Administration of therapeutics with anti-FCRH5 / anti-CD3 bispecific antibodies

Abstract

The present invention provides methods of administration directed to the treatment of cancer, such as multiple myeloma, with an anti-crystallizable fragment receptor-like 5 (FcRH5) / anti-cluster of differentiation 3 (CD3) bispecific antibody and lenalidomide.

Description

[0001] Sequence Listing

[0002] This application contains a sequence listing, which has been submitted electronically in XML format and is incorporated herein by reference in its entirety. The XML copy was created on July 17, 2023, named 50474-302WO3_Sequence_Listing_7_17_23_.XML, and is 41,595 bytes in size. Technical Field

[0003] The present invention relates to the treatment of cancers such as B-cell proliferative disorders. More specifically, the present invention relates to the specific treatment of human patients with multiple myeloma (MM) using an anti-crystallizable fragment receptor-like 5 (FcRH5) / anti-cluster of differentiation 3 (CD3) bispecific antibody and lenalidomide. Background Art

[0004] Cancer remains one of the most deadly threats to human health, striking more than 1.7 million new patients each year and being the second leading cause of death after heart disease in the United States, accounting for approximately one in four deaths.

[0005] In particular, blood cancers are the second leading cause of cancer-related death. Blood cancers include multiple myeloma (MM), a tumor characterized by the proliferation and accumulation of malignant plasma cells. Approximately 110,000 people are diagnosed with MM each year worldwide. Despite advances in treatment, MM remains incurable, with an estimated median survival of 8 to 10 years for standard-risk myeloma and 2 to 3 years for high-risk disease, even with autologous stem cell transplantation (ASCT). Although patient survival has improved significantly over the past 20 years, only 10% to 15% of patients meet or exceed expected survival compared to the matched general population.

[0006] Therefore, there is a need for improved treatment options for MM and other hematological cancers. Summary of the invention

[0007] Provided herein, inter alia, are methods of treating a subject having cancer (eg, MM), compositions for use, and related articles of manufacture.

[0008] In one aspect, provided herein is a method of treating a subject having multiple myeloma (MM) having a high-risk cytogenetic characteristic, the method comprising administering to the subject (i) a bispecific antibody that binds to fragment crystallizable receptor-like 5 (FcRH5) and cluster of differentiation 3 (CD3) and (ii) lenalidomide.

[0009] In another example, provided herein is a bispecific antibody that binds to FcRH5 and CD3, the bispecific antibody being used for treating a subject having MM with high-risk cytogenetic characteristics, the treatment comprising administering the bispecific antibody and lenalidomide to the subject.

[0010] In some aspects, the subject experiences a partial remission (PR) or better following induction therapy.

[0011] In some aspects, the subject has received autologous stem cell transplantation (ASCT) within 100 days of initiating the method or treatment and / or does not have progressive disease.

[0012] In some aspects, the bispecific antibody and lenalidomide are administered to the patient as post-transplant maintenance therapy.

[0013] In some aspects, the high-risk cytogenetic profile includes one or more of the following: translocation events t(4;14) or t(14;16), del(17p), or 1q gain.

[0014] In some aspects, the subject has a high-risk cytogenetic profile at the time of diagnosis of MM.

[0015] In some aspects, the bispecific antibody and lenalidomide are administered to the subject at a dosing regimen comprising: (i) a first phase comprising one or more dosing cycles, wherein the first phase comprises administering the bispecific antibody to the subject every two weeks (Q2W); (ii) a second phase comprising one or more dosing cycles, wherein the second phase comprises administering the bispecific antibody to the subject every four weeks (Q4W).

[0016] In some aspects, each dosing cycle of the first phase and / or the second phase is a 28-day dosing cycle.

[0017] In some aspects, the method or treatment further comprises a preliminary phase comprising one or more dosing cycles prior to the first phase, wherein the preliminary phase comprises administering the bispecific antibody to the subject weekly (QW).

[0018] In some aspects, each dosing cycle of the preliminary phase is a 28-day dosing cycle.

[0019] In some aspects, the preliminary phase includes one dosing cycle (C1).

[0020] In some aspects, the preliminary phase comprises administering the bispecific antibody to the subject on Day 1, Day 8, and Day 15 of C1.

[0021] In some aspects, a target dose of the bispecific antibody is administered to the subject in a preliminary phase for each administration.

[0022] In some aspects, the preliminary phase comprises administering to the subject a first step-escalating dose of the bispecific antibody.

[0023] In some aspects, the subject is administered a first step-escalating dose on day 1 of C1.

[0024] In some aspects, the target dose is administered to the subject on Days 8 and 15 of C1.

[0025] In some aspects, the preliminary phase comprises administering to the subject a first step-escalating dose and a second step-escalating dose of the bispecific antibody.

[0026] In some aspects, the subject is administered a first step-escalating dose on day 1 of C1 and the subject is administered a second step-escalating dose on day 8 of C1.

[0027] In some aspects, the target dose is administered to the subject on day 15 of C1.

[0028] In some aspects, the first stepped-escalating dose is 3.6 mg.

[0029] In some aspects, the first stepped-up dose is 0.3 mg and the second stepped-up dose is 3.6 mg.

[0030] In some aspects, the first phase includes at least two dosing cycles, at least three dosing cycles, at least four dosing cycles, or at least five dosing cycles.

[0031] In some aspects, the first phase includes a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), and a fifth dosing cycle (C5).

[0032] In some aspects, the first phase comprises administering the bispecific antibody to the subject on Day 1 and Day 15 of C1, C2, C3, C4, and / or C5.

[0033] In some aspects, the subject is administered a target dose of the bispecific antibody for each administration during the first stage.

[0034] In some aspects, the second phase includes at least two dosing cycles, at least three dosing cycles, at least four dosing cycles, at least five dosing cycles, at least six dosing cycles, or at least seven dosing cycles.

[0035] In some aspects, the second phase includes a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), a fifth dosing cycle (C5), a sixth dosing cycle (C6), and a seventh dosing cycle (C7).

[0036] In some aspects, the second phase comprises administering the bispecific antibody to the subject on day 1 of C1, C2, C3, C4, C5, C6, and / or C7.

[0037] In some aspects, the subject is administered a target dose of the bispecific antibody for each administration during the second stage.

[0038] In some aspects, the target dose is between 90 mg and 198 mg, inclusive.

[0039] In some aspects, the target dose is 90 mg.

[0040] In some aspects, the target dose is 132 mg.

[0041] In some aspects, the target dose is 160 mg.

[0042] In some aspects, the bispecific antibody is administered intravenously to the subject.

[0043] In some aspects, lenalidomide is administered to the subject on day 1 to day 21 of each dosing cycle in the first phase and / or the second phase.

[0044] In some aspects, lenalidomide is administered to the subject on day 1 to day 21 of each dosing cycle in the pre-period.

[0045] In some aspects, lenalidomide is administered to the subject at a dose of about 10 mg to about 20 mg.

[0046] In some aspects, lenalidomide is administered to the subject at a dose of about 10 mg.

[0047] In some aspects, lenalidomide is administered to the subject at a dose of about 15 mg.

[0048] In some aspects, lenalidomide is administered orally to the subject.

[0049] In some aspects, the method or treatment further comprises administering a corticosteroid to the subject.

[0050] In some aspects, the method or treatment further comprises administering a corticosteroid to the subject during the first phase and / or the second phase.

[0051] In some aspects, the subject is administered a corticosteroid during the first phase on days 1 and 15 of C1 of the first phase.

[0052] In some aspects, if the subject experienced a cytokine release syndrome (CRS) event with a previous dose, a corticosteroid is administered to the subject in C2, C3, C4, and / or C5 of the first phase.

[0053] In some aspects, if the subject experienced a CRS event at a previous dose, a corticosteroid is administered to the subject in the second phase C1, C2, C3, C4, C5, C6, and / or C7.

[0054] In some aspects, the method or treatment further comprises administering a corticosteroid to the subject during the pre-period.

[0055] In some aspects, the subject is administered a corticosteroid on Day 1, Day 8, and Day 15 of C1 during the pre-phase.

[0056] In some aspects, the corticosteroid is administered to the subject intravenously or orally.

[0057] In some aspects, the corticosteroid is administered intravenously to the subject.

[0058] In some aspects, a corticosteroid is administered intravenously to the subject prior to administration of the bispecific antibody.

[0059] In some aspects, the subject is administered a corticosteroid intravenously about 1 hour prior to administration of the bispecific antibody.

[0060] In some aspects, the corticosteroid is dexamethasone or methylprednisolone.

[0061] In some aspects, the corticosteroid is dexamethasone.

[0062] In some aspects, dexamethasone is administered to the subject at a dose of about 20 mg.

[0063] In some aspects, methylprednisolone is administered to the subject at a dose of about 80 mg.

[0064] In some aspects, the bispecific antibody comprises an anti-FcRH5 arm comprising a first binding domain comprising the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence of RFGVH (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of VIWRGGSTDYNAAFVS (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of HYYGSSDYALDN (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of KASQDVRNLVV (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of SGSYRYS (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QQHYSPPYT (SEQ ID NO: 6).

[0065] In some aspects, the bispecific antibody comprises an anti-FcRH5 arm comprising a first binding domain, the first binding domain comprising: (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:7; (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:8; or (c) a VH domain as in (a) and a VL domain as in (b).

[0066] In some aspects, the first binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO:7; and a VL domain comprising the amino acid sequence of SEQ ID NO:8.

[0067] In some aspects, the bispecific antibody comprises an anti-CD3 arm comprising a second binding domain comprising the following six HVRs: (a) HVR-H1 comprising the amino acid sequence of SYYIH (SEQ ID NO:9); (b) HVR-H2 comprising the amino acid sequence of WIYPENDNTKYNEKFKD (SEQ ID NO:10); (c) HVR-H3 comprising the amino acid sequence of DGYSRYYFDY (SEQ ID NO:11); (d) HVR-L1 comprising the amino acid sequence of KSSQSLLNSRTRKNYLA (SEQ ID NO:12), (e) HVR-L2 comprising the amino acid sequence of WTSTRKS (SEQ ID NO:13); and (f) HVR-L3 comprising the amino acid sequence of KQSFILRT (SEQ ID NO:14).

[0068] In some aspects, the bispecific antibody comprises an anti-CD3 arm comprising a second binding domain, the second binding domain comprising: (a) a VH domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 15; (b) a VL domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16; or (c) a VH domain as in (a) and a VL domain as in (b).

[0069] In some aspects, the second binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO:15; and a VL domain comprising the amino acid sequence of SEQ ID NO:16.

[0070] In some aspects, the bispecific antibody comprises: an anti-FcRH5 arm comprising a heavy chain polypeptide (H1) and a light chain polypeptide (L1); and an anti-CD3 arm comprising a heavy chain polypeptide (H2) and a light chain polypeptide (L2); and wherein: (a) H1 comprises the amino acid sequence of SEQ ID NO:35; (b) L1 comprises the amino acid sequence of SEQ ID NO:36; (c) H2 comprises the amino acid sequence of SEQ ID NO:37; and (d) L2 comprises the amino acid sequence of SEQ ID NO:38.

[0071] In some aspects, the bispecific antibody comprises an aglycosylation site mutation.

[0072] In some aspects, the aglycosylation site mutation reduces the effector function of the bispecific antibody.

[0073] In some aspects, the deglycosylation site mutation is a substitution mutation.

[0074] In some aspects, the bispecific antibody comprises a substitution mutation in the Fc region that reduces effector function.

[0075] In some aspects, the bispecific antibodies are monoclonal antibodies.

[0076] In some aspects, the bispecific antibodies are humanized antibodies.

[0077] In some aspects, the bispecific antibodies are chimeric antibodies.

[0078] In some aspects, the bispecific antibody is an antibody fragment that binds FcRH5 and CD3.

[0079] In some aspects, the antibody fragment is selected from the group consisting of: Fab, Fab'-SH, Fv, scFv and (Fab') 2 Fragment.

[0080] In some aspects, the bispecific antibodies are full-length antibodies.

[0081] In some aspects, the bispecific antibody is an IgG antibody.

[0082] In some aspects, the IgG antibody is IgG 1 Antibody.

[0083] In some aspects, 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, the first CH2 (CH2 1 ) domain, the first CH3 (CH3 1 ) domain, the second CH1 (CH1 2) domain, the second CH2 (CH2 2 ) domain and the second CH3 (CH3 2 ) domain.

[0084] In some aspects, at least one of the one or more heavy chain constant domains is paired with another heavy chain constant domain.

[0085] In some aspects, CH3 1 Domain and CH3 2 The domains each contain a protrusion or a cavity, and wherein CH3 1 The protrusion or cavity in the domain can be located at the CH3 2 In a cavity or protrusion in a domain.

[0086] In some aspects, CH3 1 Domain and CH3 2 The domains meet at the interface between the protrusion and the cavity.

[0087] In some aspects, CH2 1 Domain and CH2 2 The domains each contain a protrusion or a cavity, and wherein CH2 1 The protrusion or cavity in the domain can be located at the CH2 2 In a cavity or protrusion in a domain.

[0088] In some aspects, CH2 1 Domain and CH2 2 The domains meet at the interface between the protrusion and the cavity.

[0089] In some aspects, the anti-FcRH5 arm comprises a protrusion and the anti-CD3 arm comprises a cavity.

[0090] In some aspects, the CH3 domain of the anti-FcRH5 arm comprises a protrusion comprising a T366W amino acid substitution mutation (EU numbering), and the CH3 domain of the anti-CD3 arm comprises a cavity comprising T366S, L368A, and Y407V amino acid substitution mutations (EU numbering).

[0091] In some aspects, the bispecific antibody is cevostamab.

[0092] In some aspects, the bispecific antibody and lenalidomide are administered to the subject simultaneously with one or more additional therapeutic agents.

[0093] In some aspects, the bispecific antibody and / or lenalidomide is administered to the subject prior to administration of one or more additional therapeutic agents.

[0094] In some aspects, the bispecific antibody and / or lenalidomide is administered to the subject after administration of one or more additional therapeutic agents.

[0095] In some aspects, the one or more additional therapeutic agents comprises an effective amount of tocilizumab.

[0096] In some aspects, the subject experiences a CRS event, and the method further comprises treating symptoms of the CRS event while suspending treatment with the bispecific antibody.

[0097] In some aspects, the method or treatment further comprises administering to the subject an effective amount of tocilizumab to treat the CRS event.

[0098] In some aspects, the CRS event does not resolve, or worsens, within 24 hours of treating symptoms of the CRS event, and the method further comprises administering one or more additional doses of tocilizumab to the subject to manage the CRS event.

[0099] In some aspects, tocilizumab is administered to the subject by intravenous infusion.

[0100] In some aspects: (i) the subject weighs ≥30 kg and tocilizumab is administered to the subject at a dose of 8 mg / kg; or (b) the subject weighs <30 kg and tocilizumab is administered to the subject at a dose of 12 mg / kg.

[0101] In some aspects, tocilizumab is administered to the subject 2 hours prior to administration of the bispecific antibody.

[0102] In some aspects, the one or more additional therapeutic agents include an effective amount of a B cell maturation antigen (BCMA) directed therapy, an additional immunomodulatory agent (IMiD), a CD38 directed therapy, or a combination of any of the foregoing.

[0103] In some aspects, the one or more additional therapeutic agents comprises an effective amount of acetaminophen or paracetamol.

[0104] In some aspects, acetaminophen or paracetamol is administered to the subject at a dose of between about 500 mg to about 1000 mg.

[0105] In some aspects, acetaminophen or paracetamol is orally administered to the subject.

[0106] In some aspects, the one or more additional therapeutic agents comprises an effective amount of diphenhydramine.

[0107] In some aspects, diphenhydramine is administered to the subject at a dose of between about 25 mg to about 50 mg.

[0108] In some aspects, diphenhydramine is administered orally to the subject.

[0109] In another aspect, provided herein is a method of treating a subject having MM with a high-risk cytogenetic profile, the method comprising administering to the subject silvostomumab and lenalidomide, wherein: (i) the subject experiences a PR or better after induction therapy; (ii) the subject receives ASCT within 100 days of initiation of the method and / or does not have progressive disease; (iii) silvostomumab and lenalidomide are administered to the patient as post-transplant maintenance therapy; and (iv) the high-risk cytogenetic profile comprises one or more of the following: translocation events t(4;14) or t(14;16), del(17p), or 1q gain.

[0110] In another aspect, provided herein is silvositumomab for use in treating a subject with MM having a high-risk cytogenetic profile, the treatment comprising administering silvositumomab and lenalidomide to the subject, wherein: (i) the subject experiences a PR or better after induction therapy; (ii) the subject receives ASCT within 100 days of initiation of the method and / or does not have progressive disease; (iii) silvositumomab and lenalidomide are administered to the patient as post-transplant maintenance therapy; and (iv) the high-risk cytogenetic profile comprises one or more of the following: translocation events t(4;14) or t(14;16), del(17p), or 1q gain.

[0111] On the other hand, provided herein is a method for treating a subject having MM with a high-risk cytogenetic feature, the method comprising administering silvostomumab and lenalidomide to the subject at a dosing regimen comprising: (i) a preliminary phase comprising a 28-day dosing cycle (C1); (ii) a first phase after the preliminary phase comprising a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), and a fifth dosing cycle (C5), wherein each dosing cycle of the first phase is a 28-day dosing cycle; and (iii) a second phase after the first phase comprising a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), a fifth dosing cycle (C5), a sixth dosing cycle (C6), and a seventh dosing cycle (C7), wherein each dosing cycle of the second phase is 28 days. The subject is administered silvostomumab as follows: (i) at a first step-escalating dose on day 1 of C1 during the pre-phase, and at a second step-escalating dose on day 8 of C1 during the pre-phase; (ii) at a target dose on day 15 of C1 during the pre-phase; (iii) at a target dose on days 1 and 15 of C1, C2, C3, C4, and C5 during the first phase; and (iv) at a target dose on day 1 of C1, C2, C3, C4, C5, C6, and C7 during the second phase; and wherein lenalidomide is administered to the subject as follows: (i) on days 1 to 21 of C1 during the pre-phase; (ii) on days 1 to 21 of C1, C2, C3, C4, and C5 during the first phase; and (iii) on days 1 to 21 of C1, C2, C3, C4, C5, C6, and C7 during the second phase.

[0112] On the other hand, provided herein is silvositumomab for use in treating a subject with MM having a high-risk cytogenetic feature, the treatment comprising administering silvositumomab and lenalidomide to the subject at a dosing regimen comprising: (i) a preliminary phase comprising a 28-day dosing cycle (C1); (ii) a first phase following the preliminary phase comprising a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), and a fifth dosing cycle (C5), wherein each dosing cycle of the first phase is a 28-day dosing cycle; and (iii) a second phase following the first phase comprising a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), a fifth dosing cycle (C5), a sixth dosing cycle (C6), and a seventh dosing cycle (C7), wherein each dosing cycle of the second phase is a 28-day dosing cycle. The phase is a 28-day dosing cycle, wherein silvostomumab is administered to the subject as follows: (i) at a first step-escalating dose on Day 1 of C1 during the pre-phase, and at a second step-escalating dose on Day 8 of C1 during the pre-phase; (ii) at a target dose on Day 15 of C1 during the pre-phase; (iii) at a target dose on Days 1 and 15 of C1, C2, C3, C4, and C5 during the first phase; and (iv) at a target dose on Day 1 of C1, C2, C3, C4, C5, C6, and C7 during the second phase; and wherein lenalidomide is administered to the subject as follows: (i) on Day 1 to Day 21 of C1 during the pre-phase; (ii) on Day 1 to Day 21 of C1, C2, C3, C4, and C5 during the first phase; and (iii) on Day 1 to Day 21 of C1, C2, C3, C4, C5, C6, and C7 during the second phase.

[0113] In some aspects: (i) the subject experiences a PR or better after induction therapy; (ii) the subject receives ASCT within 100 days of initiation of the approach and / or does not have progressive disease; (iii) the patient is administered silvostomumab and lenalidomide as post-transplant maintenance therapy; and (iv) the high-risk cytogenetic profile includes one or more of the following: translocation events t(4;14) or t(14;16), del(17p), or 1q gain.

[0114] In some aspects: (i) the first step-escalating dose of silvositumomab is 0.3 mg; (ii) the second step-escalating dose of silvositumomab is 3.6 mg; (iii) the target dose of silvositumomab is between 90 mg and 198 mg, inclusive; and (iv) lenalidomide is administered at a dose of 10 mg or 15 mg.

[0115] In some aspects, the target dose is 90 mg.

[0116] In some aspects, the target dose is 132 mg.

[0117] In some aspects, the target dose is 160 mg. BRIEF DESCRIPTION OF THE DRAWINGS

[0118] Figure 1 Schematic diagram of the cevos + lenalidomide ("Len") substudy for CO43923. D, day. DETAILED DESCRIPTION

[0119] I. Definitions

[0120] As used herein, the term "about" refers to the common error range for the corresponding value that is readily known to those skilled in the art. Reference herein to "about" a value or parameter includes (and describes) aspects relating to the value or parameter itself.

[0121] It should be understood that aspects of the invention described herein include "comprising," "consisting of," and "consisting essentially of" aspects.

[0122] As used herein, the term "FcRH5" or "anti-crystallizable fragment receptor-like 5" refers to any native FcRH5 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise indicated, and encompasses "full-length," unprocessed FcRH5, as well as any form of FcRH5 produced by processing in cells. The term also encompasses naturally occurring variants of FcRH5, including, for example, splice variants or allelic variants. FcRH5 includes, for example, human FcRH5 protein (UniProtKB / Swiss-Prot ID: Q96RD9.3), which is 977 amino acids in length.

[0123] The terms "anti-FcRH5 antibody" and "antibody that binds to FcRH5" refer to an antibody that is capable of binding to FcRH5 with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent targeting FcRH5. In one embodiment, the extent of binding of an anti-FcRH5 antibody to an unrelated, non-FcRH5 protein is less than about 10% of the extent of binding of the antibody to FcRH5, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the antibody that binds to FcRH5 has a binding affinity of ≤1 μM, ≤250 nM, ≤100 nM, ≤15 nM, ≤10 nM, ≤6 nM, ≤4 nM, ≤2 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or ≤0.001 nM (e.g., 10 -8 M or smaller, e.g. 10 -8 M to 10-13 M, for example 10 -9 M to 10 -13 The dissociation constant (K D In certain embodiments, the anti-FcRH5 antibody binds to an epitope of FcRH5 that is conserved among FcRH5 from different species.

[0124] Unless otherwise indicated, the term "cluster of differentiation 3" or "CD3" as used herein refers to any native CD3 from any vertebrate source, including, for example, primates (e.g., humans) and rodents (e.g., mice and rats), including, for example, CD3ε, CD3γ, CD3α, and CD3β chains. The term encompasses "full-length" unprocessed CD3 (e.g., unprocessed or unmodified CD3ε or CD3γ), as well as any form of CD3 produced by processing in cells. The term also encompasses naturally occurring variants of CD3, including, for example, splice variants or allelic variants. CD3 includes, for example, a human CD3ε protein (NCBI RefSeq No. NP_000724) having a length of 207 amino acids and a human CD3γ protein (NCBI RefSeq No. NP_000064) having a length of 182 amino acids.

[0125] The terms "anti-CD3 antibody" and "antibody that binds to CD3" refer to an antibody that is capable of binding to CD3 with sufficient affinity such that the antibody can be used as a diagnostic and / or therapeutic agent targeting CD3. In one embodiment, the extent of binding of the anti-CD3 antibody to an unrelated, non-CD3 protein is less than about 10% of the extent of binding of the antibody to CD3, as measured, for example, by a radioimmunoassay (RIA). In certain embodiments, the antibody that binds to CD3 has a binding affinity of ≤1 μM, ≤250 nM, ≤100 nM, ≤15 nM, ≤10 nM, ≤5 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or ≤0.001 nM (e.g., 10 -8 M or smaller, e.g. 10 -8 M to 10 -13 M, for example 10 -9 M to 10 -13 The dissociation constant (K D In certain embodiments, the anti-CD3 antibody binds to an epitope of CD3 that is conserved in CD3 from different species.

[0126] For the purposes of this article, "Sivositumab" is also referred to as BFCR4350A or RO7187797, which is an Fc-engineered, humanized, full-length aglycosylated IgG1κ T cell-dependent bispecific antibody (TDB) that binds to FcRH5 and CD3 and comprises an anti-FcRH5 arm comprising a heavy chain polypeptide sequence of SEQ ID NO:35 and a light chain polypeptide sequence of SEQ ID NO:36, and an anti-CD3 arm comprising a heavy chain polypeptide sequence of SEQ ID NO:37 and a light chain polypeptide sequence of SEQ ID NO:38. Sivositumab contains an amino acid substitution of threonine to tryptophan at position 366 of the anti-FcRH5 arm (T366W) using EU numbering of amino acid residues in the Fc region, and three amino acid substitutions on the heavy chain of the anti-CD3 arm (Y407V, T366S, and L368A) using EU numbering of amino acid residues in the Fc region (a tyrosine to valine substitution at position 407, a threonine to serine substitution at position 366, and a leucine to alanine substitution at position 368) to drive heterodimerization of the two arms (half antibody). Sivositumab also contains an amino acid substitution (N297G) at position 297 on each heavy chain using EU numbering of amino acid residues in the Fc region, which results in a minimal aglycosylated antibody that binds to Fc (Fcγ) receptors and thereby prevents Fc effector function. Sivostomumab is also described in the following document: WHO Drug Information (International Nonproprietary Names), Proposed INN: List 84, Volume 34, Issue 3, published in 2020 (see page 701).

[0127] The term "antibody" herein is used in the broadest sense and covers various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) and antibody fragments (e.g., bis-Fab), so long as they exhibit the desired antigen-binding activity.

[0128] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be expressed by the dissociation constant (K D ) represents. Affinity can be measured by conventional methods known in the art, including those described herein. Specific illustrative and exemplary aspects for measuring binding affinity are described below.

[0129] An "affinity matured" antibody is one with one or more alterations in one or more hypervariable regions (HVRs) which result in an improvement in the affinity of the antibody for antigen, compared to a parent antibody which does not possess such alterations.

[0130] The terms "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein to refer to an antibody having a structure substantially similar to a native antibody structure or having heavy chains that contain an Fc region as defined herein.

[0131] "Antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody and binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to: bis-Fab; Fv; Fab; Fab, Fab'-SH; F(ab') 2 ; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv, ScFab); and multispecific antibodies formed from antibody fragments.

[0132] "Single domain antibody" refers to an antibody fragment that contains all or part of the heavy chain variable domain or all or part of the light chain variable domain of an antibody. In certain aspects, a single domain antibody is a human single domain antibody (see, e.g., U.S. Patent No. 6,248,516B1). Examples of single domain antibodies include, but are not limited to, VHH.

[0133] The "Fab" fragment is an antigen-binding fragment produced by papain digestion of an antibody and consists of an intact L chain as well as the variable region domain of the H chain (VH) and the first constant domain (CH1) of one heavy chain. Papain digestion of an antibody produces two identical Fab fragments. Pepsin treatment of an antibody produces a single large F(ab') 2 Fragments are roughly equivalent to two Fab fragments connected by disulfide bonds with divalent antigen binding activity and still capable of cross-linking antigens. The difference between Fab' fragments and Fab fragments is that Fab' fragments have some residues added to the carboxyl terminus of the CH1 domain, which contain one or more cysteines from the antibody hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine ​​residues of the constant domains carry free thiol groups. F(ab') 2 Antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them.Other chemical couplings of antibody fragments are also known.

[0134] "Fv" consists of a tight, non-covalently associated dimer of a heavy chain variable region domain and a light chain variable region domain. Six hypervariable loops (3 loops each from the H chain and L chain) are generated by the folding of these two domains, which contribute amino acid residues to achieve antigen binding and provide the antibody with antigen binding specificity. However, even a single variable domain (or half of an Fv, containing only three CDRs specific for an antigen) has the ability to recognize and bind to an antigen, although its affinity is often lower than that of the complete binding site.

[0135] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain, which includes a native sequence Fc region and a variant Fc region. Although the boundaries of an immunoglobulin heavy chain Fc region may vary, the human IgG heavy chain Fc region is generally defined as extending from the amino acid residue at position Cys226 or from Pro230 to the carboxyl terminus of the heavy chain. The C-terminal lysine in the Fc region (residue 447 according to the EU numbering system) can be removed in, for example, the production or purification process of an antibody or by recombinantly designing a nucleic acid encoding an antibody heavy chain. Therefore, the composition of a complete antibody can include an antibody population removing all Lys447 residues, an antibody population not removing Lys447 residues, and an antibody population with and without an antibody mixture containing Lys447 residues.

[0136] A "functional Fc region" possesses the "effector functions" of a native sequence Fc region. Exemplary "effector functions" include C1q binding; CDC; Fc receptor binding; ADCC; phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor; BCR), etc. Such effector functions generally require an Fc region in combination with a binding domain (e.g., an antibody variable domain), and can be assessed using various assays disclosed, for example, in the definitions herein.

[0137] A "native sequence Fc region" comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature. Native sequence human Fc regions include native sequence human IgG 1 Fc regions (non-A and A allotypes); native sequence human IgG2 Fc regions; native sequence human IgG3 Fc regions; and native sequence human IgG4 Fc regions and naturally occurring variants thereof.

[0138] A "variant Fc region" comprises an amino acid sequence that differs from a native sequence Fc region by virtue of at least one amino acid modification, preferably one or more amino acid substitutions. Preferably, the variant Fc region has at least one amino acid substitution compared to the native sequence Fc region or the Fc region of a parent polypeptide, for example, from about one to about ten amino acid substitutions in the native sequence Fc region or the Fc region of a parent polypeptide, and preferably from about one to about five amino acid substitutions. The variant Fc regions described herein preferably have at least about 80% homology with the native sequence Fc region and / or with the Fc region of a parent polypeptide, preferably at least about 90% homology therewith, or preferably at least about 95% homology therewith.

[0139] As used herein, "Fc complex" refers to the CH3 domains of two Fc regions interacting together to form a dimer, or in certain aspects, two Fc regions interacting to form a dimer, wherein the cysteine ​​residues in the hinge region and / or the CH3 domains interact through bonds and / or forces (e.g., van der Waals forces, hydrophobic forces, hydrogen bonds, electrostatic forces, or disulfide bonds).

[0140] As used herein, "Fc component" refers to the hinge region, CH2 domain or CH3 domain of the Fc region.

[0141] The "hinge region" is generally defined as stretching from about residues 216 to 230 of IgG (EU numbering), from about residues 226 to 243 of IgG (Kabat numbering), or from about residues 1 to 15 of IgG (IMGT unique numbering).

[0142] The "lower hinge region" of an Fc region is generally defined as the stretch of residues immediately C-terminal to the hinge region, ie, residues 233 to 239 (EU numbering) of the Fc region.

[0143] A "variant Fc region" comprises an amino acid sequence that differs from a native sequence Fc region by virtue of at least one amino acid modification, preferably one or more amino acid substitutions. Preferably, the variant Fc region has at least one amino acid substitution compared to the native sequence Fc region or the Fc region of a parent polypeptide, for example, from about one to about ten amino acid substitutions in the native sequence Fc region or the Fc region of a parent polypeptide, and preferably from about one to about five amino acid substitutions. The variant Fc region described herein preferably has at least about 80% homology with the native sequence Fc region and / or with the Fc region of a parent polypeptide, and preferably has at least about 90% homology therewith, more preferably has at least about 95% homology therewith.

[0144] "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. The preferred FcR is a native sequence human FcR. In addition, the preferred FcR is an FcR that binds to IgG antibodies (a gamma receptor), and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced ​​forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), which have similar amino acid sequences, the main difference being their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (see Annu. Rev. Immunol. 15:203-234, 1997 (reviewed in M.). For reviews of FcRs, see: Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). The term "FcR" herein encompasses other FcRs, including those to be identified in the future. The term also includes the neonatal receptor FcRn, which is responsible for the transfer of maternal IgG to the fetus (Guyer et al., J. Immunol. 117:587 (1976); and Kim et al., J. Immunol. 24:249 (1994)).

[0145] As described herein, the term "knob-into-hole" or "KnH" technology refers to a technique for guiding the pairing of two polypeptides together in vivo or in vitro by introducing a protrusion (knob) into one polypeptide and a cavity (hole) into the other polypeptide at their interactive interface. For example, KnH has been introduced into the Fc:Fc interaction interface, CL:CH1 interface, or VH / VL interface of antibodies (e.g., US2007 / 0178552; WO 96 / 027011; WO 98 / 050431; and Zhu et al. (1997) Protein Science 6:781-788). This is particularly useful for driving two different heavy chains to pair together during the preparation of multispecific antibodies. For example, a multispecific antibody having a KnH in its Fc region can further comprise a single variable domain connected to each Fc region, or further comprise different heavy chain variable domains paired with the same, similar or different light chain variable domains. KnH technology can also be used to pair together two different receptor extracellular domains or any other polypeptide sequences containing different target recognition sequences.

[0146] "Framework" or "FR" refers to the variable domain residues other than the hypervariable region (HVR) residues. The FR of the variable domain is usually composed of the following four FR domains: FR1, FR2, FR3 and FR4. Therefore, HVR and FR sequences usually appear in the following sequence in VH (or VL): FR1-H1 (L1)-FR2-H2 (L2)-FR3-H3 (L3)-FR4.

