a fragment thereof.

[0196] The term "vector" or "expression vector" is synonymous with "expression construct" and refers to a DNA molecule for introducing a specific gene and directing the expression of that gene, the DNA molecule being operably linked to the gene in a target cell . The term includes vectors that are self-replicating nucleic acid structures as well as vectors that are incorporated into the genome of the host cell that has been introduced. The presentation carrier of the present invention includes a presentation box. Expression vectors transcribe large amounts of stable mRNA. Once the expression vector enters the target cell, the ribonucleic acid molecule or protein encoded by the gene is produced by the cellular transcription and / or translation mechanism. In one embodiment, the expression vector of the present invention comprises an expression cassette comprising a polynucleotide sequence encoding the bispecific antigen-binding molecule of the present invention or a fragment thereof.

[0197] As used herein, the term "about" refers to the usual error range of the respective value readily known to those skilled in the art. Reference herein to "about" a value or parameter includes (and describes) embodiments that are specific to that value or parameter per se.

[0198] "B-cell proliferative disorder" means a disease in which the number of B cells in a patient is increased compared to that in a healthy individual, particularly in which the increased number of B cells is a cause or hallmark of the disease. A "CD20-positive B-cell proliferative disorder" is a B-cell proliferative disorder in which B cells, particularly malignant B cells (in addition to normal B cells), express CD20.

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

[0200] "Refractory disease" was defined as incomplete response to first-line therapy. In one embodiment, refractory disease is defined as non-response or relapse to prior therapy within 6 months. In one embodiment, refractory disease is characterized by one or more of the following: progressive disease (PD) as best response to first-line therapy, stable disease (SD) as best response to first-line therapy for at least 4 cycles (e.g. , 4 cycles of rituximab, cyclophosphamide, doxorubicin hydrochloride (hydroxydaunorubicin), vincristine sulfate (Oncovin) and prednisone (also abbreviated as R-CHOP) followed by the final Best response, or partial response (PR) as best response after at least 6 cycles, and biopsy-proven residual disease or disease progression after partial response. "Relapsed disease" was defined as complete response to first-line therapy. In one embodiment, disease recurrence is confirmed by biopsy. In one embodiment, the patient has relapsed after at least two prior systemic treatment regimens (including at least one prior regimen containing anthracyclines, and at least one prior regimen containing anti-CD20-directed therapy) or on Unresponsive to systemic therapy.

[0201] A "subject" or "individual" is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cattle, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats) mouse). Preferably, the subject or individual is human. In one case, each individual in a group of individuals is a person. In one instance, each individual in a reference group of individuals is a person.

[0202] "Ineligible for transplantation" individuals or "ineligible for autologous stem cell transplantation (SCT)" individuals are individuals who are not eligible for autologous stem cell transplantation, are not recommended, cannot receive or refuse autologous stem cell transplantation. Examples of preferred individual characteristics include age ≤ 65 years, Karnofsky performance status (KPS; Karnofsky et al. Cancer. 1948; 1(4): 634-656) > 60, forced expiratory volume in 1 second (FEV 1 ) > 60 % predicted value, lung diffusing capacity (DLCO) > 60% predicted value, left ventricular ejection fraction > 45%, normal heart rhythm, serum bilirubin ≤ 2 mg / 100 mL, alanine aminotransferase (ALT) / day Aspartate aminotransferase (AST) < 2 × normal, serum creatinine ≤ 1.5 mg / 100 mL, creatinine clearance > 60 mL / min, no second active malignancy, no pregnancy, and no uncontrolled infections (including dental) (Hamadani M et al. Bone Marrow Transplant. 2010; 45:1259–68).

[0203] As used herein, "treatment" (and its grammatical variants, such as "the course of treatment" or "in treatment"), refers to a clinical intervention that seeks to alter the natural course of the disease in the individual being treated, and may be performed either prophylactically or during the course of clinical pathology. in the implementation. Desired therapeutic effects include, but are not limited to, prevention of occurrence or recurrence of disease, relief of symptoms, relief of any direct or indirect pathological consequences of disease, prevention of metastasis, slowing of disease progression, amelioration or alleviation of disease state, remission or improved prognosis. In some embodiments, the methods of the invention are used to delay the development of a disease or slow the progression of a disease.

[0204] As used herein, "delaying progression" of a disorder or disease means delaying, hindering, slowing, delaying, stabilizing and / or delaying the development of a disease or disorder (e.g. CD20 positive B cell proliferative disorder, e.g. NHL, e.g. DLBCL). This delay can be of varying lengths, depending on the medical history and / or the individual being treated. As will be apparent to those skilled in the art, a sufficient or substantial delay may in fact encompass prophylaxis such that the individual does not develop the disease. For example, in advanced cancer, the development of central nervous system (CNS) metastases may be delayed.

[0205] "Reduce" or "inhibit" means the ability to cause an overall reduction of 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% or greater. For clarity, the term also includes reduction to zero (or below the detection limit of the analytical method), ie complete abolition or elimination. In certain embodiments, reduction or inhibition may refer to no change relative to the target dose of the bispecific antibody following treatment with the anti-CD20 / anti-CD3 bispecific antibody using the escalating dosing regimen of the invention , pre-doser, reduction or suppression of undesired events, such as cytokine-driven toxicity (eg, 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. In other embodiments, reducing or inhibiting may refer to antibody effector functions mediated by the Fc region of the antibody, such effector functions specifically including complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC), and antibody-dependent cytotoxicity. Sex cell phagocytosis (ADCP). In other embodiments, reduction or inhibition may refer to a CD20 positive B cell proliferative disorder being treated (e.g. NHL (e.g. DLBCL), FL (e.g. relapsed and / or refractory FL or transformed FL), MCL, hyperplastic malignant B-cell lymphoma or PMLBCL), the presence or size of metastases, or the size of the primary tumor.

[0206] As used herein, "administering" means administering to an individual a given dose of a compound (e.g., anti-CD20 / anti-CD3 bispecific antibody) or composition (e.g., a pharmaceutical composition, e.g., comprising an anti-CD20 / anti-CD3 bispecific antibody) Antibody pharmaceutical composition) method. Compounds and / or compositions utilized in the methods disclosed herein can be administered intravenously (eg, by intravenous infusion).

[0207] A "fixed" or "uniform" dose of a therapeutic agent (eg, bispecific antibody) herein refers to the dose administered to a patient regardless of the patient's body weight or body surface area (BSA). Thus, fixed or flat doses are not given in mg / kg or mg / m2 doses, but in absolute amounts (eg, mg) of the therapeutic agent.

[0208] The "target dose" herein refers to the dose of the anti-CD20 / anti-CD3 bispecific antibody that achieves the therapeutic effect, that is, the desired clinical efficacy. The possible target doses of grafitumumab were found to be 16 mg or 30 mg.

[0209] "Constant or preset dosing with target dose" and "regimen without step-up dosing regimen" refer to dosing using the same dose in the first and second cycles and any subsequent treatment cycles, as appropriate. As opposed to step-up dosing, step-up dosing uses lower doses in the first few treatment cycles and reaches the target dose only in the second or subsequent treatment cycles.

[0210] As used herein, the term "treatment cycle" or "cycle" (abbreviation: "C" refers to a course of one or more doses of anti-CD20 / anti-CD3 bispecific antibody repeated on a regular schedule, as appropriate, There is a rest period (no treatment) in between. In one aspect of the invention, the first treatment cycle comprises a first dose of anti-CD20 / anti-CD3 bispecific antibody and a second dose, followed by a rest period. In one such embodiment , the first treatment cycle consisted of the first dose of the anti-CD20 / anti-CD3 bispecific antibody on day 1 of the first cycle and the second dose of the anti-CD20 / anti-CD3 bispecific antibody on day 8 of the first cycle. dose, followed by a 12-day break. In one embodiment, the second and any subsequent cycle comprises administering one dose of an anti-CD20 / anti-CD3 bispecific antibody on the first day of the cycle, followed by a 20-day rest. In one embodiment In another embodiment, a treatment cycle includes 21 days. In another embodiment, a treatment cycle includes 14 days. A treatment cycle comprising one or more doses of an anti-CD20 / anti-CD3 bispecific antibody may further comprise one or more One or more doses of other therapeutic agents (such as anti-CD20 antibodies, especially obinutuzumab). The treatment regimen according to the invention may comprise 2 or more treatment cycles, or 3, 4, 5, 6 , 7, 8, 9, 10, 11, especially 12 treatment cycles.

[0211] "Individual response" or "response" can be assessed using any endpoint indicative of benefit to the individual, including, but not limited to, (1) some degree of inhibition of disease progression (e.g., CD20-positive B-cell proliferative disorder, e.g., non-Hodgkin lymphoma (NHL) progression); including slowing and complete arrest; (2) reduction in tumor size; (3) inhibition (i.e., reduction, slowing, or complete cessation) of cancer cell migration to adjacent peripheral organs and / or Tissue infiltration; (4) inhibition (i.e., reduction, slowing, or complete cessation) of metastasis; (5) partial alleviation of one or more of the conditions associated with CD20-positive B-cell proliferative disorders (eg, B-cell proliferative dysregulation) (6) increase or prolong the length of survival, including overall survival and disease progression-free survival; and / or (7) reduce mortality at a given time point after treatment.

[0212] As used herein, "complete response" or "CR" refers to the disappearance of all target lesions. In one embodiment, standard NHL response criteria are assessed to determine CR. (Lugano classification, Cheson et al. J Clin Oncol. 2014 Sep 20; 32(27): 3059–3067.).

[0213] 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 with reference to baseline SLD, or at least a 30% reduction in the sum of target lesion diameters (SPD) ) is reduced by at least 50%.

[0214] "Sustained response" refers to the continued reduction in tumor growth after cessation of treatment. For example, the size of the tumor can remain constant or decrease compared to the size at the beginning of the administration period. In some embodiments, the duration of the sustained response is at least as long as the duration of treatment, or at least 1.5 times, 2.0 times, 2.5 times, or 3.0 times the duration of treatment.

[0215] An "effective response" in a subject or a "response" in a subject to drug treatment and similar terms refers to a clinical or therapeutic benefit conferred on a subject having or suffering from a risk of a disease or disorder, such as cancer. In one embodiment, such benefits include one or more of: prolonging survival (including overall survival and progression-free survival); producing an objective response (including complete or partial response); or improving signs of cancer or symptoms.

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

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

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

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

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

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

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

[0223] As used herein, "no change in disease" or "SD" refers to the minimum SLD since the start of treatment as a reference, and the target symptom has neither shrunk to meet the requirements of PR nor increased to meet the requirements of PD.

[0224] As used herein, "disease progression" or "PD" refers to an increase in the SLD of the target lesion of at least 20% with reference to the smallest SLD recorded since the start of treatment, or to the smallest SLD recorded since the start of treatment. SPD of target lesions increased by at least 50%, or one or more new lesions appeared.

[0225] As used herein, an "infusion-related reaction," "IRR," or infusion-related adverse event is a reaction to a patient's or individual's response to a drug (e.g., anti-CD20 / anti-CD3 bispecific antibody, e.g., grifiduzumab; or anti-CD79b Adverse events occurring during or within 24 hours of administration of an antibody-drug conjugate (e.g., parotuzumab). IRR can be graded on a scale of 1-5 according to eg NCI CTCAE v.4.

[0226] The term "PD-1 axis binding antagonist" refers to a molecule that inhibits the interaction of a PD-1 axis binding partner with one or more of its binding partners, thereby eliminating T cell Dysfunction, the result of which is restoration or enhancement of T cell function (eg, 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.

