RU2848338C1 - Methods, therapies and uses for treatment of malignant neoplasm - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to oncology, particularly to a method of treating multiple myeloma in an individual. Method provides a combined treatment involving the introduction of the bispecific anti-BCMA antibody – elranatamab, and the anti-PD-1 antibody sasanlimab, or the immunomodulatory agent – lenalidomide or pomalidomide.

EFFECT: method demonstrates higher efficacy in treating multiple myeloma than treatment with any therapeutic agent separately.

46 cl, 8 dwg, 19 tbl, 16 ex

Description

FIELD OF INVENTION

The present invention relates to monotherapy and combination therapy methods that can be used to treat malignant neoplasms and/or tumor-associated diseases. In particular, the invention relates to monotherapy and combination therapy methods comprising a bispecific antibody against BCMA×CD3.

LEVEL OF TECHNOLOGY

B-cell maturation antigen (BCMA, CD269, or TNFRSF17) is a member of the tumor necrosis factor receptor (TNFR) superfamily. BCMA was identified in a malignant human T-cell lymphoma containing the t(4;16) translocation. The gene is selectively expressed in the B-cell lineage, with highest expression in plasmablasts and plasma cells, antibody-secreting cells. BCMA binds two ligands, B-cell activating factor (BAFF) (also called B-lymphocyte stimulator (BLyS) and APOL-related leukocyte-expressed ligand (TALL-1)) and proliferation-inducing ligand (APRIL), with affinities of 1 μM and 16 nM, respectively. Binding of APRIL or BAFF to BCMA stimulates a signaling cascade involving NF-kappa B, Elk-1, c-Jun N-terminal kinase, and p38 mitogen-activated protein kinase, which produces signals for cell survival and proliferation. BCMA is also expressed on malignant B cells and several malignancies involving B lymphocytes, including multiple myeloma, plasmacytoma, Hodgkin lymphoma, and chronic lymphocytic leukemia. In autoimmune diseases involving plasmablasts, such as systemic lupus erythematosus (SLE) and rheumatoid arthritis, BCMA-expressing antibody-producing cells secrete autoantibodies that attack the body itself. BCMA is also found in a soluble form (i.e., soluble BCMA or sBCMA) in the peripheral blood of patients with multiple myeloma and can lead to the failure of BCMA-specific therapies. Several BCMA-specific therapies are currently in development; however, multiple myeloma remains an incurable disease, and almost all patients develop resistance to these therapies and eventually relapse.

Programmed death receptor 1 (PD-1) and PD-1 ligands 1 and 2 (PD-L1 and PD-L2, respectively) play key roles in regulating the immune system. Expressed on activated T cells, PD-1 is upregulated by PD-L1 (also known as B7-H1) and PD-L2 expressed by stromal cells, tumor cells, or both, initiating T cell death and localized immunosuppression (Dong et al., Nat Med 1999; 5:1365–69; Freeman et al. J Exp Med 2000; 192:1027–34), potentially providing an immunotolerant environment for tumor development and growth. Conversely, inhibition of this interaction can enhance local T-cell responses and mediate antitumor activity in nonclinical animal models (Iwai Y, et al. Proc Natl Acad Sci USA 2002; 99:12293-97). Several antibodies that inhibit the interaction between PD-1 and one or both of its ligands, PD-L1 and PD-L2, are currently in development for the treatment of malignancies.

The Notch pathway is a conserved signaling pathway involved in cell fate determination, proliferation, angiogenesis, and apoptosis. A unique characteristic of the Notch pathway is that both the ligands (Jagged-1, 2, and Delta-1, 3, 4) and the receptors (Notch-1, 2, 3, 4) are type I membrane proteins. Following direct cell-cell contact, Notch receptors are cleaved by γ-secretase, releasing the intracellular domain (NICD), which translocates to the nucleus to modulate transcription. γ-secretase inhibitors (GSIs) have been developed for a number of diseases, such as Alzheimer's disease and malignancies.

There remains a need for improved therapies for the treatment of malignancies and/or tumor-associated diseases, such as multiple myeloma. Furthermore, there is a need for therapies that are more effective than existing therapies. Preferred combination therapies of the present invention demonstrate greater efficacy than treatment with either therapeutic agent alone.

ESSENCE OF THE INVENTION

The present invention relates to methods of therapy, including combination therapy methods, for the treatment of malignant neoplasms and/or tumor-associated diseases. The present invention relates to methods of treating malignant neoplasms and/or tumor-associated diseases in an individual. The present invention also relates to methods of inhibiting tumor growth or progression in an individual having malignant cells. The present invention also relates to methods of inhibiting the metastasis of malignant cells in an individual. The present invention also relates to methods of inducing tumor regression in an individual having malignant cells.

The present invention relates to methods for treating a malignant neoplasm and/or a tumor-associated disease in an individual, comprising subjecting the individual to a combination treatment comprising a first therapeutic agent and a second therapeutic agent. The present invention further relates to a medicament comprising the first therapeutic agent and the second therapeutic agent for use in treating a malignant neoplasm and/or a tumor-associated disease in an individual. The present invention further relates to a first therapeutic agent for use in treating a malignant neoplasm and/or a tumor-associated disease in an individual, wherein the first therapeutic agent is administered in combination with the second therapeutic agent.

In some aspects, the first therapeutic agent is a therapeutic agent specific for the B-cell maturation antigen (BCMA). In some aspects, the second therapeutic agent is an anti-PD-1 antibody, an anti-PD-L1 antibody, an immunomodulatory agent, or a gamma-secretase inhibitor (GSI).

In some aspects, the first therapeutic agent is a bispecific anti-BCMA antibody. In some aspects, the second therapeutic agent is an anti-PD-1 antibody. In another aspect, the second therapeutic agent is an anti-PD-L1 antibody. In another aspect, the second therapeutic agent is an immunomodulatory agent. In another aspect, the second therapeutic agent is a GSI.

In some aspects, the first therapeutic agent is a bispecific anti-BCMA antibody and the second therapeutic agent is an anti-PD-1 antibody. In another aspect, the first therapeutic agent is a bispecific anti-BCMA antibody and the second therapeutic agent is an anti-PD-L1 antibody. In another aspect, the first therapeutic agent is a bispecific anti-BCMA antibody and the second therapeutic agent is an immunomodulatory agent. In another aspect, the first therapeutic agent is a bispecific anti-BCMA antibody and the second therapeutic agent is a GSI.

In some aspects, the combination treatment further includes a third, fourth, or fifth therapeutic agent. In some aspects, the combination treatment further includes a chemotherapeutic agent. In some aspects, the therapeutic agents are administered to the individual simultaneously, separately, or sequentially.

In some aspects, the anti-BCMA bispecific antibody is PF-06863135, the anti-PD-1 antibody is sasanlimab, the immunomodulatory agent is lenalidomide or pomalidomide, and/or GSI is nirogacestat or a pharmaceutically acceptable salt thereof. In one aspect, the anti-BCMA bispecific antibody is PF-06863135. In one aspect, the anti-PD-1 antibody is sasanlimab. In one aspect, the immunomodulatory agent is lenalidomide. In another aspect, the immunomodulatory agent is pomalidomide. In one aspect, GSI is nirogacestat or a pharmaceutically acceptable salt thereof.

In some aspects, at least one of the therapeutic agents is administered to the individual in an intravenous (IV), subcutaneous (SC), or oral dose.

In some aspects, at least one of the therapeutic agents is administered to the individual at a dose of about 0.01 μg/kg, 0.02 μg/kg, 0.03 μg/kg, 0.04 μg/kg, 0.05 μg/kg, 0.06 μg/kg, 0.07 μg/kg, 0.08 μg/kg, 0.09 μg/kg, 0.1 μg/kg, 0.2 μg/kg, 0.3 μg/kg, 0.4 μg/kg, 0.5 μg/kg, 0.6 μg/kg, 0.7 μg/kg, 0.8 μg/kg, 0.9 μg/kg, 1 μg/kg, 2 μg/kg, 3 μg/kg, 4 μg/kg, 5 mcg/kg, 6 mcg/kg, 7 mcg/kg, 8 mcg/kg, 9 mcg/kg, 10 mcg/kg, 15 mcg/kg, 20 mcg/kg, 25 mcg/kg, 30 mcg/kg, 35 mcg/kg, 40 mcg/kg, 45 mcg/kg, 50 mcg/kg, 60 mcg/kg, 70 mcg/kg, 80 mcg/kg, 90 mcg/kg, 100 mcg/kg, 110 mcg/kg, 120 mcg/kg, 130 mcg/kg, 140 mcg/kg, 150 mcg/kg, 200 mcg/kg, 250 mcg/kg, 300 mcg/kg, 400 mcg/kg, 500 mcg/kg, 600 mcg/kg, 700 mcg/kg, 800 mcg/kg, 900 mcg/kg, 1000 mcg/kg, 1200 mcg/kg, 1400 mcg/kg or more.