[0147] A "CH1 region" or "CH1 domain" comprises a stretch of residues extending from about residue 118 to residue 215 of IgG (EU numbering), from about residue 114 to 223 of IgG (Kabat numbering), or from about residue 1.4 to residue 121 of IgG (IMGT unique numbering) (Lefranc et al., the international ImMunoGeneTics information 25 years on. Nucleic Acids Res. 2015 Jan; 43 (Database issue): D413-22).

[0148] The "CH2 domain" of the human IgG Fc region generally extends from about 244 to about 360 (Kabat numbering) of IgG, from about 231 to about 340 (EU numbering) of IgG, or from about 1.6 to about 125 (IGMT unique numbering) of IgG. The uniqueness of the CH2 domain is that it is not tightly paired with another domain. In contrast, two N-connected branched carbohydrate chains are between the two CH2 domains of the complete native IgG molecule. It is speculated that carbohydrates may provide alternatives for domain-domain pairing and help stabilize the CH2 domain. Burton, Molec. Immunol. 22: 161-206 (1985).

[0149] A “CH3 domain” comprises a stretch of residues extending C-terminally of the CH2 domain in the Fc region (i.e., from about amino acid residue 361 to about amino acid residue 478 of IgG (Kabat numbering), from about amino acid residue 341 to about amino acid residue 447 of IgG (EU numbering), or from about amino acid residue 1.4 to about amino acid residue 130 of IgG (IGMT unique numbering)).

[0150] The "CL domain" or "constant light domain" comprises a stretch of residues extending from the C-terminus of the light chain variable domain (VL). The light chain of an antibody may be a kappa (κ) ("Cκ") or lambda (λ) ("Cλ") light chain region. The Cκ region typically extends from about residue 108 to residue 214 of IgG (Kabat or EU numbering) or from about residue 1.4 to residue 126 of IgG (IMGT unique numbering). The Cλ residues typically extend from about residue 107a to residue 215 (Kabat numbering) or from about residue 1.5 to residue 127 (IMGT unique numbering) (Lefranc et al., the international ImMunoGeneTics information 25 years on. Nucleic Acids Res. 2015 Jan; 43 (Database issue): D413-22).

[0151] The light chains (LC) derived from any vertebrate can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequence of their constant domains. Immunoglobulins can be assigned to different classes or isotypes based on the amino acid sequence of their heavy chain constant domains (CH). There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM have heavy chains called α, δ, γ, ε, and μ, respectively. The γ and α classes are further divided into subclasses based on relatively minor differences in CH sequence and function, for example, humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0152] The term "chimeric" antibody refers to an antibody in which a portion of the heavy and / or light chain is derived from a particular source or species, while the remainder of the heavy and / or light chain is derived from a different source or species.

[0153] The "class" of an antibody refers to the type of constant domain or region possessed by the heavy chain of the antibody. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and some of these antibodies can be further divided into subclasses (isotypes), such as IgG 1 IgG 2 IgG 3 IgG 4 , IgA 1 and IgA 2 The heavy-chain constant domains that correspond to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0154] A "human antibody" is an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human or human cell, or derived from an antibody of non-human origin using a full repertoire of human antibodies or other human antibody coding sequences. This definition of a human antibody specifically excludes humanized antibodies comprising non-human antigen binding residues. Human antibodies can be produced using various techniques known in the art, including phage display libraries. Hoogenboom and Winter. J. Mol. Biol. 227: 381, 1991; Marks et al. J. Mol. Biol. 222: 581, 1991. Methods for preparing human monoclonal antibodies can also be described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147 (1): 86-95, 1991. See also van Dijk and van de Winkel. Curr. Opin. Pharmacol. 5:368-74, 2001. Human antibodies can be prepared by administering an antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge but whose endogenous loci have been disabled, for example, by immunizing a xenogeneic mouse (see, e.g., XENOMOUSE TM Nos. 6,075,181 and 6,150,584 of the Human B Cell Hybridoma Technology). See also, e.g., Li et al. Proc. Natl. Acad. Sci. USA 103:3557-3562, 2006 for human antibodies generated via human B cell hybridoma technology.

[0155] A "human consensus framework" is a framework that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. In general, the selection of human immunoglobulin VL or VH sequences is from a subset of variable domain sequences. In general, the subset of sequences is a subset as described in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda MD (1991), Volumes 1-3. In one aspect, for VL, the subgroup is subgroup κI as described in Kabat et al., cited above. In one aspect, for VH, the subgroup is subgroup III as described in the literature of Kabat et al., cited above.

[0156] "Humanized" antibody refers to such chimeric antibody, which includes amino acid residues from non-human HVR and amino acid residues from human FR. In some aspects, humanized antibody will substantially include all at least one, usually two variable domains, wherein all or substantially all HVR (e.g., CDR) correspond to the HVR of non-human antibodies, and all or substantially all FRs correspond to the FRs of human antibodies. In some aspects where all or substantially all FRs of humanized antibodies correspond to the FRs of human antibodies, any FR of humanized antibodies may contain one or more amino acid residues from non-human FRs (e.g., one or more vernier position residues of FR). Humanized antibodies may optionally include at least a portion of an antibody constant region derived from a human antibody. "Humanized form" antibodies, such as non-human antibodies, refer to antibodies that have undergone humanization.

[0157] The term "variable region" or "variable domain" refers to the domain of an antibody heavy chain or light chain that participates in the binding of an antibody to an antigen. The variable domains of the heavy and light chains of natural antibodies (VH and VL, respectively) generally have similar structures, wherein each domain comprises four conserved framework regions (FRs) and three hypervariable regions (HVRs). (See, e.g., Kindt et al., Kuby Immunology, 6th edition, WH Freeman and Co., p. 91 (2007).) A single VH or VL domain may be sufficient to confer antigen binding specificity. In addition, antibodies that bind to a specific antigen can be isolated using the VH or VL domains from antibodies that bind to the antigen, respectively, to screen libraries of complementary VL or VH domains. See, e.g., Portolano et al. J. Immunol. 150: 880-887, 1993; Clarkson et al. Nature 352: 624-628, 1991.

[0158] As used herein, the term "hypervariable region" or "HVR" refers to each region of an antibody variable domain that is hypervariable in sequence (complementarity determining region or CDR). Typically, an antibody comprises six CDRs; three in VH (CDR-H1, CDR-H2, CDR-H3), and three in VL (CDR-L1, CDR-L2, CDR-L3). Exemplary CDRs herein include:

[0159] (a) CDRs 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);

[0160] (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 Ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991)); and

[0161] (c) Antigenic contact points 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).

[0162] Unless otherwise indicated, HVR residues and other residues in the variable domain (eg, FR residues) are numbered herein according to Kabat et al., supra.

[0163] "Single-chain Fv", also abbreviated as "sFv" or "scFv", is an antibody fragment comprising VH and VL antibody domains linked into a single polypeptide chain. Preferably, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains, allowing the scFv to form a desired antigen binding structure. For a review of scFv, see Pluckthun's The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds., Springer-Verlag, New York, pp. 269-315 (1994); Malmborg et al., J. Immunol. Methods 183: 7-13, 1995.

[0164] "Targeting domain" means a portion of a compound or molecule that specifically binds to a target epitope, antigen, ligand, or receptor. Targeting domains include, but are not limited to, antibodies (e.g., monoclonal antibodies, polyclonal antibodies, recombinant antibodies, humanized antibodies, and chimeric antibodies), antibody fragments or portions thereof (e.g., bi-Fab fragments, Fab fragments, Fab' 2, scFab, scFv antibodies, SMIPs, single domain antibodies, diabodies, minibodies, scFv-Fc, affibodies, nanobodies, and VH and / or VL domains of antibodies), receptors, ligands, aptamers, peptide targeting domains (e.g., cysteine ​​knot proteins (CKPs)) and other molecules with identified binding partners. The targeting domain can target, block, excite or antagonize the antigen bound to it.

[0165] The term "monoclonal antibody" as used herein refers to an antibody obtained from a substantially homogeneous antibody population, that is, except for possible variant antibodies (e.g., containing naturally occurring mutations or produced during the production of monoclonal antibody preparations, such variants are usually present in small amounts), the individual antibodies comprising the population have identity and / or bind to the same epitope. Contrary to the polyclonal antibody preparations that generally include different antibodies for different determinants (epitopes), each monoclonal antibody in the monoclonal antibody preparation is directed to a single determinant on the antigen. Therefore, the modifier "monoclonal" represents that the feature of the antibody is obtained from a substantially homogeneous antibody population, and should not be interpreted as requiring antibody to be produced by any ad hoc method. For example, the monoclonal antibody to be used according to the present invention can be prepared by a variety of techniques, including but not limited to hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of human immunoglobulin loci, such methods and other exemplary methods for preparing monoclonal antibodies are described herein.

[0166] The term "multispecific antibody" is used in the broadest sense and specifically encompasses antibodies with multi-epitope specificity. In one aspect, a multispecific antibody binds to two different targets (e.g., a bispecific antibody). Such multispecific antibodies include, but are not limited to, antibodies comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH / VL unit has multi-epitope specificity; antibodies having two or more VL and VH domains, wherein each VH / VL unit binds to a different epitope; antibodies having two or more single variable domains, wherein each single variable domain binds to a different epitope; full-length antibodies; antibody fragments such as Fab, Fv, dsFv, scFv, diabodies, bispecific diabodies and triabodies, covalently or non-covalently linked antibody fragments. "Multi-epitope specificity" refers to the ability to specifically bind to two or more different epitopes on the same or different targets. "Monospecificity" refers to the ability to bind only one antigen. In one aspect, a monospecific bi-epitope antibody binds to two different epitopes on the same target / antigen. In one aspect, a monospecific multi-epitope antibody binds to multiple different epitopes of the same target / antigen. According to one aspect, a multispecific antibody is an IgG antibody that binds to each epitope with an affinity of 5 μM to 0.001 pM, 3 μM to 0.001 pM, 1 μM to 0.001 pM, 0.5 μM to 0.001 pM, or 0.1 μM to 0.001 pM.

[0167] A "naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (eg, a cytotoxic moiety) or a radiolabel. Naked antibodies can be present in pharmaceutical preparations.

[0168] "Natural antibodies" refer to naturally occurring immunoglobulin molecules with different structures. For example, natural IgG antibodies are heterotetrameric glycoproteins of about 150,000 daltons, consisting of two identical light chains and two identical heavy chains bonded by disulfide bonds. From the N-terminus to the C-terminus, each heavy chain has a variable region (VH), also referred to as a variable heavy chain domain or a heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3). Similarly, from the N-terminus to the C-terminus, each light chain has a variable region (VL), also referred to as a variable light chain domain or a light chain variable domain, followed by a constant light chain (CL) domain. The light chain of an antibody can be classified into one of two types based on the amino acid sequence of its constant domain, which are called kappa (κ) and lambda (λ).

[0169] As used herein, the term "immunoadhesin" refers to a molecule that combines the binding specificity of a heterologous protein ("adhesin") with the effector function of an immunoglobulin constant domain. Structurally, an immunoadhesin comprises a fusion of an amino acid sequence with a desired binding specificity and an immunoglobulin constant domain sequence (e.g., CH2 and / or CH3 sequence of IgG), which is not an antigen recognition and binding site of an antibody (i.e., "heterologous" compared to the constant region of an antibody). Adhesin and immunoglobulin constant domains may optionally be separated by an amino acid spacer sequence. Exemplary adhesin sequences include continuous amino acid sequences that include a portion of a receptor or ligand that binds to a target protein. Adhesin sequences may also be sequences that bind to a target protein, but are not receptor or ligand sequences (e.g., adhesin sequences in peptibodies). Such polypeptide sequences may be selected or identified by various methods, including phage display technology and high-throughput sorting methods. The immunoglobulin constant domain sequences in the immunoadhesins may be obtained from any immunoglobulin, such as IgG1, IgG2, IgG3 or IgG4 subtypes, IgA (including IgA1 and IgA2), IgE, IgD or IgM.

[0170] "Chemotherapeutic agents" include chemical compounds that can be used to treat cancer. Examples of chemotherapeutic agents include erlotinib ( Genentech / OSI Pharm.), bortezomib ( Millennium Pharm.), disulfiram, epigallocatechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol, lactate dehydrogenase A (LDH-A), fulvestrant ( AstraZeneca), sunitib ( Pfizer / Sugen), letrozole ( Novartis), imatinib mesylate ( Novartis、finasunate( Novartis), oxaliplatin ( Sanofi), 5-FU (5-fluorouracil), folinic acid, rapamycin (Sirolimus, Wyeth), lapatinib ( GSK572016, Glaxo Smith Kline), Lonafamib (SCH 66336), Sorafenib ( Bayer Labs), gefitinib ( AstraZeneca), AG1478, alkylating agents such as thiotepa and Cyclophosphamide; alkyl sulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylmelamines, including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylomelamine; acetogen ins (particularly bullatacin and bullatacinone); camptothecins (including topotecan and irinotecan); bryostatin; callystatin; CC-1065 (including its synthetic analogs adozelesin, carzelesin, and bizelesin); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); adrenocortical steroids (including prednisone and prednisolone); cyproterone acetate; 5α-reductase inhibitors (including finasteride and dutasteride); vorinostat, romidepsin, panbicin, valproic acid, mocetinostat, dolastatin, and valproic acid. dolastatin; aldesleukin, talcum duocarcin (including synthetic analogs KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards, such as chlorambucil, chlomaphazine, clofosamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine hydrochloride, melphalan, neomycin, phenylephrine, prednimustine, trofosfamide, uracil mustard, mustard); nitrosoureas, for example carmustine, chlorzoxazone, fermustine, lomustine, nimustine and ranimnustine; antibiotics, for example enediyne antibiotics (such as calicheamicin, in particular calicheamicin γ1I and calicheamicin ω1I (Angew Chem. Intl. Ed. Engl.1994 33:183-186); dynemicins, including dynemicin A; bisphosphonates, such as clodronate; esperamicin; and the new carcinogens and related chromophore proteins (enediyne antibiotic chromophores), aclacnomysins, actinomycins, authramycins, azaserines, bleomycins, cactinomycins, carabicins, caminomycins, carzinophilins, chromomycins, dactinomycins, daunomycins, detorubicin, 6-diazo-5-oxo-L-norleucine, (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolidine-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogamycin, olivetomycin, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozotocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denofolate, pterin), methotrexate, pteropterin, trimetrexate; purine analogs, such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens, such as calestrol, drostanolone propionate, cyclothiodol, melastosane, testolactone; antiadreners, such as aminoglutamine, mitotane, trilostane; folic acid supplements, such as folic acid; aceglatone; aldophosphamide glycosides, such as dapoxetine; glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids, such as maytansine and ansamitocins;Mitoguazone; mitoxantrone; mopidamnol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazine; benzylhydrazine; Polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A) A) and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; cytarabine ("Ara-C"); cyclophosphamide; thiotepa; taxanes, such as TAXOL (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ), (without Cremophor), an albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and (docetaxel, docetaxel; Sanofi-Aventis); chlorambucil; (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vinblastine; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; (Changchunorelbine); novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine Ibandronate; CPT-11; topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinols, such as retinoic acid; and any pharmaceutically acceptable salts, acids and derivatives thereof.

[0171] Chemotherapeutic agents also include: (i) antihormonal agents that act to modulate or inhibit the effects of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including tamoxifen (including Tamoxifen citrate), raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and (toremifine citrate); (ii) aromatase inhibitors that inhibit aromatase, an enzyme that regulates estrogen production by the adrenal glands, such as 4(5)-imidazole, aminoglutethimide, (Megestrol acetate), (exemestane; Pfizer), formestanie, fadrozole, (vorozole), (letrozole; Novartis) and (anastrozole; AstraZeneca); (iii) antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, triptorelin, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, all trans-retinoic acid, fenretinide, and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly those that inhibit the expression of genes in signal transduction pathways involved in abnormal cell proliferation, such as PKC-α, Ralf, and H-Ras; (vii) ribozymes, such as inhibitors of VEGF expression (e.g. ) and HER2 expression inhibitors; (viii) vaccines, such as gene therapy vaccines, such as and rIL-2; topoisomerase 1 inhibitors, e.g. rmRH; and (ix) any of the pharmaceutically acceptable salts, acids and derivatives of the foregoing.

[0172] Chemotherapeutic agents also include antibodies, such as alemtuzumab (Campath), bevacizumab ( Genentech; Cetuximab ( Imclone; Panitumumab ( Amgen), rituximab ( Genentech / Biogen Idec), Pertuzumab ( 2C4, Genentech), trastuzumab ( Genentech), tositumomab (Bexxar, Corixia) and antibody-drug conjugates, gemtuzumab ozogamicin ( Other humanized monoclonal antibodies with therapeutic potential for use as agents in combination with the compounds of the present invention include: apolizumab, aselizumab, atlizumab, bapineuzumab, bivatuzumab mertansine, cantuzumab mertansine, cedilizumab, certolizumab pegol-conjugated pegol), cidfusituzumab, cidtuzumab, daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab, felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin), ipilimumab, labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab, motovizumab, natalizumab, nimotuzumab, nolovizumab, numavizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, pecfusituzumab, pectuzumab, pexelizumab, ralivizumab, ranibizumab umab), reslivizumab, reslizumab, resyvizumab, rovelizumab, ruplizumab, sibrotuzumab, siplizumab, sontuzumab, tacatuzumabtetraxetan, tadocizumab, talizumab, tefibazumab, tocilizumab, toralizumab, tucotuzumabcelmoleukin), tucusituzumab, umavizumab, urtoxazumab, ustekinumab, visilizumab, and anti-IL-12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories), a recombinant, fully human sequence-specific IgG1λ antibody genetically modified to recognize the IL-12p40 protein.

[0173] Chemotherapeutic agents also include "EGFR inhibitors", which refer to compounds that bind to or directly interact with EGFR and prevent or reduce its signaling activity, and are alternatively referred to as "EGFR antagonists". Examples of such agents include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb 579 (ATCC CRL HB 8506), MAb 455 (ATCC CRL HB8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see, U.S. Pat. No. 4,943,533, Mendelsohn et al.) and variants thereof, such as chimeric 225 (C225 or cetuximab; ) and remodeled human 225 (H225) (see, WO 96 / 40210, Imclone Systems Inc.); IMC-11F8, a fully human antibody targeting EGFR (Imclone); antibodies that bind to type II mutant EGFR (U.S. Pat. No. 5,212,290); humanized and chimeric antibodies that bind to EGFR as described in U.S. Pat. No. 5,891,996; and human antibodies that bind to EGFR, such as ABX-EGF or panitumumab (see WO 98 / 50433, Abgenix / Amgen); EMD55900 (Stragliotto et al. Eur. J. Cancer 32A:636-640 (1996)); EMD7200 (matuzumab), a humanized EGFR antibody against EGFR that competes with EGF and TGF-α for binding to EGFR (EMD / Merck); human EGFR antibody, HuMax-EGFR (GenMab); fully human antibodies, designated E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3 and E7.6.3, and described in US 6,235,883; MDX-447 (Medarex Inc); and mAb 806 or humanized mAb 806 (Johns et al., J. Biol. Chem. 279(29):30375-30384 (2004)). Anti-EGFR antibodies can be conjugated to cytotoxic agents to generate immunoconjugates (see, e.g., EP659,439A2, Merck Patent GmbH). EGFR antagonists include small molecules, such as U.S. Pat. Nos. 5,616,582, 5,457,105, 5,475,001, 5,654,307, 5,679,683, 6,084,095, 6,265,410, 6,455,534, 6,521,620, 6,596,726, 6,713,484, 5,770,599, 6,140,332, 5, 866,572, 6,399,602, 6,344,459, 6,602,863, 6,391,874, 6,344,455, 5,760,041, 6,002,008 and 5,747,498, and the following PCT publications: WO98 / 14451, WO98 / 50038, WO99 / 09016 and WO99 / 24037. Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, Genentech / OSI Pharmaceuticals); PD 183805 (CI 1033, 2-acrylamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-, dihydrochloride, Pfizer); ZD1839, gefitinib 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca); ZM 105180 (6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-(1-methyl-piperidin-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim Ingelheim); PKI-166 ((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide); 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); dual EGFR / HER2 tyrosine kinase inhibitors such as lapatinib ( GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[(2-methylsulfonyl)ethyl]amino]methyl]-2-furyl]-4-quinazolinamine).

[0174] Chemotherapeutic agents also include "tyrosine kinase inhibitors", including the EGFR targeted drugs described in the previous paragraph; small molecule HER2 tyrosine kinase inhibitors, such as TAK165 available from Takeda Pharmaceuticals; 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 to EGFR but simultaneously inhibits cells overexpressing HER2 and EGFR; lapatinib (GSK572016; available from GlaxoSmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor; PKI-166 (available from Novartis); pan-HER inhibitors, such as canertinib (CI-1033; Pharmacia); Raf-1 inhibitors, such as ISIS-5132, an antisense agent that inhibits Raf-1 signaling, available from ISIS Pharmaceuticals; non-HER targeted TK inhibitors, such as imatinib mesylate ( available from GlaxoSmithKline); multitargeted tyrosine kinase inhibitors such as sunitinib ( VEGF receptor tyrosine kinase inhibitors such as vatalanib (PTK787 / ZK222584, available from Novartis / Schering AG); MAPK extracellular regulated kinase I inhibitor CI-1040 (available from Pharmacia); quinazolines such as PD 153035, 4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyrimidopyrimidines; pyrrolopyrimidines such as CGP 59326, CGP60261 and CGP 62706; pyrazolopyrimidines, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine; curcumin (difluoroformylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tyrosine tyrosine containing a nitrothiophene moiety; PD-0183805 (Warner-Lamber); antisense molecules (e.g., molecules that bind to HER encoding nucleic acids); quinoxalines (U.S. Pat. No. 5,804,396); tyrosine phosphorylation inhibitors (U.S. Pat. No. 5,804,396); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering AG); pan-HER inhibitors, such as CI-1033 (Pfizer); Affinitac (ISIS 3521; Isis / Lilly); imatinib mesylate PKI 166 (Novartis); GW2016 (GlaxoSmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); seramatinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering); INC-1C11 (Imclone), rapamycin (sirolimus, ); or any of the following patent publications: U.S. Pat. No. 5,804,396, WO 1999 / 09016 (American Cyanamid), WO 1998 / 43960 (American Cyanamid), WO 1997 / 38983 (Warner Lambert), WO 1999 / 06378 (Warner Lambert), WO 1999 / 06396 (Warner Lambert), WO 1996 / 30347 (Pfizer, Inc), WO 1996 / 33978 (Zeneca), WO 1996 / 3397 (Zeneca), and WO 1996 / 33980 (Zeneca).

[0175] Chemotherapeutic agents also include dexamethasone, interferon, colchicine, metoprine, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostine, arsenic trioxide, asparaginase, live BCG, bevacizumab, bexarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane, epoetin alfa, erlotinib, filgrastim, histrelin acetate, acetate), ibritumomab, interferon alfa-2a, interferon alfa-2b, lenalidomide, levamisole, mesna, methoxsalen, nandrolone, nelarabine, nofetumomab, oprelvekin, palifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium sodium), quinacrine, rasburicase, sargramostim, temozolomide, VM-26, 6-TG, toremifene, tretinoin, ATRA, valrubicin, zoledronate and zoledronic acid and pharmaceutically acceptable salts thereof.

[0176] Chemotherapeutic agents also include hydrocortisone, hydrocortisone acetate, cortisone acetate, thiocortisone pivalate, triamcinolone acetonide, triamcinolone acetonide alcohol, mometasone, amcinonide, budesonide, desonide, fluocinolone acetate, fluocinolone, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortolone, hydrocortisone-17-butyrate, hydrocortisone-17-valerate, aclometasone dipropionate, and phenoxyethanol. dipropionate), betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, clobetasol-17-propionate, fluocortolone hexanoate, fluocortolone valerate, and flupredniidine acetate; immunoselective anti-inflammatory peptides (ImSAIDs), such as phenylalanine-glutamine-glycine (FEG) and its D-isomer form (feG) (IMULAN Biotherapeutics Ltd. BioTherapeutics, LLC); antirheumatic drugs such as azathioprine, cyclosporine (cyclosporine A), D-penicillamine, gold salts, hydroxychloroquine, leflunomide, minocycline, sulfasalazine; tumor necrosis factor alpha (TNFα) blockers such as etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira), certolizumab pegol (Cimzia), golimumab (Simponi); interleukin 1 (IL-1) blockers such as anakinra (Kineret); T-cell costimulation blockers such as abatacept (Orencia); interleukin 6 (IL-6) blockers such as tocilizumab Interleukin 13 (IL-13) blockers, such as lebrikizumab; interferon alpha (IFN) blockers, such as lonavirin; β7 integrin blockers, such as rhuMAb β7; IgE pathway blockers, such as anti-M1 primers; secreted homotrimeric LTa3 and membrane-bound heterotrimeric LTa1 / β2 blockers, such as anti-lymphotoxin alpha (LTa); radioisotopes (e.g. At 211 ,I 131 ,I 125 , Y 90 、Re 186 、Re 188 、Sm 153 、Bi 212 , P 32 , Pb 212and radioisotopes of Lu); various experimental agents such as thioplatin, PS-341, phenylbutyrate, ET-18-OCH3 or farnesyl transferase inhibitors (L-739749, L-744832); polyphenols such as quercetin, resveratrol, picrophenol, epigallocatechin gallate, theaflavins, flavanols, proanthocyanidins, betulinic acid and its derivatives; autophagy inhibitors such as chloroquine; delta-9-tetrahydrocannabinol (dronabinol, ); β-lapachone; lapaol; colchicine; betulinic acid; acetylcamptothecin, scopolectin, and 9-aminocamptothecin); podophyllotoxin; tegafur Bexarotene Bisphosphonates, such as clodronate (e.g. or ), etidronate NE-58095, Zoledronic Acid / Zoledronate Alendronate Pamidronate Tirodronate Risedronate and epidermal growth factor receptor (EGF-R); vaccines such as Vaccines; Perifosine; COX-2 inhibitors (e.g., celecoxib or etoricoxib); proteosome inhibitors (e.g., PS341); CCI-779; Tipifarnib (R11577); Orafenib, ABT510; Bcl-2 inhibitors such as oblimersen sodium Pixantrone; farnesyl transferase inhibitors such as lonafarnib (SCH 6636, SARASAR TM ); and pharmaceutically acceptable salts, acids or derivatives of any of the above; and combinations of two or more of the above, such as CHOP (an abbreviation for the combination therapy of cyclophosphamide, doxorubicin, vincristine and prednisolone); and FOLFOX (oxaliplatin (ELOXATIN TM ) is an abbreviation for the combination therapy with 5-FU and folinate).

[0177] Chemotherapeutic agents also include nonsteroidal anti-inflammatory drugs with analgesic, antipyretic and anti-inflammatory effects.NSAID includes non-selective inhibitors of cyclooxygenase.Specific examples of NSAID include aspirin, propionic acid derivatives (such as ibuprofen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin and naproxen), acetic acid derivatives (such as indomethacin, sulindac, etodolac, diclofenac), enolic acid derivatives (such as piroxicam, meloxicam, tenoxicam, droxicam and lornoxicam), fenamic acid derivatives (such as mefenamic acid, meclofenamic acid, flufenamic acid, toluenesulfonic acid) and COX-2 inhibitors (such as celecoxib, etoricoxib, lumixib, parecoxib, rofecoxib and valdecoxib). NSAIDs may be useful for relieving symptoms of conditions such as rheumatoid arthritis, osteoarthritis, inflammatory arthritis, ankylosing spondylitis, psoriatic arthritis, Reiter's syndrome, acute gout, dysmenorrhea, metastatic bone pain, headaches and migraines, postoperative pain, mild to moderate pain due to inflammation and tissue damage, fever, intestinal obstruction, and renal colic.

[0178] As used herein, the term "cytotoxic agent" refers to a substance that inhibits or prevents cellular function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioactive isotopes (e.g., At 211 ,I 131 ,I 125 , Y 90 、Re 186 、Re 188 、Sm 153 、Bi 212 , P 32 , Pb 212 and radioactive isotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitors; enzymes and fragments thereof such as nucleolytic enzymes; antibiotics; toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof; and various anti-tumor or anti-cancer drugs disclosed below.

[0179] A "disorder" is any condition that would benefit from treatment, including but not limited to chronic and acute disorders or diseases, including those pathological conditions that predispose the mammal to the disorder. In one aspect, the disorder is cancer, such as multiple myeloma (MM).

[0180] The terms "cell proliferative disorder" and "proliferative disorder" refer to a disorder associated with some degree of abnormal cell proliferation. In one aspect, the cell proliferative disorder is cancer. In one aspect, the cell proliferative disorder is a tumor.

[0181] As used herein, the term "tumor" refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all precancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor" are not mutually exclusive herein.

[0182] The terms "cancer" and "cancerous" refer to or describe a physiological condition in mammals that is generally characterized by uncontrolled cell growth / proliferation. Aspects of cancer include solid tumor cancers and non-solid tumor cancers. Examples of cancer include, but are not limited to, B cell proliferative disorders such as multiple myeloma (MM), which may be relapsed or refractory MM. MM may be, for example, a typical MM (e.g., immunoglobulin G (IgG) MM, IgA MM, IgD MM, IgE MM, or IgM MM), a light chain MM (LCMM) (e.g., λ light chain MM or κ light chain MM), or a non-secretory MM. MM may be a newly diagnosed MM (NDMM).

[0183] MM may have one or more cytogenetic features. In some instances, the cytogenetic features are "high-risk cytogenetic features", for example, t(4; 14), t(11; 14), t(14; 16) and / or del(17p), as described in Table 1 and Sonneveld et al., Blood, 127(24): 2955-2962, 2016, International Myeloma Working Group (IMWG) criteria provided, and / or 1q21, as described in Chang et al., Bone Marrow Transplantation, 45: 117-121, 2010, which are incorporated herein by reference in their entirety. In some instances, high-risk cytogenetic features include one or more of the following: (i) translocation events: t(4; 14), t(14; 16) (IMWG criteria); deletion (del) (17p) (IMWG criteria); or gain in chromosome 1q. Cell occurrence features can be detected, for example, using fluorescent in situ hybridization (FISH).

[0184] Table 1. Cytogenetic characteristics of MM

[0185]

[0186] The term "B cell proliferative disorder" or "B cell malignancy" refers to a disorder associated with some degree of abnormal B cell proliferation, and includes, for example, lymphoma, leukemia, myeloma, and myelodysplastic syndrome. In one embodiment, the B cell proliferative disorder is a lymphoma, such as non-Hodgkin's lymphoma (NHL), including, for example, diffuse large B cell lymphoma (DLBCL) (e.g., relapsed or refractory DLBCL). In another embodiment, the B cell proliferative disorder is a leukemia, such as chronic lymphocytic leukemia (CLL). Other specific examples of cancer include germinal center B cell-like (GCB) diffuse large B cell lymphoma (DLBCL), activated B cell-like (ABC) DLBCL, follicular lymphoma (FL), mantle cell lymphoma (MCL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), marginal zone lymphoma (MZL), small lymphocytic leukemia (SLL), lymphoplasmacytic lymphoma (LL), Waldenstrom's macroglobulinemia (WM), central nervous system lymphoma (CNSL), Burkitt's lymphoma (BL), precursor B cell lymphocytic leukemia, splenic marginal zone lymphoma, hairy cell leukemia, splenic lymphoma / leukemia (unclassifiable), splenic diffuse red pulp small B cell lymphoma, variant hairy cell leukemia, heavy chain disease (alpha heavy chain disease, gamma heavy chain disease, mu heavy chain disease), plasma cell myeloma, solitary plasmacytoma of bone, extraosseous plasmacytoma, mucosal leukemia, Extranodal marginal zone lymphoma of lymphoid tissue (MALT lymphoma), nodal marginal zone lymphoma, pediatric nodal marginal zone lymphoma, pediatric follicular lymphoma, primary cutaneous follicle center lymphoma, T-cell / histiocyte-rich large B-cell lymphoma, primary DLBCL of the CNS, primary cutaneous DLBCL (leg type), EBV-positive DLBCL of the elderly, DLBCL associated with chronic inflammation, lymphomatoid granuloma, primary mediastinal (thymic) large B-cell lymphoma, intravascular large B-cell lymphoma, ALK-positive large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma due to HHV8-associated multicentric Castleman disease, primary effusion lymphoma: B-cell lymphoma (unclassifiable, with features intermediate between DLBCL and Burkitt's lymphoma), and B-cell lymphoma (unclassifiable, with features intermediate between DLBCL and classical Hodgkin's lymphoma). Other examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies, including B-cell lymphoma.More specific examples of such cancers include, but are not limited to: low-grade / follicular NHL; small lymphocytic (SL) NHL; intermediate-grade / follicular NHL; intermediate-grade diffuse NHL; high-grade immunoblastic NHL; high-grade lymphoblastic NHL; high-grade small non-truncated cell NHL; bulky disease NHL; AIDS-related lymphoma; and acute lymphoblastic leukemia (ALL); chronic myeloid leukemia; and post-transplant lymphoproliferative disorder (PTLD). Examples of solid tumors include squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer (including small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma), peritoneal cancer, hepatocellular carcinoma, gastric cancer (including gastrointestinal cancer and gastrointestinal stromal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, urinary tract cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, hepatic carcinoma, anal cancer, penile cancer, melanoma, superficial spreading melanoma, tonsillar malignant melanoma, acral lentiginous melanoma, nodular melanoma, and abnormal vascular proliferation associated with nevus hamartomas, edema (such as associated with brain tumors), Meigs syndrome, brain cancer, and head and neck cancer and related metastases. In certain embodiments, cancers suitable for treatment by the antibodies disclosed herein include breast cancer, colorectal cancer, rectal cancer, non-small cell lung cancer, glioblastoma, non-Hodgkin lymphoma (NHL), renal cell carcinoma, prostate cancer, liver cancer, pancreatic cancer, soft tissue sarcoma, Kaposi's sarcoma, carcinoid cancer, head and neck cancer, ovarian cancer, and mesothelioma.