[0227] The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with the interaction between PD-1 and its one or more binding partners (such as PD-L1, PD-L2). Interaction-induced message transduction. In some embodiments, a PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to its one or more binding partners. In specific aspects, a PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1 and / or PD-L2. For example, PD-1 binding antagonists include anti-PD-1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and agents that reduce, block, inhibit, eliminate or interfere with the interaction between PD-1 and PD-L1 and / or Or other molecules of signal transduction caused by the interaction of PD-L2. In one embodiment, a PD-1 binding antagonist reduces negative co-stimulatory signals (signals mediated by PD-1) that are mediated by or expressed by cell surface proteins expressed on T lymphocytes, thereby alleviating Dysfunction Dysfunction of T cells (eg, enhancing effector responses to antigen recognition). In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody. In a specific aspect, the PD-1 binding antagonist is MDX-1106 (nivolumab). In another specific aspect, the PD-1 binding antagonist is pembrolizumab (formerly known as lambrolizumab (MK-3475)). In another specific aspect, the PD-1 binding antagonist is AMP-224. In some embodiments, the PD-1 binding antagonist is MDX-1106 (nivolumab). In some embodiments, the PD-1 binding antagonist is MK-3475 (pembrolizumab). In some embodiments, the PD-1 binding antagonist is MED1-0680. In some instances, the PD-1 binding antagonist is PDR001 (spartalizumab). In some instances, the PD-1 binding antagonist is REGN2810 (cemiplimab). In some examples, the PD-1 binding antagonist is BGB-108. In other cases, the anti-PD-1 binding antagonist antibody is prolgolimab, camrelizumab, sintilimab, tislelizumab or toripalimab.

[0228] Further examples of PD-1 axis binding antagonists include simiprizumab, prologumab, camrelizumab, sintilimab, tislelizumab, toripalimab , Dostarlimab, Retifanlimab, Spartakizumab, Sasanlimab, Penpulimab, CS1003, HLX10, SCT-I10A, SHR-1316, CS1001, Envafolimab, TQB2450, ZKAB001, LP-002, Zimberelimab, Balstilimab, Genolimzumab, BI 754091, Cetrelimab , YBL-006, BAT1306, HX008, CX-072, IMC-001, KL-A167, Budigalimab, AMG 404, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, APL-502, Cosibelimab, lodapolimab, GS-4224, INCB086550, FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, MAX-10181, RC98, BION-004, AM0001, 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.

[0229] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with the interaction of PD-L1 with one or more of its binding partners (e.g., PD-1 or B7-1). Action-induced signal transduction. In some embodiments, a PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partner. In specific aspects, a PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1 and / or B7-1. In some embodiments, PD-L1-binding antagonists include anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and agents that reduce, block, inhibit, eliminate, or interfere with the binding of PD-L1 to one of them. Other molecules for signal transduction caused by the interaction of one or more binding partners (such as PD-1 or B7-1). In one embodiment, a PD-L1 binding antagonist reduces negative co-stimulatory signals (signals mediated by PD-L1) that are mediated by or expressed by cell surface proteins expressed on T lymphocytes, thereby alleviating Dysfunction Dysfunction of T cells (eg, enhancing effector responses 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 disclosed herein. In another specific embodiment, the anti-PD-L1 antibody is MDX-1105 disclosed herein. In yet another specific aspect, the anti-PD-L1 antibody is MEDI4736 disclosed herein.

[0230] As used herein, the term "atezolizumab" refers to drugs with an International Nonproprietary Name (INN) list112 (WHO Medicines Information, Vol. 28, No. 4, 2014, p. 488) or a CAS registry number. 1380723-44-3 Anti-PD-L1 antagonist antibody.

[0231] The term "PD-L2 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with the interaction of PD-L2 with any one or more of its binding partners, such as PD-1 signal transduction. In some embodiments, a PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to its one or more binding partners. In specific aspects, a PD-L2 binding antagonist inhibits 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 agents that reduce, block, inhibit, eliminate, or interfere with the expression of PD-L2 and one of them. Or other molecules of signal transduction caused by the interaction of multiple binding partners (such as PD-1). In one embodiment, a PD-L2 binding antagonist reduces negative co-stimulatory signals (signals mediated by PD-L2) that are mediated by or expressed by cell surface proteins expressed on T lymphocytes, thereby alleviating Dysfunction Dysfunction of T cells (eg, enhancing effector responses to antigen recognition). In some embodiments, the PD-L2 binding antagonist is an immunoadhesin.

[0232] 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., relapsed or refractory B-cell proliferative disorder) , eg, non-Hodgkin's lymphoma (NHL; eg, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL; eg, grade 1 FL, grade 2 FL, grade 3 FL (eg, grade 3a FL, grade 3b FL), or transformed FL), mantle cell lymphoma (MCL) or marginal zone lymphoma (MZL)), e.g., relapsed or refractory NHL (e.g., relapsed or refractory DLBCL , relapsed or refractory FL, relapsed or refractory MCL, or marginal zone lymphoma (MZL))). Examples of chemotherapeutic agents include: EGFR inhibitors (including small molecule inhibitors (e.g., erlotinib (TARCEVA®, Genentech / OSI Pharm.)); PD 183805 ( CI 1033, 2-acrylamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-𠰌linyl)propoxy]-6-quinoline Azolinyl]-, dihydrochloride, Pfizer Inc.); ZD1839, gefitinib (IRESSA®) 4-(3'-chloro-4'-fluoroanilino)-7 -Methoxy-6-(3-(N-𠰌linyl)propoxy)quinazoline, AstraZeneca); ZM 105180 (6-amino-4-(3-methylbenzene -amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-(1-methyl-piperidine-4- base)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166((R)-4-[4-[(1-benzene (Ethyl)amino]-1H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-benzene (ethylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine);CL-387785(N-[4-[(3-bromophenyl)amino]-6-quinazolinyl] -2-butyramide); 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 dual EGFR / HER2 tyrosine kinase Inhibitors such as lapatinib (TYKERB®, GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6-[5-[[[2 -methylsulfonyl)ethyl]amino]methyl]-2-furyl]-4-quinazolinamine)); tyrosine kinase inhibitors (e.g., EGFR inhibitors; small molecule HER2 tyramine Acid 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 binds EGFR but inhibits both HER2-overexpressing cells and EGFR-overexpressing cells; PKI-166 (Novartis); pan-HER inhibitors such as Canertinib ( canertinib ) (CI-1033; Pharmacia); Raf-1 inhibitors such as the antisense agent ISIS-5132 (ISIS Pharmaceuticals), which inhibits Raf-1 signaling; non-HER-targeted tyrosine Kinase inhibitors such as imatinib mesylate (GLEEVEC®, Glaxo SmithKline); multi-targeted tyrosine kinase inhibitors such as sunitinib (SUTENT ®, Pfizer); VEGF receptor tyrosine kinase inhibitors such as valatanib (PTK787 / ZK222584, Novartis / Schering AG); MAPK extracellular regulated kinase I inhibitor CI- 1040 (Pharmacia); quinazolines such as PD 153035, 4-(3-chloroanilino)quinazolines; pyridopyrimidines; pyrimidopyrimidines; pyrrolopyrimidines such as CGP 59326, CGP 60261 and CGP 62706; pyrazolopyrimidines, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidines; curcumin (diaferuloylmethane, 4,5-bis(4- fluoroanilino)phthalimide); tyrosine phosphorylation inhibitors containing a nitrothiophene moiety; PD-0183805 (Warner-Lamber); antisense molecules (e.g., those with HER-encoded nucleic acid binders); quinolines (US Patent No. 5,804,396); tyrosine phosphorylation inhibitors (US Patent No. 5,804,396); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering Pharmaceuticals); pan-HER inhibitors such as Ci-1033 (Pfizer); Affinitac (ISIS 3521; Isis / Lilly); PKI 166 (Novartis); SuSmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Sematinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering pharmaceuticals); INC-1C11 (Imclone); and rapamycin (sirolimus, RAPAMUNE®); proteasome inhibitors such as bortezomib (VELCADE ®, Millennium Pharm.)); disulfiram; epigallocatechin gallate; marine actinomycin (salinosporamide) A; carfilzomib; 17-AAG (geldanamycin (geldanamycin)); radicicol; lactate dehydrogenase A (LDH-A); fulvestrant (FASLODEX®, AstraZeneca); ritrozole (FEMARA ®, Novartis); finasunate (VATALANIB®, Novartis); oxaliplatin (ELOXATIN®, Sanofi); 5-FU (5-fluorouracil); formyltetrahydrofolate (leucovorin); lonafamib (SCH 66336); sorafenib (NEXAVAR®, Bayer Labs); AG1478, alkylating agents such as thiotepa and CYTOXAN ® Cyclophosphamide; Alkylsulfonates such as busulfan, improsulfan, and piposulfan; Acridines such as benzodopa, carbosulfan Carboquone, meturedopa and uredopa; ethyleneimines and methylamelamines, including altretamine, triethylenemelamine ( triethylenemelamine), triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; acetogenins (especially burrata) Bullatacin and bullatacinone); camptothecin (including topotecan and irinotecan); bryostatin; callystatin ); CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); nostocins (especially nostocin 1 and nostoc 8); adrenocorticosteroids (including prednisone and penicortisol); cyproterone acetate; 5α-reductase, including finasteride and dutasteride ); vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat, dolastatin; Aldesleukin, talc duocarmycin (including synthetic analogues KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyn ; spongistatin; nitrogen mustards, such as chlorambucil, chlomaphazine, chlorophosphamide, estramustine, everamide, methyl dimethicone (Chloroethyl)amine, Methyldi(Chloroethyl)amine Oxide Hydrochloride, Moldfarin, Novembichin, Phenesterine, Prednimustine , trofosfamide, uracil mustard; nitrosoureas, such as dichloroethylnitrosourea, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics, such as enediyne antibiotics (e.g., calicheamicin, especially calicheamicin calicheamicin gamma 1 and calicheamicin omega 1); dynemicins, including dannamycin A; bisphosphonates, such as clodronate; esperamicin; and neocarcinogens Neocarzinostatin (neocarzinostatin) chromophore and related chromoprotein enediyne antibiotic chromophore), aclacinomysin, actinomycin (authramycin), nitrogen serine , cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, detorubicin (detorubicin), 6-diazo-5-oxo-L-norleucine, N-? (N-pyrrolinyl)-doxorubicin and doxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin ), mitomycins such as mitomycin C, mycophenolic acid, nogalamycin (nogalamycin), olivine, peplomycin (peplomycin), bofferomycin, puromycin, tri-iron quelamycin, rodorubicin, streptavidin, streptozocin, tubercidin, ubenimex, zinostatin, Zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pterotin (pteropterin), trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamipriine, thioguanine; pyrimidine analogs such as amcitabine (ancitabine), azacitidine, 6-azuridine, carmofur, cytarabine, dideoxyuridine, doxifluridine , enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane ), testolactone; anti-adrenergics such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as folinic acid; aceglatone;aldophosphamide glycoside;aminolevulinic acid;eniluracil;amsacrine;bestrabucil; Bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate );Epothilone;Etoglucid;Gallium nitrate;Hydroxyurea;Entinan;Lonidainine;Maytansinoids (maytansinoid), such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidamnol; nitraerine); pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazine ( ethylhydrazide); procarbazine; PSK® polysaccharide complex (JHS Natural Products); razoxane; rhizoxin; sizofuran; spirogermanium); tenuazonic acid; triaziquone; 2,2',2''-trichlorotriethylamine; trichothecenes (especially T- 2 toxins, verracurin A, roridin A and anguidine); urethane; vindesine; dacarbazine; Mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; cytarabine ("Ara-C") ); thiotepa; chloranmbucil; GEMZAR® (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; Etoposide (VP-16); Efumide; Mitoxantrone; Mitoxantrone; Teniposide; Edatrexate; Daunomycin; aminopterin; capecitabine (XELODA®); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; amino acid (DMFO); vitamin A acids such as tretinoin; and pharmaceutically acceptable salts, acids, prodrugs and derivatives of any of the above. .