In some aspects, at least one of the therapeutic agents is administered to the individual at a dose of from about 1 mg/kg to about 1000 mg/kg, from about 2 mg/kg to about 900 mg/kg, from about 3 mg/kg to about 800 mg/kg, from about 4 mg/kg to about 700 mg/kg, from about 5 mg/kg to about 600 mg/kg, from about 6 mg/kg to about 550 mg/kg, from about 7 mg/kg to about 500 mg/kg, from about 8 mg/kg to about 450 mg/kg, from about 9 mg/kg to about 400 mg/kg, from about 5 mg/kg to about 200 mg/kg, from about 2 mg/kg to about 150 mg/kg, from about 5 mg/kg to about 100 mg/kg, from about 10 mg/kg to about 100 mg/kg or from about 10 mg/kg to about 60 mg/kg; or

In some aspects, at least one of the therapeutic agents is administered to the individual at a fixed dose of about 0.05 mcg, 0.2 mcg, 0.5 mcg, 1 mcg, 10 mcg, 100 mcg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 75 mg, 80 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 350 mg, 700 mg, 750 mg, 800 mg, 900 mg, 1000 mg, 1500 mg or more.

In some aspects, at least one of the therapeutic agents is administered to the individual at least once per day, once per day, twice per day, three times per day, four times per day, once every two days, once every three days, once per week, once every two weeks, once every three weeks, once every four weeks, once every 30 days, once every five weeks, once every six weeks, once per month, once every two months, once every three months, or once every four months.

In some aspects, the malignancy and/or tumor-associated disease is a B-cell malignancy and/or tumor-associated disease. In some aspects, the B-cell malignancy and/or tumor-associated disease is selected from multiple myeloma, malignant plasma cell neoplasia, lymphoma, Hodgkin's lymphoma, nodular Hodgkin's lymphoma with lymphocyte predominance, Kahler disease and myelomatosis, plasma cell leukemia, bone and extramedullary plasmacytoma with multiple myeloma, solitary bone and extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering myeloma, light chain amyloidosis, osteosclerotic myeloma, B-cell prolymphocytic leukemia, hairy cell leukemia, B-cell non-Hodgkin's lymphoma (NHL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL), chronic myeloid leukemia (CML), follicular lymphoma, Burkitt lymphoma, marginal zone cell lymphoma, mantle cell lymphoma, large cell lymphoma, progenitor B-cell lymphoblastic lymphoma, myeloid leukemia, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma, mucosal lymphoid tissue lymphoma, small lymphocytic lymphoma, primary mediastinal (thymic) large B-cell lymphoma, lymphoplasmacytic lymphoma, marginal zone B-cell lymphoma, splenic marginal zone cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, Hodgkin's lymphoma, T-cell/histiocytic large B-cell lymphoma, primary central nervous system lymphoma, primary cutaneous Diffuse large B-cell lymphoma (foot type), EBV-positive diffuse large B-cell lymphoma of the elderly, inflammation-associated diffuse large B-cell lymphoma, ALK-positive large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma arising in HHV8-associated multicentric Castleman disease, undifferentiated B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and Burkitt's lymphoma, undifferentiated B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin's lymphoma, and other B-cell lymphomas. In some aspects, the B-cell malignancy is multiple myeloma. In some aspects, multiple myeloma is relapsed/refractory multiple myeloma.

The present invention also relates to methods for treating multiple myeloma in an individual, comprising subjecting the individual to a combination treatment comprising a first therapeutic agent and a second therapeutic agent, wherein the first therapeutic agent is a bispecific antibody against a B-cell maturation antigen (BCMA), and the second therapeutic agent is an anti-PD-1 antibody, an anti-PD-L1 antibody, an immunomodulatory agent, or a gamma-secretase inhibitor (GSI). The present invention also relates to a first therapeutic agent for use in methods for treating multiple myeloma in an individual, wherein the first therapeutic agent is a bispecific antibody against a B-cell maturation antigen (BCMA), and is administered in combination with a second therapeutic agent selected from an anti-PD-1 antibody, an anti-PD-L1 antibody, an immunomodulatory agent, or a gamma-secretase inhibitor (GSI). In some aspects, the first therapeutic agent is a bispecific anti-BCMA antibody and the second therapeutic agent is an anti-PD-1 antibody. In another aspect, the first therapeutic agent is a bispecific anti-BCMA antibody and the second therapeutic agent is an anti-PD-L1 antibody. In another aspect, the first therapeutic agent is a bispecific anti-BCMA antibody and the second therapeutic agent is an immunomodulatory agent. In another aspect, the first therapeutic agent is a bispecific anti-BCMA antibody and the second therapeutic agent is a GSI.

The present invention also relates to methods of treating multiple myeloma in an individual, comprising subjecting the individual to a combination treatment comprising a first therapeutic agent and a second therapeutic agent, wherein the first therapeutic agent is PF-06863135 and the second therapeutic agent is sasanlimab.

The present invention also relates to methods of treating multiple myeloma in an individual, comprising subjecting the individual to a combination treatment comprising a first therapeutic agent and a second therapeutic agent, wherein the first therapeutic agent is PF-06863135 and the second therapeutic agent is lenalidomide.

The present invention also relates to methods of treating multiple myeloma in an individual, comprising subjecting the individual to a combination treatment comprising a first therapeutic agent and a second therapeutic agent, wherein the first therapeutic agent is PF-06863135 and the second therapeutic agent is pomalidomide.

The present invention also relates to methods of treating multiple myeloma in an individual, comprising subjecting the individual to a combination treatment comprising a first therapeutic agent and a second therapeutic agent, wherein the first therapeutic agent is PF-06863135 and the second therapeutic agent is nirogacestat.

The present invention also relates to methods of treating a malignancy in an individual comprising administering PF-06863135 to the individual according to a dosing regimen.

In some embodiments, the administration regimen is:

(a) 0.1, 0.3, 1, 3, 10, 30, 50, or 100 mcg/kg once weekly (Q1W) intravenously (IV).

(b) 0.1, 0.3, 1, 3, 10, 30, 50, or 100 mcg/kg every two weeks (Q2W) IV;

(c) approximately 0.5 to 10 mg Q1W IV or Q2W IV;

(d) approximately 0.5, 1, 2, 3, 4, 5, 6, 7, 7.5, or 8 mg Q1W IV or Q2V IV.

(e) a priming administration of a single priming dose of approximately 0.5, 1, 2, 3, 4, 5, 6, 7.5, or 8 mg Q1W IV for one week followed by a first treatment administration of approximately 6, 7, 7.5, 8, 9, or 10 mg Q1W IV or Q2W IV, where the priming dose is less than the single treatment dose; or

(f) a priming administration of a single priming dose of about 0.5, 1, 2, 3, 4, 5, 6, 7, 7.5, or 8 mg Q1W for one week, followed by a first treatment administration of about 6, 7, 7.5, 8, 9, or 10 mg Q1W IV for 2 to 20, 21, 22, 23, 24, 25 to 46, 47, or 48 weeks, followed by a second treatment administration of about 6, 7, 7.5, 8, 9, or 10 Q2W IV, where the priming dose is less than the single dose in the first treatment administration.

In another aspect of the invention, the administration regimen is:

(a) 80, 130, 215, 360, 600, or 1000 mcg/kg Q1W subcutaneously (SC);

(b) 80, 130, 215, 360, 600, or 1000 μg/kg Q2W SC;

(c) approximately 16 to 80 mg Q1W SC or Q2W SC;

(d) approximately 16 to 20, 40 to 44, or 76 to 80 mg Q1W SC;

(e) approximately 16 to 20, 40 to 44, or 76 to 80 mg Q2W SC;

(f) approximately 40 mg Q1W SC or Q2W SC;

(g) approximately 44 mg Q1W SC or Q2W SC;

(h) approximately 76 mg Q1W SC or Q2W SC;

(i) approximately 80 mg Q1W SC or Q2W SC;

(j) a priming dose of approximately 44 mg Q1W SC for 1-4 weeks or a priming dose of approximately 32 mg Q1W SC for 1-4 weeks followed by a first treatment dose of approximately 76 mg Q1W SC or Q2W SC;

(k) a priming dose of approximately 40 mg Q1W SC for 1-4 weeks followed by a first treatment dose of approximately 80 mg Q1W SC or Q2W SC;

(l) a priming administration of approximately 44 mg Q1W SC for 1-4 weeks or a priming administration of approximately 32 mg Q1W SC for 1-4 weeks followed by a first treatment administration of approximately 76 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47 or 48 weeks followed by a second treatment administration of approximately 76 mg Q2W SC;

(m) a priming administration of approximately 40 mg Q1W SC for 1-4 weeks followed by a first treatment administration of approximately 80 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47 or 48 weeks followed by a second treatment administration of approximately 80 mg Q2W SC;

(n) a priming dose of approximately 44 mg Q1W SC for 1 week followed by a first treatment dose of approximately 76 mg Q1W SC or Q2W SC;

(o) a priming dose of approximately 32 mg Q1W SC for 1 week followed by a first treatment dose of approximately 76 mg Q1W SC or Q2W SC;

(p) a priming dose of approximately 40 mg Q1W SC for 1 week followed by a first treatment dose of approximately 80 mg Q1W SC or Q2W SC;

(q) a priming administration of approximately 44 mg Q1W SC for 1 week followed by a first treatment administration of approximately 76 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47 or 48 weeks followed by a second treatment administration of approximately 76 mg Q2W SC;

(r) a priming dose of approximately 44 mg Q1W SC for 1 week followed by a first treatment dose of approximately 76 mg Q1W SC for 23 weeks followed by a second treatment dose of approximately 76 mg Q2W SC;

(s) a priming dose of approximately 44 mg Q1W SC for 1 week followed by a first treatment dose of approximately 76 mg Q1W SC for 24 weeks followed by a second treatment dose of approximately 76 mg Q2W

(t) a priming administration of approximately 32 mg Q1W SC for 1 week followed by a first treatment administration of approximately 76 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47 or 48 weeks followed by a second treatment administration of approximately 76 mg Q2W SC;

(u) a priming administration of approximately 40 mg Q1W SC for 1 week followed by a first treatment administration of approximately 80 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47, or 48 weeks followed by a second treatment administration of approximately 80 mg Q2W SC; or

(v) a priming dose of approximately 40 mg Q1W SC for 1 week, followed by a first treatment dose of approximately 80 mg Q1W for 23 or 24 weeks, followed by a second treatment dose of approximately 80 mg Q2W SC.