[0187] The term "FcRH5-positive cells" refers to a cancer comprising cells expressing FcRH5 on their surface. For the purpose of determining whether a cell expresses FcRH5 on the surface, FcRH5 mRNA expression is considered to be associated with FcRH5 expression on the cell surface. In some embodiments, the expression of FcRH5 mRNA is determined by a method selected from in situ hybridization and RT-PCR (including quantitative RT-PCR). Alternatively, the expression of FcRH5 on the cell surface can be determined, for example, using antibodies against FcRH5 in methods such as immunohistochemistry, FACS, etc. In some embodiments, FcRH5 is one or more of FcRH5a, FcRH5b, FcRH5c, UniProt identifier Q96RD9-2 and / or FcRH5d. In some embodiments, FcRH5 is FcRH5c. For example, a FcRH5-positive cancer may be a FcRH5-positive MM.

[0188] "Effector functions" refer to those biological activities attributable to the Fc region of an antibody, which vary with the antibody isotype. Examples of antibody effector functions include: C1q binding and complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor); and B cell activation.

[0189] "Complement-dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to antibodies (of the appropriate subclass) bound to its cognate antigen. To assess complement activation, a CDC assay may be performed, e.g., as described in Gazzano-Santoro et al., J. Immunol. Methods 202: 163 (1996).

[0190] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which secreted Ig binds to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., natural killer (NK) cells, neutrophils, and macrophages), allowing these cytotoxic effector cells to specifically bind to target cells bearing antigens and subsequently kill the target cells with cytotoxic agents. Antibodies "arm" cytotoxic cells and are required for such killing. NK cells, the main cells that mediate ADCC, express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression on hematopoietic cells is summarized in Table 3 of Ravetch and Kinet.Annu.Rev.Immunol.9:457-92, 1991, page 464. To assess the ADCC activity of a molecule of interest, an in vitro ADCC assay, such as that described in U.S. Pat. No. 5,500,362 or 5,821,337, may be performed. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively or additionally, the ADCC activity of the target molecule can be assessed in vivo, for example, in an animal model (such as that disclosed in Clynes et al. Proc. Natl. Acad. Sci. USA. 95: 652-656, 1998).

[0191] As used herein, "complex" or "complexed" refers to the association of two or more molecules that interact with each other through bonds and / or forces other than peptide bonds (e.g., van der Waals forces, hydrophobic forces, hydrophilic forces). In one aspect, the complex is a heteromultimer. It should be understood that the term "protein complex" or "polypeptide complex" as used herein includes complexes of non-protein entities (e.g., including but not limited to chemical molecules such as toxins or detection agents) conjugated to proteins in the protein complex.

[0192] As used herein, "delaying progression" of a condition or disease means slowing, hindering, slowing, delaying, stabilizing, and / or postponing the development of a disease or condition (e.g., a cell proliferative disorder, e.g., cancer). Such delays may be of varying lengths of time, depending on the medical history and / or the individual to be treated. It will be apparent to one skilled in the art that a substantial or significant delay may actually encompass prevention, in that the individual will not develop the disease. For example, the development of an advanced cancer, such as metastasis, may be delayed.

[0193] An "effective amount" of a compound, e.g., an anti-FcRH5 / anti-CD3 T cell-dependent bispecific antibody (TDB) disclosed herein, lenalidomide, or a composition thereof (e.g., a pharmaceutical composition) (e.g., a pharmaceutical composition comprising an anti-FcRH5 / anti-CD3 TDB and / or lenalidomide disclosed herein) is at least the minimum amount required to achieve a desired therapeutic or preventive result, such as a measurable improvement or prevention of a particular disorder (e.g., a cell proliferative disorder, e.g., cancer, e.g., MM, e.g., MM with a high-risk cytogenetic profile). The effective amount herein may vary according to factors such as the disease state, age, sex, and weight of the patient, and the ability of the antibody to elicit a desired response in an individual. An effective amount is also an amount in which any toxic or deleterious effects of the treatment are outweighed by the beneficial effects of the treatment. For prophylactic uses, beneficial or desired results include, for example, elimination or reduction of risk, lessening of severity, or delay of disease onset, including biochemical, histological, and / or behavioral symptoms of the disease, its complications, and intermediate pathological phenotypes that occur during the course of disease progression. For therapeutic use, beneficial or expected results include clinical results, such as reducing one or more symptoms caused by the disease, improving the quality of life of the patient, reducing the dose of other drugs required to treat the disease, enhancing the effect of other drugs (such as by targeting, delaying disease progression and / or prolonging survival). In the case of cancer or tumors, an effective amount of the drug may reduce the number of cancer cells; reduce tumor size; inhibit (i.e., slow down or expect to stop to some extent) cancer cell infiltration into surrounding organs; inhibit (i.e., slow down and expect to stop to some extent) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with the condition to some extent. The effective amount can be administered once or multiple times. For the purposes of the present invention, the effective amount of a drug, compound or pharmaceutical composition is an amount sufficient to prevent or treat directly or indirectly. As understood in the clinical context, an effective amount of a drug, compound or pharmaceutical composition may or may not be achieved in combination with another drug, compound or pharmaceutical composition. Therefore, an "effective amount" can be considered in the context of administering one or more therapeutic agents, and if the expected result can be obtained or achieved in combination with one or more other agents, an effective amount of a single agent can be considered.

[0194] As used herein, "overall survival" or "OS" refers to the percentage of individuals in a group of individuals who are likely to survive after a specific period of time.

[0195] As used herein, “objective response rate” (ORR) refers to the sum of stringent complete response (sCR), complete response (CR), very good partial response (VGPR), and partial response (PR) rates determined using the International Myeloma Working Group response criteria (Table 8).

[0196] The term "epitope" refers to a specific site on an antigen molecule to which an antibody binds. In some aspects, the specific site on an antigen molecule to which an antibody binds is determined by hydroxyl radical footprinting. In some aspects, the specific site on an antigen molecule to which an antibody binds is determined by crystallography.

[0197] As used herein, "growth inhibitor" refers to a compound or composition that inhibits cell growth in vitro or in vivo. In one aspect, a growth inhibitor is a growth inhibitory antibody that prevents or reduces the proliferation of cells expressing the antigen to which the antibody binds. On the other hand, a growth inhibitor can be an inhibitor that significantly reduces the percentage of cells in the S phase. Aspects of growth inhibitors include agents that block cell cycle progression (in places other than the S phase), such as agents that induce G1 arrest and M phase arrest. Classical M phase blockers include periwinkle (vincristine and vinblastine), taxanes, and topoisomerase II inhibitors (e.g., doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin). Those agents that block G1 also overflow into S phase arrest, such as DNA alkylating agents, such as tamoxifen, prednisone, dacarbazine, nitrogen mustard, cisplatin, methotrexate, 5-fluorouracil, and ara-C. Further information can be found in Chapter 1 by Murakami et al., entitled "Cell cycle regulation, oncogenes, and antineoplastic drugs" in Mendelsohn and Israel, eds., The Molecular Basis of Cancer (WB Saunders, Philadelphia, 1995), e.g., page 13. Taxanes (paclitaxel and docetaxel) are both anticancer drugs derived from yew. Docetaxel ( Rhone-PoulencRorer) is derived from European yew and is a semi-synthetic analog of paclitaxel ( Bristol-Myers Squibb). Paclitaxel and docetaxel promote microtubule assembly of tubulin dimers and stabilize microtubules by preventing depolymerization, thereby inhibiting cell mitosis.

[0198] An "immunoconjugate" is an antibody conjugated to one or more heterologous molecules, including but not limited to a cytotoxic agent.

[0199] The term "immunomodulator" or "IMiD" refers to a class of molecules that alter the immune system response or immune system function. Immunomodulators include, but are not limited to, PD-1 axis binding antagonists, thalidomide (α-N-phthalimide-glutarimide) and its analogs, (Apremilast), (lenalidomide) and (Pomalidomide) and its pharmaceutically acceptable salts or acids.

[0200] 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). In certain aspects, the subject or individual is a human. In certain aspects, the subject or individual is a patient, e.g., a human patient.

[0201] An "isolated" protein or peptide is one that has been separated from a component of its natural environment. In some aspects, the protein or peptide is purified to a purity greater than 95% or 99%, as determined by, for example, electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reversed phase HPLC).

[0202] An "isolated" nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that normally contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0203] 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 to eliminate T cell dysfunction caused by signaling on the PD-1 signaling axis, resulting in restoration or enhancement of T cell function (e.g., proliferation, cytokine production, and / or target cell killing). As used herein, PD-1 axis binding antagonists include PD-L1 binding antagonists, PD-1 binding antagonists, and PD-L2 binding antagonists. In some cases, PD-1 axis binding antagonists include PD-L1 binding antagonists or PD-1 binding antagonists. In a preferred aspect, the PD-1 axis binding antagonist is a PD-L1 binding antagonist.

[0204] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with the signal transduction generated by the interaction of PD-L1 with one or more of its binding partners (such as PD-1 and / or B7-1). In some cases, the PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partner. In a specific aspect, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1 and / or B7-1. In some cases, the PD-L1 binding antagonist includes anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that reduce, block, inhibit, eliminate or interfere with the signal transduction generated by the interaction of PD-L1 with one or more of its binding partners (such as PD-1 and / or B7-1). In one instance, the PD-L1 binding antagonist reduces the negative co-stimulatory signal mediated by signaling through PD-L1 mediated by or through cell surface proteins expressed on T lymphocytes, thereby rendering the dysfunctional T cells less dysfunctional (e.g., improving the response of effectors to antigen recognition). In some instances, the PD-L1 binding antagonist binds to PD-L1. In some instances, the PD-L1 binding antagonist is an anti-PD-L1 antibody (e.g., an anti-PD-L1 antagonist antibody). Exemplary anti-PD-L1 antagonist antibodies include atezolizumab, MDX-1105, MEDI4736 (durvalumab), MSB0010718C (avelumab), SHR-1316, CS1001, envafolimab, TQB2450, ZKAB001, LP-002, CX-072, IMC-001 , KL-A167, APL-502, cosibelimab, lodapolimab, FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, RC98, PDL-GEX, KD036, KY1003, YBL-007, and HS-636. In some aspects, the anti-PD-L1 antibody is atezolizumab, MDX-1105, MEDI4736 (durvalumab), or MSB0010718C (avetumab). In one specific aspect, the PD-L1 binding antagonist is MDX-1105. In another specific aspect, the PD-L1 binding antagonist is MEDI4736 (durvalumab). In another specific aspect, the PD-L1 binding antagonist is MSB0010718C (avelumab).In other aspects, the PD-L1 binding antagonist can be a small molecule, for example, GS-4224, INCB086550, MAX-10181, INCB090244, CA-170 or ABSK041, which can be administered orally in some cases. Other exemplary PD-L1 binding antagonists include AVA-004, MT-6035, VXM10, LYN192, GB7003 and JS-003. In a specific aspect, the PD-L1 binding antagonist is atezolizumab.

[0205] The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with the signal transduction generated by the interaction of PD-1 with one or more of its binding partners (such as PD-L1 and / or PD-L2). PD-1 (programmed death 1) is also referred to as "programmed cell death 1", "PDCD1", "CD279" and "SLEB2" in the art. Exemplary human PD-1 is shown in UniProtKB / Swiss-Prot accession number Q15116. In some cases, a PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to one or more of its binding partners. In a specific aspect, 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 and antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that reduce, block, inhibit, eliminate or interfere with the signal transduction generated by the interaction of PD-1 with PD-L1 and / or PD-L2. In one instance, a PD-1 binding antagonist reduces negative co-stimulatory signals mediated by signaling through PD-1 mediated by or through cell surface proteins expressed on T lymphocytes, thereby rendering dysfunctional T cells less dysfunctional (e.g., improving effector responses to antigen recognition). In some instances, a PD-1 binding antagonist binds to PD-1. In some instances, a PD-1 binding antagonist is an anti-PD-1 antibody (e.g., an anti-PD-1 antagonist antibody). Exemplary anti-PD-1 antagonist antibodies include nivolumab, pembrolizumab, MEDI-0680, PDR001 (spartalizumab), REGN2810 (cemiprilimab), BGB-108, palolizumab, carrelizumab, sintilimab, tislelizumab, toripalimab, dotalimumab, rivulimab, sazanlimab, penampalimab, CS1003, HLX10, SCT-I10A, sepalizumab, batilizumab, genoluzumab, BI 754091, silimumab, YBL-006, BAT1306, HX008, brigalimumab, AMG 404, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, AM0001, ENUM 244C8, ENUM 388D4, STI-1110, AK-103 and hAb21. In one specific aspect, the PD-1 binding antagonist is MDX-1106 (nivolumab). In another specific aspect, the PD-1 binding antagonist is MK-3475 (pembrolizumab). In another specific aspect, the PD-1 binding antagonist is a PD-L2 fusion protein, for example, AMP-224. In another specific aspect, the PD-1 binding antagonist is MED1-0680.In another specific aspect, the PD-1 binding antagonist is PDR001 (spartalizumab). In another specific aspect, the PD-1 binding antagonist is REGN2810 (simiprilimab). In another specific aspect, the PD-1 binding antagonist is BGB-108. In another specific aspect, the PD-1 binding antagonist is palolizumab. In another specific aspect, the PD-1 binding antagonist is carrelizumab. In another specific aspect, the PD-1 binding antagonist is sindilimab. In another specific aspect, the PD-1 binding antagonist is tislelizumab. In another specific aspect, the PD-1 binding antagonist is toripalizumab. Other additional exemplary PD-1 binding antagonists include BION-004, CB201, AUNP-012, ADG104, and LBL-006.

[0206] The term "PD-L2 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with the signal transduction generated by the interaction of PD-L2 with one or more of its binding partners (such as PD-1). PD-L2 (programmed death ligand 2) is also referred to in the art as "programmed cell death 1 ligand 2", "PDCD1LG2", "CD273", "B7-DC", "Btdc" and "PDL2". An exemplary human PD-L2 is shown in UniProtKB / Swiss-Prot accession number Q9BQ51. In some cases, a PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to one or more of its binding partners. In a specific aspect, a PD-L2 binding antagonist inhibits the binding of PD-L2 to PD-1. Exemplary PD-L2 antagonists include anti-PD-L2 antibodies, antigen binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, eliminate, or interfere with the signal transduction generated by the interaction of one or more of PD-L2 and its binding partners (such as PD-1). On the one hand, the PD-L2 binding antagonist reduces the negative co-stimulatory signal mediated by or expressed by the cell surface protein expressed on T lymphocytes, and the surface protein makes the dysfunctional T cell dysfunction less (e.g., improve the response of the effector to antigen recognition) through the signal transduction mediated by PD-L2. In some aspects, the PD-L2 binding antagonist binds to PD-L2. In some aspects, the PD-L2 binding antagonist is an immunoadhesin. In other aspects, the PD-L2 binding antagonist is an anti-PD-L2 antagonist antibody.

[0207] Unless otherwise indicated, the term "protein" as used herein refers to any native protein from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term includes "full-length" unprocessed proteins, as well as any form of proteins produced by processing in cells. The term also encompasses naturally occurring protein variants, such as splice variants or allelic variants.

[0208] "Amino acid sequence identity percentage (%)" relative to a reference polypeptide sequence is defined as the percentage of amino acid residues in the candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence after aligning the candidate sequence with the reference polypeptide sequence and introducing spaces (if necessary) to achieve maximum sequence identity percentage, and for the purpose of alignment without considering any conservative substitutions as components of sequence identity. Alignment for determining amino acid sequence identity percentage can be achieved in various ways within the technical scope of the art, for example, using publicly available computer software, such as BLAST, BLAST-2, Clustal W, Megalign (DNASTAR) software or FASTA program package. Those skilled in the art can determine the appropriate parameters for aligning sequences, including any algorithm required for achieving maximum alignment over the full length of the compared sequence. Alternatively, the sequence comparison computer program ALIGN-2 can be used to generate identity percentage values. The ALIGN-2 sequence comparison computer program was written by Genentech, Inc. and the source code has been filed with user documentation in the US Copyright Office, Washington DC, 20559, where it is registered under US Copyright Registration No. TXU510087 and is described in WO 2001 / 007611.

[0209] Unless otherwise indicated, for the purposes of this article, the values ​​of percent amino acid sequence identity are generated using the BLOSUM50 comparison matrix using the ggsearch program of the FASTA package version 36.3.8c or higher. The FASTA package is by W.R. Pearson and D.J. Lipman (1988), "Improved Tools for Biological Sequence Analysis", PNAS 85:2444-2448; W.R. Pearson (1996) "Effective protein sequence comparison" Meth. Enzymol. 266:227-258; and Pearson et al. (1997) Genomics 46:24-36 creation and can be obtained from www.fasta.bioch.virginia.edu / fasta_www2 / fasta_down.shtml or www.ebi.ac.uk / Tools / sss / fasta. Alternatively, sequences can be compared using a public server accessible at fasta.bioch.virginia.edu / fasta_www2 / index.cgi, using the ggsearch (global protein:protein) program and default options (BLOSUM50; open: -10; ext: -2; Ktup = 2) to ensure that a global rather than a local alignment is performed. The amino acid identity percentage is given in the output alignment header.

[0210] The term "pharmaceutical formulation" refers to a preparation that is in a form that permits the biological activity of the active ingredient contained therein to be effective, and that contains no additional components that are unacceptably toxic to a subject to which the preparation would be administered.

[0211] "Pharmaceutically acceptable carrier" refers to a component of a pharmaceutical preparation other than the active ingredient that is non-toxic to a subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0212] "Radiotherapy" refers to the use of directed gamma or beta rays to induce sufficient damage to cells to limit the ability of the cells to function normally or to completely destroy the cells. It will be understood that there are many known methods in the art to determine the dosage and duration of treatment. Typical treatment is a single administration, with a typical dosage range of 10 to 200 units (Gray) per day.

[0213] As used herein, "treatment" (and grammatical variations thereof, such as "treat" or "treating") refers to clinical intervention in an attempt to alter the natural course of the individual being treated, and may be performed for prevention or during the course of clinical pathology. Desired effects of treatment include, but are not limited to, preventing the occurrence or recurrence of disease, alleviating symptoms, diminishing any direct or indirect pathological consequences of disease, preventing metastasis, reducing the rate of disease progression, ameliorating or palliating the disease state, and alleviating or improving prognosis. In some aspects, the antibodies disclosed herein (e.g., the anti-FcRH5 / anti-CD3 TDB disclosed herein) and / or lenalidomide are used to delay the development of disease or slow the progression of disease.

[0214] "Reduce" or "inhibit" means the ability to cause an overall decrease, for example, an overall decrease of 20% or more, 50% or more, or 75%, 85%, 90%, 95% or more. In certain aspects, reduction or inhibition can refer to the effector function of an antibody mediated by the Fc region of the antibody, which effector function specifically includes complement dependent cytotoxicity (CDC), antibody dependent cellular toxicity (ADCC) and antibody dependent cellular phagocytosis (ADCP).

[0215] According to the present invention, the term "vaccine" refers to a pharmaceutical preparation (pharmaceutical composition) or product that induces an immune response after administration, particularly a cellular immune response that recognizes and attacks pathogens or diseased cells, such as cancer cells. Vaccines can be used to prevent or treat diseases. The vaccine can be a cancer vaccine. As used herein, a "cancer vaccine" is a composition that stimulates an immune response in a subject against cancer. Cancer vaccines are typically composed of cancer-related materials or cell (antigen) sources that may be autologous (from oneself) or allogeneic (from others) for the subject, together with other ingredients (e.g., adjuvants) to further stimulate and promote an immune response to the antigen. Cancer vaccines can result in stimulation of the subject's immune system to produce antibodies against one or more specific antigens, and / or produce killer T cells to attack cancer cells with those antigens.

[0216] As used herein, "administering" refers to a method of giving a dose of a compound (e.g., an anti-FcRH5 / anti-CD3 TDB of the invention (e.g., silvostomumab) or lenalidomide) to a subject. In some aspects, the composition used in the methods herein is administered intravenously. The composition used in the methods described herein can be administered, for example, intramuscularly, intravenously, intradermally, percutaneously, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctivally, intracapsularly, mucosally, intrapericardially, intraumbilically, intraocularly, orally, topically, topically, by inhalation, by injection, by infusion, by continuous infusion, by local perfusion directly into target cells, by catheter, by lavage, as an emulsion or in the form of a lipid composition. The method of administration can vary depending on a variety of factors (eg, the compound or composition being administered and the severity of the condition, disease or disorder being treated).

[0217] As used herein, "CD38" refers to the CD38 glycoprotein found on the surface of many immune cells (including CD4+, CD8+, B lymphocytes and natural killer (NK) cells), including any native CD38 from any vertebrate source, including mammals, such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise indicated. Compared with normal lymphoid and myeloid cells, the expression level of CD38 on myeloma cells is higher and more uniform. The term includes "full-length" unprocessed CD38, as well as any form of CD38 produced by processing in cells. The term also encompasses naturally occurring variants of CD38, such as splice variants or allelic variants. CD38 is also referred to in the art as cluster of differentiation 38, ADP-ribosyl cyclase 1, cADPr hydrolase 1, and cyclic ADP-ribose hydrolase 1. CD38 is encoded by the CD38 gene. The nucleic acid sequence of an exemplary human CD38 is set forth in the following NCBI Reference Sequence: NM_001775.4 or in SEQ ID NO: 33. The amino acid sequence of an exemplary human CD38 protein encoded by CD38 is set forth in UniProt Accession No. P28907 or in SEQ ID NO:34.

[0218] The term "anti-CD38 antibody" encompasses all antibodies that bind to CD38 with sufficient affinity so that the antibody can be used as a therapeutic agent targeting cells expressing the antigen and does not significantly cross-react with other proteins (such as negative control proteins) in the assays described below. For example, anti-CD38 antibodies can bind to CD38 on the surface of MM cells and mediate cell lysis by activating complement-dependent cytotoxicity, ADCC, antibody-dependent cellular phagocytosis (ADCP), and Fc cross-linking-mediated apoptosis, leading to the consumption of malignant cells and a reduction in the overall cancer burden. Anti-CD38 antibodies can also modulate CD38 enzymatic activity by inhibiting ribosyl cyclase activity and stimulating the cyclic adenosine diphosphate ribose (cADPR) hydrolase activity of CD38. In certain aspects, the dissociation constant (K) of an anti-CD38 antibody bound to CD38 is D ) is ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM or ≤0.001 nM (e.g., 10 -8 M or smaller, e.g. 10 -8 M to 10 -13 M, for example 10 -9 M to 10 - 13 M). In certain aspects, the anti-CD38 antibody can bind to both human CD38 and chimpanzee CD38. Anti-CD38 antibodies also include anti-CD38 antagonist antibodies. Bispecific antibodies in which one arm of the antibody binds CD38 are also contemplated. This definition of anti-CD38 antibodies also includes functional fragments of the foregoing antibodies. Examples of antibodies that bind to CD38 include: Daratumumab (U.S. Patent No.: 7,829,673 and U.S. Publication No.: 20160067205A1); "MOR202" (U.S. Patent No.: 8,263,746); and isatuximab (SAR-650984).

[0219] II. Methods of Treatment and Compositions for Use

[0220] The present invention is based, in part, on methods of treating subjects having cancer, e.g., multiple myeloma (MM) (e.g., MM with high-risk cytogenetic characteristics), with an anti-crystallizable fragment receptor-like 5 (FcRH5) / anti-cluster of differentiation 3 (CD3) bispecific antibody and lenalidomide, e.g., using a fractionated, dose-escalating dosing regimen disclosed herein.

[0221] At present, there is no curative treatment for MM, and almost all patients will eventually relapse.Lenalidomide is currently the only approved maintenance therapy for autologous stem cell transplantation (ASCT) to postpone relapse and prolong survival.So far, maintenance therapy is usually performed as a monotherapy, and patients with low-risk features of cytogenetics can obtain survival benefits.However, patients with high-risk features of cytogenetics still maintain a high degree of unmet medical needs, wherein the survival benefits maintained by a single agent are very poor, and compared to patients in low-risk categories, the death hazard ratio is between 6 and 15 times higher.In high-risk groups, it is expected that dual-agent maintenance with silvostomumab and lenalidomide as described herein will improve and deepen response, thereby increasing survival while maintaining quality of life.

[0222] A. Dosage regimen

[0223] i. Dosing regimens for treating cancers with high-risk cytogenetic characteristics

[0224] The present disclosure provides methods and compositions for treating cancers having high-risk cytogenetic characteristics, eg, hematological cancers, eg, B-cell proliferative disorders (eg, MM).

[0225] For example, provided herein is a method of treating a subject having a cancer having a high-risk cytogenetic characteristic, e.g., a hematological cancer, e.g., a B-cell proliferative disorder (e.g., MM), comprising administering to the subject (i) a bispecific antibody that binds to fragment crystallizable receptor-like 5 (FcRH5) and cluster of differentiation 3 (CD3) and (ii) lenalidomide.

[0226] In another example, provided herein is a bispecific antibody that binds to FcRH5 and CD3 for use in treating a subject having a cancer with a high-risk cytogenetic profile, e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM)), the treatment comprising administering to the subject the bispecific antibody and lenalidomide.

[0227] In some examples, the subject experiences a partial remission (PR) or better following induction therapy.

[0228] In some examples, the subject receives an autologous stem cell transplant (ASCT) and / or does not have progressive disease within 100 days (e.g., within 100 days, within 95 days, within 90 days, within 85 days, within 80 days, within 75 days, within 70 days, within 65 days, within 60 days, within 55 days, within 50 days, within 45 days, within 40 days, within 35 days, within 30 days, within 25 days, within 20 days, within 15 days, within 10 days, within 5 days, within 4 days, within 3 days, within 2 days, or within 1 day) of the start of the method or treatment (e.g., first administration of the bispecific antibody).

[0229] In some examples, the bispecific antibody and lenalidomide are administered to the patient as post-transplant maintenance therapy.

[0230] The patient can have any suitable high-risk cytogenetic features or combinations thereof. In some examples, the high-risk cytogenetic features include one or more of the following: translocation events t(4;14) or t(14;16), del(17p), or 1q gain.

[0231] In some examples, the subject has a high-risk cytogenetic profile at the time of diagnosis of cancer (eg, a hematological cancer (eg, MM)).

[0232] In some examples, the bispecific antibody and lenalidomide are administered to the subject at a dosing regimen comprising: (i) a first phase comprising one or more dosing cycles, wherein the first phase comprises administering the bispecific antibody to the subject weekly (Q1W), every two weeks (Q2W), every three weeks (Q3W), or every four weeks (Q4W); and (ii) a second phase comprising one or more dosing cycles, wherein the second phase comprises administering the bispecific antibody to the subject weekly (Q1W), every two weeks (Q2W), every three weeks (Q3W), or every four weeks (Q4W).

[0233] In a specific example, the bispecific antibody and lenalidomide are administered to the subject at a dosing regimen comprising: (i) a first phase comprising one or more dosing cycles, wherein the first phase comprises administering the bispecific antibody to the subject every two weeks (Q2W); and (ii) a second phase comprising one or more dosing cycles, wherein the second phase comprises administering the bispecific antibody to the subject every four weeks (Q4W).

[0234] Each dosing cycle of the first stage and / or the second stage can have any suitable length. In some instances, each dosing cycle of the first stage and the second stage is a 28-day dosing cycle. However, in other instances, each dosing cycle of the first stage and / or the second stage can be a 7-day dosing cycle, a 14-day dosing cycle, or a 21-day dosing cycle. It should be understood that dosing cycles do not necessarily all have the same length.

[0235] In some examples, the method or treatment further comprises a preliminary phase comprising one or more dosing cycles prior to the first phase, wherein the preliminary phase comprises administering the bispecific antibody to the subject weekly (QW), every two weeks (Q2W), every three weeks (Q3W), or every four weeks (Q4W).

[0236] In one embodiment, the method or treatment further comprises a preliminary phase comprising one or more dosing cycles prior to the first phase, wherein the preliminary phase comprises administering the bispecific antibody to the subject weekly (QW).

[0237] Each dosing cycle in the preliminary phase may have any suitable length. In some instances, each dosing cycle in the preliminary phase is a 28-day dosing cycle. However, in other instances, each dosing cycle in the preliminary phase may be a 7-day dosing cycle, a 14-day dosing cycle, or a 21-day dosing cycle. It should be understood that the dosing cycles do not necessarily all have the same length.

[0238] The preliminary phase can include any suitable number of dosing cycles, for example, one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen or more dosing cycles.

[0239] In certain examples, the preliminary phase includes one administration cycle (C1).

[0240] The preliminary phase can comprise administering the bispecific antibody to the subject on any suitable day of a dosing cycle (e.g., C1) (e.g., day 1, day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11, day 12, day 13, day 14, day 15, day 16, day 17, day 18, day 19, day 20, day 21, day 22, day 23, day 24, day 25, day 26, day 27 and / or day 28 of the dosing cycle).

[0241] In one specific example, the pre-phase comprises administering the bispecific antibody to the subject on day 1, day 8, and day 15 of C1. In another example, the pre-phase comprises administering the bispecific antibody to the subject on day 1, day 2, and day 15 of C1.

[0242] In some examples, the target dose of the bispecific antibody is administered to the subject for each administration in the preliminary phase. In other words, the preliminary phase may not utilize step-up dosing.

[0243] In other examples, the preliminary phase includes administering to the subject a first step-up dose of the bispecific antibody. In some examples, the preliminary phase includes a single step-up dose of the bispecific antibody. Any of the single step-up dosing regimens described in Subsection II below can be used.

[0244] The first step-escalating dose can be administered to the subject on any suitable day of a dosing cycle (e.g., C1) (e.g., day 1, day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11, day 12, day 13, day 14, day 15, day 16, day 17, day 18, day 19, day 20, day 21, day 22, day 23, day 24, day 25, day 26, day 27 and / or day 28 of the dosing cycle).

[0245] In a specific example, the subject is administered a first step-escalating dose on Day 1 of C1.

[0246] In a single step-up dosing regimen, a target dose may be administered on any suitable day after the first step-up dose (e.g., day 1, day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11, day 12, day 13, day 14, day 15, day 16, day 17, day 18, day 19, day 20, day 21, day 22, day 23, day 24, day 25, day 26, day 27, and / or day 28 of a dosing cycle (e.g., C1). In a specific example, a target dose is administered to a subject on days 8 and 15 of C1.

[0247] In some examples, the preliminary phase includes administering to the subject a first step-up dose and a second step-up dose of the bispecific antibody. Any of the dual step-up dosing regimens described in Subsection III below can be used.

[0248] The first step-escalating dose and / or the second step-escalating dose can be administered to the subject on any suitable day of the dosing cycle (e.g., C1) (e.g., day 1, day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11, day 12, day 13, day 14, day 15, day 16, day 17, day 18, day 19, day 20, day 21, day 22, day 23, day 24, day 25, day 26, day 27 and / or day 28 of the dosing cycle).

[0249] In a specific example, the subject is administered a first step-escalating dose on day 1 of C1 and the subject is administered a second step-escalating dose on day 8 of C1.

[0250] In a dual step-up dosing regimen, the target dose may be administered on any suitable day after the second step-up dose (e.g., on day 1, day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11, day 12, day 13, day 14, day 15, day 16, day 17, day 18, day 19, day 20, day 21, day 22, day 23, day 24, day 25, day 26, day 27, and / or day 28 of the dosing cycle). In a specific example, the target dose is administered to the subject on day 15 of C1.

[0251] Any suitable dose can be used for the first step-up dose, including any of the doses described in Subsections II and III below. In some examples, the first step-up dose is 3.6 mg.

[0252] Any suitable dose can be used for the first step-up dose, including any of the doses described in Subsection III below. In some examples, the first step-up dose is 0.3 mg and the second step-up dose is 3.6 mg. In other examples, the first step-up dose is 0.3 mg and the second step-up dose is 3.3 mg.

[0253] The first phase can include any suitable number of dosing cycles (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more dosing cycles). In some instances, the first phase includes at least two dosing cycles, at least three dosing cycles, at least four dosing cycles, or at least five dosing cycles. In some instances, the first phase consists of five dosing cycles.

[0254] In some examples, the first phase includes a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), and a fifth dosing cycle (C5).

[0255] In some examples, the first phase comprises administering the bispecific antibody to the subject on Day 1 and Day 15 of C1, C2, C3, C4, and / or C5.