[0233] Chemotherapeutic agents also include (i) antihormonal agents that have hormone modulating or suppressing effects on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including NOLVADEX ®; tamoxifen citrate), raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, trioxifene , keoxifene, LY117018, onapristone, and FARESTON® (toremifine citrate); (ii) aromatase inhibitors that inhibit the enzyme aromatase, which Modulation of estrogen production by the adrenal glands, for example, 4(5)-imidazole, aminoglutarimide, MEGASE® (megestrol acetate), AROMASIN® (Ixemestane; Pfizer), Formestanie, fadrozole, RIVISOR® (vorozole), FEMARA® (ritorazole; Novartis), and ARIMIDEX® (anastrozole; AstraZeneca ); (iii) antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; Buserelin, tripterelin, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluroxymethyltestosterone, all transretinoic acids, fentanyl (fenretinide) and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; ( vi) antisense oligonucleotides, especially those that inhibit expression of genes in signaling pathways associated with abnormal cell proliferation, such as PKC-α, Ralf, and H-Ras; (vii) ribozymes, such as VEGF expression inhibitors (e.g., ANGIOZYME®) and HER2 expression inhibitors; (viii) vaccines, such as gene therapy vaccines, such as ALLOVECTIN®, LEUVECTIN®, and VAXID®; (ix) growth inhibitors, including vinca alkaloids (e.g., Vinca vinblastine), NAVELBINE® (vinorelbine), taxanes (e.g., paclitaxel, nab-paclitaxel, and docetaxel), topoisomerase II inhibitors (e.g., albino doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin) and DNA alkylating agents (e.g., tamoxifen (tamoxigen, dacarbazine, methylbis(chloroethyl)amine, cisplatin, methotrexate, 5-fluorouracil, and ara-C); and (x) any pharmaceutically acceptable Accepted salts, acids and derivatives.

[0234] The term "cytotoxic agent" as used herein refers to a substance that inhibits or prevents cellular function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioactive isotopes (for example, radioisotopes of 211At, 131I, 125I, 90Y, 186Re, 188Re, 153Sm, 212Bi, 32P, 212Pb, and Lu); chemotherapeutic agents or drugs (such as methotrexate methotrexate, adriamicin, or vinca alkaloids (vincristine, vinblastine, or etoposide), doxorubicin, mold melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitors; enzymes and fragments thereof, such as nucleases ; Antibiotics; Toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof; and various anti-tumor or anti-cancer agents disclosed below. [III.] [Therapeutic agent used in the method of the present invention] [] A. Anti-CD20 / anti-CD3 bispecific antibody

[0235] The present invention provides novel dosages of anti-CD20 / anti-CD3 bispecific antibodies. In one embodiment, the antibody is a monoclonal antibody. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is a polyclonal antibody. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is a human antibody. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is a humanized antibody. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is a chimeric antibody. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is a full length antibody. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is an IgG class antibody, especially an IgG1 subclass antibody. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is a recombinant antibody.

[0236] In certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody comprises an antibody fragment. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab')2, Fv and scFv fragments and others described below. For a review of certain antibody fragments, see Hudson et al., Nat. Med. 9:129-134 (2003). For a review of scFv fragments see, e.g., Plückthun, 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 US Patent Nos. 5,571,894 and 5,587,458. See U.S. Patent No. 5,869,046 for a discussion of Fab and F(ab')2 fragments comprising salvage receptor-binding epitope residues with increased in vivo half-life. In one embodiment, the antibody fragment is a Fab fragment or a scFv fragment.

[0237] Diabodies are antibody fragments that have two antigen-binding sites (which may be bivalent or bispecific). See, eg, EP 404,097; WO 1993 / 01161; Hudson et al., Nat. Med. 9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA90: 6444-6448 (1993). Trifunctional and tetrafunctional antibodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003).

[0238] A single domain antibody is an antibody fragment comprising all or part of the heavy chain variable domain of an antibody or all or part of the light chain variable domain of an antibody. In certain embodiments, the single domain antibody is a human single domain antibody (Domantis, Inc., Waltham, MA; see, eg, US Patent No. 6,248,516 B1).

[0239] Antibody fragments can be produced by various techniques including, but not limited to, proteolytic digestion of intact antibodies as disclosed herein and production by recombinant host cells (eg, E. coli or phage).

[0240] In certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody is a chimeric antibody. 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 non-human variable regions (eg, variable regions derived from mouse, rat, hamster, rabbit, or a non-human primate such as monkey) and human constant regions. In yet another example, a chimeric antibody is a "class-switched" antibody, in which the class or subclass has been altered compared to its parental antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0241] In certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody is a humanized antibody. Typically, non-human antibodies are humanized to reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. Typically, a humanized antibody comprises one or more variable domains in which HVRs such as CDRs (or portions thereof) are derived from non-human antibodies and FRs (or portions thereof) are derived from human antibody sequences. A humanized antibody optionally will comprise at least a portion of a human constant region. In some embodiments, some FR residues in a humanized antibody are substituted with corresponding residues from a non-human antibody (e.g., an antibody from which HVR residues are derived), e.g., to restore or improve antibody specificity or affinity.

[0242] Humanized antibodies and methods for their preparation are reviewed, e.g., in Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and further described, e.g., in Riechmann et al., Nature 332:323-329 (1988); Queen et al. Al, Proc. Nat'l Acad. Sci. USA86:10029-10033 (1989); US Patent Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods36:25-34 (2005) (specifically describing the decision region (SDR) grafting); Padlan, Mol. Immunol.28:489-498 (1991) (describing "resurfacing"); Dall'Acqua et al., Methods36:43-60 (2005) (describing " FR reorganization"); Osbourn et al., Methods 36:61-68 (2005); and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing the "guided selection" method for FR reorganization).

[0243] Human framework regions that can be used for humanization include, but are not limited to: framework regions selected using the "best fit" approach (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); The framework region of the consensus sequence of human antibodies of a particular subgroup of chain variable regions (see for example: 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 or human germline framework (see e.g. Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and derived from screening FR The framework region of the library (see, e.g., Baca et al., J. Biol. Chem. 272:10678-10684 (1997); and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996)).

[0244] In certain embodiments, 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).

[0245] Human antibodies can be prepared by administering an immunogen to a transgenic animal that has been modified to produce fully human antibodies or fully antibodies with human variable regions in response to antigenic challenge. Such animals typically contain all or part of the human immunoglobulin loci, which replace the endogenous immunoglobulin loci, or are present extrachromosomally or integrated randomly into the animal's chromosomes. In such transgenic mice, endogenous immunoglobulin loci have generally been inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125 (2005). See also, for example: U.S. Patent Nos. 6,075,181 and 6,150,584 (describing XENOMOUSE™ technology); U.S. Patent No. 5,770,429 (describing HuMab® technology); U.S. Patent No. 7,041,870 (describing K-M MOUSE® technology); and U.S. Patent Application Publication No. US 2007 / 0061900 (describing VelociMouse® technology). The human variable regions derived from intact antibodies produced by such animals may be further modified, for example, by binding to different human constant regions.

[0246] Human antibodies can also be produced by fusionoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for the production of human monoclonal antibodies have been described. (See e.g.: Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al. , J. Immunol., 147: 86 (1991).) Human antibodies produced by human B cell fusion tumor technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) middle. Other methods include those described in, for example, U.S. Pat. No. 7,189,826 (describing production of monoclonal human IgM antibodies by hybridoma cell lines), and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) ( Human-human hybridomas are described). Human hybridoma technology (Trioma technology) is also described in: Vollmers and Brandlein, Histology and Histopathology, 20(3):927-937 (2005); and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27 (3):185-91 (2005).

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

[0248] Binding domains comprised in anti-CD20 / anti-CD3 bispecific antibodies can be isolated by screening combinatorial libraries of binding moieties with the desired activity. For example, a variety of methods are known in the art for generating phage display libraries and screening such libraries for antibodies with desired binding properties. Such methods are reviewed, e.g., in Hoogenboom et al., in Methods in Molecular Biology 178:1-37 (eds. O'Brien et al., Human Press, Totowa, NJ, 2001), and are further described, e.g., in McCafferty et al. Nature 348 :552-554; Clackson et al. Nature 352: 624-628 (1991); Marks et al. J. Mol. Biol.222: 581-597 (1992); Marks and Bradbury, included in Methods in Molecular Biology 248:161-175 ( Lo, ed., Human Press, Totowa, NJ, 2003); Sidhu et al. J. Mol. Biol. 338(2): 299-310 (2004); Lee et al. J. Mol. Biol. 340(5): 1073- 1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA101(34): 12467-12472 (2004); and Lee et al. J. Immunol. Methods 284(1-2): 119-132 (2004).

[0249] In some phage display methods, VH and VL gene libraries are cloned separately by polymerase chain reaction (PCR) and randomly recombined in the phage library, and antigen-binding phage can then be screened as described in: Winter et al., Ann. Rev. Immunol., 12: 433-455 (1994). Phage typically display antibody fragments as single-chain Fv (scFv) fragments or Fab fragments. Libraries from immunized sources provide high-affinity antibodies to the immunogen without the need to construct fusionomas. Alternatively, natural lineages (e.g., from humans) can be bred without any immunization to provide a single source of antibodies to various nonself as well as self antigens, as Griffiths et al. in EMBO J.12: 725-734 (1993). Finally, natural libraries can also be synthesized by selecting unrearranged V gene fragments in stem cells and using PCR primers containing random sequences to encode the highly variable CDR3 region and rearranging in vitro, such as Hoogenboom and Winter Described in J. Mol. Biol., 227: 381-388 (1992). Patent publications describing human antibody phage libraries include, for example: US Pat. 6 and 2009 / 0002360.

[0250] Antibodies or antibody fragments isolated from human antibody repertoires are considered human antibodies or human antibody fragments herein.

[0251] Techniques for making bispecific antibodies include, but are not limited to, recombinant coexpression of two immunoglobulin heavy chain-light chain pairs with different specificities (see: Milstein et al., Nature 305: 537 (1983); WO 93 / 08829; and Traunecker et al., EMBO J.10: 3655, 1991) and "pestle and mortar" engineering (see eg US Patent No. 5,731,168). Multispecific antibodies can also be prepared by: engineering electrostatic steering effects for the preparation of antibody Fc-heterodimer molecules (WO 2009 / 089004A1); cross-linking two or more antibodies or fragments (see, e.g. , U.S. Patent No. 4,676,980; and Brennan et al., Science, 229: 81 (1985)); use of leucine zippers to generate bispecific antibodies (see, e.g., Kostelny et al., J. Immunol., 148(5) : 1547-1553 (1992)); use of "diabody" technology to prepare diabody fragments (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and Using single-chain Fv (scFv) dimers (see, e.g., Gruber et al., J. Immunol., [] 152:5368 (1994)); and prepare trispecific antibodies according to the method disclosed in, for example, Tutt et al., J. Immunol. 147: 60 (1991).

[0252] Also included herein are engineered antibodies having three or more antigen combining sites, including "Octopus antibodies" (see, eg, US 2006 / 0025576A1).

[0253] Anti-CD20 / anti-CD3 bispecific antibodies herein also include "dual-acting FAbs" or "DAFs", which comprise an antigen-binding site that binds to two different antigens (see, for example, US 2008 / 0069820).

[0254] "Crossmab" antibodies are also included herein (see eg WO2009080251, WO2009080252, WO2009080253, WO2009080254).