In some embodiments, the priming administration is carried out for only one week as a single priming dose of 44 mg Q1W SC, 40 mg Q1W SC, or 32 Q1W SC.

The present invention also relates to methods of treating a cancer in an individual, comprising administering to the individual PF06863135, (a) a single priming dose of about 32 mg SC or about 44 mg SC at week 1, or both a first priming dose of about 12 mg SC and a second priming dose of about 32 mg SC at week 1, and (b) a first treatment administration of about 76 mg Q1W SC beginning at week 2, where week 1, week 2, and any subsequent weeks refer to the first, second, and any subsequent weeks that the individual is administered PF06863135, and PF06863135 is administered to the individual as a pharmaceutical formulation containing PF06863135.

In some embodiments, the individual is administered PF06863135 as a single priming dose of approximately 44 mg SC at week 1. In some embodiments, the individual is administered a first priming dose of approximately 12 mg SC on day 1 of week 1 and a second priming dose of approximately 32 mg SC on day 4 of week 1.

In some embodiments, the method further comprises administering to the individual PF06863135 in a second treatment administration of approximately 76 mg Q2W SC, beginning at week 25 or the first week of cycle 7, wherein PF06863135 in the first treatment administration is administered until the end of week 24 or the end of cycle 6, wherein the cycle is 28 days, and cycle 1, cycle 2 and subsequent cycles refer to the first, second and subsequent cycles that the individual is administered PF06863135.

In some embodiments, the individual is administered PF06863135 in a first therapeutic administration of approximately 76 mg Q1W SC and, after at least 23 weeks of such first therapeutic administration, the individual is administered PF06863135 in a second therapeutic administration of 76 mg Q2W or continues to be administered PF06863135 in the first therapeutic administration. In some embodiments, the individual is administered PF06863135 in a second therapeutic administration after at least 23 weeks of the first therapeutic administration in accordance with the relevant regulatory labeling of the pharmaceutical product or in accordance with the individual's response. In some embodiments, the individual continues to receive PF06863135 in the first treatment administration after at least 23 weeks of the first treatment administration, if the individual does not demonstrate an IMWG response in the form of a partial or better response, wherein the response is maintained for at least one month, at least two months, at least three months, at least one cycle, at least two cycles, or at least three cycles after at least six treatment cycles have been completed, and each cycle is 28 days long, and the first cycle begins on the day the individual is administered a single priming dose or the first priming dose of PF06863135.

The present invention also relates to methods of treating a malignancy in an individual comprising administering to the individual PF-06863135 according to the administration regimen:

(a) a priming dose of approximately 32 mg Q1W SC for 1 week followed by a first treatment dose of approximately 44 mg Q1W SC;

(b) a priming dose of approximately 32 mg Q1W SC for 1 week followed by a first treatment dose of approximately 44 mg Q2W SC;

(c) a priming administration of approximately 32 mg Q1W SC for 1 week followed by a first treatment administration of approximately 44 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47, or 48 weeks and a second treatment administration of approximately 44 mg Q2W SC; or

(d) a priming administration of approximately 32 mg Q1W SC for 1 week, followed by a first treatment administration of approximately 44 mg Q1W SC for 23 or 24 weeks and a second treatment administration of approximately 44 mg Q2W SC.

In some embodiments, PF-06863135 is administered to the individual as part of a priming administration of about 32 mg Q1W SC for 1 week, followed by a first treatment administration of about 44 mg Q1W SC. In some embodiments, PF-06863135 is administered to the individual as part of a priming administration of about 32 mg Q1W SC for 1 week, followed by a first treatment administration of about 44 mg Q1W SC for 23 or 24 weeks, followed by a second treatment administration of about 44 mg Q2W SC.

The present invention also relates to methods of treating a malignant neoplasm in an individual comprising subcutaneously administering PF-06863135 to the individual, a first treatment administration for 23, 24 or 25 weeks followed by a second treatment administration.

In some embodiments, the first therapeutic administration is about 4 mg Q1W, and the second therapeutic administration is about 4 mg Q1W or about 4 mg Q2W. In some embodiments, the first therapeutic administration is about 12 mg Q1W, and the second therapeutic administration is about 12 mg Q1W or about 12 mg Q2W. In some embodiments, the first therapeutic administration is about 24 mg Q1W, and the second therapeutic administration is about 24 mg Q1W or about 24 mg Q2W. In some embodiments, the first therapeutic administration is about 32 mg Q1W, and the second therapeutic administration is about 32 mg Q1W or about 32 mg Q2W. In some embodiments, the first therapeutic administration is about 44 mg Q1W, and the second therapeutic administration is about 44 mg Q1W or about 44 mg Q2W. In some embodiments, the first therapeutic administration is about 76 mg Q1W, and the second therapeutic administration is about 76 mg Q1W or about 76 mg Q2W. In some embodiments, the first therapeutic administration is about 4 mg Q1W, and the second therapeutic administration is about 4 mg Q2W. In some embodiments, the first therapeutic administration is about 12 mg Q1W, and the second therapeutic administration is about 12 mg Q2W. In some embodiments, the first therapeutic administration is about 24 mg Q1W, and the second therapeutic administration is about 24 mg Q2W. In some embodiments, the first therapeutic administration is about 32 mg Q1W, and the second therapeutic administration is about 32 mg Q2W. In some embodiments, the first therapeutic administration is about 44 mg Q1W, and the second therapeutic administration is about 44 mg Q2W. In some embodiments, the first therapeutic administration is about 76 mg Q1W and the second therapeutic administration is about 76 mg Q2W.

In some embodiments, if the amount of the dose within the first therapeutic administration is 32 mg or more, the method further comprises administering PF06863135 to the individual as part of a priming administration, and the priming administration is carried out for one week, the first dose within the first therapeutic administration is administered in the week immediately following the week in which the priming administration is carried out. In some embodiments, the priming administration is a single reconciliation dose, and the single priming dose is approximately 24 mg. In some embodiments, the priming administration includes a first reconciliation dose of approximately 4 mg and a second reconciliation dose of approximately 20 mg, and the two priming doses are administered on two different days, and the first reconciliation dose is administered before the second priming dose. In some embodiments, the priming administration comprises a first reconciliation dose of approximately 8 mg and a second reconciliation dose of approximately 16 mg, and the two priming doses are administered on two different days, and the first reconciliation dose is administered before the second priming dose. In some embodiments, the priming administration comprises a first reconciliation dose of approximately 12 mg and a second reconciliation dose of approximately 12 mg, and the two priming doses are administered on two different days, and the first reconciliation dose is administered before the second priming dose. In some embodiments, the priming administration comprises a first reconciliation dose of approximately 8 mg and a second reconciliation dose of approximately 24 mg, and the two priming doses are administered on two different days, and the first reconciliation dose is administered before the second priming dose. In some embodiments, the priming administration comprises a first priming dose of approximately 4 mg and a second priming dose of approximately 28 mg, and the two priming doses are administered on two different days and the first priming dose is administered before the second priming dose.

In some embodiments, the individual is administered PF06863135 in a second therapeutic administration for 6 to 18 cycles, wherein a cycle is 21 days or 28 days, then the individual is subjected to a third therapeutic administration of PF06863135 subcutaneously. In some embodiments, the third therapeutic administration is about 4 mg Q2W or about 4 mg Q4W. In some embodiments, the third therapeutic administration is about 12 mg Q2W or about 12 mg Q4W. In some embodiments, the third therapeutic administration is about 24 mg Q2W or about 24 mg Q4W. In some embodiments, the third therapeutic administration is about 32 mg Q2W or about 32 mg Q4W. In some embodiments, the third therapeutic administration is about 44 mg Q2W, about 44 mg Q4W. In some embodiments, the third therapeutic administration is about 76 mg Q2W or about 76 mg Q4W.

In some embodiments, the first therapeutic administration is about 4 mg Q1W, the second therapeutic administration is about 4 mg Q2W, and the third therapeutic administration is about 4 mg Q4W. In some embodiments, the first therapeutic administration is about 12 mg Q1W, the second therapeutic administration is about 12 mg Q2W, and the third therapeutic administration is about 12 mg Q4W. In some embodiments, the first therapeutic administration is about 24 mg Q1W, the second therapeutic administration is about 24 mg Q2W, and the third therapeutic administration is about 32 mg Q4W. In some embodiments, the first therapeutic administration is about 32 mg Q1W, the second therapeutic administration is about 32 mg Q2W, and the third therapeutic administration is about 24 mg Q4W. In some embodiments, the first therapeutic administration is about 44 mg Q1W, the second therapeutic administration is about 44 mg Q2W, and the third therapeutic administration is about 44 mg Q4W. In some embodiments, the first therapeutic administration is about 76 mg Q1W, the second therapeutic administration is about 76 mg Q2W, and the third therapeutic administration is about 76 mg Q4W.