[0256] In some examples, a target dose of the bispecific antibody is administered to the subject for each administration during the first phase.Any suitable target dose can be used, including any dose described in Subsections II and / or III below. In some instances, the target dose is between 20 mg and 600 mg (e.g., between 30 mg and 500 mg, 40 mg and 400 mg, 60 mg and 350 mg, 80 mg and 300 mg, 100 mg and 200 mg, or 140 mg and 180 mg, for example, 20 mg, 40 mg, 60 mg, 80 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg, 280 mg, 300 mg, 320 mg, 340 mg, 360 mg, 380 mg, 400 mg, 420 mg, 440 mg, 460 mg, 480 mg, 500 mg, 520 mg, 540 mg, 560 mg, 580 mg, or 600 mg). In some aspects, C1D3 is between 80 mg and 300 mg. In some aspects, the target dose is about 90 mg. In some aspects, the target dose is about 132 mg. In some target doses, C1D3 is about 160 mg. In some target doses, C1D3 is about 198 mg.

[0257] The second phase can include any suitable number of dosing cycles (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more dosing cycles). In some instances, the second phase includes at least two dosing cycles, at least three dosing cycles, at least four dosing cycles, at least five dosing cycles, at least six dosing cycles, or at least seven dosing cycles. In some instances, the second phase consists of seven dosing cycles.

[0258] In some examples, the second phase includes a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), a fifth dosing cycle (C5), a sixth dosing cycle (C6), and a seventh dosing cycle (C7).

[0259] In some examples, the second phase comprises administering the bispecific antibody to the subject on day 1 of C1, C2, C3, C4, C5, C6, and / or C7.

[0260] In some examples, a target dose of the bispecific antibody is administered to the subject for each administration during the second phase.Any suitable target dose can be used, including any dose described in Subsections II and / or III below. In some instances, the target dose is between 20 mg and 600 mg (e.g., between 30 mg and 500 mg, 40 mg and 400 mg, 60 mg and 350 mg, 80 mg and 300 mg, 100 mg and 200 mg, or 140 mg and 180 mg, for example, 20 mg, 40 mg, 60 mg, 80 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg, 280 mg, 300 mg, 320 mg, 340 mg, 360 mg, 380 mg, 400 mg, 420 mg, 440 mg, 460 mg, 480 mg, 500 mg, 520 mg, 540 mg, 560 mg, 580 mg, or 600 mg). In some aspects, C1D3 is between 80 mg and 300 mg. In some aspects, the target dose is about 90 mg. In some aspects, the target dose is about 132 mg. In some target doses, C1D3 is about 160 mg. In some target doses, C1D3 is about 198 mg.

[0261] In some instances, the target dose is between 90 mg and 198 mg, inclusive. In some instances, the target dose is 90 mg. In some instances, the target dose is 132 mg. In some instances, the target dose is 160 mg.

[0262] The bispecific antibody can be administered by any suitable route of administration. In some instances, the bispecific antibody is administered intravenously to the subject. In other instances, the bispecific antibody is administered subcutaneously to the subject.

[0263] Lenalidomide may be administered on any suitable day of the dosing cycle (e.g., on day 1, day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11, day 12, day 13, day 14, day 15, day 16, day 17, day 18, day 19, day 20, day 21, day 22, day 23, day 24, day 25, day 26, day 27, and / or day 28 of the dosing cycle). In a specific example, lenalidomide is administered to the subject on day 1 to day 21 of each dosing cycle in the first phase and / or the second phase. In some examples, lenalidomide is administered to the subject on day 1 to day 21 of each dosing cycle in the preliminary phase.

[0264] Any suitable dose of lenalidomide (e.g., about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, or about 30 mg) can be used. In some instances, lenalidomide is administered to a subject at a dose of about 10 mg to about 20 mg. In some instances, lenalidomide is administered to a subject at a dose of 10 mg to 20 mg.

[0265] In some instances, lenalidomide is administered to a subject at a dose of about 10 mg. In some instances, lenalidomide is administered to a subject at a dose of 10 mg. In other instances, lenalidomide is administered to a subject at a dose of about 15 mg. In other instances, lenalidomide is administered to a subject at a dose of 15 mg. For example, lenalidomide may be administered at a dose of 15 mg after three cycles (e.g., the first three cycles may include administration of lenalidomide at a dose of 10 mg, and then the dose may be increased to 15 mg, e.g., at the discretion of a clinician).

[0266] Lenalidomide can be administered by any suitable route of administration. In some examples, lenalidomide is administered orally to the subject.

[0267] In some examples, the method or treatment further comprises administering a corticosteroid to the subject. Any suitable corticosteroid can be used, such as dexamethasone or methylprednisolone.

[0268] In some examples, the method or treatment further comprises administering a corticosteroid to the subject during the first phase and / or the second phase.

[0269] Corticosteroids may be administered on any suitable date during the dosing cycle in the first and / or second phases (e.g., on day 1, day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11, day 12, day 13, day 14, day 15, day 16, day 17, day 18, day 19, day 20, day 21, day 22, day 23, day 24, day 25, day 26, day 27, and / or day 28 of the dosing cycle). Corticosteroids may be administered on the same date as the bispecific antibody or on a different date from the bispecific antibody (e.g., one or more days before or after the administration of the bispecific antibody). In some instances, corticosteroids are administered to subjects on day 1 and day 15 of C1 in the first phase.

[0270] In some instances, if the subject experiences cytokine release syndrome (CRS) with a previous dose, a corticosteroid is administered to the subject. In some instances, if the subject experiences a CRS event with a previous dose, a corticosteroid is administered to the subject in C2, C3, C4, and / or C5 of the first phase.

[0271] In some examples, if the subject experienced a CRS event at a previous dose, a corticosteroid is administered to the subject in the second phase C1, C2, C3, C4, C5, C6, and / or C7.

[0272] In some examples, the method or treatment further comprises administering a corticosteroid to the subject during the pre-period.

[0273] The corticosteroid may be administered on any suitable day during the dosing cycle in the pre-phase (e.g., on day 1, day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11, day 12, day 13, day 14, day 15, day 16, day 17, day 18, day 19, day 20, day 21, day 22, day 23, day 24, day 25, day 26, day 27, and / or day 28 of the dosing cycle). In some examples, the corticosteroid is administered to the subject on day 1, day 8, and day 15 of C1 during the pre-phase.

[0274] The corticosteroid can be administered by any suitable route of administration. In some instances, the corticosteroid is administered intravenously or orally to the subject. In some instances, the corticosteroid is administered intravenously to the subject.

[0275] In some examples, a corticosteroid is administered intravenously to the subject prior to administration of the bispecific antibody.

[0276] The corticosteroid can be administered at any suitable amount of time prior to administration of the bispecific antibody (e.g., about 1 min, 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 35 min, 40 min, 45 min, 50 min, 55 min, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, or 24 hours prior to administration of the bispecific antibody). In some examples, the corticosteroid is administered intravenously to the subject about 1 hour prior to administration of the bispecific antibody.

[0277] In some examples, the corticosteroid is dexamethasone or methylprednisolone.

[0278] In some examples, the corticosteroid is dexamethasone.

[0279] Dexamethasone can be administered at any suitable dose (e.g., 1 mg to 100 mg). In some examples, dexamethasone is administered to a subject at a dose of about 20 mg.

[0280] Methylprednisolone can be administered at any suitable dose, such as 1 mg to 400 mg. In some examples, methylprednisolone is administered to a subject at a dose of about 80 mg.

[0281] Any suitable bispecific antibody can be used, such as any bispecific antibody disclosed herein (eg, in Section H below).

[0282] In some examples, the bispecific antibody is sivostatin.

[0283] In some examples, the bispecific antibody and lenalidomide are administered to the subject simultaneously with one or more additional therapeutic agents.Any suitable additional therapeutic agent can be used, including any therapeutic agent disclosed herein.

[0284] In some examples, the bispecific antibody and / or lenalidomide is administered to the subject prior to administration of one or more additional therapeutic agents.

[0285] In some examples, the bispecific antibody and / or lenalidomide is administered to the subject after administration of one or more additional therapeutic agents.

[0286] In some examples, the one or more additional therapeutic agents include an effective amount of tocilizumab.

[0287] In some examples, the subject experiences a CRS event, and the method further comprises treating symptoms of the CRS event while suspending treatment with the bispecific antibody.

[0288] In some examples, the method or treatment further comprises administering to the subject an effective amount of tocilizumab to treat the CRS event.

[0289] In some examples, the CRS event does not resolve, or worsens, within 24 hours of treating symptoms of the CRS event, and the method further comprises administering one or more additional doses of tocilizumab to the subject to manage the CRS event.

[0290] In some examples, tocilizumab is administered to the subject by intravenous infusion.

[0291] In some instances: (i) the subject weighs ≥30 kg and tocilizumab is administered to the subject at a dose of 8 mg / kg; or (b) the subject weighs <30 kg and tocilizumab is administered to the subject at a dose of 12 mg / kg.

[0292] In some examples, tocilizumab is administered to the subject 2 hours prior to administration of the bispecific antibody.

[0293] In some examples, the one or more additional therapeutic agents include an effective amount of a B cell maturation antigen (BCMA) directed therapy, an additional immunomodulatory agent (IMiD), a CD38 directed therapy, or a combination of any of the foregoing.

[0294] In some examples, the one or more additional therapeutic agents include an effective amount of acetaminophen or paracetamol.

[0295] Any suitable dose of acetaminophen or paracetamol may be used. In some examples, acetaminophen or paracetamol is administered to a subject at a dose of between about 500 mg to about 1000 mg.

[0296] Acetaminophen or paracetamol can be administered by any suitable route of administration, including any route of administration disclosed herein. In some examples, acetaminophen or paracetamol is administered orally to the subject.

[0297] In some examples, the one or more additional therapeutic agents includes an effective amount of diphenhydramine.

[0298] Any suitable dose of diphenhydramine can be used. In some examples, diphenhydramine is administered to a subject at a dose between about 25 mg to about 50 mg.

[0299] Diphenhydramine can be administered by any suitable route of administration, including any route of administration disclosed herein. In some examples, diphenhydramine is administered orally to the subject.

[0300] In another example, provided herein is a method of treating a subject having a cancer with a high-risk cytogenetic profile (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM))), the method comprising administering to the subject silvostomumab and lenalidomide, wherein: (i) the subject experiences a PR or better after induction therapy; (ii) the subject receives ASCT within 100 days of initiation of the method and / or does not have progressive disease; (iii) silvostomumab and lenalidomide are administered to the patient as post-transplant maintenance therapy; and (iv) the high-risk cytogenetic profile comprises one or more of the following: translocation events t(4;14) or t(14;16), del(17p), or 1q gain.

[0301] In another example, provided herein is silvositumomab for use in treating a subject having a cancer with a high-risk cytogenetic profile (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM))), the treatment comprising administering silvositumomab and lenalidomide to the subject, wherein: (i) the subject experiences a PR or better after induction therapy; (ii) the subject receives ASCT within 100 days of initiation of the method and / or does not have progressive disease; (iii) silvositumomab and lenalidomide are administered to the patient as post-transplant maintenance therapy; and (iv) the high-risk cytogenetic profile comprises one or more of the following: translocation events t(4;14) or t(14;16), del(17p), or 1q gain.

[0302] In another example, provided herein is a method for treating a subject having a cancer with a high-risk cytogenetic characteristic (e.g., a blood cancer (B-cell proliferative disorder (e.g., MM))), the method comprising administering to the subject silvostomumab and lenalidomide at a dosing regimen comprising: (i) a preliminary phase comprising a 28-day dosing cycle (C1); (ii) a first phase after the preliminary phase comprising a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), and a fifth dosing cycle (C5), wherein each dosing cycle in the first phase is a 28-day dosing cycle; and (iii) a second phase after the first phase comprising a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), a fifth dosing cycle (C5), a sixth dosing cycle (C6), and a seventh dosing cycle (C7), wherein the Each dosing cycle of the second phase is a 28-day dosing cycle, in which silvostomumab is administered to the subject as follows: (i) at a first step-escalating dose on day 1 of C1 during the pre-phase, and at a second step-escalating dose on day 8 of C1 during the pre-phase; (ii) at a target dose on day 15 of C1 during the pre-phase; (iii) at a target dose on days 1 and 15 of C1, C2, C3, C4, and C5 during the first phase; and (i v) at the target dose on Day 1 of C1, C2, C3, C4, C5, C6, and C7 during the second phase; and wherein lenalidomide is administered to the subject as follows: (i) on Day 1 to Day 21 of C1 during the pre-phase; (ii) on Day 1 to Day 21 of C1, C2, C3, C4, and C5 during the first phase; and (iii) on Day 1 to Day 21 of C1, C2, C3, C4, C5, C6, and C7 during the second phase.

[0303] In another example, provided herein is silvostomumab for use in treating a subject having a cancer with a high-risk cytogenetic characteristic (e.g., a blood cancer (e.g., a B-cell proliferative disorder (e.g., MM))), the treatment comprising administering silvostomumab and lenalidomide to the subject at a dosing regimen comprising: (i) a preliminary phase comprising a 28-day dosing cycle (C1); (ii) a first phase after the preliminary phase comprising a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), and a fifth dosing cycle (C5), wherein each dosing cycle in the first phase is a 28-day dosing cycle; and (iii) a second phase after the first phase comprising a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), a fifth dosing cycle (C5), a sixth dosing cycle (C6), and a seventh dosing cycle (C7). ), wherein each dosing cycle of the second phase is a 28-day dosing cycle, wherein silvostomumab is administered to the subject as follows: (i) at a first step-escalating dose on day 1 of C1 during the pre-phase, and at a second step-escalating dose on day 8 of C1 during the pre-phase; (ii) at a target dose on day 15 of C1 during the pre-phase; (iii) at a target dose on days 1 and 15 of C1, C2, C3, C4, and C5 during the first phase; and (iv) at the target dose on Day 1 of C1, C2, C3, C4, C5, C6, and C7 during the second phase; and wherein lenalidomide is administered to the subject as follows: (i) on Day 1 to Day 21 of C1 during the pre-phase; (ii) on Day 1 to Day 21 of C1, C2, C3, C4, and C5 during the first phase; and (iii) on Day 1 to Day 21 of C1, C2, C3, C4, C5, C6, and C7 during the second phase.

[0304] In some examples: (i) the subject experiences a PR or better after induction therapy; (ii) the subject receives ASCT within 100 days of initiation of the approach and / or does not have progressive disease; (iii) the patient is administered silvostomumab and lenalidomide as post-transplant maintenance therapy; and (iv) the high-risk cytogenetic profile includes one or more of the following: translocation events t(4;14) or t(14;16), del(17p), or 1q gain.

[0305] In some examples: (i) the first step-escalating dose of silvosetumab is 0.3 mg; (ii) the second step-escalating dose of silvosetumab is 3.6 mg; (iii) the target dose of silvosetumab is between 90 mg and 198 mg, inclusive; and (iv) lenalidomide is administered at a dose of 10 mg or 15 mg.

[0306] In some examples, the target dose is 90 mg.

[0307] In some instances, the target dose is 132 mg.

[0308] In some examples, the target dose is 160 mg.

[0309] ii. Single step-up dosing regimen

[0310] In some aspects, the invention provides a method of treating a subject having cancer (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM (e.g., MM with high-risk cytogenetic characteristics)))), the method comprising administering to the subject a bispecific antibody that binds to FcRH5 and CD3 and lenalidomide in a single step-up dosing regimen.

[0311] In some aspects, the invention provides a method of treating a subject having MM (e.g., MM with high-risk cytogenetic characteristics), the method comprising administering to the subject a bispecific antibody that binds to FcRH5 and CD3 and lenalidomide in a dosing regimen that includes at least a first dosing cycle, wherein the first dosing cycle includes a first dose (C1D1) and a second dose (C1D2) of the bispecific antibody, wherein C1D1 is between about 0.05 mg and about 180 mg (e.g., between about 0.1 mg and about 160 mg, between about 0.5 mg and about 140 mg, between about 1 mg and about 120 mg, between about 1.5 mg and about 100 mg, between about 2.0 mg and about 80 mg, between about 2.5 mg and about 50 mg, between about 3.0 mg and about 25 mg, between about 3.0 mg and about 15 mg, between about 3.0 mg and about 10 mg, or between about 3.0 mg and about 5 mg). ), and C1D2 is between about 0.15 mg and about 1000 mg (e.g., between about 0.5 mg and about 800 mg, between about 1 mg and about 700 mg, between about 5 mg and about 500 mg, between about 10 mg and about 400 mg, between about 25 mg and about 300 mg, between about 40 mg and about 200 mg, between about 50 mg and about 100 mg, between about 75 mg and about 100 mg, or between about 85 mg and about 100 mg), and C1D2 is between about 0.15 mg and about 1000 mg (e.g., between about 0.5 mg and about 800 mg, between about 1 mg and about 700 mg, between about 5 mg and about 500 mg, between about 10 mg and about 400 mg, between about 25 mg and about 300 mg, between about 50 mg and about 250 mg, between about 100 mg and about 225 mg, or between about 150 mg and about 200 mg).

[0312] In some aspects, the present invention provides a method of treating a subject having cancer (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM (e.g., MM with high-risk cytogenetic characteristics)))), the method comprising administering to the subject a bispecific antibody that binds to FcRH5 and CD3 and lenalidomide in 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; cycle 1, dose 1) and a second dose (C1D2; cycle 1, dose 2), wherein wherein C1D1 is less than C1D2, and wherein the C1D1 is between about 0.05 mg and about 180 mg (e.g., between about 0.1 mg and about 160 mg, between about 0.5 mg and about 140 mg, between about 1 mg and about 120 mg, between about 1.5 mg and about 100 mg, between about 2.0 mg and about 80 mg, between about 2.5 mg and about 50 mg, between about 3.0 mg and about 25 mg, between about 3.0 mg and about 15 mg, between about 3.0 mg and about 10 mg, or between about 3.0 mg and about 5 mg). and (b) the second dosing cycle comprises a single dose of a bispecific Antibody (C2D1; Cycle 2, Dose 1), wherein C2D1 is equal to or greater than C1D2 and is between about 0.15 mg to about 1000 mg (e.g., between about 0.5 mg to about 800 mg, between about 1 mg to about 700 mg, between about 5 mg to about 500 mg, between about 10 mg to about 400 mg, between about 25 mg to about 300 mg, between about 40 mg to about 200 mg, between about 50 mg to about 100 mg, between about 75 mg to about 100 mg, or between about 85 mg to about 100 mg).

[0313] In some aspects, (a) C1D1 is between about 0.5 mg and about 19.9 mg (e.g., between about 1 mg and about 18 mg, between about 2 mg and about 15 mg, between about 3 mg and about 10 mg, between about 3.3 mg and about 6 mg, or between about 3.4 mg and about 4 mg, for example, about 3 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.6 mg, 3.8 mg, 4 mg, 4.2 mg, 4.4 mg, 4.6 mg, 4.8 mg, 5 mg, 5.2 mg, 5.6 mg, 5.8 mg, 6 mg, 6.2 mg, 6.4 mg, 6.6mg, 6.8mg, 7mg, 7.2mg, 7.4mg, 7.6mg, 7.8mg, 8mg, 8.2mg, 8.4mg, 8.6mg, 8.8mg, 9mg, 9.2mg, 9.4mg, 9.6mg, 9.8mg, 10mg, 1 0.2mg, 10.4mg, 10.6mg, 10.8mg, 11mg, 11.2mg, 11.4mg, 11.6mg, 11.8mg, 12mg, 12.2mg, 12.4mg, 12.6mg, 12.8mg, 13mg, 13.2m or 19.8 mg), and (b) C1D2 is between about 20 mg and about 20 mg. or about 600 mg (e.g., between about 30 to 500 mg, 40 to 400 mg, 60 to 350 mg, 80 to 300 mg, 100 to 200 mg, or 140 to 180 mg, for example, about 20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, 400, 420, 440, 460, 480, 500, 520, 540, 560, 580, or 600 mg).

[0314] In some aspects, C1D1 is between about 1.2 mg and about 10.8 mg and C1D2 is between about 80 mg and about 300 mg. In some aspects, C1D1 is about 3.6 mg and C1D2 is about 198 mg. In some aspects, C1D1 is between 1.2 mg and 10.8 mg and C1D2 is between 80 mg and 300 mg. In some aspects, C1D1 is 3.6 mg and C1D2 is 90 mg. In some aspects, C1D1 is 3.6 mg and C1D2 is 132 mg. In some aspects, C1D1 is 3.6 mg and C1D2 is 160 mg. In some aspects, C1D1 is 3.6 mg and C1D2 is 198 mg.

[0315] In other aspects, C1D1 is 3.3 mg. In some aspects, C1D1 is 3.3 mg and C1D2 is between 90 mg and 198 mg, for example, 90 mg, 132 mg, 160 mg, or 198 mg.

[0316] In some cases, the above methods may include a first dosing cycle of three weeks or 21 days. In some cases, the methods may include administering C1D1 and C1D2 to the subject on or around day 1 and day 8, respectively, of the first dosing cycle.

[0317] iii. Double step-up dosing regimen

[0318] In other aspects, the invention provides a method of treating a subject having cancer (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM (e.g., MM with high-risk cytogenetic characteristics))))), the method comprising administering to the subject a bispecific antibody that binds to FcRH5 and CD3 in a dual step-up dosing regimen.

[0319] In some aspects, the disclosure features a method of treating a subject having cancer (e.g., MM (e.g., MM with a high-risk cytogenetic profile)), the method comprising administering to the subject a bispecific antibody that binds to FcRH5 and CD3 and lenalidomide in a dosing regimen that includes at least a first dosing cycle, wherein the first dosing cycle includes a first dose (C1D1), a second dose (C1D2), and a third dose (C1D3) of the bispecific antibody, wherein C1D1 is between about 0.2 mg and about 0.4 mg In some aspects, C1D1 is about 0.3 mg.

[0320] In some embodiments, C1D1 is between 0.2 mg and 0.4 mg (e.g., 0.20 mg, 0.21 mg, 0.22 mg, 0.23 mg, 0.24 mg, 0.25 mg, 0.26 mg, 0.27 mg, 0.28 mg, 0.29 mg, 0.30 mg, 0.31 mg, 0.32 mg, 0.33 mg, 0.34 mg, 0.35 mg, 0.36 mg, 0.37 mg, 0.38 mg, 0.39 mg, or 0.40 mg). In some aspects, C1D1 is 0.3 mg.

[0321] In some aspects, the present disclosure provides a method of treating a subject having cancer (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM (e.g., MM with high-risk cytogenetic characteristics)))), the method comprising administering to the subject a bispecific antibody that binds to FcRH5 and CD3 and lenalidomide in a dosing regimen that includes at least a first dosing cycle, wherein the first dosing cycle includes a first dose (C1D1), a second dose (C1D2), and a third dose (C1D3) of the bispecific antibody, wherein C1D1 is between about 0.01 mg and about 2.9 mg, C1D2 is between about 3 mg and about 19.9 mg, and C1D3 is between about 20 mg and about 600 mg.

[0322] In some aspects, the invention provides a method of treating a subject having cancer (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM (e.g., MM with high-risk cytogenetic characteristics)))), the method comprising administering to the subject a bispecific antibody that binds to FcRH5 and CD3 and lenalidomide in a dosing regimen comprising at least a first dosing cycle and a second dosing cycle, wherein (a) the first dosing cycle comprises a first dose (C1D1), a second dose (C1D2), and a third dose (C1D3) of the bispecific antibody, wherein C1D1 and C1D2 are each less than C1D3, and wherein C1D1 is between about 0.01 mg and about 2.9 mg, C1D2 is between about 3 mg and about 19.9 mg, and C1D3 is between about 20 mg and about 600 mg; and (b) the second dosing cycle comprises a single dose (C2D1) of the bispecific antibody, wherein C2D1 is equal to or greater than C1D3 and is between about 20 mg and about 600 mg.

[0323] In some aspects, C1D1 is between about 0.05 mg to about 2.5 mg, about 0.1 mg to about 2 mg, about 0.2 mg to about 1 mg, or about 0.2 mg to about 0.4 mg (e.g., about 0.01 mg, 0.05 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.9 mg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, or 2.9 mg). In some aspects, C1D1 is about 0.3 mg.

[0324] In some aspects, C1D1 is between 0.05 mg and 2.5 mg, 0.1 mg and 2 mg, 0.2 mg and 1 mg, or 0.2 mg and 0.4 mg (e.g., 0.01 mg, 0.05 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.9 mg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, or 2.9 mg). In some aspects, C1D1 is 0.3 mg.

[0325] In some aspects, C1D2 is between about 3 mg to about 19.9 mg (e.g., between about 3 mg to about 18 mg, between about 3.1 mg to about 15 mg, between about 3.2 mg to about 10 mg, between about 3.3 mg to about 6 mg, or between about 3.4 mg to about 4 mg, for example, about 3 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.6 mg, 3.8 mg, 4 mg, 4.2 mg, 4.4 mg, 4.6 mg, 4.8 mg, 5 mg, 5.2 mg, 5.6 mg, 5.8 mg, 6 mg, 6.2 mg, 6.4 mg, 6.6 mg, 6.8 mg, 7 mg, 7.2 mg, 7.4 mg, 7.6 mg, 7.8 mg, 8 mg, 8.2 mg, 8.4 mg, 8.6 mg, 8.8 mg, 9 mg, 9.2 mg, 9.4 mg, 9.6 mg, 9.8 mg, 8mg, 10mg, 10.2mg, 10.4mg, 10.6mg, 10.8mg, 11mg, 11.2mg, 11.4mg, 11.6mg, 11.8mg, 12mg, 12.2mg, 12.4mg, 12.6mg, 12.8mg, 13mg, 13.2mg, 13.4mg, 13.6mg, 13.8mg, 14mg, 14.2mg, 14. In some aspects, C1D2 is between about 3.2 mg and about 10 mg. In some aspects, C1D2 is about 3.6 mg. In other aspects, C1D2 is about 3.3 mg.

[0326] In some aspects, C1D2 is between 3 mg and 19.9 mg (e.g., between 3 mg and 18 mg, between 3.1 mg and 15 mg, between 3.2 mg and 10 mg, between 3.3 mg and 6 mg, or between 3.4 mg and 4 mg, for example, 3 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.6 mg, 3.8 mg, 4 mg, 4.2 mg, 4.4 mg, 4.6 mg, 4.8 mg, 5 mg, 5.2 mg, 5.6 mg, 5.8 mg, 6 mg, 6.2 mg, 6.4 mg, 6.6 mg, 6.8 mg, 7 mg, 7.2 mg, 7.4 mg, 7.6 mg, 7.8 mg, 8 mg, 8.2 mg, 8.4 mg, 8.6 mg, 8.8 mg, 9 mg, 9.2 mg, 9.4 mg, 9.6 mg, 9.8 mg, 10 mg, 10.2mg, 10.4mg, 10.6mg, 10.8mg, 11mg, 11.2mg, 11.4mg, 11.6mg, 11.8mg, 12mg, 12.2m g, 12.4mg, 12.6mg, 12.8mg, 13mg, 13.2mg, 13.4mg, 13.6mg, 13.8mg, 14mg, 14.2mg, 14.4mg In some aspects, C1D2 is between 3.2 mg and 10 mg. In some aspects, C1D2 is 3.6 mg. In other aspects, C1D2 is 3.3 mg.

[0327] In some aspects, C1D3 is between about 20 mg to about 600 mg (e.g., between about 30 mg to about 500 mg, about 40 mg to about 400 mg, about 60 mg to about 350 mg, about 80 mg to about 300 mg, about 100 mg to about 200 mg, or about 140 mg to about 180 mg, for example, about 20 mg, 40 mg, 60 mg, 80 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg, 280 mg, 300 mg, 320 mg, 340 mg, 360 mg, 380 mg, 400 mg, 420 mg, 440 mg, 460 mg, 480 mg, 500 mg, 520 mg, 540 mg, 560 mg, 580 mg, or 600 mg). In some aspects, C1D3 is between about 80 mg and about 300 mg. In some aspects, C1D3 is about 90 mg. In some aspects, C1D3 is about 132 mg. In some aspects, C1D3 is about 160 mg. In some aspects, C1D3 is about 198 mg.

[0328] In some aspects, C1D3 is between 20 mg and 600 mg (e.g., between 30 mg and 500 mg, 40 mg and 400 mg, 60 mg and 350 mg, 80 mg and 300 mg, 100 mg and 200 mg, or 140 mg and 180 mg, for example, 20 mg, 40 mg, 60 mg, 80 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg, 280 mg, 300 mg, 320 mg, 340 mg, 360 mg, 380 mg, 400 mg, 420 mg, 440 mg, 460 mg, 480 mg, 500 mg, 520 mg, 540 mg, 560 mg, 580 mg, or 600 mg). In some aspects, C1D3 is between 80 mg and 300 mg. In some aspects, C1D3 is about 90 mg. In some aspects, C1D3 is about 132 mg. In some aspects, C1D3 is about 160 mg. In some aspects, C1D3 is about 198 mg.

[0329] In some aspects, the method comprises only a single dosing cycle (e.g., a dosing cycle comprising C1D1, C1D2, and C1D3). In other aspects, the dosing regimen further comprises a second dosing cycle comprising at least a single dose (C2D1) of the bispecific antibody. In some aspects, C2D1 is equal to or greater than C1D3 and is between about 20 mg to about 600 mg (e.g., between about 30 mg to about 500 mg, about 40 mg to about 400 mg, about 60 mg to about 350 mg, about 80 mg to about 300 mg, about 100 mg to about 200 mg, or about 140 mg to about 180 mg, for example, about 20 mg, 40 mg, 60 mg, 80 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg, 280 mg, 300 mg, 320 mg, 340 mg, 360 mg, 380 mg, 400 mg, 420 mg, 440 mg, 460 mg, 480 mg, 500 mg, 520 mg, 540 mg, 560 mg, 580 mg, or 600 mg). In some aspects, C2D1 is between about 80 mg and about 300 mg. In some aspects, C2D1 is about 90 mg. In some aspects, C2D1 is about 132 mg. In some aspects, C2D1 is about 160 mg. In some aspects, C2D1 is about 198 mg.

[0330] In some aspects, C2D1 is between 20 mg and 600 mg (e.g., between 30 mg and 500 mg, 40 mg and 400 mg, 60 mg and 350 mg, 80 mg and 300 mg, 100 mg and 200 mg, or 140 mg and 180 mg, for example, 20 mg, 40 mg, 60 mg, 80 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg, 280 mg, 300 mg, 320 mg, 340 mg, 360 mg, 380 mg, 400 mg, 420 mg, 440 mg, 460 mg, 480 mg, 500 mg, 520 mg, 540 mg, 560 mg, 580 mg, or 600 mg). In some aspects, C2D1 is between 80 mg and 300 mg. In some aspects, C2D1 is 160 mg. In some aspects, C2D1 is 159 mg.

[0331] Alternatively, in any of the above embodiments, C1D1 can be between about 0.01 mg and about 60 mg (e.g., between about 0.05 mg and about 50 mg, between about 0.01 mg and about 40 mg, between about 0.1 mg and about 20 mg, between about 0.1 mg and about 10 mg, between about 0.1 mg and about 5 mg, between about 0.1 mg and about 2 mg, between about 0.1 mg and about 1.5 mg, between about 0.1 mg and about 1.2 mg, between about 0.1 mg and about 0.5 mg, or between about 0.2 mg and about 0.4 mg, for example, about 0.3 mg, such as 0.3 mg), C1D2 can be between about 0.05 mg and about 180 mg (e.g., between about 0.1 mg and about 160 mg, between about 0.5 mg and about 140 mg, between about 1 mg and about 120 mg, between about 1.5 mg and about 100 mg, between about 2.0 mg and about 80 mg, between about 2.5 mg and about 50 mg, between about 3.0 mg and about 25 mg, between about 3.0 mg and about 15 mg, between about 3.0 mg and about 10 mg, between about 3.0 mg and about 5 mg, or between about 3.0 mg and about 4.0 mg, e.g., about 3.6 mg, e.g., 3.6 mg), and C1D3 may be between about 0.15 mg and about 1000 mg (e.g., between about 0.5 mg and about 800 mg, between about 1 mg and about 700 mg, between about 5 mg and about 500 mg, between about 10 mg and about 400 mg, between about 25 mg and about 300 mg, between about 40 mg and about 200 mg, between about 50 mg and about 190 mg, between about 140 mg and about 180 mg, or between about 150 mg and about 170 mg, e.g., about 160 mg, e.g., 1 60 mg); and in aspects comprising a second dosing cycle, C2D1 can be between about 0.15 mg to about 1000 mg (e.g., between about 0.5 mg to about 800 mg, between about 1 mg to about 700 mg, between about 5 mg to about 500 mg, between about 10 mg to about 400 mg, between about 25 mg to about 300 mg, between about 40 mg to about 200 mg, between about 50 mg to about 190 mg, between about 140 mg to about 180 mg, or between about 150 mg to about 170 mg, e.g., about 160 mg, e.g., 160 mg).

[0332] In some cases, the length of the first dosing cycle is four weeks or 28 days. In other cases, the length of the first dosing cycle is three weeks or 21 days. In some cases, the method may include administering C1D1, C1D2, and C1D3 to the subject on or around day 1, day 8, and day 15 of the first dosing cycle, respectively.