[0255] Another technique for preparing bispecific antibody fragments is the "bispecific T cell engager" or BiTE® approach (see eg WO2004 / 106381, WO2005 / 061547, WO2007 / 042261 and WO2008 / 119567). This approach utilizes two antibody variable domains arrayed on a single polypeptide. For example, a single polypeptide chain consists of two single-chain Fv (scFv) fragments, each with variable heavy (VH) and variable light (VL) domains, by lengths sufficient to allow molecular separation between the two domains. Intra-associated polypeptide linkers are separated. This single polypeptide further includes a polypeptide spacer sequence located between the two scFv fragments. Each scFv recognizes a different epitope, and these epitopes may be specific for different cell types, so when each scFv is joined to its cognate epitope, cells of two different cell types will be in close proximity or bound. A specific embodiment of the method comprises a scFv that recognizes a cell surface antigen expressed by an immune cell, such as a CD3 polypeptide on a T cell, and another scFv that recognizes a cell surface antigen expressed by a target cell, such as a malignant or tumor cell. A scFv linkage.

[0256] Since it is a single polypeptide, any prokaryotic or eukaryotic cell expression system known in the art, such as CHO cell lines, can be used to express bispecific T cell engagers. However, specific purification techniques (see, e.g., EP1691833) may be required to separate monomeric bispecific T cell engagers from other multimeric species that may have different biological activities than the expected activity of the monomers . In an exemplary purification scheme, the solution containing the secreted polypeptide is first subjected to metal affinity chromatography, and the polypeptide is eluted using an imidazole concentration gradient. This eluate was further purified using anion exchange chromatography, and the peptide was eluted using a sodium chloride concentration gradient. Finally, the eluate was subjected to size sizing chromatography to separate monomers from polymers.

[0257] In certain embodiments, anti-CD20 / anti-CD3 bispecific antibodies can be further modified to include additional non-protein moieties known in the art and readily available. Moieties suitable for derivatization of anti-CD20 / anti-CD3 bispecific antibodies include, but are not limited to, water soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, Poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymer, polyamino acid (homopolymer or random copolymer) and dextran or poly (n-vinylpyrrolidone) polyethylene glycol, polypropylene glycol homopolymer, polypropylene oxide / ethylene oxide copolymer, polyoxyethylenated polyols (eg glycerol), polyvinyl alcohol and mixtures thereof. Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water. The polymer can be of any molecular weight and can be branched or unbranched. The number of polymers attached to the antibody can vary, and if more than one polymer is attached, they can be the same or different molecules. In general, the amount and / or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the specific properties or functions of the antibody to be improved, whether the antibody derivative will be used in a given condition The next treatment is moderate.

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

[0259] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises an antibody-drug conjugate (ADC), wherein the antibody is conjugated to one or more drugs, including but not limited to maytansinoids ) (see U.S. Patent No. 5,208,020, No. 5,416,064 and European Patent EP 0 425 235 B1); No. 5,635,483 and No. 5,780,588 and No. 7,498,298); dolastatin; calicheamicin or its derivatives (see U.S. Patent No. 5,712,374, No. 5,714,586, No. 5,739,116, No. 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. 58:2925-292 8 ( 1998) ); anthracyclines such as daunomycin or doxorubicin (see Kratz et al., Current Med. Chem. 13:477-523 (2006); Jeffrey et al., Bioorganic & Med. Chem. Letters16:358-362 (2006);Torgov et al., Bioconj. Chem.16:717-721 (2005);Nagy et al., Proc.Natl.Acad.Sci.USA97:829-834 (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); Pterin; vindesine; taxanes such as docetaxel, paclitaxel, larotaxel, tesetaxel, and ortataxel ; trichothecene; and CC1065.

[0260] In another embodiment, the anti-CD20 / anti-CD3 bispecific antibody is bound to an enzymatically active toxin or a fragment thereof, which includes but not limited to diphtheria A chain, non-binding active fragment of diphtheria toxin, exotoxin A chain (derived from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modinus A chain, α-sarcinia, tung protein, carnation toxin, pokeweed protein (PAPI, PAPII and PAP-S), bitter gourd inhibitors, curcumin, crotonin, soapwort inhibitors, gelonin, mitocetins, limited aspergillus, phenomycin, ionomycin and single-ended Sporane toxins.

[0261] In another embodiment, the anti-CD20 / anti-CD3 bispecific antibody is bound to a radioactive atom to form a radioconjugate. A variety of radioactive isotopes are available to produce radioactive conjugates. Examples include At 211, I 131, I 125, Y 90, Re 186, Re 188, Sm 153, Bi 212, P 32, Pb 212, and radioactive isotopes of Lu. When the radioconjugate is used for detection, it may contain radioactive atoms such as Tc 99m or I 123 for scintigraphy studies, or for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, mri). Spin labels such as iodine-123, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese, or iron.

[0262] Conjugates of anti-CD20 / anti-CD3 bispecific antibodies and cytotoxic agents can be prepared using a variety of bifunctional protein coupling agents such as N-succinimidyl-3-(2- Pyridyl dithio)propionate (SPDP), succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), iminosulfur alkanes (IT), bifunctional derivatives of imidic acid esters (e.g. dimethyl adipate HCl), active esters (e.g. disuccinimidyl suberic acid), aldehydes (e.g. glutaraldehyde), bis- Nitrogen compounds (e.g. bis(p-azidobenzoyl)hexamethylenediamine), dinitrogen derivatives (e.g. bis-(p-diazobenzoyl)-ethylenediamine), diisocyanates (e.g. toluene 2, 6-diisocyanate) and bis-reactive fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et al., Science 238:1098 (1987). An exemplary chelating agent for conjugating radionucleotides to antibodies is carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA). See WO94 / 11026. The linker may be a "cleavable linker" that facilitates the release of the cytotoxic drug in the cell. For example, acid-labile linkers, peptidase-sensitive linkers, photolabile linkers, dimethyl linkers, or disulfide-containing linkers can be used (Chari et al., Cancer Res. 52:127 -131 (1992); US Patent No. 5,208,020).

[0263] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is useful for treating cancer. In one embodiment, the cancer is a B cell proliferative disorder. In one embodiment, the cancer is a CD20 positive B cell proliferative disorder. In one embodiment, the cancer is non-Hodgkin's lymphoma (NHL). In one embodiment, NHL is diffuse large B-cell lymphoma (DLBCL), high-grade B-cell lymphoma (HGBCL), DLBCL derived from FL [transformed FL; trFL], primary mediastinal large B-cell lymphoma tumor (PMBCL) or marginal zone lymphoma (MZL). MZL can be divided into splenic, nodal, and extranodal MZL. In one embodiment, the NHL is mantle cell lymphoma (MCL). In one embodiment, the NHL is grade 1-3a follicular lymphoma (FL). In one embodiment, the CD20 positive B cell proliferative disorder is a relapsed or refractory B cell proliferative disorder. In one embodiment, the relapsed or refractory B-cell proliferative disorder is relapsed or refractory NHL (e.g., relapsed or refractory DLBCL, relapsed or refractory FL, or relapsed or refractory sexual MCL).

[0264] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody specifically binds CD3ɛ.

[0265] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody can be combined with antibody H2C (PCT Publication No. WO2008 / 119567), antibody V9 (Rodrigues et al., Int J Cancer Suppl. 7, 45-50 (1992) and US Patent No. 6,054,297), antibody FN18 (Nooij et al., Eur J Immunol. 19, 981-984 (1986)), antibody SP34 (Pessano et al., EMBO J. 4, 337-340 (1985)), antibody OKT3 ( Kung et al., Science 206, 347-349 (1979)), antibody WT31 (Spits et al., JImmunol. 135, 1922 (1985)), antibody UCHT1 (Burns et al., JImmunol.129, 1451-1457 (1982)) , antibody 7D6 (Coulie et al., Eur J Immunol. 21, 1703-1709 (1991 )) or antibody Leu-4 competed for binding. In some embodiments, the anti-CD20 / anti-CD3 bispecific antibody may also comprise an antigen-binding portion that specifically binds to CD3 as disclosed in: WO 2005 / 040220, WO 2005 / 118635, WO 2007 / 042261, WO 2008 / 119567, WO 2008 / 119565, WO 2012 / 162067, WO 2013 / 158856, WO 2013 / 188693, WO 2013 / 186613, WO 2014 / 110601, WO 2014 / 145806, WO 2014 / 19 1113, WO 2014 / 047231, WO 2015 / 095392, WO 2015 / 181098, WO 2015 / 001085, WO 2015 / 104346, WO 2015 / 172800, WO 2016 / 020444 or WO 2016 / 014974.

[0266] In some embodiments, the anti-CD20 / anti-CD3 bispecific antibody can comprise rituximab, obinutuzumab, ocrelizumab, ofatumumab, ocatuzumab ( ocaratuzumab), veltuzumab, and the antibody or antigen-binding portion of ublituximab.

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

[0268] In some embodiments, anti-CD20 / anti-CD3 bispecific antibodies may comprise generic, biosimilar or non-comparable biological versions of the antibodies named herein.

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

[0270] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain specifically binding to CD20, which comprises a heavy chain variable region sequence and a light chain sequence. The region sequence is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 7, and the light chain variable region sequence is identical to SEQ ID NO: 8 The sequences are at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical. In a further embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain specifically binding to CD20, which comprises the heavy chain variable region sequence of SEQ ID NO: 7 and SEQ ID NO: 8 The sequence of the light chain variable region.

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

[0272] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain specifically binding to CD3, which comprises a heavy chain variable region sequence and a light chain sequence. The region sequence is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 15, and the light chain variable region sequence is identical to SEQ ID NO: 16 sequences that are at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical. In a further embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain specifically binding to CD3, which comprises the heavy chain variable region sequence of SEQ ID NO: 15 and SEQ ID NO: 16 The sequence of the light chain variable region.

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

[0274] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises (i) at least one antigen binding domain specifically binding to CD20 comprising the heavy chain variable region sequence of SEQ ID NO: 7 and the light chain variable region sequence of SEQ ID NO: 8, and (ii) at least one antigen-binding domain specifically binding to CD3 comprising the heavy chain variable region sequence of SEQ ID NO: 15 and the light chain variable region sequence of SEQ ID NO: 16.

[0275] In one embodiment, the antigen-binding domain that specifically binds to CD3 of the anti-CD20 / anti-CD3 bispecific antibody is an antibody fragment, particularly a Fab molecule or scFv molecule, more particularly a Fab molecule. In specific embodiments, the antigen-binding domain that specifically binds to CD3 of the anti-CD20 / anti-CD3 bispecific antibody is an exchanged Fab molecule, wherein the variable or constant domains of the Fab heavy chain and the Fab light chain are exchanged ( i.e. replace each other).

[0276] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises at least one antigen-binding domain that specifically binds to CD20 and one antigen-binding domain that specifically binds to CD3. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises a first antigen-binding domain that specifically binds to CD3 and a second and a third antigen-binding domain that specifically bind to CD20. In one embodiment, the first antigen binding domain is an exchanged Fab molecule and the second and third antigen binding domains are each conventional Fab molecules. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody further comprises an Fc domain. Anti-CD20 / anti-CD3 bispecific antibodies may comprise modifications as disclosed herein in the Fc region and / or in the antigen binding domain. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises an IgG1 Fc domain comprising one or more amino acid substitutions that reduce binding to Fc receptors and / or effector function. In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises an IgG1 Fc domain comprising the amino acid substitutions L234A, L235A and P329G (EU numbering).

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

[0278] In a specific embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises a) a first Fab molecule which specifically binds to CD3 (in particular CD3ε) and wherein the variable domains VL and VH of the Fab light chain and the Fab heavy chain replace each other; b) A second Fab and a third Fab molecule that specifically binds to CD20, wherein in the constant domain CL of the second and third Fab molecules the amino acid at position 124 is replaced by lysine (K) substituted (according to Kabat numbering), and the amino acid at position 123 is substituted (according to Kabat numbering) by lysine (K) or arginine (R), especially arginine (R), and, wherein In the constant domain CH1 of the second and third Fab molecules, the amino acid at position 147 was replaced by glutamic acid (E) (EU numbering), and the amino acid at position 213 was replaced by glutamic acid (EU numbering). E) supersedes (EU number); and c) Fc domain, which consists of a first unit and a second unit capable of stable association.