The present invention also relates to methods of treating a malignant neoplasm in an individual comprising administering to the individual PF-06863135

(a) the first treatment administration of approximately 32 mg to approximately 76 mg Q1W SC, beginning at week 1; or

(b) a priming administration during week 1, and a first treatment administration beginning in week 2, wherein the priming administration is (i) a first reconciliation dose of from about 4 mg SC to about 32 mg SC and a second reconciliation dose of from about 12 mg SC to about 44 mg SC, wherein the first reconciliation dose and the second reconciliation dose are administered sequentially in week 1, or (ii) a single reconciliation dose of from about 24 mg to about 44 mg SC, and wherein the first treatment administration is from about 32 mg to about 76 mg Q1W SC or from about 32 mg to about 152 mg Q2W SC, beginning in week 2, and wherein the dose amount within the first treatment administration is higher than the dose amount of each of the respective single priming dose, the first priming dose, and the second priming dose;

where week 1, week 2 and any subsequent weeks refer to the first, second and any subsequent weeks that PF06863135 is administered to the individual, respectively, and PF06863135 is administered to the individual in the form of a pharmaceutical preparation containing PF06863135.

In some embodiments, the individual is given a matched administration of a single priming dose of about 24 mg SC, about 32 mg SC, or about 44 mg SC at week 1. In some embodiments, the individual is given a matched administration of a first priming dose of about 12 mg SC and a second priming dose of about 32 mg SC at week 1. In some embodiments, the individual is given a matched administration of a single priming dose of about 4 mg, about 8 mg, about 12 mg, or about 24 mg at week 1. In some embodiments, the individual is given a matched administration of a first priming dose and a second priming dose. In some embodiments, the first priming dose is about 4 mg and the second priming dose is about 20 mg. In some embodiments, the first priming dose is about 8 mg and the second priming dose is about 16 mg. In some embodiments, the first priming dose is approximately 12 mg and the second priming dose is approximately 12 mg. In some embodiments, the first priming dose is approximately 8 mg and the second priming dose is approximately 24 mg.

In some embodiments, the first therapeutic administration is about 32 mg Q1W SC or about 32 mg Q2W SC. In some embodiments, the first therapeutic administration is about 44 mg Q1W SC or about 44 mg Q2W SC. In some embodiments, the individual is subjected to the first therapeutic administration until at least the end of cycle 1 or at least the end of cycle 6, where the cycle is 21 days or 28 days, cycle 1 begins on day 1 of week 1, day 1 of week 2, or day 1 of week 3, and cycle 1, cycle 2, and subsequent cycles refer to the first, second, and subsequent cycles when PF06863135 is administered to the individual, respectively.

In some embodiments, the method further comprises administering to the individual PF06863135 in a second therapeutic administration from about 32 mg to about 152 mg Q2W SC, from about 32 mg to about 152 mg Q3W SC, or from about 32 mg to about 152 mg Q4W SC, after which the individual is no longer in the period of the first therapeutic administration, wherein the second therapeutic administration is carried out at a frequency less than the frequency of the corresponding first therapeutic administration, or the second therapeutic administration has a smaller dose amount than the first therapeutic administration. In some embodiments, where after subjecting the individual to the first treatment administration, at least until the end of cycle 6, the individual is subjected to a second treatment administration of PF06863135 instead of the first treatment administration, or the individual can continue to be subjected to the first treatment administration, and where the second treatment administration is from about 32 mg to about 152 mg Q2W SC, from about 32 mg to about 152 mg Q3W SC, or from about 32 mg to about 152 mg Q4W SC, where the second treatment administration is carried out at a frequency less than the frequency of the first treatment administration, or the second treatment administration has a smaller dose amount than the first treatment administration. In some embodiments, wherein (i) the first therapeutic administration is about 32 mg Q1W SC, and the second therapeutic administration is about 32 mg Q2W SC, 32 mg Q3W SC, 32 mg Q4W SC, 44 mg Q2W SC, 44 mg Q3W SC, 44 mg Q4W SC, 76 mg Q3W SC, 76 mg Q4W SC, 116 mg Q4W SC, or 152 mg Q4W SC, or (ii) the first therapeutic administration is about 32 Q2W SC, and the second therapeutic administration is about 32 mg Q3W SC, 32 mg Q4W SC, 44 mg Q3W SC, 44 mg Q4W SC, 76 mg Q3W SC, 76 mg Q4W SC, 116 mg Q4W SC, or 152 mg Q4W SC. In some embodiments, wherein (i) the first therapeutic administration is about 44 mg Q1W SC and the second therapeutic administration is about 44 mg Q2W SC, 44 mg Q3W SC, 44 mg Q4W SC, 76 mg Q2W SC, 76 mg Q3W SC, 76 mg Q4W SC, 116 mg Q4W SC, or about 152 mg Q4W SC, or (ii) the first therapeutic administration is about 44 mg Q2W SC and the second therapeutic administration is about 32 mg Q2W SC, 44 mg Q3W SC, 76 mg Q3W SC, 116 mg Q3W SC, 152 mg Q3W SC, 32 mg Q4W SC, 44 mg Q4W SC, 76 mg Q4W SC, 116 mg Q4W SC, or about 152 mg Q4W SC. In some embodiments, the individual is subjected to a second therapeutic administration in accordance with the relevant regulatory labeling of the pharmaceutical product. In some embodiments, the individual is subjected to the second therapeutic administration in accordance with the individual's response to the first therapeutic administration. In some embodiments, the individual continues to be subjected to the first therapeutic administration if the individual does not demonstrate an IMWG response in the form of a partial or better response, and the response is maintained for at least one month, at least two months, at least three months, at least one cycle, at least two cycles, or at least three cycles while the individual is in the period of the first therapeutic administration.

In some embodiments, the first therapeutic administration is (i) about 76 mg Q1W SC, (ii) about 76 mg Q2W SC, or (iii) about 76 mg Q1W SC for three weeks followed by about 116 mg Q1W SC, or (iv) about 76 mg Q1W SC for three weeks followed by about 152 mg Q1W SC. In some embodiments, the individual is subjected to the first therapeutic administration until at least the end of cycle 1, at least the end of cycle 3, or at least the end of cycle 6, where the cycle is 21 days or 28 days, and cycle 1 begins on day 1 of week 1, day 1 of week 2, or day 1 of week 3, and cycle 1, cycle 2, and subsequent cycles refer to the first, second, and subsequent cycles when PF06863135 is administered to the individual, respectively. In some embodiments, the method further comprises administering to the individual PF06863135 in a second therapeutic administration from about 44 mg to about 152 mg Q2W SC, from about 44 mg to about 152 mg Q3W SC, or from about 44 mg to about 152 mg Q4W SC, after which the individual is no longer in the period of the first therapeutic administration, wherein the second therapeutic administration is carried out at a frequency less than the frequency of the first therapeutic administration, or the second therapeutic administration has a smaller dose amount than the first therapeutic administration. In some embodiments, where after subjecting the individual to the first therapeutic administration, at least until the end of cycle 6, the individual is subjected to a second therapeutic administration of from about 44 mg to about 152 mg Q2W SC, from about 44 mg to about 152 mg Q3W SC, or from about 44 mg to about 152 mg Q4W SC, instead of the first therapeutic administration, or the individual may continue to be subjected to the first therapeutic administration, where the second therapeutic administration is carried out at a frequency less than the frequency of the corresponding first therapeutic administration, or the second therapeutic administration has a smaller dose amount than the first therapeutic administration. In some embodiments, the first therapeutic administration is about 76 mg Q1W SC, and the second therapeutic administration is about 44 mg Q2W SC, about 76 mg Q2W SC, about 116 mg Q2W SC, about 152 mg Q2W SC, about 44 mg Q3W SC, about 76 mg Q3W SC, about 116 mg Q3W SC, about 152 mg Q3W SC, about 44 mg Q4W SC, about 76 mg Q4W SC, about 116 mg Q4W SC, or about 152 mg Q4W SC. In some embodiments, the first therapeutic administration is about 76 mg Q2W SC, and the second therapeutic administration is about 44 mg Q2W SC, about 44 mg Q3W SC, about 76 mg Q3W SC, about 116 mg Q3W SC, about 152 mg Q3W SC, about 44 mg Q4W SC, about 76 mg Q4W SC, about 116 mg Q4W SC, or about 152 mg Q4W SC. In some embodiments, the first therapeutic administration is about 76 mg Q1W, the second therapeutic administration is about 76 mg Q2W. In some embodiments, the first therapeutic administration is about 76 mg Q2W, the second therapeutic administration is about 76 mg Q4W. In some embodiments, the individual is subjected to the second therapeutic administration in accordance with the relevant regulatory labeling of the pharmaceutical product. In some embodiments, the individual is subjected to a second therapeutic administration based on the individual's response to the first therapeutic administration. In some embodiments, the individual is subjected to the second therapeutic administration if the individual exhibits an IMWG response in the form of a partial or better response, wherein the response is maintained for at least one month, at least two months, at least three months, at least one cycle, at least two cycles, or at least three cycles while the individual is in the period of the first therapeutic administration.