[0333] iv. Additional dosing cycles

[0334] In some cases, the above methods may include a second dosing cycle of four weeks or 28 days. In other cases, the length of the first dosing cycle is one week or 7 days; two weeks or 14 days; or three weeks or 21 days. In some cases, the method may include administering C2D1 to the subject on or around day 1 of the second dosing cycle.

[0335] In some cases where the method includes at least a second dosing cycle, the method may include one or more additional dosing cycles. In some cases, the dosing regimen includes 1 to 17 additional dosing cycles (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 additional dosing cycles, for example, 1 to 3 additional dosing cycles, 1 to 5 additional dosing cycles, 3 to 8 additional dosing cycles, 5 to 10 additional dosing cycles, 8 to 12 additional dosing cycles, 10 to 15 additional dosing cycles, 12 to 17 additional dosing cycles, or 15 to 17 additional dosing cycles, i.e., the dosing regimen includes one or more additional dosing cycles C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, C32, C33, C34, C35, C36, C37, C38, C39, C40, C41, C42, C43, C44, C45, C46, ​​C47, C48, C49, C50, C51, C52, C53, C54, C55, C56, C57, C58, C59, C6 5, C16, C17, C18 and C19. In some embodiments, the length of each of the one or more additional dosing cycles is 7 days, 14 days, 21 days or 28 days. In some embodiments, the length of each of the one or more additional dosing cycles is between 5 days and 30 days, for example, between 5 days and 9 days, between 7 days and 11 days, between 9 days and 13 days, between 11 days and 15 days, between 13 days and 17 days, between 15 days and 19 days, between 17 days and 21 days, between 19 days and 23 days, between 21 days and 25 days, between 23 days and 27 days, or between 25 days and 30 days. In some cases In some cases, the length of each of the one or more additional dosing cycles is three weeks or 21 days. In some cases, each of the one or more additional dosing cycles comprises a single dose of the bispecific antibody. In some aspects, the dose of the bispecific antibody in the one or more additional dosing cycles is equal to C2D1, for example, between about 20 mg to about 600 mg (e.g., between about 30 mg to about 500 mg, about 40 mg to about 400 mg, about 60 mg to about 350 mg, about 80 mg to about 300 mg, about 100 mg to about 200 mg, or about 140 mg to about 180 mg, for example, about 20 mg, 40 mg In some aspects, the dosage of the bispecific antibody in one or more additional dosing cycles is about 90 mg. In some aspects, the dosage of the bispecific antibody in one or more additional dosing cycles is about 132 mg.In some aspects, the dose of the bispecific antibody in one or more additional dosing cycles is about 160 mg. In some aspects, the dose of the bispecific antibody in one or more additional dosing cycles is about 198 mg. In some aspects, the dose of the bispecific antibody in one or more additional dosing cycles is equal to C2D1, e.g., between 20 mg and 600 mg (e.g., between 30 mg and 500 mg, 40 mg and 400 mg, 60 mg and 350 mg, 80 mg and 300 mg, 100 mg and 200 mg, or 140 mg and 180 mg, e.g., 20 mg, 40 mg, 60 mg, 80 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg, 280 mg, 300 mg, 320 mg, 340 mg, 360 mg, 380 mg, 400 mg, 420 mg, 440 mg, 460 mg, 480 mg, 500 mg, 520 mg, 540 mg, 560 mg, 580 mg, or 600 mg). In some aspects, the dosage of the bispecific antibody in one or more additional dosing cycles is 90 mg. In some aspects, the dosage of the bispecific antibody in one or more additional dosing cycles is 132 mg. In some aspects, the dosage of the bispecific antibody in one or more additional dosing cycles is 160 mg. In some aspects, the dosage of the bispecific antibody in one or more additional dosing cycles is 198 mg. In some aspects, the method includes administering a single dose of the bispecific antibody to a subject on or about the first day of the one or more additional dosing cycles.

[0336] In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody is administered to a subject as a monotherapy.

[0337] B. Combination Therapy with Additional Therapeutic Agents

[0338] In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide are administered to a subject along with one or more additional therapeutic agents, including any additional therapeutic agents disclosed herein.

[0339] i. Anti-CD38 Antibody

[0340] In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide are administered to a subject in combination with an anti-CD38 antibody. The anti-CD38 antibody can be administered to a subject by any suitable route of administration (e.g., intravenous (IV) or subcutaneous (SC)). In some aspects, the anti-CD38 antibody is daratumumab (e.g., daratumumab / rHuPH20). The amount of lenalidomide administered to a subject can be from about 900 mg to about 3600 mg (e.g., about 900 mg, about 950 mg, about 1000 mg, about 1100 mg, about 1200 mg, about 1300 mg, about 1400 mg, about 1500 mg, about 1600 mg, about 1650 mg, about 1700 mg, about 1750 mg, about 1800 mg, about 1850 mg, about 1900 mg, about 1950 mg, about 20 In some aspects, daratumumab is administered to a subject at a dosage of about 1000 mg, about 2100 mg, about 2200 mg, about 2300 mg, about 2400 mg, about 2500 mg, about 2600 mg, about 2700 mg, about 2800 mg, about 2900 mg, about 3000 mg, about 3100 mg, about 3200 mg, about 3300 mg, about 3400 mg, about 3500 mg or about 3600 mg).Daratumumab can be administered to a subject at a dosage of about 1800 mg.In some aspects, daratumumab is administered by intravenous infusion (for example, infused over 3 hours to 5 hours) with a dosage of 16 mg / kg once a week, once every two weeks or once every four weeks.In some aspects, daratumumab is administered by intravenous infusion (for example, infused over 3 hours to 5 hours) with a dosage of 16 mg / kg. In some aspects, daratumumab is administered subcutaneously. In other aspects, the anti-CD38 antibody is isatuximab. In some aspects, an anti-CD38 antibody (e.g., daratumumab or isatuximab) is administered to a subject prior to administration of a bispecific anti-FcRH5 / anti-CD3 antibody (e.g., one day prior to administration of a bispecific anti-FcRH5 / anti-CD3 antibody). In some aspects, an anti-CD38 antibody (e.g., daratumumab or isatuximab) is administered to a subject simultaneously with administration of a bispecific anti-FcRH5 / anti-CD3 antibody.

[0341] ii. Corticosteroids

[0342] In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide are administered to a subject in combination with a corticosteroid. Corticosteroids can be administered orally to a subject. Corticosteroids can be administered to a subject by any suitable route of administration (e.g., intravenous or subcutaneous). Any suitable corticosteroid can be used, such as dexamethasone, methylprednisolone, prednisone, prednisolone, betamethasone, hydrocortisone, etc. In some aspects, the corticosteroid is methylprednisolone. Methylprednisolone can be administered to a subject at a dosage of about 80 mg. In other aspects, the corticosteroid is dexamethasone. Dexamethasone can be administered to a subject at a dosage of about 20 mg. In some aspects, a corticosteroid (e.g., methylprednisolone or dexamethasone) is administered to a subject before administering bispecific anti-FcRH5 / anti-CD3 antibodies and / or lenalidomide (e.g., one hour before administering bispecific anti-FcRH5 / anti-CD3 antibodies and / or lenalidomide). In some aspects, a corticosteroid (e.g., methylprednisolone or dexamethasone) is administered to the subject about one day prior to administration of the bispecific anti-FcRH5 / anti-CD3 antibody and / or lenalidomide. In some aspects, a corticosteroid (e.g., methylprednisolone or dexamethasone) is administered to the subject simultaneously with administration of the bispecific anti-FcRH5 / anti-CD3 antibody and / or lenalidomide.

[0343] iii. Immunomodulatory drugs (IMiDs)

[0344] In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide are administered to a subject in combination with an additional immunomodulatory drug (IMiD). IMiDs may be administered to a subject by any suitable route of administration (e.g., oral). IMiDs may be administered intravenously to a subject. IMiDs may be administered subcutaneously to a subject. In some aspects, IMiDs are pomalidomide. Pomalidomide may be administered to a subject at a dose of about 4 mg. In some aspects, IMiDs (e.g., pomalidomide) are administered to a subject before administering bispecific anti-FcRH5 / anti-CD3 antibodies and / or lenalidomide (e.g., one hour before administering bispecific anti-FcRH5 / anti-CD3 antibodies and / or lenalidomide). In some aspects, IMiDs (e.g., pomalidomide) are administered to a subject while administering bispecific anti-FcRH5 / anti-CD3 antibodies and / or lenalidomide. In some aspects, an IMiD (eg, pomalidomide) is administered daily between doses of the bispecific anti-FcRH5 / anti-CD3 antibody and / or lenalidomide.

[0345] iv. Tocilizumab and treatment of CRS

[0346] In one embodiment, the additional therapeutic agent is an effective amount of tocilizumab In some cases, a subject has a cytokine release syndrome (CRS) event (for example, a CRS event occurs after treatment with a bispecific antibody, such as a CRS event occurs after treatment with a bispecific antibody of C1D1, C1D2, C1D3, C2D1 or an additional dose), and the method further includes treating the symptoms of the CRS event (for example, by applying an effective amount of tocilizumab to the subject to treat the CRS event), while suspending treatment with the bispecific antibody. In some aspects, tocilizumab is administered intravenously to the subject in a single dose of about 8mg / kg. In some aspects, the CRS event does not subside or worsen within 24 hours of treating the symptoms of the CRS event, and the method further includes administering one or more additional doses of tocilizumab to the subject to manage the CRS event, for example, administering one or more additional doses of tocilizumab to the subject intravenously at a dosage of about 8mg / kg.

[0347] In some aspects, treating the symptoms of a CRS event further comprises treatment with a high-dose vasopressor (e.g., norepinephrine, dopamine, phenylephrine, epinephrine, or vasopressin and norepinephrine), e.g., as described in Table 2A and Table 2B.

[0348] In other cases, tocilizumab is administered as a premedication, e.g., administered to a subject prior to administration of the bispecific anti-FcRH5 / anti-CD3 antibody. In some cases, tocilizumab is administered as a premedication in cycle 1, e.g., prior to the first dose (C1D1) of the bispecific antibody, the second dose (C1D2) of the bispecific antibody, and / or the third dose (C1D3) of the bispecific anti-FcRH5 / anti-CD3 antibody. In some aspects, tocilizumab is administered intravenously to a subject at a single dose of about 8 mg / kg.

[0349] v.CRS symptoms and grading

[0350] CRS can be graded according to a modified cytokine release syndrome grading system established by Lee et al., Blood, 124: 188-195, 2014 or Lee et al., Biol Blood Marrow Transplant, 25(4): 625-638, 2019, as described in Table 2A. In addition to the diagnostic criteria, recommended CRS management based on its severity (including early intervention with corticosteroids and / or anti-cytokine therapy) is also provided and mentioned in Tables 2A and 2B.

[0351] Table 2A. Cytokine release syndrome grading system

[0352]

[0353] Lee 2014 criteria: Lee et al., Blood, 124: 188-195, 2014.

[0354] ASTCT consensus grade: Lee et al., Biol Blood Marrow Transplant, 25(4):625-638, 2019.

[0355] a Low-dose vasopressor: A single vasopressor at a dose lower than the dose shown in Table 2B.

[0356] b High-dose vasopressors: As defined in Table 2B.

[0357] * Fever was defined as a temperature ≥ 38°C not attributable to any other cause. In patients who had CRS and subsequently received antipyretic or anticytokine therapy (such as tocilizumab or steroids), fever was no longer required to grade the subsequent severity of CRS. In this setting, CRS grading was driven by hypotension and / or hypoxia.

[0358] The grade of CRS is determined by the more severe event: hypotension or hypoxia that is not attributable to any other cause. For example, a patient with a temperature of 39.5°C, hypotension requiring 1 vasopressor, and hypoxia requiring a low-flow nasal cannula is classified as grade 3 CRS.

[0359] Low flow nasal cannula is defined as oxygen delivered at ≤6 L / min. Low flow also includes insufflation oxygen delivery, which is sometimes used in pediatrics. High flow nasal cannula is defined as oxygen delivered at >6 L / min.

[0360] Table 2B. High-dose vasopressors

[0361]

[0362] min = minutes; VASST = vasopressin and septic shock trial.

[0363] a VASST vasopressor equivalent equation: Norepinephrine equivalent dose = [norepinephrine (μg / min)] + [dopamine (μg / kg / min) ÷ 2] + [epinephrine (μg / min)] + [phenylephrine (μg / min) ÷ 10].

[0364] Mild to moderate CRS and / or infusion-related reactions (IRR) may include symptoms such as fever, headache, and myalgia, and may be treated symptomatically with analgesics, antipyretics, and antihistamines as needed. Severe or life-threatening manifestations of CRS and / or IRR, such as hypotension, tachycardia, dyspnea, or chest discomfort, should be treated aggressively with supportive and resuscitative measures as indicated, including the use of high-dose corticosteroids, IV fluids, intensive care unit admission, and other supportive measures. Severe CRS may be associated with other clinical sequelae, such as disseminated intravascular coagulation, capillary leak syndrome, or macrophage activation syndrome (MAS). Standards of care for severe or life-threatening CRS resulting from immune-based therapies have not been established; case reports and recommendations for the use of anti-cytokine therapies such as tocilizumab have been published (Teachey et al., Blood, 121:5154-5157, 2013; Lee et al., Blood, 124:188-195, 2014; Maude et al., New Engl J Med, 371:1507-1517, 2014).

[0365] As shown in Table 2A, even subjects with extensive comorbidities who develop moderate manifestations of CRS should be closely monitored, and admission to the intensive care unit and administration of tocilizumab should be considered.

[0366] vi. Tocilizumab as a premedication

[0367] In some aspects, an effective amount of an interleukin-6 receptor (IL-6R) antagonist (e.g., an anti-IL-6R antibody such as tocilizumab) is administered to ) is used as a pro-medication (prevention) administration, for example, it is administered to a subject before the administration of the bispecific antibody (for example, administered about 2 hours before the administration of the bispecific antibody). The administration of tocilizumab as a pro-medication can reduce the frequency or severity of CRS. In some aspects, tocilizumab is administered as a pro-medication in the first cycle, for example, before the first dose (C1D1; Cycle 1, Dose 1), the second dose (C1D2; Cycle 1, Dose 2) and / or the third dose (C1D3; Cycle 1, Dose 3) of the bispecific antibody. In some aspects, tocilizumab is administered intravenously to a subject at a single dose of about 1 mg / kg to about 15 mg / kg (for example, about 4 mg / kg to about 10 mg / kg, for example, about 6 mg / kg to about 10 mg / kg, for example, about 8 mg / kg). In some aspects, tocilizumab is administered intravenously to a subject at a single dose of about 8 mg / kg. In some aspects, tocilizumab is administered intravenously to a subject at a single dose of about 8 mg / kg (maximum 800 mg) for patients weighing 30 kg or more, and at a dose of about 12 mg / kg for patients weighing less than 30 kg. Other anti-IL-6R antibodies that can be used in combination with tocilizumab include sarilumab, vobarilizumab (ALX-0061), SA-237, and variants thereof.

[0368] For example, in one embodiment, a bispecific antibody and tocilizumab Co-administration, where tocilizumab is administered first to the subject The bispecific antibody is then administered alone (e.g., the subject is treated with tocilizumab pre-treatment).

[0369] In some aspects, the incidence of CRS (e.g., grade 1 CRS, grade 2 CRS, and / or grade 3+ CRS) is reduced in patients treated with tocilizumab as a premedication, relative to patients not treated with tocilizumab as a premedication. In some aspects, less intervention is required to treat CRS (e.g., less need for additional tocilizumab, IV infusion, steroids, or O) in patients treated with tocilizumab as a premedication, relative to patients not treated with tocilizumab as a premedication. 2 In some aspects, the severity of CRS symptoms is reduced (e.g., limited to fever and chills) in patients treated with tocilizumab as a prior medication relative to patients not treated with tocilizumab as a prior medication.

[0370] vii. Administration of Tocilizumab to Treat CRS

[0371] In some aspects, the subject is treated with a therapeutic bispecific antibody and an effective amount of an IL-6R antagonist (e.g., an anti-IL-6R antibody such as tocilizumab A CRS event was experienced during treatment, and an effective amount of tocilizumab was administered to manage the CRS event.

[0372] In some aspects, the subject develops a CRS event (e.g., a CRS event develops after treatment with the bispecific antibody, e.g., a CRS event develops after a first dose or a subsequent dose of the bispecific antibody), and the method further comprises treating a symptom of the CRS event while treatment with the bispecific antibody is suspended.

[0373] In some aspects, the subject experiences a CRS event, and the method further comprises administering to the subject an effective amount of an interleukin-6 receptor (IL-6R) antagonist (e.g., an anti-IL-6R antibody, such as tocilizumab) while treatment with the bispecific antibody is suspended. ) to manage CRS events. In some aspects, an IL-6R antagonist (e.g., tocilizumab) is administered intravenously to a subject at a single dose of about 1 mg / kg to about 15 mg / kg (e.g., about 4 mg / kg to about 10 mg / kg, for example, about 6 mg / kg to about 10 mg / kg, for example, about 8 mg / kg). In some aspects, tocilizumab is administered intravenously to a subject at a single dose of about 8 mg / kg. Other anti-IL-6R antibodies that can be used in combination with tocilizumab include sarilumab, vobarilizumab (ALX-0061), SA-237, and variants thereof.

[0374] In some aspects, the CRS event does not subside or worsen within 24 hours of treating the symptoms of the CRS event, and the method further includes administering to the subject one or more additional doses of an IL-6R antagonist (e.g., an anti-IL-6R antibody, such as tocilizumab) to manage the CRS event, for example, at a dose of about 1 mg / kg to about 15 mg / kg, for example, about 4 mg / kg to about 10 mg / kg, for example, about 6 mg / kg to about 10 mg / kg, for example, about 8 mg / kg to the subject intravenously administering one or more additional doses of tocilizumab. In some aspects, one or more additional doses of tocilizumab are administered intravenously to the subject at a single dose of about 8 mg / kg.

[0375] In some aspects, the method further includes administering an effective amount of a corticosteroid to the subject. The corticosteroid can be administered intravenously to the subject. In other examples, the corticosteroid can be administered subcutaneously to the subject. In some aspects, the corticosteroid is methylprednisolone. In some cases, methylprednisolone is administered at a dosage of about 1 mg / kg to about 5 mg / kg per day, for example, about 2 mg / kg per day. In some cases, the corticosteroid is dexamethasone. In some cases, dexamethasone is administered at a dosage of about 10 mg (e.g., administered intravenously with a single dose of about 10 mg) or at a dosage of about 0.5 mg / kg / day.

[0376] If IL-6R antagonists (e.g., tocilizumab) are used alone and CRS events cannot be managed, corticosteroids such as methylprednisolone or dexamethasone may be administered to the subject. In some aspects, the symptoms of treating CRS events further include treatment with high-dose vasopressors (e.g., norepinephrine, dopamine, phenylephrine, epinephrine, or vasopressin and norepinephrine), for example, as described in Table 2A and Table 2B. Table 3A and Table 3B further provide detailed information on tocilizumab treatment of severe or life-threatening CRS.

[0377] viii. Manage CRS events by level

[0378] Management of CRS events can be tailored based on the grade of CRS (Tables 2A and 3A) and the presence of comorbidities. Table 3A provides suggestions for the management of CRS syndrome by grade. Table 3B provides suggestions for the management of IRR syndrome by grade.

[0379] Table 3A. Recommendations for the management of cytokine release syndrome (CRS)

[0380]

[0381]

[0382]

[0383]

[0384]

[0385] BiPAP = biphasic positive airway pressure; CPAP = continuous positive airway pressure; CRS = cytokine release syndrome; HLH = hemophagocytic lymphohistiocytosis; ICU = intensive care unit; IV = intravenous; MAS = macrophage activation syndrome.

[0386] a A complete description of the symptom scale is provided in Table 2A.

[0387] b Guidelines for the management of CRS are based on Lee et al., Biol Blood Marrow Transplant, 25(4):625-638, 2019 and Riegler et al. (2019).

[0388] c Treat patients with acetaminophen and antihistamines (e.g., diphenhydramine) if they have not been treated with these medications within the previous 4 hours. Treat bronchospasm, urticaria, or dyspnea according to institutional practice. Treat fever and neutropenia as needed; consider broad-spectrum antibiotics and / or G-CSF if indicated.

[0389] d Tocilizumab should be administered at a dose of 8 mg / kg IV (8 only for patients weighing ≥30 kg).

[0390] mg / kg; 12 mg / kg for patients weighing <30 kg; infusion exceeding 800 mg / kg per infusion is not recommended

[0391] mg dose); repeat every 8 hours as needed (up to 4 doses).

[0392] e If the patient does not experience CRS during the next infusion at a 50% reduced rate, the infusion rate can be increased to the initial rate in subsequent cycles. However, if the patient experiences another CRS event, the infusion rate should be reduced by 25% to 50%, depending on the severity of the event.

[0393] Table 3B. Recommendations for the management of silvostomumab infusion-related reactions (IRRs)

[0394]

[0395]

[0396] ICU = intensive care unit; NCI CTCAE = National Cancer Institute Common Terminology Criteria for Adverse Events.

[0397] a Symptom classification refers to NCI CTCAE v5.0.

[0398] bSupportive care: If the patient has not been treated with acetaminophen / paracetamol and antihistamines such as diphenhydramine within the first 4 hours, treat with these drugs. Intravenous fluids (e.g., normal saline) may be administered as clinically indicated. For bronchospasm, urticaria, or dyspnea, antihistamines, oxygen, corticosteroids (e.g., 100 mg IV prednisolone or equivalent), and / or bronchodilators may be administered according to institutional practice. Fluid and vasopressor support is provided for hypotension, if necessary.

[0399] c Subsequent silvostomumab infusions can be started at the original rate.

[0400] ix. Management of Level 2 CRS Events

[0401] If a subject develops a Grade 2 CRS event after administration of a therapeutic bispecific antibody (e.g., a Grade 2 CRS event occurs in the absence of comorbidities or in the presence of minimal comorbidities), the method may further include treating the symptoms of the Grade 2 CRS event while suspending treatment with the bispecific antibody. If a Grade 2 CRS event then resolves to a Grade 1 CRS event for at least three consecutive days, the method may further include resuming treatment with the bispecific antibody without changing the dose. On the other hand, if a Grade 2 CRS event does not resolve within 24 hours of treating the symptoms of a Grade 2 CRS event, or deteriorates to a Grade 3 CRS event, the method may further include administering to the subject an effective amount of an interleukin-6 receptor (IL-6R) antagonist (e.g., an anti-IL-6R antibody, such as tocilizumab ) to manage grade 2 or ≥ grade 3 CRS events. In some cases, tocilizumab is administered intravenously to a subject at a single dose of about 8 mg / kg. Other anti-IL-6R antibodies that can be used in combination with tocilizumab include sarilumab, vobarilizumab (ALX-0061), SA-237, and variants thereof.

[0402] If the subject develops a Grade 2 CRS event in the presence of extensive comorbidities after administration of the therapeutic bispecific antibody, the method may further comprise: administering to the subject an IL-6R antagonist (e.g., an anti-IL-6R antibody, e.g., tocilizumab ) to manage the grade 2 CRS event, while suspending treatment with the bispecific antibody. In some cases, the first dose of tocilizumab is administered intravenously to the subject at a dose of about 8 mg / kg. Other anti-IL-6R antibodies that can be used in combination with tocilizumab include sarilumab, vobarilizumab (ALX-0061), SA-237 and variants thereof. In some cases, if the grade 2 CRS event resolves to ≤grade 1 CRS event within two weeks, the method further includes resuming treatment with the bispecific antibody at a reduced dose. In some cases, if the event occurs during an infusion or within 24 hours thereof, the smaller dose is 50% of the initial infusion rate of the previous cycle. On the other hand, if a grade 2 CRS event does not subside or worsen to a grade 3 CRS event within 24 hours of treating the symptoms of the grade 2 CRS event, the method may further include: administering to the subject one or more (e.g., one, two, three, four, or five or more) additional doses of an IL-6R antagonist (e.g., an anti-IL-6R antibody, such as tocilizumab) to manage a grade 2 or grade 3 CRS event. In some specific cases, a grade 2 CRS event does not subside within 24 hours of treating the symptoms of the grade 2 CRS event, or worsens to a grade 3 CRS event, and the method may further include administering to the subject one or more additional doses of tocilizumab to manage a grade 2 or grade 3 CRS event. In some cases, one or more additional doses of tocilizumab are administered intravenously to the subject at a dose of about 1 mg / kg to about 15 mg / kg (e.g., about 4 mg / kg to about 10 mg / kg, for example, about 6 mg / kg to about 10 mg / kg, for example, about 8 mg / kg). In some cases, the method further includes administering an effective amount of a corticosteroid to the subject. A corticosteroid may be administered before, after, or simultaneously with one or more additional doses of tocilizumab or another anti-IL-6R antibody. In some cases, a corticosteroid is administered intravenously to the subject. In some cases, the corticosteroid is methylprednisolone. In some cases, methylprednisolone is administered at a dose of about 1 mg / kg to about 5 mg / kg per day, for example, about 2 mg / kg per day. In some cases, the corticosteroid is dexamethasone. In some cases, dexamethasone is administered at a dose of about 10 mg (e.g., administered intravenously with a single dose of about 10 mg) or at a dose of about 0.5 mg / kg / day.

[0403] Management of Level 3 CRS Events

[0404] If the subject develops a Grade 3 CRS event after administration of the therapeutic bispecific antibody, the method may further comprise: administering to the subject an IL-6R antagonist (e.g., an anti-IL-6R antibody, e.g., tocilizumab) ) to manage the grade 3 CRS event while suspending treatment with the bispecific antibody. In some cases, the first dose of tocilizumab is administered intravenously to the subject at a dose of about 8 mg / kg. Other anti-IL-6R antibodies that can be used in combination with tocilizumab include sarilumab, vobarilizumab (ALX-0061), SA-237 and variants thereof. In some cases, the subject recovers within 8 hours after treatment with the bispecific antibody (e.g., no fever and vasopressors are discontinued), and the method further includes resuming treatment with the bispecific antibody at a reduced dose. In some cases, if the event occurs during an infusion or within 24 hours thereof, the smaller dose is 50% of the initial infusion rate of the previous cycle. In other cases, if the grade 3 CRS event does not subside or worsens to a grade 4 CRS event within 24 hours of treating the symptoms of the grade 3 CRS event, the method may further include: administering to the subject one or more (e.g., one, two, three, four, or five or more) additional doses of an IL-6R antagonist (e.g., an anti-IL-6R antibody, such as tocilizumab) to manage a grade 3 or 4 CRS event. In some specific cases, the grade 3 CRS event does not subside or worsens to a grade 4 CRS event within 24 hours of treating the symptoms of the grade 3 CRS event, and the method further includes administering to the subject one or more additional doses of tocilizumab to manage a grade 3 or 4 CRS event. In some cases, one or more additional doses of tocilizumab are administered intravenously to the subject at a dose of about 1 mg / kg to about 15 mg / kg (e.g., about 4 mg / kg to about 10 mg / kg, for example, about 6 mg / kg to about 10 mg / kg, for example, about 8 mg / kg). In some cases, the method further includes administering an effective amount of a corticosteroid to the subject. A corticosteroid may be administered before, after, or simultaneously with one or more additional doses of tocilizumab or another anti-IL-6R antibody. In some cases, a corticosteroid is administered intravenously to the subject. In some cases, the corticosteroid is methylprednisolone. In some cases, methylprednisolone is administered at a dose of about 1 mg / kg to about 5 mg / kg per day, for example, about 2 mg / kg per day. In some cases, the corticosteroid is dexamethasone. In some cases, dexamethasone is administered at a dose of about 10 mg (e.g., administered intravenously with a single dose of about 10 mg) or at a dose of about 0.5 mg / kg / day.

[0405] xi. Management of Level 4 CRS Events

[0406] If the subject develops a Grade 4 CRS event after administration of the therapeutic bispecific antibody, the method may further comprise: administering to the subject an IL-6R antagonist (e.g., an anti-IL-6R antibody, e.g., tocilizumab) ) to manage the Grade 4 CRS event, and permanently discontinue treatment with the bispecific antibody. In some cases, the first dose of tocilizumab is administered intravenously to the subject at a dose of about 8 mg / kg. Other anti-IL-6R antibodies that can be used in combination with tocilizumab include sarilumab, vobarilizumab (ALX-0061), SA-237 and variants thereof. In some cases, a Grade 4 CRS event may subside within 24 days after treating the symptoms of the Grade 4 CRS event. If the Grade 4 CRS event does not subside within 24 hours of treating the symptoms of the Grade 4 CRS event, the method may further include administering an IL-6R antagonist (e.g., an anti-IL-6R antibody, such as tocilizumab) to the subject. ) to manage grade 4 CRS events. In some specific cases, grade 4 CRS events do not subside within 24 hours of treating the symptoms of the grade 4 CRS events, and the method further includes administering one or more (e.g., one, two, three, four, or five or more) additional doses of tocilizumab to the subject to manage grade 4 CRS events. In some cases, one or more additional doses of tocilizumab are administered intravenously to the subject at a dose of about 1 mg / kg to about 15 mg / kg (e.g., about 4 mg / kg to about 10 mg / kg, for example, about 6 mg / kg to about 10 mg / kg, for example, about 8 mg / kg). In some cases, the method further includes administering an effective amount of corticosteroids to the subject. Corticosteroids can be administered before, after, or simultaneously with one or more additional doses of tocilizumab or another anti-IL-6R antibody. In some cases, corticosteroids are administered intravenously to the subject. In some cases, the corticosteroid is methylprednisolone. In some cases, methylprednisolone is administered at a dose of about 1 mg / kg per day to about 5 mg / kg per day, e.g., about 2 mg / kg per day. In some cases, the corticosteroid is dexamethasone. In some cases, dexamethasone is administered at a dose of about 10 mg (e.g., administered intravenously in a single dose of about 10 mg) or at a dose of about 0.5 mg / kg / day.

[0407] xii. Acetaminophen or paracetamol

[0408] In another embodiment, the additional therapeutic agent is an effective amount of acetaminophen or paracetamol. The acetaminophen or paracetamol can be administered orally to the subject, for example, at a dose of between about 500 mg and about 1000 mg. In some aspects, the acetaminophen or paracetamol is administered to the subject as a premedication, for example, prior to administration of the bispecific anti-FcRH5 / anti-CD3 antibody.

[0409] xiii. Diphenhydramine

[0410] In another instance, the additional therapeutic agent is an effective amount of diphenhydramine. Diphenhydramine can be administered orally to the subject, for example, at a dose of between about 25 mg and about 50 mg. In some aspects, diphenhydramine is administered to the subject as a premedication, for example, prior to administration of the bispecific anti-FcRH5 / anti-CD3 antibody.

[0411] xiv. Anti-myeloma agents

[0412] In another case, the additional therapeutic agent is an effective amount of an anti-myeloma agent, such as an anti-myeloma agent that enhances and / or supplements T cell-mediated killing of myeloma cells. The anti-myeloma agent can be, for example, pomalidomide, daratumumab, and / or B cell maturation antigen (BCMA) directed therapy (e.g., antibody-drug conjugates (BCMA-ADC) targeting BCMA). In some aspects, the anti-myeloma agent is administered in a four-week cycle.

[0413] xv. Other combination therapies

[0414] In some aspects, the one or more additional therapeutic agents include a PD-1 axis binding antagonist, an immunomodulatory agent, an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, a cell-based therapy, or a combination thereof.

[0415] xvi. PD-1 axis binding antagonists

[0416] In some aspects, the one or more additional therapeutic agents include a PD-1 axis binding antagonist. The PD-1 axis binding antagonist may include a PD-L1 binding antagonist, a PD-1 binding antagonist, and a PD-L2 binding antagonist. Any suitable PD-1 axis binding antagonist may be used.

[0417] In some cases, the PD-L1 binding antagonist inhibits the binding of PD-L1 to one or more of its ligand binding partners. In other cases, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1. In some other cases, the PD-L1 binding antagonist inhibits the binding of PD-L1 to B7-1. In some cases, the PD-L1 binding antagonist inhibits the binding of PD-L1 to both PD-1 and B7-1. The PD-L1 binding antagonist can be, but is not limited to, an antibody, an antigen binding fragment thereof, an immunoadhesin, a fusion protein, an oligopeptide or a small molecule. In some cases, the PD-L1 binding antagonist is a small molecule that inhibits PD-L1 (e.g., GS-4224, INCB086550, MAX-10181, INCB090244, CA-170 or ABSK041). In some cases, the PD-L1 binding antagonist is a small molecule that inhibits PD-L1 and VISTA. In some cases, the PD-L1 binding antagonist is CA-170 (also known as AUPM-170). In some cases, the PD-L1 binding antagonist is a small molecule that inhibits PD-L1 and TIM3. In some cases, the small molecule is a compound described in WO 2015 / 033301 and WO 2015 / 033299.