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

[0280] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody is bivalent to CD20 and monovalent to CD3.

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

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

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

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

[0285] In a particular embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises: a polypeptide at least 95%, 96%, 97%, 98% or 99% identical to the sequence of SEQ ID NO: 17; The sequence of SEQ ID NO: 18 is at least 95%, 96%, 97%, 98% or 99% identical to a polypeptide; and the sequence of SEQ ID NO: 19 is at least 95%, 96%, 97%, 98% or 99% identical a polypeptide; and a polypeptide that is at least 95%, 96%, 97%, 98% or 99% identical to the sequence of SEQ ID NO: 20. In a further embodiment, the bispecific antibody comprises the polypeptide sequence of SEQ ID NO:17, the polypeptide sequence of SEQ ID NO:18, the polypeptide sequence of SEQ ID NO:19 and the polypeptide sequence of SEQ ID NO:20. In a further specific embodiment, the bispecific antibody comprises a polypeptide chain comprising SEQ ID NO: 17, a polypeptide chain comprising SEQ ID NO: 18, two polypeptide chains comprising SEQ ID NO: 19, and a polypeptide chain comprising SEQ ID NO: 19 Polypeptide chain of ID NO: 20.

[0286] Specific anti-CD20 / anti-CD3 bispecific antibodies are described in PCT Publication No. WO 2016 / 020309 and European Patent Application Nos. EP15188093 and EP16169160 (each incorporated herein by reference in its entirety). Gefitumumab

[0287] In one embodiment, the anti-CD20 / anti-CD3 bispecific antibody useful in the methods provided herein is grifiduzumab. Grafitumumab (Proposed INN: List 121 WHO Drug Information, Vol. 33, No. 2, 2019, p. 276, also known as CD20-TCB, RO7082859, or RG6026; CAS #: 2229047-91-8) It is a novel T cell engaging bispecific (TCB) full-length antibody with a 2:1 molecular configuration, which can bivalently bind to CD20 on B cells and bind to CD3 (especially CD3ε) on T cells chain (CD3e)) for monovalent binding. Its CD3-binding domain is fused to one of the CD20-binding domains via a flexible linker in a head-to-tail fashion. This structure conferred superior in vitro potency of grafitumumab to other CD20-CD3 bispecific antibodies with a 1:1 configuration and produced significant antitumor efficacy in preclinical DLBCL models. CD20 bivalence preserves this potency in the presence of competing anti-CD20 antibodies, opening up opportunities for pretreatment or co-treatment with these agents. Griffithumab contains an engineered heterodimeric Fc region that completely abolishes binding to FcgR and C1q. By simultaneously binding to CD3e on tumor cells expressing human CD20 and T cell receptor (TCR) complexes on T cells, it induces tumor cell lysis in addition to T cell activation, proliferation and cytokine release. B cell lysis mediated by grafitumumab was CD20 specific and did not occur in the absence of CD20 expression or in the absence of simultaneous association (crosslinking) of T cells with CD20 expressing cells. In addition to killing, T cells undergo activation due to CD3 cross-linking, such as by T cell activation markers (CD25 and CD69), cytokine release (IFNγ, TNFα, IL-2, IL-6, IL-10), cell Toxic granule release (granzyme B) and T cell proliferation detected. The schematic diagram of the molecular structure of grafitumumab is shown in Figure 2. The sequences of grafitumumab are summarized in Table 2. [surface] [2.] [Grafitumumab sequence] [ID] [Grafitumumab sequence] [ID] CD3 heavy chain CD3 light chain SEQ ID NO: illustrate SEQ ID NO: illustrate 9 HVR-H1 (Kabat) 12 HVR-L1 (Kabat) 10 HVR-H2 (Kabat) 13 HVR-L2 (Kabat) 11 HVR-H3 (Kabat) 14 HVR-L3 (Kabat) 15 VH 16 VL CD20 heavy chain CD20 light chain 1 HVR-H1 (Kabat) 4 HVR-L1 (Kabat) 2 HVR-H2 (Kabat) 5 HVR-L2 (Kabat) 3 HVR-H3 (Kabat) 6 HVR-L3 (Kabat) 7 VH 8 VH full length antibody 17 HC-pestle 18 HC-mortar 19 LC-CD3 20 LC-CD20 B. Anti-CD79b Antibody Drug Conjugates

[0288] Can be used in the methods disclosed herein (e.g., for the treatment of CD20 positive cell proliferative disorders, such as B cell proliferative disorders (e.g., NHL (e.g., relapsed and / or refractory NHL, DLBCL (e.g., relapsed and / or refractory NHL) or refractory DLBCL), FL (e.g., relapsed and / or refractory FL or transformed FL) or MCL (e.g., relapsed or refractory MCL)) or CNSL) anti-CD79b antibody-drug conjugates including US Any of the anti-CD79b antibody drug conjugates disclosed in Patent No. 8,088,378, which is incorporated herein by reference in its entirety. In some cases, the anti-CD79b antibody drug conjugates include an anti-CD79b binding domain comprising an At least one, two, three, four, five, or six hypervariable regions (HVR): (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR -H2, which comprises the amino acid sequence of SEQ ID NO: 22; (c) HVR-H3, which comprises the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1, which comprises the amino acid sequence of SEQ ID NO: 24 (e) HVR-L2, which comprises the amino acid sequence of SEQ ID NO: 25; and (f) HVR-L3, which comprises the amino acid sequence of SEQ ID NO: 26. In some cases Here, the anti-CD79b antibody drug conjugate comprises an anti-CD79b binding domain comprising all six of the following HVRs: (a) HVR-H1 comprising the amino acid sequence of GYTFSSYWIE (SEQ ID NO: 21); (b) HVR-H2, which comprises the amino acid sequence of GEILPGGGDTNYNEIFKG (SEQ ID NO: 22); (c) HVR-H3, which comprises the amino acid sequence of TRRVPIRLDY (SEQ ID NO: 23); (d) HVR-L1, It comprises the amino acid sequence of KASQSVDYEGDSFLN (SEQ ID NO: 24); (e) HVR-L2, it comprises the amino acid sequence of AASNLES (SEQ ID NO: 25); and (f) HVR-L3, it comprises QQSNEDPLT (SEQ ID NO: 26). In some cases, the anti-CD79b antibody drug conjugate comprises heavy chain framework regions FR-H1, FR, respectively comprising the sequences of SEQ ID NO: 29 to SEQ ID NO: 32. -At least one of -H2, FR-H3 and FR-H4 (such as 1, 2, 3 or 4), and / or respectively comprise the light chain framework region of the sequence of SEQ ID NO: 33 to SEQ ID NO: 36 At least one of FR-L1, FR-L2, FR-L3 and FR-L4 (such as 1, 2, 3 or 4). In some instances, an anti-CD79b antibody drug conjugate comprises: (a) a heavy chain variable (VH) domain comprising or having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 27 (e.g. , at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) amino acid sequence; (b) light chain variable (VL) domain , which comprises the amino acid sequence of SEQ ID NO: 28 or has at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% to the sequence) %, or 99% sequence identity); 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: 27; and a VL domain comprising the amino acid sequence of SEQ ID NO: 28.

[0289] In some instances, an anti-CD79b antibody drug conjugate comprises: (a) a heavy chain comprising or having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 37 (e.g., at least 91%, 92% %, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity); (b) a light chain comprising the amino acid of SEQ ID NO: 38 sequence or an amino group having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the sequence 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: 37; and a VL domain comprising the amino acid sequence of SEQ ID NO: 38.

[0290] The sequences of the anti-CD79b antibodies to parotuzumab are summarized in Table 3 below. [surface] [3.] [antibody to Parotuzumab] [CD79b] [Sequence of Antibody] [ID] heavy chain light chain SEQ ID NO: illustrate SEQ ID NO: illustrate twenty one HVR-H1 twenty four HVR-L1 twenty two HVR-H2 25 HVR-L2 twenty three HVR-H3 26 HVR-L3 27 VH 28 VL 37 heavy chain 38 light chain

[0291] In some instances, the anti-CD79b antibody is linked to a toxin such as monomethyl auristatin E (MMAE, ie, vedotin). In some cases, the anti-CD79b antibody-drug conjugate is parotuzumab (immunoglobulin G1-κ auristatin E conjugate, anti-[Homo sapiens CD79b (immunoglobulin-related CD79β)], Humanized monoclonal antibody bound to Auristatin E; γ1 heavy chain (1-447) [Humanized VH (Homo sapiens IGHV3-23*04 (76.50%)-(IGHD)-IGHJ4*01)[8.8 .10] (1-117) – Homo sapiens IGHG1*03 (CH1 R120>K (214) (118-215), Hinge (216-230), CH2 (231-340), CH3 (341-445), CHS (446-447))(118-447)], (220-218')-disulfide bond with κ light chain (1'-218') [humanized V-KAPPA (Homo sapiens IGKV1-39*01 (85.90%) -IGKJ1*01)[10.3.9] (1'-111') -Homo sapiens IGKC*01 (112'-218')]; dimer (226-226":229-229") - a double disulfide bond; via a cleavable maleimidocapryl-pentyl-citrulline-p-aminobenzoyloxycarbonyl (mc-val-cit-PABC) type linker, Conjugated to monomethyl auristatin E (MMAE), also known as RG-7596, or RO5541077-000)) with an average of 3 to 4 cysteamine groups, as International Pharmaceutical Substance Nonproprietary Name (INN) As defined in List 110 (WHO Drug Information, Vol. 27, No. 4, 2016, p. 443). Parotuzumab is also known as IUPHAR / BPS Number 8404, KEGG Number D10761 or CAS Registry Number 1313206-42-6. Parotuzumab-piiq is also referred to interchangeably as "Parotuzumab-piiq", "huMA79bv28-MC-vc-PAB-MMAE", or "DCDS4501A". In some embodiments, an anti-CD79b antibody (e.g., an anti-CD79b ADC) comprises the heavy chain sequence of SEQ ID NO: 37 and the light chain sequence of SEQ ID NO: 38.

[0292] In some instances, the anti-CD79b antibody drug conjugate comprises the formula: Wherein Ab is an anti-CD79b antibody, which comprises: (i) hypervariable region-H1 (HVR-H1), which comprises the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2, which comprises SEQ ID NO : the amino acid sequence of 22; (iii) HVR-H3, it comprises the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1, it comprises the amino acid sequence of SEQ ID NO: 24; (v ) HVR-L2, which comprises the amino acid sequence of SEQ ID NO: 25; and (vi) HVR-L3, which comprises the amino acid sequence of SEQ ID NO: 26, and wherein p is between 1 and 8.

[0293] In some embodiments, the antibody drug conjugate comprises an anti-CD79b antibody comprising: (a) a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from: (i) HVR- H1, which comprises the amino acid sequence of SEQ ID NO: 21, (ii) HVR-H2, which comprises the amino acid sequence of SEQ ID NO: 22, and (iii) HVR-H3, which comprises the amino acid sequence selected from SEQ ID NO and (b) a VL domain comprising at least one, at least two or all three VL HVR sequences selected from: (i) HVR-L1 comprising the sequence of SEQ ID NO: 24 Amino acid sequence, (ii) HVR-L2, which comprises the amino acid sequence of SEQ ID NO: 25, and (iii) HVR-L3, which comprises the amino acid sequence of SEQ ID NO: 26. In some embodiments, the antibody drug conjugate comprises an anti-CD79b antibody comprising at least one of: (i) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23, and / or (ii) HVR -L1, which comprises the amino acid sequence of SEQ ID NO: 24. In some embodiments, the antibody drug conjugate comprises an anti-CD79b antibody comprising: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 comprising SEQ ID NO: the amino acid sequence of 22; (c) HVR-H3, it comprises the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1, it comprises the amino acid sequence of SEQ ID NO: 24; ( e) HVR-L2, which comprises the amino acid sequence of SEQ ID NO: 25; and (f) HVR-L3, which comprises the amino acid sequence of SEQ ID NO: 26.