In some embodiments, the individual is administered PF06863135 in a first treatment administration until the end of cycle 1, followed by a second treatment administration, where the cycle is 21 days or 28 days, cycle 1 begins on day 1 of week 1, or day 1 of week 2, or day 1 of week 3, and cycle 1, cycle 2, and subsequent cycles refer to the first, second, and subsequent cycles when the individual is administered PF06863135, respectively. In some embodiments, the second treatment administration is carried out at least until the end of cycle 6, and then the individual is subjected to a third treatment administration of about 76 mg to about 152 mg of Q3W SC or about 76 mg to about 152 mg of Q4W SC instead of the second treatment administration, or the individual continues to be subjected to the second treatment administration. In some embodiments, the second therapeutic administration is carried out at least until the end of cycle 6, and then a third therapeutic administration of from about 76 mg to about 152 mg Q3W SC or from about 76 mg to about 152 mg Q4W SC is carried out. In some embodiments, the second therapeutic administration is carried out until the end of cycle 6, the first dose in the third therapeutic administration begins in cycle 7, and the third therapeutic administration is 116 mg Q4W SC or 152 mg Q4W SC. In some embodiments, PF06863135 is administered to the individual in the third therapeutic administration after the second therapeutic administration has been carried out at least until cycle 6 in accordance with the relevant regulatory labeling of the pharmaceutical product or in accordance with the individual's response. In some embodiments, the individual continues to receive PF06863135 in a second treatment administration until at least cycle 6 if the individual does not demonstrate an IMWG response of a partial or better response, wherein the response is maintained for at least one month, at least two months, at least three months, at least one cycle, at least two cycles, or at least three cycles while the individual is in the second treatment administration period. In some embodiments, the first treatment administration is about 76 mg Q1W SC, the second treatment administration is about 116 mg Q2W SC, and the third treatment administration is about 116 mg Q4W SC. In some embodiments, the first treatment administration is about 76 mg Q1W SC, the second treatment administration is about 152 mg Q2W SC, and the third treatment administration is about 152 mg Q4W SC.

In some embodiments, the method comprises subjecting the individual to a first therapeutic administration of about 32 mg Q1W for 23, 24, or 25 weeks, followed by a second therapeutic administration of about 32 mg Q1W or about 32 mg Q2W for 6 to 18 cycles, followed by a third therapeutic administration of about 32 mg Q2W or about 32 mg Q4W, wherein the cycle is 21 or 28 days. In some embodiments, the second therapeutic administration is about 32 mg Q2W, and the third therapeutic administration is about 32 mg Q4W.

In some embodiments, the method comprises subjecting the individual to a first therapeutic administration of about 44 mg Q1W for 23, 24, or 25 weeks, followed by a second therapeutic administration of about 44 mg Q1W or about 44 mg Q2W for 6 to 18 cycles, followed by a third therapeutic administration of about 44 mg Q2W or about 44 mg Q4W, wherein the cycle is 21 or 28 days. In some embodiments, the second therapeutic administration is about 44 mg Q2W, and the third therapeutic administration is about 44 mg Q4W.

In some embodiments, the method comprises subjecting the individual to a first therapeutic administration of about 76 mg Q1W for 23, 24, or 25 weeks, followed by a second therapeutic administration of about 76 mg Q1W or about 76 mg Q2W for 6 to 18 cycles, followed by a third therapeutic administration of about 76 mg Q2W or about 76 mg Q4W, wherein the cycle is 21 or 28 days. In some embodiments, the second therapeutic administration is about 76 mg Q2W, and the third therapeutic administration is about 76 mg Q4W.

In some embodiments, the method comprises subjecting the individual to a first therapeutic administration of about 116 mg Q1W for 23, 24, or 25 weeks, followed by a second therapeutic administration of about 116 mg Q1W or about 116 mg Q2W for 6 to 18 cycles, followed by a third therapeutic administration of about 116 mg Q2W or about 116 mg Q4W, wherein the cycle is 21 or 28 days. In some embodiments, the second therapeutic administration is about 116 mg Q2W, and the third therapeutic administration is about 116 mg Q4W.

In some embodiments, the method comprises subjecting the individual to a first therapeutic administration of about 152 mg Q1W for 23, 24, or 25 weeks, followed by a second therapeutic administration of about 152 mg Q1W or about 152 mg Q2W for 6 to 18 cycles, followed by a third therapeutic administration of about 152 mg Q2W or about 152 mg Q4W, wherein the cycle is 21 or 28 days. In some embodiments, the second therapeutic administration is about 152 mg Q2W, and the third therapeutic administration is about 152 mg Q4W.

In some embodiments, the cycle is 21 days if the frequency of administration of PF06863135 to the individual is Q1W or Q3W, and the cycle is 28 days if the frequency of administration of PF06863135 to the individual is Q2W or Q4W. In some embodiments, the cycle is 28 days if the frequency of administration of PF06863135 to the individual is not Q3W. In some embodiments, the cycle is 21 days in cycle 1 and until the end of the last cycle if the individual is in the first therapeutic administration period.

The present invention also relates to methods for treating a malignancy, comprising administering to an individual elranatamab (PF06863135) according to a dosing regimen as shown below, and wherein the dosing regimen is described by a week number, a dose amount, and a frequency of administration corresponding to each number of weeks:

(a)

Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-24 32; 44; 76; 116 or 152 Weekly 25 and beyond 32; 44; 76; 116 or 152 Weekly; once every two weeks; once every three weeks or once every four weeks

(b)

Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-25 32; 44; 76; 116 or 152 Weekly 26 and further 32; 44; 76; 116 or 152 Weekly; once every two weeks; once every three weeks or once every four weeks

(c)

Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-26 44 or 76 Weekly 27 and further 44 or 76 Weekly; once every two weeks; once every three weeks or once every four weeks

(d)

Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-24 32; 44; 76; 116 or 152 Once every two weeks 25 and beyond 32; 44; 76; 116 or 152 Once every two weeks; once every three weeks or once every four weeks

(e)

Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-25 32; 44; 76; 116 or 152 Once every two weeks 26 and further 32; 44; 76; 116 or 152 Once every two weeks; once every three weeks or once every four weeks

or (f)

Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-26 32; 44; 76; 116 or 152 Once every two weeks 27 and further 32; 44; 76; 116 or 152 Once every two weeks; once every three weeks or once every four weeks

where if the dose amount is 12 mg and 32 mg during week 1, the dose amount of 12 mg is administered on one day, then the dose amount of 32 mg is administered on another day, where A and B are 4 (A) and 20 (B), 8 (A) and 16 (B), 12 (A) and 12 (B), or 8 (A) and 24 (B), and where if the dose amount is A mg and B mg during week 1, the dose amount of A mg is administered on one day, then the dose amount of B mg is administered on another day.

In some embodiments, the individual is administered elranatamab (PF06863135) according to the administration regimen shown below.

(a)

Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-24 76 Weekly 25 and beyond 76 Weekly; once every two weeks; once every three weeks or once every four weeks

(b)

Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-25 76 Weekly 26 and further 76 Weekly; once every two weeks; once every three weeks or once every four weeks

(c)

Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-26 76 Weekly 27 and further 76 Weekly; once every two weeks; once every three weeks or once every four weeks

(d)

Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-24 76 Once every two weeks 25 and beyond 76 Once every two weeks; once every three weeks or once every four weeks

(e)

Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-25 76 Once every two weeks 26 and further 76 Once every two weeks; once every three weeks or once every four weeks

or (f)

Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-26 76 Once every two weeks 27 and further 76 Once every two weeks; once every three weeks or once every four weeks

In some embodiments, the individual is administered PF06863135 according to the administration schedule (a), (b), or (c), and the frequency of administration during week 25 onward, week 26 onward, and week 27 onward in the administration schedule (a), (b), and (c), respectively, is (i) weekly, (ii) once every two weeks, (iii) once every three weeks; (iv) once every four weeks; (v) weekly or once every two weeks; (vi) weekly or once every three weeks, or (vii) weekly or once every four weeks. In some embodiments, the individual is administered PF06863135 according to the administration schedule (d), (e), or (f), and the frequency of administration during week 25 onward, week 26 onward, and week 27 onward in the administration schedule (d), (e), and (f), respectively, is (i) once every two weeks, (ii) once every three weeks, (iii) once every four weeks, (iv) once every two weeks or once every three weeks, or (v) once every two weeks or once every four weeks.

In some embodiments, the individual is administered elranatamab (PF06863135) according to the administration regimen shown below.

(a)

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-24 76 Weekly 25 and beyond 76 Weekly; once every two weeks; once every three weeks or once every four weeks

(b)

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-25 76 Weekly 26 and further 76 Weekly; once every two weeks; once every three weeks or once every four weeks

(c)

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-26 76 Weekly 27 and further 76 Weekly; once every two weeks; once every three weeks or once every four weeks

(d)

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-24 76 Once every two weeks 25 and beyond 76 Once every two weeks; once every three weeks or once every four weeks

(e)

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-25 76 Once every two weeks 26 and further 76 Once every two weeks; once every three weeks or once every four weeks

or (f)

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-26 76 Once every two weeks 27 and further 76 Once every two weeks; once every three weeks or once every four weeks

In some embodiments, the individual is administered 12 mg of elranatamab on day 1 of week 1 followed by 32 mg of elranatamab on day 4 of week 1. In some embodiments, the individual is administered PF06863135 according to the administration regimen (a), (b), or (c), and the frequency of administration during week 25 onward, week 26 onward, and week 27 onward in the administration regimen (a), (b), and (c), respectively, is (i) weekly, (ii) once every two weeks, (iii) once every three weeks; (iv) once every four weeks; (v) weekly or once every two weeks; (vi) weekly or once every three weeks, or (vii) weekly or once every four weeks. In some embodiments, the individual is administered PF06863135 according to the administration schedule (d), (e), or (f), and the frequency of administration during week 25 onward, week 26 onward, and week 27 onward in the administration schedule (d), (e), and (f), respectively, is (i) once every two weeks, (ii) once every three weeks, (iii) once every four weeks, (iv) once every two weeks or once every three weeks, or (v) once every two weeks or once every four weeks.