[0418] In some cases, the PD-L1 binding antagonist is an anti-PD-L1 antibody. A variety of anti-PD-L1 antibodies are contemplated and described herein. In any case herein, the isolated anti-PD-L1 antibody can bind to human PD-L1 (e.g., human PD-L1 shown in UniProtKB / Swiss-Prot Accession No. Q9NZQ7-1, or a variant thereof). In some cases, the anti-PD-L1 antibody is capable of inhibiting the binding between PD-L1 and PD-1 and / or between PD-L1 and B7-1. In some cases, the anti-PD-L1 antibody is a monoclonal antibody. In some cases, the anti-PD-L1 antibody is selected from Fab, Fab'-SH, Fv, scFv and (Fab') 2In some cases, the anti-PD-L1 antibody is a humanized antibody. In some cases, the anti-PD-L1 antibody is a human antibody. Exemplary anti-PD-L1 antibodies include atezolizumab, MDX-1105, MEDI4736 (durvalumab), MSB0010718C (avelumab), SHR-1316, CS1001, envolimab, TQB2450, ZKAB001, LP-002, CX-072, IMC-001, KL-A167, APL-502, coxilimab, lodalimab, FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, RC98, PDL-GEX, KD036, KY1003, YBL-007, and HS-636. In some instances, the anti-PD-L1 antibody is atezolizumab. Examples of anti-PD-L1 antibodies useful in the methods of the invention and methods for their preparation are described in International Patent Application Publication No. WO 2010 / 077634 and U.S. Patent No. 8,217,149, each of which is incorporated herein by reference in its entirety.

[0419] In some instances, the anti-PD-L1 antibody is avelumab (CAS Registry Number: 1537032-82-8). Avelumab, also known as MSB0010718C, is a human monoclonal IgG1 anti-PD-L1 antibody (Merck KGaA, Pfizer).

[0420] In some instances, the anti-PD-L1 antibody is durvalumab (CAS Registry Number: 1428935-60-7). Durvalumab, also known as MEDI4736, is an Fc-optimized human monoclonal IgG1 kappa anti-PD-L1 antibody described in WO 2011 / 066389 and US 2013 / 034559 (MedImmune, AstraZeneca).

[0421] In some instances, the anti-PD-L1 antibody is MDX-1105 (Bristol Myers Squibb). MDX-1105, also known as BMS-936559, is an anti-PD-L1 antibody described in WO 2007 / 005874.

[0422] In some instances, the anti-PD-L1 antibody is LY3300054 (Eli Lilly).

[0423] In some instances, the anti-PD-L1 antibody is STI-A1014 (Sorrento). STI-A1014 is a human anti-PD-L1 antibody.

[0424] In some cases, the anti-PD-L1 antibody is KN035 (Suzhou Alphamab). KN035 is a single domain antibody (dAB) generated from a camelid phage display library.

[0425] In some cases, the anti-PD-L1 antibody comprises a cleavable portion or linker that, when cleaved (e.g., by a protease in the tumor microenvironment), activates the antibody antigen-binding domain to bind its antigen, e.g., by removing a non-binding steric portion. In some cases, the anti-PD-L1 antibody is CX-072 (CytomX Therapeutics).

[0426] In some instances, the anti-PD-L1 antibody comprises six HVR sequences (e.g., three heavy chain HVRs and three light chain HVRs) and / or heavy chain variable domains and light chain variable domains from an anti-PD-L1 antibody described in US20160108123, WO 2016 / 000619, WO 2012 / 145493, U.S. Pat. No. 9,205,148, WO 2013 / 181634, or WO 2016 / 061142.

[0427] In some cases, the PD-1 axis binding antagonist is a PD-1 binding antagonist. For example, in some cases, the PD-1 binding antagonist inhibits the binding of PD-1 to one or more of its ligand binding partners. In some cases, the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1. In other cases, the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L2. In some other cases, the PD-1 binding antagonist inhibits the binding of PD-1 to both PD-L1 and PD-L2. The PD-1 binding antagonist can be, but is not limited to, an antibody, an antigen binding fragment thereof, an immunoadhesin, a fusion protein, an oligopeptide or a small molecule. In some cases, the PD-1 binding antagonist is an immunoadhesin (e.g., an extracellular or PD-1 binding portion of a PD-L1 or PD-L2 fused to a constant region (e.g., an Fc region of an immunoglobulin sequence)). For example, in some cases, the PD-1 binding antagonist is an Fc fusion protein. In some cases, the PD-1 binding antagonist is AMP-224. AMP-224 is also called B7-DCIg, which is a PD-L2-Fc fusion soluble receptor described in WO 2010 / 027827 and WO 2011 / 066342. In some cases, the PD-1 binding antagonist is a peptide or a small molecule compound. In some cases, the PD-1 binding antagonist is AUNP-12 (Pierre Fabre / Aurigene). See, for example, WO 2012 / 168944, WO 2015 / 036927, WO2015 / 044900, WO 2015 / 033303, WO 2013 / 144704, WO 2013 / 132317, and WO 2011 / 161699. In some instances, a PD-1 binding antagonist is a small molecule that inhibits PD-1.

[0428] In some cases, the PD-1 binding antagonist is an anti-PD-1 antibody. A variety of anti-PD-1 antibodies can be utilized in the methods and uses disclosed herein. In any case herein, the PD-1 antibody can bind to human PD-1 or a variant thereof. In some cases, the anti-PD-1 antibody is a monoclonal antibody. In some cases, the anti-PD-1 antibody is an antibody fragment selected from the group consisting of: Fab, Fab', Fab'-SH, Fv, scFv and (Fab') 2Fragment. In some cases, the anti-PD-1 antibody is a humanized antibody. In other cases, the anti-PD-1 antibody is a human antibody. Exemplary anti-PD-1 antagonist antibodies include nivolumab, pembrolizumab, MEDI-0680, PDR001 (spartalizumab), REGN2810 (cemiprilimab), BGB-108, palolizumab, carrelizumab, sintilimab, tislelizumab, toripalimab, dotalimumab, rivulimab, sazanlimab, penampalimab, CS1003, HLX10, SCT-I10A, sepalimab, batilizumab, genomumab, BI 754091, silimab, YBL-006, BAT1306, HX008, buglilimab, AMG 404, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, AM0001, ENUM 244C8, ENUM 388D4, STI-1110, AK-103, and hAb21.

[0429] In some instances, the anti-PD-1 antibody is nivolumab (CAS Registry Number: 946414-94-4). Nivolumab (Bristol-Myers Squibb / Ono Pharmaceutical (Ono)), also known as MDX-1106-04, MDX-1106, ONO-4538, BMS-936558 and It is the anti-PD-1 antibody described in WO 2006 / 121168.

[0430] In some instances, the anti-PD-1 antibody is pembrolizumab (CAS Registry Number: 1374853-91-4). Pembrolizumab (Merck), also known as MK-3475, Merck 3475, pembrolizumab, SCH-900475, and It is the anti-PD-1 antibody described in WO 2009 / 114335.

[0431] In some instances, the anti-PD-1 antibody is MEDI-0680 (AMP-514; AstraZeneca). MEDI-0680 is a humanized IgG4 anti-PD-1 antibody.

[0432] In some cases, the anti-PD-1 antibody is PDR001 (CAS Registry No. 1859072-53-9; Novartis). PDR001 is a humanized IgG4 anti-PD-1 antibody that blocks the binding of PD-L1 and PD-L2 to PD-1.

[0433] In some instances, the anti-PD-1 antibody is REGN2810 (Regeneron). REGN2810 is a human anti-PD-1 antibody.

[0434] In some instances, the anti-PD-1 antibody is BGB-108 (BeiGene).

[0435] In some instances, the anti-PD-1 antibody is BGB-A317 (BeiGene).

[0436] In some cases, the anti-PD-1 antibody is JS-001 (Shanghai Junshi Biosciences). JS-001 is a humanized anti-PD-1 antibody.

[0437] In some instances, the anti-PD-1 antibody is STI-A1110 (Sorrento). STI-A1110 is a human anti-PD-1 antibody.

[0438] In some instances, the anti-PD-1 antibody is INCSHR-1210 (Incyte). INCSHR-1210 is a human IgG4 anti-PD-1 antibody.

[0439] In some instances, the anti-PD-1 antibody is PF-06801591 (Pfizer).

[0440] In some instances, the anti-PD-1 antibody is TSR-042 (also known as ANB011; Tesaro / AnaptysBio).

[0441] In some instances, the anti-PD-1 antibody is AM0001 (ARMO Biosciences).

[0442] In some cases, the anti-PD-1 antibody is ENUM 244C8 (Enumeral Biomedical Holdings). ENUM 244C8 is an anti-PD-1 antibody that inhibits the function of PD-1 without preventing the binding of PD-L1 to PD-1.

[0443] In some cases, the anti-PD-1 antibody is ENUM 388D4 (Enumeral Biomedical Holdings). ENUM 388D4 is an anti-PD-1 antibody that competitively inhibits the binding of PD-L1 to PD-1.

[0444] In some cases, the anti-PD-1 antibody comprises six HVR sequences (e.g., three heavy chain HVRs and three light chain HVRs) and / or heavy chain variable domains and light chain variable domains from the anti-PD-1 antibodies described in WO 2015 / 112800, WO 2015 / 112805, WO 2015 / 112900, US20150210769, WO 2016 / 089873, WO 2015 / 035606, WO 2015 / 085847, WO 2014 / 206107, WO 2012 / 145493, US 9,205,148, WO 2015 / 119930, WO 2015 / 119923, WO 2016 / 032927, WO 2014 / 179664, WO 2016 / 106160 and WO 2014 / 194302.

[0445] In some cases, the PD-1 axis binding antagonist is a PD-L2 binding antagonist. In some cases, the PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to its ligand binding partner. In specific aspects, the PD-L2 binding ligand partner is PD-1. The PD-L2 binding antagonist can be, but is not limited to, an antibody, an antigen binding fragment thereof, an immunoadhesin, a fusion protein, an oligopeptide, or a small molecule.

[0446] In some cases, the PD-L2 binding antagonist is an anti-PD-L2 antibody. In any case herein, the anti-PD-L2 antibody can bind to human PD-L2 or a variant thereof. In some cases, the anti-PD-L2 antibody is a monoclonal antibody. In some cases, the anti-PD-L2 antibody is an antibody fragment selected from the group consisting of: Fab, Fab', Fab'-SH, Fv, scFv and (Fab') 2 Fragments. In some cases, the anti-PD-L2 antibody is a humanized antibody. In other cases, the anti-PD-L2 antibody is a human antibody. In further specific aspects, the anti-PD-L2 antibody has reduced or minimal effector function. In further specific aspects, the minimal effector function comes from an "effectorless Fc mutation" or an aglycosylation mutation. In further cases, the effectorless Fc mutation is an N297A or D265A / N297A substitution in the constant region. In some cases, the isolated anti-PD-L2 antibody is aglycosylated.

[0447] xvii. Growth inhibitors

[0448] In some aspects, one or more additional therapeutic agents include growth inhibitors. Exemplary growth inhibitors include agents that block cell cycle progression outside the S phase, such as agents that induce G1 arrest (e.g., DNA alkylating agents such as tamoxifen, prednisone, dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, or ara-C) or agents that arrest in the M phase (e.g., vincristine, vinblastine, taxanes (e.g., paclitaxel and docetaxel), doxorubicin, epirubicin, daunorubicin, etoposide, or bleomycin).

[0449] xviii.Radiotherapy

[0450] In some aspects, one or more additional therapeutic agents include radiation therapy. Radiation therapy includes the use of directed gamma rays or beta rays to induce sufficient damage to cells, thereby limiting the ability of cells to function normally or completely destroying cells. Typical treatment is a single administration, and typical dosage ranges are 10 to 200 units (Gray) per day.

[0451] xix. Cytotoxic agents

[0452] In some aspects, the additional therapeutic agent is a cytotoxic agent, such as a substance that inhibits or prevents cell function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioactive isotopes (e.g., At 211 ,I 131 ,I 125 , Y 90 、Re 186 、Re 188 、Sm 153 、Bi 212 , P 32 , Pb 212 and radioactive isotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitors; enzymes and fragments thereof such as nucleolytic enzymes; antibiotics; toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof; and anti-tumor or anti-cancer drugs.

[0453] xx. Additional anticancer therapies

[0454] In some cases, the method further involves administering an effective amount of an additional therapeutic agent to the patient. In some cases, the additional therapeutic agent is selected from the group consisting of the following items: anti-tumor agents, chemotherapeutic agents, growth inhibitors, anti-angiogenic agents, radiotherapy, cytotoxic agents and combinations thereof. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with chemotherapy or chemotherapeutic agents. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with radiotherapy agents. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with targeted therapy or targeted therapeutic agents. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with immunotherapy or immunotherapeutic agents (e.g., monoclonal antibodies). In some cases, the additional therapeutic agent is an agonist for costimulatory molecules. In some cases, the additional therapeutic agent is an antagonist for costimulatory molecules.

[0455] Without wishing to be bound by theory, it is believed that enhancing T cell stimulation by promoting co-stimulatory molecules or by inhibiting co-inhibitory molecules can promote tumor cell death, thereby treating or delaying the development of cancer. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies can be co-administered with agonists for co-stimulatory molecules. In some cases, co-stimulatory molecules may include CD40, CD226, CD28, OX40, GITR, CD137, CD27, HVEM or CD127. In some cases, agonists for co-stimulatory molecules are agonist antibodies that bind to CD40, CD226, CD28, OX40, GITR, CD137, CD27, HVEM or CD127. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies can be co-administered with antagonists for co-inhibitory molecules. In some cases, the co-inhibitory molecule may include CTLA-4 (also known as CD152), TIM-3, BTLA, VISTA, LAG-3, B7-H3, B7-H4, IDO, TIGIT, MICA / B, or arginase. In some cases, the antagonist to the co-inhibitory molecule is an antagonist antibody that binds to CTLA-4, TIM-3, BTLA, VISTA, LAG-3, B7-H3, B7-H4, IDO, TIGIT, MICA / B, or arginase.

[0456] In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an antagonist to CTLA-4 (also known as CD152), such as a blocking antibody. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with ipilimumab (also known as MDX-010, MDX-101, or ). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody can be co-administered with tremelimumab (also known as ticilimumab or CP-675,206). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with an antagonist to B7-H3 (also known as CD276), such as a blocking antibody. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with MGA271. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with an antagonist to TGF-β, such as metilimumab (also known as CAT-192), fresolimumab (also known as GC1008), or LY2157299.

[0457] In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be administered in combination with a treatment comprising adoptive transfer of T cells (e.g., cytotoxic T cells or CTLs) expressing a chimeric antigen receptor (CAR). In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be administered in combination with a treatment comprising adoptive transfer of T cells comprising dominant negative TGFβ receptors, such as dominant negative TGFβ type II receptors. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be administered in combination with a treatment comprising a HERCREEM regimen (see, e.g., ClinicalTrials.gov identifier NCT00889954).

[0458] In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with agonists such as activating antibodies for CD137 (also known as TNFRSF9, 4-1BB or ILA). In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with urelumab (also known as BMS-663513). In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with agonists such as activating antibodies for CD40. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with CP-870893. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with agonists such as CD134 for OX40, such as activating antibodies. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with an anti-OX40 antibody (e.g., AgonOX). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with an agonist for CD27, such as an activating antibody. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with CDX-1127. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with an antagonist for indoleamine-2,3-dioxygenase (IDO). In some cases, the IDO antagonist is 1-methyl-D-tryptophan (also known as 1-D-MT).

[0459] In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an antibody-drug conjugate. In some cases, the antibody-drug conjugate comprises mertansine or monomethyl auristatin E (MMAE). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an anti-NaPi2b antibody-MMAE conjugate (also known as DNIB0600A or RG7599). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with trastuzumab emtansine (also known as T-DM1, ado-trastuzumab emtansine) or In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with DMUC5754A. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an antibody-drug conjugate that targets the endothelin B receptor (EDNBR), for example, a conjugate of an antibody to EDNBR with MMAE.

[0460] In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an anti-angiogenic agent. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an antibody to VEGF, such as VEGF-A. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with bevacizumab (also known as In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an antibody against angiopoietin 2 (also known as Ang2). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with MEDI3617.

[0461] In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an anti-tumor agent. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with a drug targeting CSF-1R (also known as M-CSFR or CD115). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with anti-CSF-1R (also known as IMC-CS4). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an interferon, such as interferon alpha or interferon gamma. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with Roferon-A (also known as recombinant interferon alpha-2a). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be combined with GM-CSF (also known as recombinant human granulocyte macrophage colony stimulating factor, rhu GM-CSF, sargramostim or ). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with IL-2 (also known as aldesleukin or ). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with IL-12. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an antibody targeting CD20. In some cases, the antibody targeting CD20 is obinutuzumab (also known as GA101 or ) or rituximab. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an antibody targeting GITR. In some cases, the antibody targeting GITR is TRX518.

[0462] In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with a cancer vaccine. In some cases, the cancer vaccine is a peptide cancer vaccine, which in some cases is a personalized peptide vaccine. In some cases, the peptide cancer vaccine is a multivalent long peptide, a polypeptide, a peptide mixture, a hybrid peptide, or a peptide pulsed dendritic cell vaccine (see, e.g., Yamada et al., Cancer Sci. 104: 14-21, 2013). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an adjuvant. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered with a TLR agonist such as Poly-ICLC (also known as ), LPS, MPL or CpG ODN. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with tumor necrosis factor (TNF) α. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with IL-1. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with HMGB1. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with IL-10 antagonists. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with IL-4 antagonists. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with IL-13 antagonists. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be co-administered with HVEM antagonists. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be administered in combination with ICOS agonists, for example, by administering ICOS-L or an agonistic antibody to ICOS. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be administered in combination with a treatment targeting CX3CL1. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be administered in combination with a treatment targeting CXCL9. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be administered in combination with a treatment targeting CXCL10. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be administered in combination with a treatment targeting CCL5. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be administered in combination with an LFA-1 or ICAM1 agonist. In some cases, bispecific anti-FcRH5 / anti-CD3 antibodies and lenalidomide can be administered in combination with a selectin agonist.

[0463] In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with a targeted therapy. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an inhibitor of B-Raf. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with vemurafenib (also known as In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with dabrafenib (also known as ). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with erlotinib (also known as ). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an inhibitor of MEK, such as MEK1 (also known as MAP2K1) or MEK2 (also known as MAP2K2). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with cobimetinib (also known as GDC-0973 or XL-518). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with trametinib (also known as ). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with an inhibitor of K-Ras. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with an inhibitor of c-Met. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with onartuzumab (also known as MetMAb). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with an inhibitor of Alk. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with AF802 (also known as CH5424802 or alectinib). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with an inhibitor of phosphatidylinositol 3-kinase (PI3K). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with BKM120. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with idelalisib (also known as GS-1101 or CAL-101). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with perifosine (also known as KRX-0401). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an inhibitor of Akt. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with MK2206. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody can be administered in combination with GSK690693. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with GDC-0941. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with an inhibitor of mTOR. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with sirolimus (also known as rapamycin). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with temsirolimus (also known as CCI-779 or ). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with everolimus (also known as RAD001). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be used in combination with ridaforolimus (also known as AP-23573, MK-8669 or deforolimus). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with OSI-027. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with AZD8055. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with INK128. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be co-administered with a dual PI3K / mTOR inhibitor. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with XL765. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with GDC-0980. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with anti-BEZ235 (also known as NVP-BEZ235). In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with BGT226. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with GSK2126458. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with PF-04691502. In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with anti-PF-05212384 (also known as PKI-587).

[0464] In some cases, the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can be administered in combination with a chemotherapeutic agent. A chemotherapeutic agent is a chemical compound used to treat cancer. Exemplary chemotherapeutic agents include, but are not limited to, erlotinib ( Genentech / OSIPharm.), anti-hormonal agents used to modulate or inhibit the effects of hormones on tumors (such as antiestrogens and selective estrogen receptor modulators (SERMs)), such as alemtuzumab (Campath), bevacizumab ( Genentech) antibodies; Cetuximab ( Imclone; Panitumumab ( Amgen), rituximab ( Genentech / Biogen Idec), Pertuzumab ( 2C4, Genentech) or trastuzumab ( Genentech), EGFR inhibitors (EGFR antagonists), tyrosine kinase inhibitors and chemotherapeutic agents, and also include nonsteroidal anti-inflammatory drugs (NSAIDs) with analgesic, antipyretic and anti-inflammatory effects.

[0465] In the case where the methods described herein involve combination therapy such as the specific combination therapy mentioned above, the combination therapy includes co-administration of the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide with one or more additional therapeutic agents, and such co-administration can be combined (wherein the two or more therapeutic agents are contained in the same or separate formulations) or separately, in which case the administration of the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide can occur before, simultaneously with, and / or after the administration of the one or more additional therapeutic agents. In one embodiment, the administration of the bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide and the administration of the additional therapeutic agent or exposure to radiation therapy are performed within about one month, or within about one, two, three, four, five, or six days of each other.

[0466] In some aspects, the subject does not have an increased risk of CRS (e.g., has not experienced Grade 3+ CRS during treatment with the bispecific antibody or CAR-T therapy; does not have detectable circulating plasma cells; and / or does not have extensive extramedullary disease).

[0467] C. Cancer

[0468] Any of the methods of the invention described herein can be used to treat cancer (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM (e.g., MM with a high-risk cytogenetic profile)))). In some aspects, the subject's cancer has one or more high-risk cytogenetic features. In some examples, the high-risk cytogenetic features include one or more of the following: translocation events t(4;14) or t(14;16), del(17p), or 1q gain.

[0469] Other examples of B-cell proliferative disorders / malignancies suitable for treatment with the bispecific anti-FcRH5 / anti-CD3 antibodies according to the methods described herein include, but are not limited to, non-Hodgkin lymphoma (NHL) (including diffuse large B-cell lymphoma (DLBCL), which may be relapsed or refractory DLBCL) and other cancers including: germinal center B-cell-like (GCB) diffuse large B-cell lymphoma (DLBCL), activated B-cell-like (ABC) DLBCL, follicular lymphoma (FL), mantle cell lymphoma (MCL), and leukemia. CL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), marginal zone lymphoma (MZL), small lymphocytic leukemia (SLL), lymphoplasmacytic lymphoma (LL), Waldenstrom's macroglobulinemia (WM), central nervous system lymphoma (CNSL), Burkitt's lymphoma (BL), precursor B-cell lymphocytic leukemia, splenic marginal zone lymphoma, hairy cell leukemia, splenic lymphoma / leukemia (unclassifiable), splenic diffuse red pulp small B-cell lymphoma, variant hairy cell leukemia, Waldenstrom's macroglobulinemia (WM), Oeldenstrom's macroglobulinemia, heavy chain disease (alpha heavy chain disease, gamma heavy chain disease, mu heavy chain disease), plasma cell myeloma, solitary plasmacytoma of bone, extraosseous plasmacytoma, extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma), lymph node marginal zone lymphoma, pediatric lymph node marginal zone lymphoma, pediatric follicular lymphoma, primary cutaneous follicle center lymphoma, T cell / histiocyte-rich large B cell lymphoma, primary DLBCL of the CNS, primary cutaneous DLBCL (leg type), EBV-positive DLB in the elderly CL, DLBCL associated with chronic inflammation, lymphomatoid granuloma, primary mediastinal (thymic) large B-cell lymphoma, intravascular large B-cell lymphoma, ALK-positive large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma due to HHV8-associated multicentric Castleman disease, primary effusion lymphoma: B-cell lymphoma (unclassifiable, with features intermediate between DLBCL and Burkitt's lymphoma), and B-cell lymphoma (unclassifiable, with features intermediate between DLBCL and classical Hodgkin's lymphoma). Additional examples of B-cell proliferative disorders include, but are not limited to: multiple myeloma (MM); low-grade / follicular NHL; small lymphocytic (SL) NHL; intermediate-grade / follicular NHL; intermediate-grade diffuse NHL; high-grade immunoblastic NHL; high-grade lymphoblastic NHL; high-grade small non-lytic cell NHL; bulky NHL; AIDS-related lymphoma; and acute lymphoblastic leukemia (ALL); chronic myeloid leukemia; and post-transplant lymphoproliferative disorder (PTLD). Other examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies, including B-cell lymphoma.More specific examples of such cancers include, but are not limited to: low-grade / follicular NHL; small lymphocytic (SL) NHL; intermediate-grade / follicular NHL; intermediate-grade diffuse NHL; high-grade immunoblastic NHL; high-grade lymphoblastic NHL; high-grade small non-truncated cell NHL; bulky disease NHL; AIDS-related lymphoma; and acute lymphoblastic leukemia (ALL); chronic myeloid leukemia; and post-transplant lymphoproliferative disorder (PTLD). Solid tumors that may be amenable to treatment with the bispecific anti-FcRH5 / anti-CD3 antibodies according to the methods described herein include squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer (including small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma), peritoneal cancer, hepatocellular carcinoma, gastric cancer (including gastrointestinal cancer and gastrointestinal stromal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, urinary tract cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, hepatic cancer, In some embodiments, cancers suitable for treatment by the antibodies disclosed herein include breast cancer, colorectal cancer, rectal cancer, non-small cell lung cancer, glioblastoma, non-Hodgkin lymphoma (NHL), renal cell carcinoma, prostate cancer, liver cancer, pancreatic cancer, soft tissue sarcoma, Kaposi's sarcoma, carcinoid tumor, head and neck cancer, ovarian cancer and mesothelioma.

[0470] D. Previous anticancer therapy

[0471] In some aspects, the subject has been previously treated for cancer (eg, a hematological cancer (eg, a B-cell proliferative disorder (eg, MM (eg, MM with a high-risk cytogenetic profile)))).

[0472] In some aspects, the subject has received induction therapy. Any suitable induction therapy can be used. Exemplary induction therapies for MM include, but are not limited to, the CyBorD regimen (cyclophosphamide, bortezomib, and dexamethasone); VRD regimen (bortezomib, lenalidomide, and dexamethasone); VRD lite (reduced dose and schedule of bortezomib, lenalidomide, and dexamethasone); thalidomide and dexamethasone; lenalidomide and low-dose dexamethasone; bortezomib and dexamethasone; Vd regimen (bortezomib and dexamethasone); VTD regimen (bortezomib, thalidomide, and dexamethasone); bortezomib, cyclophosphamide, and prednisone; bortezomib, doxorubicin, and dexamethasone; DARZALEX (daratumumab and hyaluronidase), bortezomib, (melphalan) and prednisone; DARZALEX (daratumumab and hyaluronidase), lenalidomide, and dexamethasone; DARZALEX (daratumumab and hyaluronidase), bortezomib, thalidomide, and dexamethasone; and liposomal doxorubicin, vincristine, and dexamethasone.

[0473] In some aspects, the subject has received autologous stem cell transplantation (ASCT). For example, in some aspects, the subject has received ASCT within about 100 days (e.g., within 100 days, within 90 days, within 80 days, within 70 days, within 60 days, within 50 days, within 40 days, within 30 days, within 20 days, within 10 days, within 5 days, or within 1 day) of the start of the method (e.g., the first administration of bispecific antibodies and / or lenalidomide). In some instances, the subject does not have a progressive disease.

[0474] In some examples, the bispecific antibody and lenalidomide are administered to the patient as post-transplant maintenance therapy.

[0475] In some aspects, the subject has received at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, or more than fifteen lines of treatment for the B-cell proliferative disorder, such as 2L+, 3L+, 4L+, 5L+, 6L+, 7L+, 8L+, 9L+, 10L+, 11L+, 12L+, 13L+, 14L+, or 15L+.

[0476] In some aspects, the subject has received at least three prior lines of treatment for cancer (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM (e.g., MM with a high-risk cytogenetic profile)))), e.g., is 4L+, e.g., has received three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, or more than fifteen lines of treatment. In some aspects, the subject has relapsed or refractory (R / R) multiple myeloma (MM), e.g., has 4L+R / R MM.

[0477] In some aspects, the prior line of therapy includes one or more of the following: a proteasome inhibitor (PI), such as bortezomib, carfilzomib, or ixazomib; an immunomodulatory drug (IMiD), such as thalidomide, lenalidomide, or pomalidomide; an autologous stem cell transplant (ASCT); an anti-CD38 agent, such as daratumumab (U.S. Patent No. 7,829,673 and U.S. Publication No. 20160067205A1), "MOR202" (U.S. Patent No. 8,263,746), isatuximab (SAR-650984); CAR-T therapy; therapy comprising bispecific antibodies; anti-SLAMF7 therapeutic agents (e.g., anti-SLAMF7 antibodies, such as elotuzumab); nuclear export inhibitors (e.g., selinexor); and histone deacetylase (HDAC) inhibitors (e.g., panobinostat). In some aspects, the previous line of treatment includes an antibody-drug conjugate (ADC). In some aspects, the previous line of treatment includes a B cell maturation antigen (BCMA) directed therapy, such as an antibody-drug conjugate (BCMA-ADC) targeting BCMA.

[0478] In some aspects, the prior line of treatment includes all three of a proteasome inhibitor (PI), an IMiD, and an anti-CD38 agent (eg, daratumumab).

[0479] In some aspects, the cancer (e.g., a hematological cancer (e.g., a B cell proliferative disorder (e.g., MM (e.g., MM with a high-risk cytogenetic profile)))) is refractory to a line of treatment, e.g., to one or more of daratumumab, a PI, an IMiD, an ASCT, an anti-CD38 agent, a CAR-T therapy, a therapy comprising a bispecific antibody, an anti-SLAMF7 therapeutic agent, a nuclear export inhibitor, an HDAC inhibitor, an ADC, or a BCMA-directed therapy. In some aspects, the B cell proliferative disorder (e.g., MM) is refractory to daratumumab.

[0480] E. Risk-Benefit Profile

[0481] The methods described herein can result in an improved benefit-risk profile for patients having cancer (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM (e.g., MM with a high-risk cytogenetic profile)))). In some cases, treatment with the methods described herein that result in administration of a bispecific anti-FcRH5 / anti-CD3 antibody and lenalidomide in the context of a fractionated, dose-escalating dosing regimen can result in a reduction in undesirable events such as cytokine-driven toxicities (e.g., cytokine release syndrome (CRS)), infusion-related reactions (IRRs), macrophage activation syndrome (MAS), neurological toxicity, severe

[00136] In some embodiments, the patient's tumor lysis syndrome (TLS), neutropenia, thrombocytopenia, liver enzyme elevations, and / or central nervous system (CNS) toxicity are reduced (e.g., 20% or greater, 25% or greater, 30% or greater, 35% or greater, 40% or greater, 45% or greater, 50% or greater, 55% or greater, 60% or greater, 65% or greater, 70% or greater, 75% or greater, 80% or greater, 85% or greater, 90% or greater, 95% or greater, 96% or greater, 97% or greater, 98% or greater, or 99% or greater) or completely inhibited (100% reduction).

[0482] F. Safety and effectiveness

[0483] i. Security

[0484] In some aspects, less than 15% (e.g., less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%) of patients treated with the methods described herein experience Grade 3 or 4 cytokine release syndrome (CRS). In some aspects, less than 5% of patients treated with the methods described herein experience Grade 3 or 4 CRS.

[0485] In some aspects, less than 10% (e.g., less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2% or less than 1%) of patients treated using the methods described herein experience 4+ CRS. In some aspects, less than 3% of patients treated using the methods described herein experience 4+ CRS. In certain aspects, no patient experiences 4+ CRS.

[0486] In some aspects, less than 10% (e.g., less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%) of patients treated with the methods described herein experience Grade 3 CRS. In some aspects, less than 5% of patients treated with the methods described herein experience Grade 3 CRS. In certain aspects, no patient experiences Grade 3 CRS.

[0487] In some aspects, a Grade 2+ CRS event occurs only in the first treatment cycle. In some aspects, a Grade 2 CRS event occurs only in the first treatment cycle. In certain aspects, no Grade 2 CRS event occurs.

[0488] In some aspects, less than 3% of patients treated using the methods described herein experienced Grade 4+ CRS, less than 5% of patients treated using the methods described herein experienced Grade 3 CRS, and Grade 2+ CRS events occurred only in the first treatment cycle.

[0489] In some aspects, no Grade 3+ CRS events occurred, and Grade 2 CRS events occurred only in the first treatment cycle.

[0490] In some respects, symptoms of immune effector cell-associated neurotoxicity syndrome (ICANS) are limited to confusion, disorientation, and expressive aphasia and resolve with steroid treatment.

[0491] In some aspects, less than 10% (e.g., less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%) of the patients treated using the methods described herein experience epileptic seizures or other grade 3+ neurological adverse events. In some aspects, less than 5% of the patients experience epileptic seizures or other grade 3+ neurological adverse events. In some aspects, no patient experiences epileptic seizures or other grade 3+ neurological adverse events.

[0492] In some aspects, all neurological symptoms are self-limited or resolve with steroid and / or tocilizumab therapy.

[0493] ii. Efficacy

[0494] In some aspects, the total response rate (ORR) of patients treated using the methods described herein is at least 25%, for example, at least 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100%. In some aspects, the ORR is at least 40%. In some aspects, the ORR is at least 45% (for example, at least 45%, 45.5%, 46%, 46.5%, 47%, 47.5%, 48%, 48.5%, 49%, 49.5% or 50%), at least 55% or at least 65%. In some aspects, the ORR is at least 47.2%. In some aspects, the ORR is about 47.2%. In some aspects, the ORR is 75% or greater. In some aspects, at least 1% of patients (e.g., at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, In some aspects, ORR is 40% to 50%, and 10% to 20% of patients have CR or VGPR. In some aspects, the ORR is at least 40%, and at least 20% of patients have a CR or VGPR.