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

[0295] In some embodiments, an anti-CD79b antibody drug conjugate comprises a humanized anti-CD79b antibody. In some embodiments, an anti-CD79b antibody comprises a HVR as in any of the embodiments provided herein, and further comprises a human acceptor framework, e.g., a human immunoglobulin framework or a human consensus framework. In some embodiments, the human acceptor framework is a human VLκ1 (VLK1) framework and / or a VH framework VHIII. In some embodiments, the humanized anti-CD79b antibody comprises: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 comprising the amine of SEQ ID NO: 22 (c) HVR-H3, which comprises the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1, which comprises the amino acid sequence of SEQ ID NO: 24; (e) HVR-L2 , which comprises the amino acid sequence of SEQ ID NO: 25; and (f) HVR-L3, which comprises the amino acid sequence of SEQ ID NO: 26. In some embodiments, the humanized anti-CD79b antibody comprises: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 comprising the amine of SEQ ID NO: 22 (c) HVR-H3, which comprises the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1, which comprises the amino acid sequence of SEQ ID NO: 24; (e) HVR-L2 , which comprises the amino acid sequence of SEQ ID NO: 25; and (f) HVR-L3, which comprises the amino acid sequence of SEQ ID NO: 26.

[0296] In some embodiments, an antibody drug conjugate (e.g., an anti-CD79b antibody drug conjugate) comprises an anti-CD79 antibody comprising a heavy chain variable domain (VH) sequence identical to SEQ ID NO: The amino acid sequence of 27 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity. In some embodiments, at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 27 The VH sequence comprises a substitution (eg, conservative substitution), insertion or deletion relative to the reference sequence, but the anti-CD79b antibody drug conjugate comprising the sequence retains the ability to bind to CD79b. In some embodiments, in SEQ ID NO: 27, a total of 1 to 10 amino acids are substituted, inserted and / or deleted. In some embodiments, in SEQ ID NO: 27, a total of 1 to 5 amino acids are substituted, inserted and / or deleted. In some embodiments, substitutions, insertions or deletions occur in regions outside of the HVRs (i.e., in FRs, e.g., SEQ ID NO: 29 to SEQ ID NO: 32). In some embodiments, the antibody drug conjugate (e.g., an anti-CD79b antibody drug conjugate) comprises the VH sequence of SEQ ID NO: 27, including post-translational modifications of that sequence. In some embodiments, the VH comprises one, two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 comprising The amino acid sequence of SEQ ID NO: 22; and (c) HVR-H3, which comprises the amino acid sequence of SEQ ID NO: 23.

[0297] In some embodiments, the antibody drug conjugate (e.g., an anti-CD79b antibody drug conjugate) comprises an anti-CD79b antibody comprising a light chain variable domain (VL) sequence identical to SEQ ID NO: The amino acid sequence of 28 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity. In certain embodiments, at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 28 The non-specific VL sequence contains substitutions (eg, conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CD79b antibody drug conjugate comprising this sequence retains the ability to bind to CD79b. In certain embodiments, in SEQ ID NO: 28, a total of 1 to 10 amino acids are substituted, inserted and / or deleted. In certain embodiments, in SEQ ID NO: 28, a total of 1 to 5 amino acids are substituted, inserted and / or deleted. In certain embodiments, substitutions, insertions or deletions occur in regions outside of the HVRs (i.e., in FRs, e.g., SEQ ID NO: 33 to SEQ ID NO: 36). In some embodiments, an anti-CD79b antibody drug conjugate comprises an anti-CD79b antibody comprising the VL sequence of SEQ ID NO: 28, including post-translational modifications of that sequence. In some embodiments, the VL comprises one, two or three HVRs selected from: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (b) HVR-L2 comprising The amino acid sequence of SEQ ID NO: 25; and (c) HVR-L3, which comprises the amino acid sequence of SEQ ID NO: 26. In some embodiments, the VL comprises one, two or three HVRs selected from: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (b) HVR-L2 comprising The amino acid sequence of SEQ ID NO: 25; and (c) HVR-L3, which comprises the amino acid sequence of SEQ ID NO: 26.

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

[0299] In some embodiments, an antibody drug conjugate (e.g., an anti-CD79b antibody drug conjugate) comprises an anti-CD79b antibody that binds to the same epitope as an anti-CD79b antibody described herein. For example, in some embodiments, the antibody drug conjugate (e.g., an anti-CD79b antibody drug conjugate) comprises the same antigenic determinant that binds to an anti-CD79b antibody comprising the VH sequence of SEQ ID NO: 27 and the VL sequence of SEQ ID NO: 28. Based anti-CD79b antibody.

[0300] In some embodiments, the antibody drug conjugate comprises an anti-CD79b antibody that is a monoclonal antibody, a chimeric antibody, a humanized antibody, or a human antibody. In some embodiments, the antibody drug conjugate comprises an antigen-binding fragment of an anti-CD79b antibody described herein, e.g., a Fv, Fab, Fab', scFv, diabody, or F(ab')2 fragment. In some embodiments, the antibody drug conjugate comprises a substantially full-length anti-CD79b antibody, e.g., an IgG1 antibody or other antibody type or isotype as described elsewhere herein. Anti-CD79b antibody drug conjugates can be manufactured using recombinant methods and compositions, for example, as disclosed in U.S. Patent No. 4,816,567.

[0301] In some cases, an anti-CD79b antibody drug conjugate according to any of the embodiments disclosed above may incorporate any of the features, alone or in combination, as disclosed below. C. Antibody Format 1. Anti-CD20 / anti-CD3 bispecific antibody

[0302] Components of anti-CD20 / anti-CD3 bispecific antibodies can be fused to each other in a variety of configurations. An example configuration is shown in Figure 1.

[0303] In certain embodiments, the antigen-binding portion comprised in the anti-CD20 / anti-CD3 bispecific antibody is a Fab molecule. In these embodiments, the first antigen binding moiety, the second antigen binding moiety, the third antigen binding moiety, etc. may be referred to herein as the first Fab molecule, the second Fab molecule, the third Fab molecule, etc., respectively. Furthermore, in certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody comprises an Fc domain consisting of first and second units capable of stable association.

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

[0305] In one such embodiment, the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first Fab molecule. In a specific such embodiment, the anti-CD20 / anti-CD3 bispecific antibody essentially consists of a first Fab molecule and a second Fab molecule, and the Fc domain consists of a first unit, a second unit and optional One or more peptide linkers, wherein a first Fab molecule is fused to the N-terminus of the first or second unit of the Fc domain at the C-terminus of the Fab heavy chain, and the second Fab molecule is at the C-terminus of the Fab heavy chain The C-terminus is fused to the N-terminus of the Fab heavy chain of the first Fab molecule. Such configurations are schematically depicted in Figures 1G and 1K. In addition, optionally, the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule can be fused to each other.

[0306] In another embodiment, the second Fab molecule is fused to the N-terminus of the first unit or the second unit of the Fc domain at the C-terminus of the Fab heavy chain. In a specific such embodiment, the antibody consists essentially of a first Fab molecule and a second Fab molecule, the Fc domain consisting of a first unit and a second unit and optionally one or more peptide linkers. A composition wherein the first Fab molecule and the second Fab molecule are each fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain. Such configurations are schematically depicted in Figures 1A and 1D. The first Fab molecule and the second Fab molecule can be fused to the Fc domain directly or via a peptide linker. In a specific embodiment, the first Fab molecule and the second Fab molecule are each fused to an Fc domain via an immunoglobulin hinge region. In a specific embodiment, the immunoglobulin hinge region is a human IgG 1 hinge region, in particular, wherein the Fc domain is an IgG 1 Fc domain.

[0307] In other embodiments, the second Fab molecule is fused to the N-terminus of the first unit or the second unit of the Fc domain at the C-terminus of the Fab heavy chain. In one such embodiment, the first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab molecule. In specific such embodiments, the antibody consists essentially of a first Fab molecule and a second Fab molecule, the Fc domain consists of a first unit and a second unit and optionally one or more peptide linkers, Wherein, the first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second Fab molecule, and the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the first unit or the second unit of the Fc domain N-terminal of the secondary unit. Such configurations are schematically depicted in Figures 1H and 1L. In addition, optionally, the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule can be fused to each other.

[0308] Fab molecules can be fused directly to the Fc domain or to each other, or to the Fc or to each other via a peptide linker comprising one or more amino acids, typically about 2 to 20 amino acids. Peptide linkers are well known in the art and are described herein. Suitable non-immunogenic peptide linkers include, for example, (G4S)n, (SG4)n, (G4S)n or G4(SG4)n peptide linkers. "N" is usually an integer from 1 to 10, especially 2 to 4. In one embodiment, the peptide linker is at least 5 amino acids in length; in one embodiment, 5 to 100 amino acids in length; in another embodiment, 10 to 50 amino acids in length amino acids. In one embodiment, the peptide linker is (GxS)n or (GxS)nGm wherein G=glycine, S=serine, and (x=3, n=3, 4, 5 or 6 , and m=0, 1, 2 or 3) or (x=4, n=2, 3, 4 or 5, and m=0, 1, 2 or 3), in one embodiment, x=4 and n=2 or 3, in another embodiment x=4 and n=2. In one embodiment, the peptide linker is (G 4S ) 2. A particularly suitable peptide linker for fusing the Fab light chains of the first and second Fab molecules to each other is (G4S)2. An exemplary peptide linker suitable for linking the Fab heavy chains of a first Fab fragment and a second Fab fragment comprises the sequence (D)-(G4S)2. Another suitable such linker comprises the sequence (G4S)4. Alternatively, the linker may comprise (part of) an immunoglobulin hinge region. In particular, in cases where the Fab molecule is fused to the N-terminus of the Fc domain subunit, the fusion may be through an immunoglobulin hinge region or a portion thereof comprising an additional peptide linker or without an additional peptide linker. .

[0309] Antibodies (e.g., as shown in Figures 1A, 1D, 1G, 1H, 1K, 1L) having a single antigen-binding portion (such as a Fab molecule) capable of specifically binding to a target cell antigen can be used, particularly in the presence of expected high Where internalization of the target cell antigen occurs following binding of the affinity antigen binding moiety. In such cases, the presence of more than one antigen-binding moiety directed against a particular target cell antigen can enhance the internalization of the target cell antigen, thereby reducing its availability.

[0310] However, in various other cases, there are antibodies comprising two or more antigen-binding portions (such as Fab molecules) specific for a particular target cell antigen (e.g., as shown in Figures 1B, 1C, 1E, 1F, 1I, 1J, 1M or 1N) would be advantageous, for example, to optimize targeting to target sites or to cross-link target cell antigens.

[0311] Accordingly, in certain embodiments, the anti-CD20 / anti-CD3 bispecific antibody comprises two anti-CD20 binding moieties, e.g., two Fab molecules targeting CD20. In one embodiment, the two CD20-targeting Fab molecules are conventional Fab molecules. In one embodiment, two CD20-targeting Fab molecules comprise the same heavy and light chain amino acid sequences and have the same domain arrangement (i.e., conventional or swapped).