In some embodiments, the individual is administered elranatamab (PF06863135) according to the administration regimen shown below.

(a)

Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-24 44 Weekly 25 and beyond 44 Weekly; once every two weeks; once every three weeks or once every four weeks

(b)

Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-25 44 Weekly 26 and further 44 Weekly; once every two weeks; once every three weeks or once every four weeks

(c)

Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-26 44 Weekly 27 and further 44 Weekly; once every two weeks; once every three weeks or once every four weeks

(d)

Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-24 44 Once every two weeks 25 and beyond 44 Once every two weeks; once every three weeks or once every four weeks

(e)

Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-25 44 Once every two weeks 26 and further 44 Once every two weeks; once every three weeks or once every four weeks

or (f)

Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-26 44 Once every two weeks 27 and further 44 Once every two weeks; once every three weeks or once every four weeks

In some embodiments, the individual is administered a single dose of 32 mg of elranatamab during week 1. In some embodiments, the individual is administered 12 mg of elranatamab on day 1 of week 1 followed by 32 mg of elranatamab on day 4 of week 1. In some embodiments, the individual is administered PF06863135 according to administration schedule (a), (b), or (c), and the frequency of administration during week 25 onward, week 26 onward, and week 27 onward in administration schedule (a), (b), and (c), respectively, is (i) weekly, (ii) every two weeks, (iii) every three weeks; (iv) every four weeks; (v) weekly or every two weeks; (vi) weekly or every three weeks, or (vii) weekly or every four weeks. In some embodiments, the individual is administered PF06863135 according to the administration schedule (d), (e), or (f), and the frequency of administration during week 25 onward, week 26 onward, and week 27 onward in the administration schedule (d), (e), and (f), respectively, is (i) once every two weeks, (ii) once every three weeks, (iii) once every four weeks, (iv) once every two weeks or once every three weeks, or (v) once every two weeks or once every four weeks.

In some embodiments, if the dose amount and the frequency of administration during week 1 are collectively referred to as a priming administration, and if the individual is administered only one dose of elranatamab as part of the priming administration, such single dose is referred to as a single remediation dose, if the individual is administered two doses of elranatamab sequentially during week 1, the two doses are referred to as the first remediation dose and the second remediation dose, respectively; the amount of dose and the frequency of administration during weeks 2-24, weeks 2-25 and weeks 2-26 at the corresponding administration doses (a) and (d), (b) and (e) and (c) and (f), respectively, in each administration schedule are collectively referred to as the first therapeutic administration, the amount of dose and the frequency of administration during week 25 and onward, week 26 and onward and week 27 and onward at the corresponding administration doses (a) and (d), (b) and (e) and (c) and (f) in each administration schedule are collectively referred to as the second therapeutic administration.

In some embodiments, the individual is subjected to a second treatment administration of PF06863135 for 6 to 18 cycles, then the individual is subjected to a third treatment administration of PF06863135 subcutaneously, wherein the third treatment administration is 32 mg Q2W, 32 mg Q4W, 44 mg Q2W, 44 mg Q4W, 76 mg Q2W, 76 mg Q4W, 116 mg Q2W, 116 mg Q4W, 152 mg Q2W, or 152 mg Q4W, wherein the cycle is 21 days or 28 days, and cycle 1 begins on day 1 of week 1, day 1 of week 2, or day 1 of week 3.

In some embodiments, the first therapeutic administration is 32 mg Q1W, the second therapeutic administration is 32 mg Q1W or 32 mg Q2W, and the third therapeutic administration is 32 mg Q2W or 32 mg Q4W. In some embodiments, the first therapeutic administration is 32 mg Q1W, the second therapeutic administration is 32 mg Q2W, and the third therapeutic administration is 32 mg Q4W. In some embodiments, the first therapeutic administration is 44 mg Q1W, the second therapeutic administration is 44 mg Q1W or 44 mg Q2W, and the third therapeutic administration is 44 mg Q2W or 44 mg Q4W. In some embodiments, the first therapeutic administration is 44 mg Q1W, the second therapeutic administration is 44 mg Q2W, and the third therapeutic administration is 44 mg Q4W. In some embodiments, the first therapeutic administration is 76 mg Q1W, the second therapeutic administration is 76 mg Q1W or 76 mg Q2W, and the third therapeutic administration is 76 mg Q2W or 76 mg Q4W. In some embodiments, the first therapeutic administration is 76 mg Q1W, the second therapeutic administration is 76 mg Q2W, and the third therapeutic administration is 76 mg Q4W. In some embodiments, the first therapeutic administration is 116 mg Q1W, the second therapeutic administration is 116 mg Q1W or 116 mg Q2W, and the third therapeutic administration is 116 mg Q2W or 116 mg Q4W. In some embodiments, the first therapeutic administration is 116 mg Q1W, the second therapeutic administration is 116 mg Q2W, and the third therapeutic administration is 116 mg Q4W. In some embodiments, the first therapeutic administration is 152 mg Q1W, the second therapeutic administration is 152 mg Q1W or 32 mg Q2W, and the third therapeutic administration is 152 mg Q2W or 152 mg Q4W.

The present invention also relates to methods for treating a malignancy, comprising administering elranatamab (PF06863135) to an individual according to a dosing regimen as shown below, and wherein the dosing regimen is described by a week number, a dose amount, and a frequency of administration corresponding to each number of weeks:

Week number Dose amount (mg) Frequency of administration 1 44; or 32; or 12 and 32; or A and B Weekly 2-4 from 44 to 152 Weekly 5-24 from 44 to 152 Weekly or biweekly 25 and beyond from 44 to 152 Once every two weeks; once every three weeks or once every four weeks

where if the dose amount is 12 mg and 32 mg during week 1, the dose amount of 12 mg is administered on one day, then the dose amount of 32 mg is administered on another day, where A and B are 4 (A) and 20 (B), 8 (A) and 16 (B), 12 (A) and 12 (B), or 8 (A) and 24 (B), and where the dose amount of A mg is administered on one day, then the dose amount of B mg is administered on another day.

In some embodiments, the individual is administered 12 mg of elranatamab on day 1 of week 1 followed by 32 mg of elranatamab on day 4 of week 1.

In some embodiments, elranatamab is administered to the individual according to the following administration regimen:

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 76 Weekly 5-24 116 Once every two weeks 25 and beyond 116 Once every two weeks; once every three weeks or once every four weeks

In some embodiments, the frequency of administration during week 25 and thereafter is once every four weeks.

In some embodiments, elranatamab is administered to the individual according to the following administration regimen:

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 76 Weekly 5-24 152 Once every two weeks 25 and beyond 152 Once every two weeks; once every three weeks or once every four weeks

In some embodiments, the frequency of administration during week 25 and thereafter is once every four weeks.

In some embodiments, the dose amount and the frequency of administration during week 1 are collectively referred to as a priming administration, and if the individual is administered only one dose of elranatamab as part of the priming administration, such a single dose is referred to as a single remediation dose, if the individual is administered two doses of elranatamab sequentially during week 1, the two doses are referred to as a first remediation dose and a second remediation dose, respectively, the dose amount and the frequency of administration during weeks 2-4 are collectively referred to as a first therapeutic administration, the dose amount and the frequency of administration during weeks 5-24 are collectively referred to as a second therapeutic administration, and the dose amount and the frequency of administration during week 25 and thereafter are collectively referred to as a third therapeutic administration.

The present invention also relates to methods for treating a malignancy, comprising administering elranatamab (PF06863135) to an individual according to a dosing regimen as shown below, and wherein the dosing regimen is described by a week number, a dose amount, and a frequency of administration corresponding to each number of weeks:

Week number Dose amount (mg) Frequency of administration 1 44; or 32; or 12 and 32; or A and B Weekly 2-4 from 44 to 76 Weekly 5-12 from 44 to 152 Weekly 13-24 from 44 to 152 Weekly or biweekly 25 and beyond from 44 to 152 Once every two weeks; once every three weeks or once every four weeks

where if the dose amount is 12 mg and 32 mg during week 1, the 12 mg dose amount is administered on one day, then the 32 mg dose amount is administered on another day, where A and B are 4 (A) and 20 (B), 8 (A) and 16 (B), 12 (A) and 12 (B), or 8 (A) and 24 (B), and where the dose amount A mg is administered on one day, then the dose amount B mg is administered on another day. In some embodiments, the individual is administered 12 mg of elranatamab on day 1 of week 1, followed by 32 mg of elranatamab on day 4 of week 1.