[0495] In some aspects, the average duration of remission (DoR) for patients treated using the methods described herein is at least two months, e.g., at least three months, at least four months, at least five months, at least six months, at least seven months, at least eight months, at least nine months, at least ten months, at least eleven months, at least one year, or more than one year. In some aspects, the average DoR is at least four months. In some aspects, the average DoR is at least five months. In some aspects, the average DoR is at least seven months.

[0496] In some aspects, the six-month progression-free survival (PFS) rate of the patient treated using the methods described herein is at least 10%, for example, at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100%. In some aspects, the six-month PFS rate is at least 25%. In some aspects, the six-month PFS rate is at least 40%. In some aspects, the six-month PFS rate is at least 55%.

[0497] G. Application Method

[0498] The methods and treatments may involve administering the bispecific anti-FcRH5 / anti-CD3 antibody, lenalidomide, and / or any additional therapeutic agent by any suitable means, including parenteral, intrapulmonary, and intranasal, and if local treatment is desired, including intralesional administration. Parenteral infusions include intravenous, subcutaneous, intramuscular, intraarterial, and intraperitoneal administration routes. In some embodiments, the bispecific anti-FcRH5 / anti-CD3 antibody is administered by intravenous infusion. In other cases, the bispecific anti-FcRH5 / anti-CD3 antibody is administered subcutaneously.

[0499] In some cases, a bispecific anti-FcRH5 / anti-CD3 antibody administered by intravenous injection exhibits less toxicity (ie, fewer adverse effects) in patients compared to the same bispecific anti-FcRH5 / anti-CD3 antibody administered by subcutaneous injection, or vice versa.

[0500] In some aspects, the bispecific anti-FcRH5 / anti-CD3 antibody is administered intravenously over 4 hours (±15 minutes), eg, the first dose of the antibody is administered over 4 hours ±15 minutes.

[0501] In some aspects, the first and second doses of the antibody are administered intravenously with a median infusion time of less than four hours (e.g., less than three hours, less than two hours, or less than one hour), and the additional doses of the antibody are administered intravenously with a median infusion time of less than 120 minutes (e.g., less than 90 minutes, less than 60 minutes, or less than 30 minutes).

[0502] In some aspects, the first and second doses of the antibody are administered intravenously with a median infusion time of less than three hours, and the additional dose of the antibody is administered intravenously with a median infusion time of less than 90 minutes.

[0503] In some aspects, the first and second doses of the antibody are administered intravenously with a median infusion time of less than three hours, and the additional doses of the antibody are administered intravenously with a median infusion time of less than 60 minutes. In some aspects, the patient is hospitalized (e.g., hospitalized for 72 hours, 48 ​​hours, 24 hours, or less than 24 hours) during one or more administrations of the anti-FcRH5 / anti-CD3 antibody, such as for C1D1 (Cycle 1, Dose 1) or C1D1 and C1D2 (Cycle 1, Dose 2). In some aspects, the patient is hospitalized for 72 hours after administering C1D1 and C1D2. In some aspects, the patient is hospitalized for 24 hours after administering C1D1 and C1D2. In some aspects, the patient is not hospitalized after administering any dose of the anti-FcRH5 / anti-CD3 antibody.

[0504] For all methods described herein, the bispecific anti-FcRH5 / anti-CD3 antibodies, lenalidomide and / or any additional therapeutic agent will be formulated, dosed and administered in a manner consistent with good medical practice. Factors to be considered in this context include the specific disorder being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the timing of administration, and other factors known to practitioners. The bispecific anti-FcRH5 / anti-CD3 antibodies, lenalidomide and / or any additional therapeutic agent need not be, but are optionally, formulated with one or more agents currently used to prevent or treat the disease in question. The effective amount of such other agents depends on the amount of the bispecific anti-FcRH5 / anti-CD3 antibodies, lenalidomide and / or any additional therapeutic agent present in the formulation, the type of disease or treatment, and the other factors described above. The bispecific anti-FcRH5 / anti-CD3 antibodies, lenalidomide and / or any additional therapeutic agent may be appropriately administered to the patient over a series of treatments.

[0505] H. Anti-FcRH5 / anti-CD3 bispecific antibody

[0506] The methods described herein include administering a bispecific antibody that binds to CD79b and CD3 (i.e., a bispecific anti-CD79b / anti-CD3 antibody) to a subject having cancer (e.g., a hematological cancer (e.g., a B-cell proliferative disorder (e.g., MM (e.g., MM with high-risk cytogenetic characteristics)))). Any suitable bispecific antibody that binds to CD79b and CD3 (i.e., a bispecific anti-CD79b / anti-CD3 antibody) can be used.

[0507] In some cases, any of the methods described herein may comprise administering a bispecific antibody comprising an anti-FcRH5 arm having a first binding domain comprising at least one, two, three, four, five or six hypervariable regions (HVRs) selected from: (a) HVR-H1 comprising the amino acid sequence of RFGVH (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of VIWRGGSTDYNAAFVS (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of HYYGSSDYALDN (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of KASQDVRNLVV (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of SGSYRYS (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QQHYSPPYT (SEQ ID NO: 6). In some aspects, the bispecific anti-FcRH5 / anti-CD3 antibody comprises: at least one (e.g., 1, 2, 3, or 4) of the heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 17 to 20, respectively; and / or at least one (e.g., 1, 2, 3, or 4) of the light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 21 to 24, respectively.

[0508] In some cases, any of the methods described herein may include administering a bispecific antibody comprising an anti-FcRH5 arm having a first binding domain comprising the following six HVRs: (a) HVR-H1 comprising the amino acid sequence of RFGVH (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of VIWRGGSTDYNAAFVS (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of HYYGSSDYALDN (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of KASQDVRNLVV (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of SGSYRYS (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QQHYSPPYT (SEQ ID NO: 6). In some aspects, the bispecific anti-FcRH5 / anti-CD3 antibody comprises: at least one (e.g., 1, 2, 3, or 4) of the heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 17 to 20, respectively; and / or at least one (e.g., 1, 2, 3, or 4) of the light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 21 to 24, respectively.

[0509] In some cases, the bispecific antibody comprises an anti-CD79b arm comprising a first binding domain comprising: (a) a heavy chain variable (VH) domain (VH) comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, or a sequence of SEQ ID NO:7; (b) a light chain variable (VL) domain (VL) comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, or a sequence of SEQ ID NO:8; or (c) a VH domain as described in (a) and a VL domain as described in (b). Thus, in some cases, the first binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO:7; and a VL domain comprising the amino acid sequence of SEQ ID NO:8.

[0510] In some cases, any of the methods described herein may comprise administering a bispecific anti-FcRH5 / anti-CD3 antibody comprising an anti-CD3 arm having a second binding domain comprising at least one, two, three, four, five or six HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SYYIH (SEQ ID NO:9); (b) HVR-H2 comprising the amino acid sequence of WIYPENDNTKYNEKFKD (SEQ ID NO:10); (c) HVR-H3 comprising the amino acid sequence of DGYSRYYFDY (SEQ ID NO:11); (d) HVR-L1 comprising the amino acid sequence of KSSQSLLNSRTRKNYLA (SEQ ID NO:12); (e) HVR-L2 comprising the amino acid sequence of WTSTRKS (SEQ ID NO:13); and (f) HVR-L3 comprising the amino acid sequence of KQSFILRT (SEQ ID NO:14). In some aspects, the anti-FcRH5 / anti-CD3 bispecific antibody comprises: at least one (e.g., 1, 2, 3, or 4) of the heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 25 to 28, respectively; and / or at least one (e.g., 1, 2, 3, or 4) of the light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 29 to 32, respectively.

[0511] In some cases, any of the methods described herein may include administering a bispecific anti-FcRH5 / anti-CD3 antibody comprising an anti-CD3 arm having a second binding domain comprising the following six HVRs: (a) HVR-H1 comprising the amino acid sequence of SYYIH (SEQ ID NO:9); (b) HVR-H2 comprising the amino acid sequence of WIYPENDNTKYNEKFKD (SEQ ID NO:10); (c) HVR-H3 comprising the amino acid sequence of DGYSRYYFDY (SEQ ID NO:11); (d) HVR-L1 comprising the amino acid sequence of KSSQSLLNSRTRKNYLA (SEQ ID NO:12), (e) HVR-L2 comprising the amino acid sequence of WTSTRKS (SEQ ID NO:13); and (f) HVR-L3 comprising the amino acid sequence of KQSFILRT (SEQ ID NO:14). In some aspects, the anti-FcRH5 / anti-CD3 bispecific antibody comprises: at least one (e.g., 1, 2, 3, or 4) of the heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 25 to 28, respectively; and / or at least one (e.g., 1, 2, 3, or 4) of the light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 29 to 32, respectively.

[0512] In some cases, the bispecific antibody comprises an anti-CD3 arm comprising a second binding domain comprising: (a) a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, or the sequence of, SEQ ID NO: 15; (b) a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, or the sequence of SEQ ID NO: 16; or (c) a VH domain as described in (a) and a VL domain as described in (b). Thus, in some cases, the second binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO:15; and a VL domain comprising the amino acid sequence of SEQ ID NO:16.

[0513] In some cases, any of the methods described herein may include administering a bispecific antibody comprising: (1) an anti-FcRH5 arm having a first binding domain comprising at least one, two, three, four, five, or six HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of RFGVH (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of VIWRGGSTDYNAAFVS (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of HYYGSSDYALDN (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of KASQDVRNLVV (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of SGSYRYS (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QQHYSPPYT (SEQ ID NO: 6). NO:6); and (2) an anti-CD3 arm having a second binding domain, the second binding domain comprising at least one, two, three, four, five or six HVRs selected from the following: (a) HVR-H1, which comprises the amino acid sequence of SYYIH (SEQ ID NO:9); (b) HVR-H2, which comprises the amino acid sequence of WIYPENDNTKYNEKFKD (SEQ ID NO:10); (c) HVR-H3, which comprises the amino acid sequence of DGYSRYYFDY (SEQ ID NO:11); (d) HVR-L1, which comprises the amino acid sequence of KSSQSLLNSRTRKNYLA (SEQ ID NO:12); (e) HVR-L2, which comprises the amino acid sequence of WTSTRKS (SEQ ID NO:13); and (f) HVR-L3, which comprises the amino acid sequence of KQSFILRT (SEQ ID NO:14).

[0514] In some cases, any of the methods described herein may include administering a bispecific antibody comprising: (1) an anti-FcRH5 arm having a first binding domain comprising the following six HVRs: (a) HVR-H1 comprising the amino acid sequence of RFGVH (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of VIWRGGSTDYNAAFVS (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of HYYGSSDYALDN (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of KASQDVRNLVV (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of SGSYRYS (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QQHYSPPYT (SEQ ID NO: 6). NO:6); and (2) an anti-CD3 arm having a second binding domain comprising the following six HVRs: (a) HVR-H1, which comprises the amino acid sequence of SYYIH (SEQ ID NO:9); (b) HVR-H2, which comprises the amino acid sequence of WIYPENDNTKYNEKFKD (SEQ ID NO:10); (c) HVR-H3, which comprises the amino acid sequence of DGYSRYYFDY (SEQ ID NO:11); (d) HVR-L1, which comprises the amino acid sequence of KSSQSLLNSRTRKNYLA (SEQ ID NO:12), (e) HVR-L2, which comprises the amino acid sequence of WTSTRKS (SEQ ID NO:13); and (f) HVR-L3, which comprises the amino acid sequence of KQSFILRT (SEQ ID NO:14).

[0515] In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody comprises: (1) at least one (e.g., 1, 2, 3, or 4) of the heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 17 to 20, respectively, and / or at least one (e.g., 1, 2, 3, or 4) of the light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 21 to 24, respectively; and (2) at least one (e.g., 1, 2, 3, or 4) of the heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 25 to 28, respectively, and / or at least one (e.g., 1, 2, 3, or 4) of the light chain framework regions FR- At least one (e.g., 1, 2, 3 or 4) of the light chain framework regions FR-L1, FR-L2, FR-L3 and FR-L4 of sequences of NO: 29 to 32. In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody comprises: (1) all four of the heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 17 to 20, respectively, and / or all four of the light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 21 to 24, respectively; and (2) all four of the heavy chain framework regions FR-H1, FR-H2, FR-H3, and FR-H4 comprising the sequences of SEQ ID NOs: 25 to 28, respectively, and / or all four (e.g., 1, 2, 3, or 4) of the light chain framework regions FR-L1, FR-L2, FR-L3, and FR-L4 comprising the sequences of SEQ ID NOs: 29 to 32, respectively.

[0516] In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody comprises: (1) an anti-FcRH5 arm comprising a first binding domain, the first binding domain comprising: (a) a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to or having an amino acid sequence of SEQ ID NO:7; (b) a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to or having an amino acid sequence of SEQ ID NO:7; NO:8 has at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity) or an amino acid sequence of that sequence; or (c) a VH domain as described in (a) and a VL domain as described in (b); and (2) an anti-CD3 arm comprising a second binding domain, the second binding domain comprising: (a) a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity) to a sequence of SEQ ID NO:15; (b) a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity) to a sequence of SEQ ID NO:16; NO:16 has at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity) or an amino acid sequence of the sequence; or (c) a VH domain as described in (a) and a VL domain as described in (b). In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody comprises: (1) a first binding domain comprising a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO:7, and the VL domain comprising the amino acid sequence of SEQ ID NO:8; and (2) a second binding domain comprising a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO:15, and the VL domain comprising the amino acid sequence of SEQ ID NO:16.

[0517] In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody comprises an anti-FcRH5 arm comprising a heavy chain polypeptide (H1) and a light chain polypeptide (L1), wherein (a) H1 comprises an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, or a sequence of SEQ ID NO:35; and (b) L1 comprises an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, or a sequence of SEQ ID NO:36.

[0518] In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody comprises an anti-FcRH5 arm comprising a heavy chain polypeptide (H1) and a light chain polypeptide (L1), wherein (a) H1 comprises the amino acid sequence of SEQ ID NO:35; and / or (b) L1 comprises the amino acid sequence of SEQ ID NO:36.

[0519] In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody comprises an anti-CD3 arm comprising a heavy chain polypeptide (H2) and a light chain polypeptide (L2), wherein (a) H2 comprises an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, or a sequence of SEQ ID NO:37; and (b) L2 comprises an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, or a sequence of SEQ ID NO:38.

[0520] In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody comprises an anti-CD3 arm comprising a heavy chain polypeptide (H2) and a light chain polypeptide (L2), wherein (a) H2 comprises the amino acid sequence of SEQ ID NO:37; and (b) L2 comprises the amino acid sequence of SEQ ID NO:38.

[0521] In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody comprises: an anti-FcRH5 arm comprising a heavy chain polypeptide (H1) and a light chain polypeptide (L1); and an anti-CD3 arm comprising a heavy chain polypeptide (H2) and a light chain polypeptide (L2), and wherein (a) H1 comprises an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a sequence of SEQ ID NO: 35, or such sequence; (b) L1 comprises an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a sequence of SEQ ID NO: 36, or such sequence; (c) H2 comprises an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a sequence of SEQ ID NO: NO:37 has at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) or an amino acid sequence of that sequence; and (d) L2 comprises an amino acid sequence that has at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a sequence of SEQ ID NO:38 or that sequence.

[0522] In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody comprises: an anti-FcRH5 arm comprising a heavy chain polypeptide (H1) and a light chain polypeptide (L1); and an anti-CD3 arm comprising a heavy chain polypeptide (H2) and a light chain polypeptide (L2), and wherein (a) H1 comprises the amino acid sequence of SEQ ID NO:35; (b) L1 comprises the amino acid sequence of SEQ ID NO:36; (c) H2 comprises the amino acid sequence of SEQ ID NO:37; and (d) L2 comprises the amino acid sequence of SEQ ID NO:38.

[0523] In some instances, the anti-FcRH5 / anti-CD3 bispecific antibody is sivostatin.

[0524] In some cases, the anti-FcRH5 / anti-CD3 bispecific antibody according to any of the above embodiments may incorporate any of the features, alone or in combination, as described in Sections 1 to 7 below.

[0525] 1. Antibody affinity

[0526] In certain embodiments, the antibodies provided herein have a molecular weight of ≤1 μM, ≤250 nM, ≤100 nM, ≤15 nM, ≤10 nM, ≤6 nM, ≤4 nM, ≤2 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or ≤0.001 nM (e.g., 10 -8M or less, for example, 10 -8 M to 10 -13 M, for example, 10 -9 M to 10 -13 The dissociation constant (K D ).

[0527] In one embodiment, K is measured by radiolabeled antigen binding assay (RIA). D In one embodiment, RIA is performed using a Fab form of the target antibody and its antigen. For example, by titrating the antigen with a minimum concentration ( 125 The solution binding affinity of Fab for antigen is measured by equilibrating Fab with 1) labeled antigen and then capturing bound antigen with a plate coated with anti-Fab antibody (see, e.g., Chen et al., J. Mol. Biol. 293:865-881 (1999)). To determine the conditions for the assay, 5 μg / ml capture anti-Fab antibody (CappelLabs) was coated in 50 mM sodium carbonate (pH 9.6). Multiwell plates (Thermo Scientific) were blocked overnight and then blocked with 2% (w / v) bovine serum albumin in PBS for two to five hours at room temperature (approximately 23°C). In non-adsorbent plates (Nunc #269620), 100 pM or 26 pM [ 125 I]-antigen is mixed with serial dilutions of the Fab of interest (e.g., following the evaluation of anti-VEGF antibody (Fab-12) in Presta et al., Cancer Res. 57:4593-4599 (1997)). The Fab of interest is then incubated overnight; however, the incubation can be continued for a longer time (e.g., about 65 hours) to ensure that equilibrium is reached. Thereafter, the mixture is transferred to a capture plate for incubation at room temperature (e.g., one hour). The solution is then removed and washed with 0.1% polysorbate 20 in PBS. The plates were washed eight times. When the plates were dry, 150 μl / well of scintillant (MICROSCINT-20 TM ; Packard), and in TOPCOUNT TM The plates were counted for tens of minutes on a gamma counter (Packard).The concentration of each Fab that gave less than or equal to 20% of maximal binding was chosen for use in the competitive binding assay.

[0528] According to another embodiment, using Surface plasmon resonance assay to measure K D For example, using -2000 or -3000 (BIAcore, Inc., Piscataway, NJ) at 37°C with an immobilized antigen CM5 chip at ~10 response units (RU). In one embodiment, a carboxymethylated dextran biosensor chip (CM5, BIACORE, Inc.) was activated with N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the supplier's instructions. The antigen was diluted to 5 μg / ml (about 0.2 μM) with 10 mM sodium acetate pH 4.8 and then injected at a flow rate of 5 μl / min to obtain approximately 10 response units (RU) of coupled protein. After the injection of the antigen, 1 M ethanolamine was injected to block unreacted groups. For kinetic measurements, at 37°C, at a flow rate of about 25 μl / min, a solution of 1% ethanolamine in a solution containing 0.05% polysorbate 20 (TWEEN-20 TM Two-fold serial dilutions (0.78 nM to 500 nM) of Fab in PBS containing PBST (PBST) were prepared using a simple one-to-one Langmuir binding model ( The association rate (k) was calculated by simultaneously fitting the association and dissociation sensorgrams. on or k a ) and dissociation rate (k off or k d ). Equilibrium dissociation constant (K D ) is calculated as the ratio k off / k on See, e.g., Chen et al., J. Mol. Biol. 293:865-881 (1999). If the association rate measured by the surface plasmon resonance above exceeds 10 6 M -1 s -1 , the association rate can be determined by using the fluorescence quenching technique, i.e., in a spectrometer such as a spectrophotometer equipped with a stop flow device (Aviv Instruments) or a 8000 series SLM-AMINCO TM The increase or decrease in fluorescence emission intensity (excitation = 295 nm; emission = 340 nm, 16 nm bandpass) of 20 nM anti-antigen antibody (Fab form) in PBS pH 7.2 at 37°C was measured in a spectrophotometer (ThermoSpectronic) using a stirred cuvette in the presence of increasing concentrations of antigen.

[0529] 2. Antibody fragments

[0530] In certain embodiments, the antibodies provided herein (e.g., anti-FcRH5 / anti-CD3 TDB) are antibody fragments that bind to FcRH5 and CD3. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab') 2 , Fv, and scFv fragments, as well as other fragments described below. For a review of certain antibody fragments, see Hudson et al. Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, e.g., Pluckthün in The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds., (Springer-Verlag, New York), pp. 269-315 (1994); see also WO 93 / 16185; and U.S. Pat. Nos. 5,571,894 and 5,587,458. For a review of Fab and F(ab') fragments comprising salvage receptor binding epitope residues and having increased in vivo half-life 2 For a discussion of fragments, see U.S. Patent No. 5,869,046.

[0531] Diabodies are antibody fragments with two antigen binding sites that can be bivalent or bispecific. See, e.g., EP 404,097; WO 1993 / 01161; Hudson et al., Nat. Med. 9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003).

[0532] Single domain antibodies are antibody fragments that contain all or part of the heavy chain variable domain or all or part of the light chain variable domain of an antibody.In certain embodiments, the single domain antibody is a human single domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U.S. Pat. No. 6,248,516 Bl).

[0533] Antibody fragments can be prepared by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies and production by recombinant host cells (eg, E. coli or phage), as described herein.

[0534] 3. Chimeric and humanized antibodies

[0535] In certain embodiments, the antibodies provided herein (e.g., anti-FcRH5 / anti-CD3 TDB) are chimeric antibodies. Certain chimeric antibodies are described in, e.g., U.S. Pat. No. 4,816,567 and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984). In one example, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate (such as a monkey)) and a human constant region. In another example, a chimeric antibody is a "class-switched" antibody in which the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0536] In certain embodiments, chimeric antibodies are humanized antibodies. Typically, non-human antibodies are humanized to reduce immunogenicity to people while retaining the specificity and affinity of the parent non-human antibody. Typically, humanized antibodies include one or more variable domains, wherein HVR (or a portion thereof) is, for example, derived from non-human antibodies, and FR (or a portion thereof) is derived from human antibody sequences. Humanized antibodies will optionally also include at least a portion of a human constant region. In certain embodiments, some FR residues in humanized antibodies are replaced by corresponding residues from non-human antibodies (e.g., antibodies from which HVR residues are derived), for example, to restore or improve antibody specificity or affinity.

[0537] 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., Proc. Natl. Acad. Sci. USA 86:10029-10033 (1989); U.S. Pat. Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (describing specificity determining region (SDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (describing "resurfacing"); Dall'Acqua et al., Methods 36:25-34 (2005) (describing specificity determining region (SDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (describing "resurfacing"); Dall'Acqua et al., Methods 36:25-34 (2005) (describing specificity determining region (SDR) grafting); 36:43-60 (2005) (describing "FR shuffling"); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing the "guided selection" method for FR shuffling).

[0538] Human framework regions that can be used for humanization include, but are not limited to, framework regions selected using the “best fit” method (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from the consensus sequence of human antibodies of a particular subgroup of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol., 151:2623 (199 3)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13: 1619-1633 (2008)); and framework regions derived from screening FR libraries (see, e.g., Baca et al., J. Biol. Chem. 272: 10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271: 22611-22618 (1996)).

[0539] 4. Human Antibodies

[0540] In certain embodiments, the antibodies provided herein (e.g., anti-FcRH5 / anti-CD3 TDB) are human antibodies. 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).

[0541] Human antibodies can be prepared by administering an immunogen to a transgenic animal that has been modified to produce complete human antibodies or complete antibodies with human variable regions in response to antigenic stimulation. Such animals typically contain all or part of a human immunoglobulin locus that replaces an endogenous immunoglobulin locus, or exists outside the chromosome of the animal or is randomly integrated into the chromosome of the animal. In such transgenic mice, the endogenous immunoglobulin locus is typically 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, description of XENOMOUSE TM US Patent Nos. 6,075,181 and 6,150,584 for the technology; description US Patent No. 5,770,429 for technology; describes KM US Patent No. 7,041,870, and describes The human variable regions from intact antibodies produced by such animals can be further modified, for example by combining with different human constant regions.

[0542] Human antibodies can also be prepared by hybridoma-based methods. Human myeloma and mouse-human hybrid myeloma cell lines for producing human monoclonal antibodies have been described. (See, for example, Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991)). Human antibodies produced via human B cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103: 3557-3562 (2006). Additional methods include, for example, those described in U.S. Pat. No. 7,189,826 (describing the production of monoclonal human IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26 (4): 265-268 (2006) (describing human-human hybridomas). Human hybridoma technology (Trioma technology) is also described in Vollmers and Brandlein, Histology and Histopathology, 20 (3): 927-937 (2005) and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27 (3): 185-91 (2005).

[0543] Human antibodies can also be produced by isolating the Fv clone variable domain sequence selected from the human phage display library. Such variable domain sequences can then be combined with the expected human constant domains. The technology of selecting human antibodies from the antibody library is described below.

[0544] 5. Multispecific Antibodies

[0545] In any of the above aspects, the anti-FcRH5 / anti-CD3 antibodies provided herein are multispecific antibodies, such as bispecific antibodies. Multispecific antibodies are antibodies (e.g., monoclonal antibodies) that have binding specificity to at least two different sites, such as antibodies that have binding specificity to immune effector cells and to cell surface antigens (e.g., tumor antigens, such as FcRH5) on target cells other than immune effector cells. In some aspects, one of the binding specificities is for FcRH5, and the other is for CD3.

[0546] In some aspects, cell surface antigen can be expressed with low copy number on target cell.For example, in some aspects, cell surface antigen is expressed or exists with less than 35,000 copies per target cell.In certain embodiments, low copy number cell surface antigen is between 100 and 35,000 copies per target cell, between 100 and 30,000 copies per target cell, between 100 and 25,000 copies per target cell, between 100 and 20,000 copies per target cell, between 100 and 15,000 copies per target cell, between 100 and 10,000 copies per target cell, between 100 and 5,000 copies per target cell, between 100 and 2,000 copies per target cell, between 100 and 1,000 copies per target cell, or between 100 and 500 copies per target cell. For example, a standard Scatchard plot can be used to determine the copy number of a cell surface antigen.

[0547] In some embodiments, bispecific antibodies can be used to localize cytotoxic agents to cells expressing tumor antigens such as FcRH5. Bispecific antibodies can be prepared as full-length antibodies or antibody fragments.

[0548] Techniques for preparing multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs with different specificities (see, Milstein and Cuello, Nature 305:537 (1983), WO 93 / 08829, and Traunecker et al., EMBO J. 10:3655 (1991)) and "knob-in-hole" engineering (see, e.g., U.S. Pat. No. 5,731,168). The "knob-in-hole" engineering of multispecific antibodies can be utilized to generate a first arm comprising a knob and a second arm comprising a hole, wherein the knob of the first arm can be bound to the hole. In one embodiment, the knob of the multispecific antibodies disclosed herein can be an anti-CD3 arm. Alternatively, in one embodiment, the knob of the multispecific antibodies disclosed herein can be an anti-target / antigen arm. In one embodiment, the hole of the multispecific antibodies disclosed herein can be an anti-CD3 arm. Alternatively, in one embodiment, the hole of the multispecific antibodies disclosed herein can be an anti-target / antigen arm.

[0549] Multispecific antibodies can also be engineered using immunoglobulin crossover (also known as Fab domain exchange or CrossMab format) technology (see, e.g., WO 2009 / 080253; Schaefer et al., Proc. Natl. Acad. Sci. USA, 108: 11187-11192 (2011)). Multispecific antibodies can also be prepared by the following techniques: engineering electrostatic manipulation effects to prepare antibody Fc-heterodimer molecules (WO 2009 / 089004A1); crosslinking two or more antibodies or fragments (see, e.g., U.S. Pat. No. 4,676,980; and Brennan et al., Science 229: 81 (1985)); using leucine zippers to produce bispecific antibodies (see, e.g., Kostelny et al., J. Immunol. 148 (5): 1547-1553 (1992)); using "diabody" technology to prepare bispecific antibodies and using single-chain Fv (sFv) dimers (see, e.g., Gruber et al., J. Immunol. 152:5368 (1994)); and preparing trispecific antibodies as described, e.g., in Tutt et al., J. Immunol. 147:60 (1991).

[0550] Engineered antibodies with three or more functional antigen binding sites, including "octopus antibodies," are also included herein (see, e.g., US 2006 / 0025576 A1).

[0551] Antibodies or antigen-binding fragments thereof may also include "dual-acting FAbs" or "DAFs," which contain antigen-binding sites that bind to CD3 as well as another, different antigen (eg, a second biomolecule) (see, eg, US 2008 / 0069820).

[0552] 6. Antibody variants

[0553] In some aspects, amino acid sequence variants of the bispecific anti-FcRH5 / anti-CD3 antibodies disclosed herein are contemplated. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody. The amino acid sequence variants of the antibody can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions, and / or insertions and / or substitutions of residues within the antibody amino acid sequence. Any combination of deletions, insertions, and substitutions may be performed to achieve the final construct, provided that the final construct has the desired characteristics, e.g., antigen binding.

[0554] a. Substitution, insertion and deletion variants

[0555] In certain embodiments, antibody variants with one or more amino acid substitutions are provided. Interested sites for substitution mutagenesis include CDR and FR. Conservative substitutions are shown under the "preferred substitutions" heading in Table 4. More substantial changes are provided under the "exemplary substitutions" heading in Table 4, and are further described below with reference to amino acid side chain categories. Amino acid substitutions can be introduced into the target antibody, and the product can be screened for desired activity (e.g., retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC).

[0556] Table 4. Exemplary and preferred amino acid substitutions

[0557]

[0558] Amino acids can be grouped according to common side chain properties:

[0559] (1) Hydrophobicity: norleucine, Met, Ala, Val, Leu, Ile;

[0560] (2) Neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln;

[0561] (3) Acidic: Asp, Glu;

[0562] (4) Basic: His, Lys, Arg;

[0563] (5) Residues that affect chain orientation: Gly, Pro;

[0564] (6) Aromatic: Trp, Tyr, Phe.

[0565] Non-conservative substitutions will entail exchanging a member of one of these classes for another class.

[0566] One type of substitution variant involves replacing one or more hypervariable region residues of a parent antibody (e.g., a humanized antibody or a human antibody). Typically, relative to the parent antibody, one or more resulting variants selected for further study will have changes (e.g., improvements) in certain biological properties (e.g., increased affinity, reduced immunogenicity) and / or will substantially retain certain biological properties of the parent antibody. Exemplary substitution variants are affinity matured antibodies, which can be conveniently generated, for example, using affinity maturation techniques based on phage display such as those described herein. In short, one or more CDR residues are mutated and the variant antibody is displayed on a phage and screened for a specific biological activity (e.g., binding affinity).

[0567] For example, changes (e.g., substitutions) can be made in the CDR to improve antibody affinity. Such changes can occur in CDR "hot spots", i.e., residues encoded by codons that undergo high-frequency mutations during somatic maturation (see, e.g., Chowdhury, Methods Mol. Biol. 207: 179-196, 2008) and / or residues in contact with the antigen (testing the binding affinity of the resulting variant VH or VL). Affinity maturation achieved by construction and reselection from a secondary library has been described, for example, by Hoogenboom et al. in Methods in Molecular Biology 178: 1-37 (O'Brien et al., Human Press, Totowa, NJ, (2001)). In some embodiments of affinity maturation, diversity is introduced into variable genes selected for maturation using any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide site-directed mutagenesis genes). A secondary library is then created. The library is subsequently screened to identify any antibody variants with the desired affinity. Another method of introducing diversity involves a CDR-directed approach in which several CDR residues (e.g., 4 to 6 residues at a time) are randomized. The CDR residues involved in antigen binding can be specifically identified, for example, using alanine scanning mutagenesis or modeling. In particular, CDR-H3 and CDR-L3 are often targeted.

[0568] In certain embodiments, substitutions, insertions or deletions may occur within one or more CDRs, as long as such changes do not substantially reduce the antigen binding ability of the antibody. For example, conservative changes (e.g., conservative substitutions as provided herein) that do not substantially reduce binding affinity may be made in the CDRs. Such changes may be, for example, outside of the antigen contact residues in the CDRs. In certain embodiments of the variant VH and VL sequences provided above, each CDR remains unchanged, or comprises no more than one, two or three amino acid substitutions.

[0569] The method that can be used to identify antibody residues or regions that can be targeted for mutagenesis is called "alanine scanning mutagenesis", as described in Cunningham and Wells (1989) Science, 244: 1081-1085. In this method, a residue or a group of target residues (e.g., charged residues, such as Arg, Asp, His, Lys and Glu) are identified and replaced with neutral or negatively charged amino acids (e.g., alanine or polyalanine) to determine whether the interaction of the antibody with the antigen is affected. Other substitutions can be introduced at amino acid positions that exhibit functional sensitivity to the initial substitution. Alternatively or additionally, the crystal structure of the antigen-antibody complex is used to identify the contact points between the antibody and the antigen. Such contact residues and adjacent residues that are candidates for substitution can be targeted or eliminated. Variants can be screened to determine whether they have the desired properties.

[0570] Amino acid sequence insertions include amino and / or carboxyl terminal fusions ranging in length from one residue to a polypeptide containing one hundred or more residues, and intrasequence insertions of one or more amino acid residues. Examples of terminal insertions include antibodies with an N-terminal methionyl residue. Other insertion variants of antibody molecules include fusions to the N-terminus or C-terminus of an antibody that increases the serum half-life of an antibody (e.g., for ADEPT) or a polypeptide.