[0312] In an alternative embodiment, the anti-CD20 / anti-CD3 bispecific antibody comprises two anti-CD3 binding moieties, e.g., two Fab molecules targeting CD3. In one such embodiment, both CD3-targeting Fab molecules are swapped Fab molecules (a Fab molecule in which the variable domains VH and VL or the constant domains CL and CH1 of the Fab heavy and light chains are exchanged for each other. / replace). In one such embodiment, two CD3-targeting Fab molecules comprise the same heavy and light chain amino acid sequences and have the same domain arrangement (i.e., conventional or swapped).

[0313] In one embodiment, the third Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of either the first unit or the second unit of the Fc domain.

[0314] In a particular embodiment, the second Fab molecule and the third Fab molecule are each fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain, and the first Fab molecule is fused at the C-terminus of the Fab heavy chain Fused to the N-terminus of the Fab heavy chain of a second Fab molecule. In specific such embodiments, the antibody consists essentially of a first Fab molecule, a second Fab molecule and a third Fab molecule, and the Fc domain consists of a first unit and a second unit and optionally one or more A peptide linker in which a first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of a second Fab molecule, and the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the end of the Fc domain N-terminus of the first unit, and wherein a third Fab molecule is fused to the N-terminus of the second unit of the Fc domain at the C-terminus of the Fab heavy chain. Such configurations are shown schematically in Figures 1B and 1E (example, where the third Fab molecule is a conventional Fab molecule and identical to the second Fab molecule) and Figures 1I and 1M (example, where the third Fab molecule is an exchanged Fab molecule and preferably the same as the first Fab molecule) is depicted in). The second Fab molecule and the third Fab molecule can be fused to the Fc domain directly or via a peptide linker. In a specific embodiment, the second Fab molecule and the third Fab molecule are each fused to an Fc domain through an immunoglobulin hinge region. In a specific embodiment, the immunoglobulin hinge region is a human IgG 1 hinge region, in particular, wherein the Fc domain is an IgG 1 Fc domain. In addition, optionally, the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule can be fused to each other.

[0315] In another embodiment, the second Fab molecule and the third Fab molecule are each fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain, and the first Fab molecule is at the C-terminus of the Fab heavy chain. fused to the N-terminus of the Fab heavy chain of a second Fab molecule. In specific such embodiments, the antibody essentially consists of a first Fab molecule, a second Fab molecule and a third Fab molecule, and the Fc domain consists of a first unit and a second unit and optionally one or more A peptide linker in which a first Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of a second Fab molecule, and the second Fab molecule is fused at the C-terminus of the Fab heavy chain to the end of the Fc domain N-terminus of the first unit, and wherein a third Fab molecule is fused to the N-terminus of the second unit of the Fc domain at the C-terminus of the Fab heavy chain. Such configurations are shown schematically in Figures 1C and 1F (example, where the third Fab molecule is a conventional Fab molecule and identical to the second Fab molecule) and Figures 1J and 1N (example, where the third Fab molecule is an exchanged Fab molecule and the same as the first Fab molecule) depicted in). The first Fab molecule and the third Fab molecule can be fused to the Fc domain directly or via a peptide linker. In a specific embodiment, the second Fab molecule and the third Fab molecule are each fused to an Fc domain through an immunoglobulin hinge region. In a specific embodiment, the immunoglobulin hinge region is a human IgG 1 hinge region, in particular, wherein the Fc domain is an IgG 1 Fc domain. In addition, optionally, the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule can be fused to each other.

[0316] In configurations of antibodies in which the Fab molecule is fused at the C-terminus of the Fab heavy chain to the N-terminus of each of the subunits of the Fc domain through the immunoglobulin hinge region, the two Fab molecules, the hinge region and the Fc domain essentially form Immunoglobulin molecule. In certain embodiments, the immunoglobulin molecule is an IgG class immunoglobulin. In an even more specific embodiment, the immunoglobulin is an IgG 1 subclass immunoglobulin. In another embodiment, the immunoglobulin is an IgG4 subclass immunoglobulin. In further specific embodiments, the immunoglobulin is a human immunoglobulin. In other embodiments, the immunoglobulin is a chimeric immunoglobulin or a humanized immunoglobulin.

[0317] In some antibodies, the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule are fused to each other, optionally via a peptide linker. According to the different configurations of the first Fab molecule and the second Fab molecule, the Fab light chain of the first Fab molecule can be fused to the N-terminus of the Fab light chain of the second Fab molecule at its C-terminus, or the Fab light chain of the second Fab molecule can The chain may be fused at its C-terminus to the N-terminus of the Fab light chain of the first Fab molecule. Fusion of the Fab light chain of the first Fab molecule to the second Fab molecule further reduces mismatches between the Fab heavy and light chains and also reduces the number of plastids required to express some antibodies.

[0318] In certain embodiments, the antibody comprises: a polypeptide wherein the Fab light chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule (i.e., the first Fab molecule comprises Swapped Fab heavy chain, in which the heavy chain variable region is replaced by the light chain variable region), which in turn shares a carboxy-terminal peptide bond with the Fc domain subunit (VL(1)-CH1(1)-CH2-CH3(- CH4)); and a polypeptide wherein the Fab heavy chain of the second Fab molecule shares a carboxy-terminal peptide bond (VH(2)-CH1(2)-CH2-CH3(-CH4)) with the Fc domain subunit. In some embodiments, the antibody further comprises: a polypeptide wherein the Fab heavy chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond (VH(1)-CL(1) with the Fab light chain constant region of the first Fab molecule )), and shares a carboxy-terminal peptide bond (VL(2)-CL(2)) with the Fab light chain polypeptide of the second Fab molecule. In certain embodiments, the polypeptides are covalently linked, eg, by disulfide bonds.

[0319] In certain embodiments, the antibody comprises: a polypeptide wherein the Fab heavy chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule (i.e., the first Fab molecule comprises Swapped Fab heavy chain, in which the heavy chain constant region is replaced by the light chain constant region), which in turn shares a carboxy-terminal peptide bond with the Fc domain subunit (VH(1)-CL(1)-CH2-CH3(-CH4) ); and a polypeptide wherein the Fab heavy chain of the second Fab molecule shares a carboxy-terminal peptide bond (VH(2)-CH1(2)-CH2-CH3(-CH4)) with the Fc domain subunit. In some embodiments, the antibody further comprises: a polypeptide wherein the Fab light chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond (VL(1)-CH1(1) with the Fab heavy chain constant region of the first Fab molecule )), and share the carboxy-terminal peptide bond (VL(2)-CL(2)) with the Fab light chain polypeptide of the second Fab molecule. In certain embodiments, the polypeptides are covalently linked, eg, via disulfide bonds.

[0320] In some embodiments, the antibody comprises a polypeptide wherein the Fab light chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule (i.e., the first Fab molecule comprises an exchanged Fab heavy chain, wherein the heavy chain variable region is replaced by a light chain variable region), which in turn shares a carboxy-terminal peptide bond with the Fab heavy chain of a second Fab molecule, which in turn shares a carboxy-terminal peptide bond with the Fc domain subunit ( VL(1)-CH1(1)-VH(2)-CH1(2)-CH2-CH3(-CH4)). In other embodiments, the antibody comprises a polypeptide wherein the Fab heavy chain of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain variable region of the first Fab molecule, which in turn is constant with the Fab heavy chain of the first Fab molecule The region shares a carboxy-terminal peptide bond (i.e., the first Fab molecule comprises an exchanged Fab heavy chain in which the heavy chain variable region is replaced by a light chain variable region), which in turn shares a carboxy-terminal peptide bond with the Fc domain subunit ( VH(2)-CH1(2)-VL(1)-CH1(1)-CH2-CH3(-CH4)).

[0321] In some of these embodiments, the antibody further comprises: an exchanged Fab light chain polypeptide of the first Fab molecule, wherein the Fab heavy chain variable region of the first Fab molecule and the Fab light chain constant region of the first Fab molecule Shared carboxy-terminal peptide bond (VH(1)-CL(1)), and shared carboxy-terminal peptide bond (VL(2)-CL(2)) with the Fab light chain polypeptide of the second Fab molecule. In others of these embodiments, the antibody further comprises: a polypeptide wherein the Fab heavy chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule, which in turn shares a The Fab light chain polypeptide of the second Fab molecule shares a carboxy-terminal peptide bond (VH(1)-CL(1)-VL(2)-CL(2)); or a polypeptide wherein the Fab light chain polypeptide of the second Fab molecule Shares a carboxy-terminal peptide bond with the Fab heavy chain variable region of the first Fab molecule, which in turn shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule (VL(2)-CL(2)-VH(1 )-CL (1)) (where appropriate).

[0322] Antibodies according to these embodiments may further comprise (i) an Fc domain subunit polypeptide (CH2-CH3(-CH4)), or (ii) a polypeptide wherein the Fab heavy chain of the third Fab molecule is shared with the Fc domain subunit Carboxy-terminal peptide bond (VH(3)-CH1(3)-CH2-CH3(-CH4)), and shares carboxy-terminal peptide bond (VL(3)-CL(3) with the Fab light chain polypeptide of the third Fab molecule ). In certain embodiments, the polypeptides are covalently linked, eg, via disulfide bonds.

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

[0324] In some of these embodiments, the antibody further comprises: an exchanged Fab light chain polypeptide of the first Fab molecule, wherein the Fab light chain variable region of the first Fab molecule and the Fab heavy chain constant region of the first Fab molecule Shared carboxy-terminal peptide bond (VL(1)-CH1(1)), and shared carboxy-terminal peptide bond (VL(2)-CL(2)) with the Fab light chain polypeptide of the second Fab molecule. In others of these embodiments, the antibody further comprises: a polypeptide wherein the Fab light chain variable region of the first Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the first Fab molecule, which in turn shares a The Fab light chain polypeptide of the second Fab molecule shares the carboxy-terminal peptide bond (VL(1)-CH1(1)-VL(2)-CL(2)); or the polypeptide, wherein the Fab light chain polypeptide of the second Fab molecule Shares a carboxy-terminal peptide bond with the Fab heavy chain variable region of the first Fab molecule, which in turn shares a carboxy-terminal peptide bond with the Fab light chain constant region of the first Fab molecule (VL(2)-CL(2)-VH(1 )-CL (1)) (where appropriate).

[0325] Antibodies according to these embodiments may further comprise (i) an Fc domain subunit polypeptide (CH2-CH3(-CH4)), or (ii) a polypeptide wherein the Fab heavy chain of the third Fab molecule is shared with the Fc domain subunit Carboxy-terminal peptide bond (VH(3)-CH1(3)-CH2-CH3(-CH4)), and shares carboxy-terminal peptide bond (VL(3)-CL(3) with the Fab light chain polypeptide of the third Fab molecule ). In certain embodiments, the polypeptides are covalently linked, eg, by disulfide bonds.

[0326] In certain embodiments, antibodies comprise polypeptides wherein the Fab heavy chain of a first Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain variable region of a second Fab molecule, which in turn shares a carboxy-terminal peptide bond with the Fab heavy chain of the second Fab molecule. The constant regions share the carboxy-terminal peptide bond (i.e., the second Fab molecule comprises an exchanged Fab heavy chain in which the heavy chain variable region is replaced by the light chain variable region) (VH(1)-CH1(1)-VL( 2)-CH1(2)). In some embodiments, the antibody step comprises: a polypeptide wherein the Fab heavy chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond (VH(2)-CL(2) with the Fab light chain constant region of the second Fab molecule )), and share the carboxy-terminal peptide bond (VL(1)-CL(1)) with the Fab light chain polypeptide of the first Fab molecule.

[0327] In certain embodiments, the antibody comprises a polypeptide wherein the Fab light chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the second Fab molecule (i.e., the second Fab molecule comprises an exchange type Fab heavy chain, wherein the heavy chain variable region is replaced by a light chain variable region), which in turn shares a carboxy-terminal peptide bond (VL(2)-CH1(2)-VH( 1)-CH1(1)). In some embodiments, the antibody step comprises: a polypeptide wherein the Fab heavy chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond (VH(2)-CL(2) with the Fab light chain constant region of the second Fab molecule )), and share the carboxy-terminal peptide bond (VL(1)-CL(1)) with the Fab light chain polypeptide of the first Fab molecule.