In some embodiments, elranatamab is administered to the individual according to the following administration regimen:

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 76 Weekly 5-12 116 Weekly 13-24 116 Weekly or biweekly 25 and beyond 116 Once every two weeks; once every three weeks or once every four weeks

In some embodiments, the frequency of administration during weeks 13-24 is once every two weeks.

In some embodiments, the frequency of administration during week 25 and thereafter is once every four weeks.

In some embodiments, elranatamab is administered to the individual according to the following administration regimen:

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 76 weekly 2-12 152 Weekly 13-24 152 Weekly or biweekly 25 and beyond 152 Once every two weeks or once every four weeks

In some embodiments, the frequency of administration during weeks 13-24 is once every two weeks.

In some embodiments, the frequency of administration during week 25 and thereafter is once every four weeks.

In some embodiments, elranatamab is administered to the individual according to the following administration regimen:

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 76 weekly 5-12 76 Weekly 13-24 76 Weekly or biweekly 25 and beyond 76 Once every two weeks; once every three weeks or once every four weeks

In some embodiments, the frequency of administration during weeks 13-24 is once every two weeks.

In some embodiments, the frequency of administration during week 25 and thereafter is once every four weeks. In some embodiments, the frequency of administration during weeks 13-24 is once every two weeks, where the frequency of administration during week 25 and thereafter is once every two weeks or once every four weeks.

In some embodiments, elranatamab is administered to the individual according to the following administration regimen:

Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 44 weekly 5-12 44 Weekly 13-24 44 Weekly or biweekly 25 and beyond 44 Once every two weeks; once every three weeks or once every four weeks

In some embodiments, the frequency of administration during weeks 13-24 is once every two weeks. In some embodiments, the frequency of administration during week 25 and thereafter is once every four weeks.

In some embodiments, the dose amount and the frequency of administration during week 1 are collectively referred to as a priming administration, and if the individual is administered only one dose of elranatamab as part of the priming administration, such a single dose is referred to as a single remediation dose, if the individual is administered two doses of elranatamab sequentially during week 1, the two doses are referred to as a first remediation dose and a second remediation dose, respectively, the dose amount and frequency of administration during weeks 2-4 and the dose amount and frequency of administration during weeks 5-12 are collectively referred to as a first therapeutic administration, the dose amount and frequency of administration during weeks 13-24 are collectively referred to as a second therapeutic administration, and the dose amount and frequency of administration during week 25 and thereafter are collectively referred to as a third therapeutic administration.

The present invention further relates to elranatamab (PF-06853135) for use in a method of treating a malignancy using the administration regimen as defined herein.

In some embodiments, the malignancy is multiple myeloma. In some embodiments, the malignancy is advanced multiple myeloma. In some embodiments, the malignancy is relapsed or refractory multiple myeloma.

In some embodiments, the malignancy is triple-class refractory multiple myeloma. In some embodiments, the individual's multiple myeloma is refractory to all three of the following therapies for multiple myeloma: (1) prior therapy for multiple myeloma including a proteasome inhibitor, (2) prior therapy for multiple myeloma including an immunomodulatory agent, and (3) prior therapy for multiple myeloma including an anti-CD38 antibody.

In some embodiments, the malignancy is dual-grade refractory multiple myeloma. In some embodiments, the individual's multiple myeloma is refractory to at least two of the following three types of multiple myeloma therapies: (1) prior multiple myeloma therapy comprising a proteasome inhibitor, (2) prior multiple myeloma therapy comprising an immunomodulatory agent, and (3) prior multiple myeloma therapy comprising an anti-CD38 antibody.

In some embodiments, the malignancy is newly diagnosed multiple myeloma. In some embodiments, the malignancy is multiple myeloma, and the individual has undergone a stem cell transplant. In some embodiments, the individual has undergone an autologous stem cell transplant. In some embodiments, the individual has undergone an autologous stem cell transplant or an allogeneic stem cell transplant. In some embodiments, the individual is positive for minimal residual disease after stem cell transplantation.

In some embodiments, the malignancy is multiple myeloma, wherein in some embodiments, the individual has progressed or is intolerant to conventional therapy for multiple myeloma. In some embodiments, the conventional therapy for multiple myeloma comprises at least one drug selected from the group consisting of a proteasome inhibitor, an immunomodulatory agent, and an anti-CD38 antibody.

In some embodiments, the malignancy is multiple myeloma, wherein the individual has been treated with at least four prior therapies, and the individual's multiple myeloma is refractory or relapsed under the conditions of (1) prior multiple myeloma therapy comprising a proteasome inhibitor, (2) prior multiple myeloma therapy comprising an immunomodulatory agent, and (3) prior multiple myeloma therapy comprising an anti-CD38 monoclonal antibody, and wherein the individual has experienced disease progression with the most recent therapy. In one aspect of these embodiments, the individual has been treated with a BCMA-targeted ADC or a BCMA-targeted CAR-T. In another aspect of these embodiments, the individual has not been treated with any prior BCMA-targeted ADC or BCMA-targeted CAR-T therapy.

In some embodiments, the malignancy is multiple myeloma, the individual has been treated with at least one, at least two, at least three, or at least four prior therapies for multiple myeloma, and the individual's multiple myeloma is refractory or relapsed under the conditions of (1) prior therapy for multiple myeloma comprising a proteasome inhibitor, (2) prior therapy for multiple myeloma comprising an immunomodulatory agent, and (3) prior therapy for multiple myeloma comprising an anti-CD38 antibody, and the individual has experienced disease progression in the most recent therapy for multiple myeloma. In one aspect of this embodiment, the individual has been treated with at least three prior therapies for multiple myeloma. In another aspect of this embodiment, the individual has been treated with at least four prior therapies for multiple myeloma.

In some embodiments, the individual's prior multiple myeloma therapies include BCMA-targeted ADC therapy or BCMA-targeted CAR-T cell therapy. In some embodiments, the individual's prior multiple myeloma therapies include BCMA-targeted therapy.

In some embodiments, the individual's prior multiple myeloma therapies do not include BCMA-targeted ADC therapy or BCMA-targeted CAR-T cell therapy. In some embodiments, the individual's prior multiple myeloma therapies do not include BCMA-targeted therapy.

In some embodiments, the malignancy is multiple myeloma, and the individual has undergone at least one or at least two prior therapies for multiple myeloma, the individual's multiple myeloma is refractory or relapsed under the conditions of (1) prior multiple myeloma therapy comprising a proteasome inhibitor, and (2) prior multiple myeloma therapy comprising an immunomodulatory agent. In some embodiments, the individual experienced disease progression during the most recent multiple myeloma therapy.

In some embodiments, the malignancy is multiple myeloma, and the individual has not undergone any prior therapies for multiple myeloma. In some embodiments, the individual has not undergone any prior therapies for multiple myeloma after being diagnosed with multiple myeloma. In some embodiments, the individual is not suitable for stem cell transplantation. In some embodiments, the malignancy is multiple myeloma, and the individual is not suitable for stem cell transplantation. In some embodiments, the individual is not suitable for autologous stem cell transplantation. In some embodiments, the individual is not suitable for allogeneic stem cell transplantation. In some embodiments, the individual is not suitable for autologous stem cell transplantation and is also not suitable for allogeneic stem cell transplantation.

In some embodiments, (i) the cycle is 21 days if the frequency of administration of PF06863135 to the individual is weekly or once every three weeks, the cycle is 28 days if the frequency of administration of PF06863135 to the individual is once every two weeks or once every four weeks; or (ii) the cycle is 28 days if the frequency of administration of PF06863135 to the patient is not once every three weeks.

In some embodiments, the method further comprises administering sasanlimab to the individual.

In some embodiments, PF-06863135 and sasanlimab are administered in a four-week treatment cycle for at least the first treatment cycle, and wherein if PF-06863135 is primed, the first treatment cycle begins on the seventh day after a single priming dose or the last dose within the priming administration, and sasanlimab is administered at a dose of 300 mg Q4W SC.

In some embodiments, the first dose of sasanlimab is administered on day 1 of the first treatment cycle. In some embodiments, the first dose of PF-06863135 in a treatment cycle is administered on day 1 of the treatment cycle.

In some embodiments, Week 1 and Cycle 1 begin on the day the individual is administered a single priming dose or the first priming dose, or if the individual is not subjected to a priming administration or is not administered a priming dose of PF06863135, Week 1 and Cycle 1 begin on the day the individual is administered the first dose of the first therapeutic administration of PF06863135, the cycle is 28 days, and sasanlimab is administered at a dose of 300 mg Q4W SC. In some embodiments, the individual is administered at least one priming dose of PF6863135 and sasanlimab is administered to the individual on Day 8 of each cycle.

In some embodiments, the method further comprises administering lenalidomide to the individual.

In some embodiments, PF-06863135 and lenalidomide are administered in a four-week treatment cycle for at least the first treatment cycle, and wherein, if PF-06863135 is primed, the first treatment cycle begins on the seventh day following administration of a single priming dose or the last dose within the priming administration, and wherein lenalidomide is administered at a dose of 25 mg orally daily on days 1-21 of each treatment cycle.

In some embodiments, lenalidomide is administered at a dose of 25 mg orally daily on days 1-21 of each treatment cycle without dexamethasone.

In some embodiments, the first dose of PF-06863135 in a treatment cycle is administered on day 1 of the treatment cycle.