[0571] b. Glycosylation variants

[0572] In certain embodiments, the bispecific anti-FcRH5 / anti-CD3 antibodies disclosed herein can be altered to increase or decrease the extent of antibody glycosylation. Addition or deletion of glycosylation sites to the anti-FcRH5 antibodies of the invention can be conveniently achieved by altering the amino acid sequence to create or remove one or more glycosylation sites.

[0573] When antibody comprises Fc district, the carbohydrate attached thereto can be changed.Native antibodies produced by mammalian cells usually comprise the biantennary oligosaccharide of side chain, and this biantennary oligosaccharide is usually connected to the Asn297 of the CH2 domain of Fc district by N key.See, for example, Wright et al. TIBTECH 15:26-32 (1997).Oligosaccharide can include various carbohydrates, for example, mannose, N-acetylglucosamine (GlcNAc), galactose and sialic acid, and the fucose of GlcNAc in the " backbone " of biantennary oligosaccharide structure.In certain embodiments, the oligosaccharide in the antibody of the present invention can be modified, so as to produce antibody variants with some improved characteristics.

[0574] In one embodiment, a bispecific anti-FcRH5 / anti-CD3 antibody variant having a carbohydrate structure lacking fucose attached (directly or indirectly) to the Fc region is provided. For example, the content of fucose in such an antibody may be 1% to 80%, 1% to 65%, 5% to 65%, or 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose at Asn297 in the sugar chain relative to the sum of all sugar structures (e.g., complex, hybrid, and high mannose structures) attached to Asn297 as measured by MALDI-TOF mass spectrometry, as described in WO 2008 / 077546. Asn297 refers to the asparagine residue located at approximately position 297 in the Fc region (EU numbering of Fc region residues); however, due to minor sequence variations in antibodies, Asn297 may also be located approximately ±3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300. Such fucosylation variants may have improved ADCC function. See, for example, US Patent Publication Nos. US2003 / 0157108 (Presta, L.); US2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd). Related "defucosylated" or "fucose-deficient" antibody variants include: US2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; US2003 / 0115614; US2002 / 0164328; US 2004 / 0093621; US2004 / 0132140; US2004 / 0110704; US2004 / 0110282; US2004 / 0109865; WO 2003 / 085119; WO 2003 / 084570; WO 2005 / 035586; WO 2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al., J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al., Biotech. Bioeng. 87:614 (2004).Examples of cell lines capable of producing defucosylated antibodies include Lec13 CHO cells deficient in protein fucosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); U.S. Patent Application No. US2003 / 0157108 A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., particularly Example 11), and knockout cell lines, such as CHO cells knocked out for the α-1,6-fucosyltransferase gene (FUT8) (see, e.g., Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO2003 / 085107).

[0575] Further provided are bispecific anti-FcRH5 / anti-CD3 antibody variants having bisected oligosaccharides, for example, wherein the biantennary oligosaccharide attached to the Fc region of the antibody is bisected by GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described, for example, in WO 2003 / 011878 (Jean-Mairet et al.), U.S. Pat. No. 6,602,684 (Umana et al.) and US2005 / 0123546 (Umana et al.). Antibody variants having at least one galactose residue in the oligosaccharide attached to the Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described, for example, in WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.).

[0576] c. Fc region variants

[0577] In certain embodiments, one or more amino acid modifications can be introduced into the Fc region of the bispecific anti-FcRH5 / anti-CD3 antibody, thereby generating an Fc region variant (see, e.g., US 2012 / 0251531). The Fc region variant can comprise a human Fc region sequence (such as a human IgG1, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid modification (such as a substitution) at one or more amino acid positions.

[0578] In certain embodiments, the present invention contemplates bispecific anti-FcRH5 / anti-CD3 antibody variants that possess some but not all effector functions, making them desirable candidates for applications where the half-life of the antibody in vivo is important and certain effector functions (such as complement and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity assays can be performed to confirm reduction / depletion of CDC and / or ADCC activity. For example, an Fc receptor (FcR) binding assay can be performed to ensure that the antibody lacks FcγR binding (and therefore may lack ADCC activity), but retains FcRn binding ability. Primary cells that mediate ADCC NK cells express only Fc(RIII), while monocytes express Fc(RI, Fc(RII, and Fc(RIII). FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991). Non-limiting examples of in vitro assays for assessing ADCC activity of target molecules are described in U.S. Pat. No. 5,500,362 (see, e.g., Hellstrom, et al., Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986)) and Hellstrom, et al., Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986)). 82:1499-1502 (1985); 5,821,337 (see Bruggemann, et al., J. Exp. Med. 166:1351-1361 (1987)). Alternatively, non-radioactive assays can be used (see, for example, ACTI for flow cytometry). TM Nonradioactive cytotoxicity assay (Cell Technology, Inc. Mountain View, CA); and CytoTox Non-radioactive cytotoxicity assay (Promega, Madison, WI). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively or additionally, ADCC activity of the target molecule can be assessed in vivo, for example, in an animal model such as disclosed in Clynes et al., Proc. Nat'l Acad. Sci. USA 95:652-656 (1998). C1q binding assays can also be performed to confirm that the antibody cannot bind to C1q and therefore lacks CDC activity. See, for example, C1q and C3c binding ELISAs in WO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, a CDC assay can be performed (see, e.g., Gazzano-Santoro et al. J. Immunol. Methods 202: 163 (1996); Cragg, et al. Blood. 101: 1045-1052 (2003); and Cragg et al. Blood. 103: 2738-2743 (2004)). FcRn binding and in vivo clearance / half-life assays can also be performed using methods known in the art (see, e.g., Petkova et al., Int'l. Immunol. 18(12): 1759-1769 (2006)).

[0579] Antibodies with reduced effector function include those with one or more substitutions of Fc region residues 238, 265, 269, 270, 297, 327, and 329 (U.S. Pat. Nos. 6,737,056 and 8,219,149). Such Fc mutants include Fc mutants with substitutions at two or more amino acids 265, 269, 270, 297, and 327, including the so-called "DANA" Fc mutants, in which residues 265 and 297 are substituted with alanine (U.S. Pat. Nos. 7,332,581 and 8,219,149).

[0580] In certain embodiments, the proline at position 329 of the wild-type human Fc region in the antibody is substituted with glycine or arginine or a sufficiently large amino acid residue to disrupt the proline sandwich within the Fc / Fcγ receptor interface formed between proline 329 of Fc and tryptophan residues Trp 87 and Trp 110 of FcgRIII (Sondermann et al. Nature. 406, 267-273, 2000). In certain embodiments, the antibody comprises at least one further amino acid substitution. In another embodiment, the additional amino acid substitution is S228P, E233P, L234A, L235A, L235E, N297A, N297D or P331S, and in yet another embodiment, the at least one additional amino acid substitution is L234A and L235A in the human IgG1 Fc region or S228P and L235E in the human IgG4 Fc region (see, e.g., US2012 / 0251531), and in yet another embodiment, the at least one additional amino acid substitution is L234A and L235A and P329G in the human IgG1 Fc region.

[0581] Certain antibody variants with improved or decreased binding to FcRs are described. (See, e.g., U.S. Pat. No. 6,737,056; WO 2004 / 056312; and Shields et al., J. Biol. Chem. 9(2):6591-6604 (2001).)

[0582] In certain embodiments, the antibody variant comprises an Fc region with one or more amino acid substitutions that improve ADCC, eg, substitutions at positions 298, 333, and / or 334 (EU numbering of residues) of the Fc region.

[0583] In some embodiments, alterations are made in the Fc region resulting in altered (ie, improved or reduced) C1q binding and / or complement dependent cytotoxicity (CDC), e.g., as described in U.S. Pat. No. 6,194,551, WO 99 / 51642, and Idusogie et al. J. Immunol. 164:4178-4184 (2000).

[0584] Antibodies with extended half-life and improved binding to the neonatal Fc receptor (FcRn) are described in US 2005 / 0014934A1 (Hinton et al.), which is responsible for the transfer of maternal IgG to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)). Those antibodies comprise an Fc region having one or more substitutions therein that improve binding of the Fc region to FcRn. Such Fc variants include those having a substitution at one or more of the following Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, ​​413, 424, or 434, such as a substitution at Fc region residue 434 (U.S. Pat. No. 7,371,826).

[0585] For other examples of Fc region variants, see also: Duncan and Winter, Nature 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821; and WO 94 / 29351.

[0586] In some aspects, the anti-FcRH5 and / or anti-CD3 antibody (e.g., a bispecific anti-FcRH5 antibody) comprises an Fc region comprising an N297G mutation (EU numbering). In some aspects, the anti-FcRH5 arm of the bispecific anti-FcRH5 antibody comprises an N297G mutation, and / or the anti-CD3 arm of the bispecific anti-FcRH5 antibody comprises an Fc region comprising an N297G mutation.

[0587] In some embodiments, an anti-FcRH5 antibody comprising an N297G mutation comprises an anti-FcRH5 arm comprising a first binding domain, wherein the first binding domain comprises the following six HVRs: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 1; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 2; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 3; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 4; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 5; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 6; and an anti-CD3 arm comprising the N297G mutation. In some embodiments, the anti-CD3 arm comprising the N297G mutation comprises the following six HVRs: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:9; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:10; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:11; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:12; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:13; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:14.

[0588] In some embodiments, the anti-FcRH5 antibody comprising the N297G mutation comprises an anti-FcRH5 arm comprising a first binding domain comprising: (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; and an anti-CD3 arm comprising the N297G mutation. In some embodiments, the anti-CD3 arm comprising the N297G mutation 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.

[0589] In some embodiments, the anti-FcRH5 antibody comprising the N297G mutation 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, the first CH2 (CH2 1 ) domain, the first CH3 (CH3 1 ) domain, the second CH1 (CH1 2 ) domain, the second CH2 (CH2 2 ) domain and the second CH3 (CH3 2) domain. In some aspects, at least one of the one or more heavy chain constant domains is paired with another heavy chain constant domain. In some aspects, CH3 1 Domain and CH3 2 The domains each contain a protrusion or a cavity, and wherein CH3 1 The protrusion or cavity in the domain can be located at the CH3 2 In some aspects, CH3 1 Domain and CH3 2 The domains meet at the interface between the protrusion and the cavity. In some aspects, CH2 1 Domain and CH2 2 The domains each contain a protrusion or a cavity, and wherein CH2 1 The protrusion or cavity in the domain can be located at the CH2 2 In other cases, CH2 1 and CH2 2 The domains meet at the interface between the protrusion and the cavity. In some aspects, the anti-FcRH5 antibody is an IgG 1 Antibody.

[0590] In some embodiments, the anti-FcRH5 antibody comprising the N297G mutation comprises an anti-FcRH5 arm comprising a first binding domain, the first binding domain comprising: (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; and an anti-CD3 arm, wherein (a) the anti-FcRH5 arm comprises T366S, L368A, Y407V, and N297G amino acid substitution mutations (EU numbering), and (b) the anti-CD3 arm comprises T366W and N297G substitution mutations (EU numbering). In some embodiments, the anti-CD3 arm comprising T366W and N297G mutations 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.

[0591] In other embodiments, the anti-FcRH5 antibody comprising the N297G mutation comprises an anti-FcRH5 arm comprising a first binding domain, the first binding domain comprising: (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; and an anti-CD3 arm, wherein (a) the anti-FcRH5 arm comprises T366W and N297G amino acid substitution mutations (EU numbering), and (b) the anti-CD3 arm comprises T366S, L368A, Y407V and N297G mutations (EU numbering). In some embodiments, the anti-CD3 arm comprising the N297G mutation 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.

[0592] d. Cysteine ​​enginee...

Claims

1. A method of treating a subject having multiple myeloma (MM) with a high-risk cytogenetic profile, the method comprising administering to the subject (i) a bispecific antibody that binds to fragment crystallizable receptor-like 5 (FcRH5) and cluster of differentiation 3 (CD3) and (ii) lenalidomide.

2. The method of claim 1, wherein the subject has experienced a partial remission (PR) or better after induction therapy.

3. The method according to claim 1 or 2, wherein the subject has undergone autologous stem cell transplantation (ASCT) within 100 days of starting the method and / or does not have progressive disease.

4. The method according to any one of claims 1 to 3, wherein the bispecific antibody and lenalidomide are administered to the patient as post-transplant maintenance therapy.

5. The method of any one of claims 1 to 4, wherein the high-risk cytogenetic features comprise one or more of the following: translocation events t(4;14) or t(14;16), del(17p), or 1q gain.

6. The method according to any one of claims 1 to 5, wherein the subject carries the high-risk cytogenetic characteristics at the time of diagnosis of MM.

7. The method according to any one of claims 1 to 6, wherein the bispecific antibody and the lenalidomide are administered to the subject in a dosing regimen comprising: (i) a first phase, which comprises one or more dosing cycles, wherein the first phase comprises administering the bispecific antibody to the subject every two weeks (Q2W); and (ii) a second phase, which comprises one or more dosing cycles, wherein the second phase comprises administering the bispecific antibody to the subject every four weeks (Q4W).

8. The method according to claim 7, wherein each dosing cycle of the first phase and / or the second phase is a 28-day dosing cycle.

9. The method according to claim 7 or 8, further comprising a preliminary phase comprising one or more dosing cycles prior to the first phase, wherein the preliminary phase comprises administering the bispecific antibody to the subject weekly (QW).

10. The method according to claim 9, wherein each dosing cycle in the preliminary phase is a 28-day dosing cycle. The method according to claim 10 , wherein the preliminary phase comprises one administration cycle ( C1 ). 12 . The method of claim 11 , wherein the preliminary phase comprises administering the bispecific antibody to the subject on Day 1 , Day 8 and Day 15 of the C1.

13. The method according to any one of claims 9 to 12, wherein in the preliminary phase a target dose of the bispecific antibody is administered to the subject for each administration.

14. The method according to any one of claims 9 to 12, wherein the preliminary phase comprises administering to the subject a first ascending dose of the bispecific antibody.

15. The method of claim 14, wherein the first ascending dose is administered to the subject on day 1 of the C1.

16. The method of claim 15, wherein the subject is administered a target dose on Day 8 and Day 15 of the C1.

17. The method according to any one of claims 9 to 12, wherein the preliminary phase comprises administering to the subject a first ascending dose and a second ascending dose of the bispecific antibody. 18 . The method of claim 17 , wherein the first ascending dose is administered to the subject on day 1 of C1 and the second ascending dose is administered to the subject on day 8 of C1.

19. The method of claim 18, wherein the target dose is administered to the subject on day 15 of the C1.

20. The method of any one of claims 15 to 19, wherein the first ascending dose is 3.6 mg.

21. The method of any one of claims 17 to 19, wherein the first ascending dose is 0.3 mg and the second ascending dose is 3.6 mg.

22. The method of any one of claims 8 to 21, wherein the first phase comprises at least two dosing cycles, at least three dosing cycles, at least four dosing cycles, or at least five dosing cycles.

23. The method according to claim 22, wherein the first phase comprises a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4) and a fifth dosing cycle (C5).

24. The method of claim 23, wherein the first phase comprises administering the bispecific antibody to the subject on Day 1 and Day 15 of the C1, the C2, the C3, the C4 and / or the C5.

25. The method of claim 24, wherein a target dose of the bispecific antibody is administered to the subject for each administration during the first stage.

26. The method of any one of claims 8 to 25, wherein the second phase comprises at least two dosing cycles, at least three dosing cycles, at least four dosing cycles, at least five dosing cycles, at least six dosing cycles, or at least seven dosing cycles.

27. The method of claim 26, wherein the second phase comprises a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), a fifth dosing cycle (C5), a sixth dosing cycle (C6) and a seventh dosing cycle (C7).

28. The method of claim 27, wherein the second phase comprises administering the bispecific antibody to the subject on day 1 of the C1, the C2, the C3, the C4, the C5, the C6 and / or the C7.

29. The method of claim 28, wherein a target dose of the bispecific antibody is administered to the subject for each administration during the second stage.

30. The method of any one of claims 13, 16, 19, 25, and 29, wherein the target dose is between 90 mg and 198 mg, inclusive.

31. The method of claim 30, wherein the target dose is 90 mg.

32. The method of claim 30, wherein the target dose is 132 mg.

33. The method of claim 30, wherein the target dose is 160 mg.

34. The method of any one of claims 1 to 33, wherein the bispecific antibody is administered intravenously to the subject.

35. The method according to any one of claims 8 to 34, wherein the subject is administered lenalidomide on day 1 to day 21 of each dosing cycle in the first phase and / or the second phase.

36. The method according to any one of claims 10 to 35, wherein the subject is administered the lenalidomide on day 1 to day 21 of each dosing cycle in the preliminary phase.

37. The method of any one of claims 1 to 36, wherein the lenalidomide is administered to the subject at a dose of about 10 mg to about 20 mg.

38. The method of claim 37, wherein the lenalidomide is administered to the subject at a dose of about 10 mg.

39. The method of claim 37, wherein the lenalidomide is administered to the subject at a dose of about 15 mg.

40. The method of any one of claims 1 to 39, wherein the lenalidomide is administered orally to the subject.

41. The method of any one of claims 1 to 40, further comprising administering a corticosteroid to the subject.

42. The method of any one of claims 7 to 41, further comprising administering a corticosteroid to the subject during the first phase and / or the second phase.

43. The method of any one of claims 24 to 42, wherein the corticosteroid is administered to the subject during the first phase on Day 1 and Day 15 of the C1 of the first phase.

44. The method of any one of claims 24 to 43, wherein the corticosteroid is administered to the subject in C2, C3, C4 and / or C5 of the first stage if the subject experienced a cytokine release syndrome (CRS) event with a previous dose.

45. The method of any one of claims 28 to 44, wherein the corticosteroid is administered to the subject in the C1, the C2, the C3, the C4, the C5, the C6 and / or the C7 of the second stage if the subject experienced a CRS event with the previous dose.

46. ​​The method of any one of claims 9 to 45, further comprising administering a corticosteroid to the subject during the preliminary period.

47. The method of any one of claims 11 to 46, wherein the corticosteroid is administered to the subject on Day 1, Day 8, and Day 15 of the C1 during the pre-phase.

48. The method of any one of claims 41 to 47, wherein the corticosteroid is administered to the subject intravenously or orally.

49. The method of claim 48, wherein the corticosteroid is administered intravenously to the subject.

50. The method of any one of claims 41 to 49, wherein the corticosteroid is administered intravenously to the subject prior to administration of the bispecific antibody.

51. The method of claim 50, wherein the corticosteroid is administered intravenously to the subject about 1 hour prior to administration of the bispecific antibody.

52. The method of any one of claims 41 to 51, wherein the corticosteroid is dexamethasone or methylprednisolone.

53. The method of claim 52, wherein the corticosteroid is dexamethasone.

54. The method of claim 52 or 53, wherein the dexamethasone is administered to the subject at a dose of about 20 mg.

55. The method of claim 52, wherein the methylprednisolone is administered to the subject at a dose of about 80 mg.

56. The method of any one of claims 1 to 55, wherein the bispecific antibody comprises an anti-CD79b arm comprising a first binding domain comprising the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence of RFGVH (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of VIWRGGSTDYNAAFVS (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of HYYGSSDYALDN (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of KASQDVRNLVV (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of SGSYRYS (SEQ ID NO:5); and (f) HVR-L3 comprising the amino acid sequence of QQHYSPPYT (SEQ ID NO: 6).

57. The method of any one of claims 1 to 56, wherein the bispecific antibody comprises an anti-FcRH5 arm comprising a first binding domain, the first binding domain comprising: (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 7; (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 8; or (c) a VH domain as in (a) and a VL domain as in (b).

58. The method of claim 57, wherein the first binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO:7; and a VL domain comprising the amino acid sequence of SEQ ID NO:

8.

59. The method of any one of claims 1 to 58, wherein the bispecific antibody comprises an anti-CD3 arm comprising a second binding domain comprising the following six HVRs: (a) HVR-H1 comprising the amino acid sequence of SYYIH (SEQ ID NO: 9); (b) HVR-H2 comprising the amino acid sequence of WIYPENDNTKYNEKFKD (SEQ ID NO: 10); (c) HVR-H3 comprising the amino acid sequence of DGYSRYYFDY (SEQ ID NO: 11); (d) HVR-L1 comprising the amino acid sequence of KSSQSLLNSRTRKNYLA (SEQ ID NO: 12); (e) HVR-L2 comprising the amino acid sequence of WTSTRKS (SEQ ID NO: 13); and (f) HVR-L3 comprising the amino acid sequence of KQSFILRT (SEQ ID NO: 14).

60. The method of any one of claims 1 to 59, wherein the bispecific antibody comprises an anti-CD3 arm comprising a second binding domain, the second binding domain comprising: (a) a VH domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 15; (b) a VL domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 15; An amino acid sequence having at least 95% sequence identity to the amino acid sequence of ID NO: 16; or (c) a VH domain as in (a) and a VL domain as in (b).

61. The method of claim 60, wherein the second binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 15; and a VL domain comprising the amino acid sequence of SEQ ID NO:

16.

62. The method of any one of claims 1 to 61, wherein the bispecific antibody comprises: an anti-FcRH5 arm comprising a heavy chain polypeptide (H1) and a light chain polypeptide (L1); and an anti-CD3 arm comprising a heavy chain polypeptide (H2) and a light chain polypeptide (L2), and wherein: (a) H1 comprises the amino acid sequence of SEQ ID NO: 35; (b) L1 comprises the amino acid sequence of SEQ ID NO: 36; (c) H2 comprises the amino acid sequence of SEQ ID NO:37; and (d) L2 comprises the amino acid sequence of SEQ ID NO:

38.

63. The method of any one of claims 1 to 62, wherein the bispecific antibody comprises an aglycosylation site mutation.

64. The method of claim 63, wherein the aglycosylation site mutation reduces the effector function of the bispecific antibody.

65. The method of claim 64, wherein the deglycosylation site mutation is a substitution mutation.

66. The method of claim 65, wherein the bispecific antibody comprises a substitution mutation in the Fc region that reduces effector function.

67. The method of any one of claims 1 to 66, wherein the bispecific antibody is a monoclonal antibody.

68. The method of any one of claims 1 to 67, wherein the bispecific antibody is a humanized antibody.

69. The method of any one of claims 1 to 68, wherein the bispecific antibody is a chimeric antibody.

70. The method of any one of claims 1 to 61 and 63 to 69, wherein the bispecific antibody is an antibody fragment that binds FcRH5 and CD3.

71. The method of claim 70, wherein the antibody fragment is selected from the group consisting of: Fab, Fab'-SH, Fv, scFv and (Fab')2 fragment.

72. The method of any one of claims 1 to 71, wherein the bispecific antibody is a full-length antibody.

73. The method of any one of claims 1 to 72, wherein the bispecific antibody is an IgG antibody.

74. The method of claim 73, wherein the IgG antibody is an IgG1 antibody.

75. The method of any one of claims 1 to 74, wherein the bispecific antibody comprises one or more heavy chain constant domains, wherein the one or more heavy chain constant domains are selected from a first CH1 (CH11) domain, a first CH2 (CH21) domain, a first CH3 (CH31) domain, a second CH1 (CH12) domain, a second CH2 (CH22) domain, and a second CH3 (CH32) domain.

76. The method of claim 75, wherein at least one of the one or more heavy chain constant domains is paired with another heavy chain constant domain.

77. The method of claim 76, wherein the CH31 domain and the CH32 domain each comprise a protrusion or a cavity, and wherein the protrusion or the cavity in the CH31 domain is positionable in the cavity or the protrusion in the CH32 domain, respectively.

78. The method of claim 77, wherein the CH31 domain and the CH32 domain meet at the interface between the protrusion and the cavity.

79. The method according to any one of claims 75 to 78, wherein the CH21 domain and the CH22 domain each comprise a protrusion or a cavity, and wherein the protrusion or the cavity in the CH21 domain is positionable in the cavity or the protrusion in the CH22 domain, respectively.

80. The method of claim 79, wherein the CH21 domain and the CH22 domain meet at an interface between the protrusion and the cavity.

81. The method of claim 80, wherein the anti-FcRH5 arm comprises the protrusion and the anti-CD3 arm comprises the cavity.

82. The method of claim 81, wherein the CH3 domain of the anti-FcRH5 arm comprises a protrusion comprising a T366W amino acid substitution mutation (EU numbering), and the CH3 domain of the anti-CD3 arm comprises a cavity comprising T366S, L368A and Y407V amino acid substitution mutations (EU numbering).

83. The method of any one of claims 1 to 69 and 72 to 82, wherein the bispecific antibody is sivostatin.

84. The method of any one of claims 1 to 83, wherein the bispecific antibody and the lenalidomide are administered to the subject simultaneously with one or more additional therapeutic agents.

85. The method of claim 84, wherein the bispecific antibody and / or the lenalidomide is administered to the subject prior to administration of one or more additional therapeutic agents.

86. The method of claim 84, wherein the bispecific antibody and / or the lenalidomide is administered to the subject after administration of one or more additional therapeutic agents.

87. The method of any one of claims 84 to 86, wherein the one or more additional therapeutic agents comprises an effective amount of tocilizumab.

88. The method of any one of claims 1 to 87, wherein the subject develops a CRS event, and the method further comprises treating a symptom of the CRS event while suspending treatment with the bispecific antibody.

89. The method of claim 88, wherein the method further comprises administering to the subject an effective amount of tocilizumab to treat the CRS event.

90. The method of claim 88, wherein the CRS event does not resolve or worsens within 24 hours of treating the symptoms of the CRS event, the method further comprising administering one or more additional doses of tocilizumab to the subject to manage the CRS event.

91. The method of any one of claims 87, 89, and 90, wherein tocilizumab is administered to the subject by intravenous infusion.

92. The method of claim 91, wherein: (a) the subject weighs ≥30 kg, and tocilizumab is administered to the subject at a dose of 8 mg / kg; or (b) the subject weighs <30 kg, and tocilizumab is administered to the subject at a dose of 12 mg / kg.

93. The method of any one of claims 87, 91 and 92, wherein tocilizumab is administered to the subject 2 hours prior to administration of the bispecific antibody.

94. The method of any one of claims 84 to 93, wherein the one or more additional therapeutic agents comprise an effective amount of a B cell maturation antigen (BCMA) directed therapy, an additional immunomodulatory agent (IMiD), a CD38 directed therapy, or a combination of any of the foregoing.

95. The method of any one of claims 84 to 94, wherein the one or more additional therapeutic agents comprises an effective amount of acetaminophen or paracetamol.

96. The method of claim 95, wherein acetaminophen or paracetamol is administered to the subject at a dose of between about 500 mg to about 1000 mg.

97. The method of claim 96, wherein acetaminophen or paracetamol is orally administered to the subject.

98. The method of any one of claims 84 to 97, wherein the one or more additional therapeutic agents comprises an effective amount of diphenhydramine.

99. The method of claim 98, wherein diphenhydramine is administered to the subject at a dose of between about 25 mg to about 50 mg.

100. The method of claim 99, wherein diphenhydramine is administered orally to the subject.

101. A method of treating a subject having MM with a high-risk cytogenetic profile, the method comprising administering to the subject silvostomumab and lenalidomide, wherein: (i) the subject experiences a PR or better after induction therapy; (ii) the subject has undergone ASCT within 100 days of starting the method and / or does not have progressive disease; (iii) administering the silvostomumab and the lenalidomide to the patient as post-transplant maintenance therapy; and (iv) the high-risk cytogenetic characteristics include one or more of the following: translocation events t(4;14) or t(14;16), del(17p) or 1q gain.

102. A method of treating a subject having MM with a high-risk cytogenetic profile, the method comprising administering to the subject silvostomumab and lenalidomide at a dosing regimen comprising: (i) a preliminary phase, which included a 28-day dosing cycle (C1); (ii) a first phase after the preliminary phase, which includes a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4) and a fifth dosing cycle (C5), wherein each dosing cycle of the first phase is a 28-day dosing cycle; as well as (iii) a second phase after the first phase, comprising a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), a fifth dosing cycle (C5), a sixth dosing cycle (C6) and a seventh dosing cycle (C7), wherein each dosing cycle of the second phase is a 28-day dosing cycle, wherein sivostomumab is administered to the subject as follows: (i) administered at a first ascending dose on day 1 of said C1 during said pre-phase, and at a second ascending dose on day 8 of said C1 during said pre-phase; (ii) administered at a target dose on day 15 of said C1 during said pre-phase; (iii) administering at a target dose on Day 1 and Day 15 of said C1, said C2, said C3, said C4 and said C5 during said first phase; and (iv) administering at a target dose on Day 1 of said C1, said C2, said C3, said C4, said C5, said C6, and said C7 during said second phase; and wherein lenalidomide is administered to the subject as follows: (i) during the pre-phase, administered on day 1 to day 21 of said C1; (ii) administering on day 1 to day 21 of said C1, said C2, said C3, said C4 and said C5 during said first phase; as well as (iii) administered on Day 1 to Day 21 of said C1, said C2, said C3, said C4, said C5, said C6 and said C7 during said second phase.

103. The method of claim 102, wherein: (i) the first increasing dose of silvostomumab is 0.3 mg; (ii) the second escalating dose of silvostomumab is 3.6 mg; (iii) the target dose of silvostomumab is between 90 mg and 198 mg, inclusive; and (iv) Lenalidomide was administered at a dose of 10 mg or 15 mg.

104. The method of claim 103, wherein the target dose is 90 mg.

105. The method of claim 103, wherein the target dose is 132 mg.

106. The method of claim 103, wherein the target dose is 160 mg.

107. The method of any one of claims 102 to 106, wherein: (i) the subject experiences a PR or better after induction therapy; (ii) the subject has undergone ASCT within 100 days of starting the method and / or does not have progressive disease; (iii) administering the silvostomumab and the lenalidomide to the patient as post-transplant maintenance therapy; and (iv) the high-risk cytogenetic characteristics include one or more of the following: translocation events t(4;14) or t(14;16), del(17p) or 1q gain.

108. A bispecific antibody that binds to FcRH5 and CD3 for use in treating a subject having MM with high-risk cytogenetic characteristics, the treatment comprising administering the bispecific antibody and lenalidomide to the subject.

109. Sivoduzumab for use in treating a subject with MM having a high-risk cytogenetic profile, the treatment comprising administering to the subject Sivoduzumab and lenalidomide, wherein: (i) the subject experiences a PR or better after induction therapy; (ii) the subject has undergone ASCT within 100 days of starting the method and / or does not have progressive disease; (iii) administering the silvostomumab and the lenalidomide to the patient as post-transplant maintenance therapy; and (iv) the high-risk cytogenetic characteristics include one or more of the following: translocation events t(4;14) or t(14;16), del(17p) or 1q gain.

110. Sivoduzumab for use in treating a subject with MM having a high-risk cytogenetic profile, the treatment comprising administering to the subject sivoduzumab and lenalidomide at a dosing regimen comprising: (i) a preliminary phase, which included a 28-day dosing cycle (C1); (ii) a first phase after the preliminary phase, which includes a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4) and a fifth dosing cycle (C5), wherein each dosing cycle of the first phase is a 28-day dosing cycle; as well as (iii) a second phase after the first phase, comprising a first dosing cycle (C1), a second dosing cycle (C2), a third dosing cycle (C3), a fourth dosing cycle (C4), a fifth dosing cycle (C5), a sixth dosing cycle (C6) and a seventh dosing cycle (C7), wherein each dosing cycle of the second phase is a 28-day dosing cycle, wherein sivostomumab is administered to the subject as follows: (i) administered at a first ascending dose on day 1 of said C1 during said pre-phase, and at a second ascending dose on day 8 of said C1 during said pre-phase; (ii) administered at a target dose on day 15 of said C1 during said pre-phase; (iii) administering at a target dose on Day 1 and Day 15 of said C1, said C2, said C3, said C4 and said C5 during said first phase; and (iv) administering at a target dose on Day 1 of said C1, said C2, said C3, said C4, said C5, said C6, and said C7 during said second phase; and wherein lenalidomide is administered to the subject as follows: (i) during the pre-phase, administered on day 1 to day 21 of said C1; (ii) administering on day 1 to day 21 of said C1, said C2, said C3, said C4 and said C5 during said first phase; as well as (iii) administered on Day 1 to Day 21 of said C1, said C2, said C3, said C4, said C5, said C6 and said C7 during said second phase.

111. Sivosutumab for use according to claim 110, wherein: (i) the first increasing dose of silvostomumab is 0.3 mg; (ii) the second escalating dose of silvostomumab is 3.6 mg; (iii) the target dose of silvostomumab is between 90 mg and 198 mg, inclusive; and; (iv) Lenalidomide was administered at a dose of 10 mg or 15 mg.

112. Sivosutumab for use according to claim 111, wherein the target dose is 90 mg.

113. Sivosutumab for use according to claim 111, wherein the target dose is 132 mg.

114. Sivosutumab for use according to claim 111, wherein the target dose is 160 mg.

115. Sivosutumab for use according to any one of claims 110 to 114, wherein: (i) the subject experiences a PR or better after induction therapy; (ii) the subject has undergone ASCT within 100 days of starting the method and / or does not have progressive disease; (iii) administering the silvostomumab and the lenalidomide to the patient as post-transplant maintenance therapy; and (iv) the high-risk cytogenetic characteristics include one or more of the following: translocation events t(4;14) or t(14;16), del(17p) or 1q gain.

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