[0328] In certain embodiments, the antibody comprises a polypeptide wherein the Fab heavy chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab light chain constant region of the second Fab molecule (i.e., the second Fab molecule comprises an exchange Type Fab heavy chain, wherein the heavy chain variable region is replaced by a light chain variable region), which in turn shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule (VH(2)-CL(2)-VH( 1)-CH1(1)). In some embodiments, the antibody step comprises: a polypeptide wherein the Fab light chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond (VL(2)-CH1(2) with the Fab heavy chain constant region of the second Fab molecule )), and share a carboxy-terminal peptide bond (VL(1)-CL(1)) with the Fab light chain polypeptide of the first Fab molecule.

[0329] In certain embodiments, the antibody comprises a polypeptide wherein the Fab heavy chain of the third Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule, which in turn shares a Fab light chain variable region of the second Fab molecule shared carboxy-terminal peptide bond, which in turn shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of a second Fab molecule (i.e., the second Fab molecule comprises an exchanged Fab heavy chain in which the heavy chain variable region is replaced by the light chain Variable region substitution) (VH(3)-CH1(3)-VH(1)-CH1(1)-VL(2)-CH1(2)). In some embodiments, the antibody step comprises: a polypeptide wherein the Fab heavy chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond (VH(2)-CL(2) with the Fab light chain constant region of the second Fab molecule )), and share a carboxy-terminal peptide bond (VL(1)-CL(1)) with the Fab light chain polypeptide of the first Fab molecule. In some embodiments, the antibody further comprises a Fab light chain polypeptide (VL(3)-CL(3)) of a third Fab molecule.

[0330] In certain embodiments, the antibody comprises a polypeptide wherein the Fab heavy chain of the third Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule, which in turn shares a Fab heavy chain variable region of the second Fab molecule A shared carboxy-terminal peptide bond, which in turn shares a carboxy-terminal peptide bond with the Fab light chain constant region of a second Fab molecule (i.e., the second Fab molecule comprises an exchanged Fab heavy chain, wherein the heavy chain constant region is replaced by the light chain constant region Substitution) (VH(3)-CH1(3)-VH(1)-CH1(1)-VH(2)-CL(2)). In some embodiments, the antibody step comprises: a polypeptide wherein the Fab light chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond (VL(2)-CH1(2) with the Fab heavy chain constant region of the second Fab molecule )), and share a carboxy-terminal peptide bond (VL(1)-CL(1)) with the Fab light chain polypeptide of the first Fab molecule. In some embodiments, the antibody further comprises a Fab light chain polypeptide (VL(3)-CL(3)) of a third Fab molecule.

[0331] In certain embodiments, the antibody comprises a polypeptide wherein the Fab light chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond with the Fab heavy chain constant region of the second Fab molecule (i.e., the second Fab molecule comprises an exchange Type Fab heavy chain, wherein the heavy chain variable region is replaced by a light chain variable region), which in turn shares a carboxy-terminal peptide bond with the Fab heavy chain of the first Fab molecule, which in turn shares a Fab heavy chain with the third Fab molecule Carboxy-terminal peptide bond (VL(2)-CH1(2)-VH(1)-CH1(1)-VH(3)-CH1(3)). In some embodiments, the antibody step comprises: a polypeptide wherein the Fab heavy chain variable region of the second Fab molecule shares a carboxy-terminal peptide bond (VH(2)-CL(2) with the Fab light chain constant region of the second Fab molecule )), and share the carboxy-terminal peptide bond (VL(1)-CL(1)) with the Fab light chain polypeptide of the first Fab molecule. In some embodiments, the antibody further comprises a Fab light chain polypeptide (VL(3)-CL(3)) of a third Fab molec...

Claims

1. The use of parotuzumab vedotin and glofitamab for the preparation of a medicament for treating an individual with CD20-positive dysplastic syndrome, wherein the treatment comprises administering parotuzumab and glofitamab to the individual in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein: (a) The first dosing cycle comprises: (i) a single dose (C1D1) of parotizumab administered on day 2 of the first dosing cycle; and (ii) a first dose (C1D1) of glimepiride administered on day 8 of the first dosing cycle and a second dose (C1D2) of glimepiride administered on day 15 of the first dosing cycle, wherein the C1D1 of glimepiride is approximately 2.5 mg and the C1D2 of glimepiride is approximately 10 mg; and (b) The second dosing cycle comprises: (i) a single dose (C2D1) of parotizumab administered on day 1 of the second dosing cycle; and (ii) a single dose (C2D1) of glimepiride administered on day 1 of the second dosing cycle, wherein the C2D1 of glimepiride is approximately 30 mg. The concentration of the parotocilizumab is approximately 1.8 mg / kg for both C1D1 and C2D1.

2. The use of parotizumab and glimetuzumab for the preparation of a medicament for treating an individual with CD20-positive dysplastic syndrome, wherein the treatment comprises administering parotizumab and glimetuzumab to the individual in a dosing regimen comprising 12 dosing cycles, wherein: (a) The first dosing cycle includes a first dose (C1D1) of glimepiride administered on day 8 of the first dosing cycle, a second dose (C1D2) of glimepiride administered on day 15 of the first dosing cycle, and a single dose (C1D1) of parotizumab administered on day 2 of the first dosing cycle, wherein the C1D1 of glimepiride is approximately 2.5 mg and the C1D2 of glimepiride is approximately 10 mg; (b) The second to sixth dosing cycles each include a single dose of glimepiride (C2D1-C6D1) and a single dose of parotizumab (C2D1-C6D1); and (c) The seventh to twelfth dosing cycles each include a single dose of glimepiride (C7D1-C12D1) and do not include the administration of parotizumab. Each single dose of glimetuzumab, C2D1-C12D1, is administered on day 1 of each dosing cycle, and each single dose of parotizumab, C2D1-C6D1, is administered on day 1 of each dosing cycle. Each single dose of glimetuzumab, C2D1-C12D1, is approximately 30 mg, and each single dose of parotizumab, C1D1-C6D1, is approximately 1.8 mg / kg.

3. As in the purpose of request item 2, wherein: (a) The dosing cycle is 21 days; (b) Griffithomab is administered intravenously; and / or (c) Parotizumab is administered intravenously.

4. As requested in claim 2, wherein the treatment further comprises administering obinutuzumab to the individual.

5. As claimed in claim 4, wherein the obbituzumab (a) is administered prior to the administration of the glimetuzumab; and / or (b) is administered in a single dose of about 1000 mg.

6. As claimed in claim 5, wherein the obbituzumab was administered approximately seven days prior to the administration of the glimetuzumab.

7. As requested in claim 2, wherein the CD20-positive cell proliferative disorder is a B-cell proliferative disorder.

8. As requested in claim 7, wherein the B-cell proliferative disorder is non-Hodgkin's lymphoma (NHL) or central nervous system lymphoma (CNSL).

9. As claimed in claim 8, wherein the NHL is: (a) relapsed and / or refractory; and / or (b) diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), high-grade B-cell lymphoma, primary mediastinal (thymic) large B-cell lymphoma (PMLBCL), diffuse B-cell lymphoma, or small lymphocyte lymphoma.

10. As claimed in claim 7, wherein the B-cell proliferative disorder is relapsed and / or refractory.

11. The use of parotizumab and glimetuzumab for the preparation of a medicament for the treatment of relapsed and / or refractory (R / R) NHL in an individual population, wherein the treatment comprises administering parotizumab and glimetuzumab to the individuals in a dosing regimen comprising 12 dosing cycles, wherein: (a) The first dosing cycle comprises: (i) a first dose (C1D1) of glimetuzumab administered on day 8 of the first dosing cycle and a second dose (C1D2) of glimetuzumab administered on day 15 of the first dosing cycle, wherein the C1D1 of glimetuzumab is approximately 2.5 mg and the C1D2 of glimetuzumab is approximately 10 mg; and (ii) a single dose (C1D1) of parotizumab administered on day 2 of the first dosing cycle; (b) The second through sixth dosing cycles each comprise a single dose (C2D1-C6D1) of glimetuzumab and a single dose (C2D1-C6D1) of parotizumab; and (c) Each of the seventh to twelfth dosing cycles includes a single dose of glimepiride (C7D1) but does not include parotizumab administration. Each single dose of glimepiride (C2D1-C12D1) is administered on day 1 of each dosing cycle, and each single dose of parotizumab (C2D1-C6D1) is administered on day 1 of each dosing cycle. Each single dose of glimepiride (C2D1-C12D1) is approximately 30 mg, and each single dose of parotizumab (C1D1-C6D1) is approximately 1.8 mg / kg.

12. As used in request item 11, wherein: (a) The complete reaction rate is at least 20%; and / or (b) The total reaction rate is at least 30%.

13. As used in request item 11, wherein: (a) The R / R NHL is an R / R MCL; and wherein: (i) the complete reaction rate is at least 60%; and / or (ii) the total reaction rate is at least 60%; or (b) The R / R NHL is an R / R DLBCL; and wherein: (i) the complete reaction rate is at least 35%; and / or (ii) the total reaction rate is at least 60%.

14. As used in request item 11, wherein: (a) The complete response rate is higher than the reference complete response rate of the reference individual population treated with combination therapy containing glimetuzumab and atezolizumab but not parotizumab; and / or (b) The objective response rate is higher than the reference objective response rate of the reference individual population treated with combination therapy containing glimetuzumab and atezolizumab but not parotizumab.

15. As claimed in claim 2, wherein the C2D1-C12D1 of the glucuronide is administered after the administration of the C2D1-C6D1 of the parotidumab.

16. For the purposes of claim 2, wherein the individual: (a) is a person; (b) has received at least two prior systemic therapies; and / or (c) is ineligible for autologous stem cell transplantation (SCT).

17. As used in request item 1, wherein: (a) The first and second dosing cycles are 21-day dosing cycles; (b) Griffithomab is administered intravenously; and / or (c) Parotizumab is administered intravenously.

18. For the purposes of claim 1, wherein the individual: (a) is a person; (b) has received at least two prior systemic therapies; and / or (c) is ineligible for autologous stem cell transplantation (SCT).

19. As claimed in claim 1, wherein the treatment further comprises administering obbituzumab to the individual.

20. As claimed in claim 19, wherein the obbituzumab (a) is administered prior to the administration of the glimetuzumab; and / or (b) is administered in a single dose of about 1000 mg.

21. As claimed in claim 20, wherein the obbituzumab was administered approximately seven days prior to the administration of the glimetuzumab.

22. As requested in claim 1, wherein the CD20-positive cell proliferative disorder is a B-cell proliferative disorder.

23. As requested in claim 22, wherein the B-cell proliferative disorder is non-Hodgkin's lymphoma (NHL) or central nervous system lymphoma (CNSL).

24. For the purposes of claim 23, wherein the NHL is: (a) relapsed and / or refractory; and / or (b) diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), high-grade B-cell lymphoma, primary mediastinal (thymic) large B-cell lymphoma (PMLBCL), diffuse B-cell lymphoma, or small lymphocyte lymphoma.

25. As claimed in claim 22, wherein the B-cell proliferative disorder is relapsed and / or refractory.

26. As claimed in claim 1, wherein the treatment further comprises administering one or more additional therapeutic agents to the individual.

27. As claimed in claim 2, wherein the treatment further comprises administering one or more additional therapeutic agents to the individual.

28. For the purposes of claim 11, wherein the individual: (a) is a person; (b) has received at least two prior systemic therapies; and / or (c) is ineligible for autologous stem cell transplantation (SCT).

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