In some embodiments, PF6863135 is administered as a primer, the cycle is 28 days, lenalidomide is administered at a daily oral dose of about 5 mg, about 10 mg, about 15 mg, about 20 mg, or about 25 mg on days 8-28 or days 15-28 of the first cycle, and then on days 1-28 of the second and third cycle, starting with the fourth cycle, lenalidomide is administered at a daily oral dose of about 5 to 10 mg more than that administered during the third cycle, or continues to be administered at the same daily oral dose as in the third cycle on days 1-28 of each cycle.

In some embodiments, PF6863135 is administered as a priming dose, with lenalidomide administered at a daily oral dose of about 10 mg or about 15 mg, beginning on day 8 of cycle 1, for at least 10 consecutive days in each cycle.

In some embodiments, priming with PF6863135 is not performed and lenalidomide is administered at a daily oral dose of about 10 mg, about 15 mg, about 20 mg, or about 25 mg for at least 10, at least 14, or at least 21 consecutive days in each cycle.

In some embodiments, the individual is administered PF06863135 in an induction phase followed by a maintenance phase, wherein the induction phase begins on the day the first dose is administered as part of a priming administration of PF06863135, or if no priming administration of PF6863135 is administered, then the induction phase begins on the day the first dose is administered as part of a first treatment administration of PF06863135, and the induction phase is completed on the last day of the last week or the last day of the last cycle, whichever is later, if the individual is in the first treatment administration period;

wherein during the induction phase, lenalidomide is administered at a daily oral induction dose of from about 5 mg to about 25 mg for at least 10 consecutive days in each cycle in the induction phase; in the maintenance phase, PF06863135 is administered in a second therapeutic administration, lenalidomide is administered at a daily oral maintenance dose of from about 5 mg to about 25 mg for at least 10 consecutive days in a cycle; wherein each cycle is 21 days or 28 days, and the induction phase lasts from 1 to 10 cycles. In some embodiments, the method further comprises administering dexamethasone to the individual during the induction phase at a daily oral dose of from about 10 mg to about 40 mg on at least day 1 and day 8 of the first cycle in the induction phase.

In some embodiments, each cycle in the induction phase is 21 days or 28 days, and cycle 1 begins on day 1 of week 3, the daily oral induction dose of lenalidomide is about 5 mg, about 10 mg, about 15 mg, about 20 mg, or about 25 mg, and is administered on days 1-14 or days 1-21 in each cycle in the induction phase, and if dexamethasone is administered, it is administered at a dose of about 20 mg daily on days 1, 8, and 15 in the first cycle and the second cycle of the induction phase; wherein each cycle in the maintenance phase is 28 days, and the daily oral maintenance dose of lenalidomide is about 5 mg, about 10 mg, or about 15 mg on days 1-28 of each cycle in the maintenance phase. In some embodiments, the induction phase is completed after 24-26 weeks. In some embodiments, the induction phase is completed after 12-14 weeks.

In some embodiments, the method further comprises administering pomalidomide to the individual. In some embodiments, PF06863135 and pomalidomide are administered in a four-week treatment cycle for at least the first treatment cycle, and if a priming administration of PF-06863135 is administered, the first treatment cycle begins on the seventh day after the administration of a single priming dose or the last dose within the priming administration, pomalidomide is administered at a dose of 4 mg daily orally on days 1-21 of each treatment cycle. In some embodiments, pomalidomide is administered at a dose of 4 mg daily, 3 mg daily, 2 mg daily, or 1 mg daily orally on days 1-21 of each treatment cycle without dexamethasone. In some embodiments, the first dose of PF-06863135 in a treatment cycle is administered on day 1 of the treatment cycle.

In some embodiments, the method further comprises administering daratumumab to the individual. In some embodiments, daratumumab is administered subcutaneously at a dose of about 1800 mg weekly, every two weeks, every three weeks, or every four weeks. In some embodiments, daratumumab administration begins with about 1800 mg weekly in Cycle 1 for about 8 doses, followed by about 1800 mg every two weeks for about 8 to about 10 doses, followed by about 1800 mg every four weeks thereafter.

In some embodiments, the method further comprises administering isatuximab to the individual. In some embodiments, isatuximab is administered at a dose of about 5 mg to about 10 mg/kg QW IV, Q2W IV, Q3W IV, or Q4W IV. In some embodiments, the dose of isatuximab at which isatuximab is administered to the individual may be the same or different when the individual is in the priming administration period, the first treatment administration, the second treatment administration, or the third treatment administration of PF06863135.

In some embodiments, the method further comprises administering to the individual at least one dose of a premedication medicament on the day the individual is administered a single compensatory dose, a first compensatory dose, a second compensatory dose, or the first dose as part of the first therapeutic administration of PF06863135, wherein the premedication medicament is acetaminophen, diphenhydramine, or dexamethasone. In some embodiments, dexamethasone is administered at a dose of about 10 mg to about 40 mg daily orally or intravenously. In some embodiments, dexamethasone is administered at a dose of about 10 mg to about 40 mg daily orally or intravenously on at least the day the individual is administered the first dose as part of the first therapeutic administration of PF06863135. In some embodiments, the dose of dexamethasone in which dexamethasone is administered to an individual as a premedication drug may be the same or different when the individual is subjected to a priming administration, a first treatment administration, a second treatment administration, or a third treatment administration of PF06863135.

In some embodiments, the method further comprises administering a second therapeutic agent to the individual. In some embodiments, the second therapeutic agent is an antineoplastic agent. In some embodiments, the second therapeutic agent is GSI. In some embodiments, the second therapeutic agent is nirogacestat or a pharmaceutically acceptable salt thereof.

In some embodiments, the method further comprises subjecting the individual to radiation therapy.

In aspects and/or embodiments relating to a method of treatment as described herein, such aspects and/or embodiments are also further aspects and/or embodiments relating to a therapeutic agent or agents for use in that method of treatment or, alternatively, the use of a particular therapeutic agent or agents for use in the manufacture of a medicament or medicaments for use in that treatment.

BRIEF DESCRIPTION OF DRAWINGS

Fig. 1 shows the induction of PD-1 expression on CD8 ⁺ T cells after treatment with the anti-BCMA×CD3 bispecific antibody.

Figures 2A and 2B show the therapeutic activity of the anti-BCMA×CD3 bispecific antibody in combination with an anti-PD1 antibody in A) the orthotopic MM.1S-Luc-PDL1 multiple myeloma model and B) the subcutaneous MM.1S-PD-L1 multiple myeloma model.

Figures 3A-3E show the upregulation of BCMA expression on the surface of multiple myeloma cells after GSI treatment.

Figures 4A-4E show the upregulation of BCMA expression on the surface of multiple myeloma cells as a function of time after GSI treatment.

Figures 5A-5E show reduced soluble BCMA (sBCMA) shedding in multiple myeloma cell lines following GSI treatment.

Figures 6A-6E show that GSI treatment enhances BCMA×CD3 bispecific antibody-mediated cell killing in multiple myeloma cell lines.

Figures 7A-7B show A) upregulation of BCMA expression on the surface of Raji lymphoma cells after GSI treatment, and B) the upregulation is time-dependent.

Fig. 8 shows that GSI treatment enhances BCMA×CD3 bispecific antibody-mediated cell killing in a lymphoma cell line.

DETAILED DESCRIPTION OF THE INVENTION

The present application relates to the treatment of a malignant neoplasm and/or a tumor-associated disease. Certain aspects relate to the treatment of an individual having a malignant neoplasm or a tumor-associated disease by subjecting the individual to combination therapy with a first therapeutic agent that is a bispecific antibody against BCMA×CD3 and a second therapeutic agent that is an antibody against PD-1, an antibody against PD-L1, or a γ-secretase inhibitor (GSI), or a pharmaceutically acceptable salt thereof.

I. Definitions

To better understand the present invention, certain technical and scientific terms are specifically defined below. Unless otherwise defined elsewhere in this description, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which the invention pertains.

For the purposes of the invention, including the claims, singular terms include corresponding plural references unless the context clearly indicates otherwise.

The term "about," when used to define a numerically defined parameter (e.g., the dose of the BCMA×CD3 bispecific antibody or the duration of treatment with the combination therapy described herein), means that the parameter may vary by as much as 10% below or above the stated numerical value of the parameter. For example, a dose of approximately 5 mg/kg may vary from 4.5 mg/kg to 5.5 mg/kg.

"An antibody" is an immunoglobulin molecule capable of specifically binding to a target, such as a carbohydrate, polynucleotide, lipid, polypeptide, etc., through at least one antigen recognition site located in the variable region of the immunoglobulin molecule. For the purposes of the invention, the term includes not only intact polyclonal or monoclonal antibodies, but also fragments thereof (such as Fab, Fab', F(ab') ₂ , Fv), single-chain (scFv) and domain antibodies (including, for example, shark and camelid antibodies), and fusion proteins containing the antibody, and any other modified configuration of the immunoglobulin molecule containing an antigen recognition site. An antibody includes an antibody of any class, such as IgG, IgA, or IgM (or a subclass thereof), and an antibody may not belong to any particular class. Depending on the amino acid sequence of the constant region of the antibody's heavy chain, immunoglobulins can be assigned to different classes. There are five main classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant regions corresponding to the different immunoglobulin classes are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of the different immunoglobulin classes are well known.

As used herein, the term "antigen-binding fragment" or "antigen-binding portion" of an antibody refers to one or more fragments of an intact antibody that retain the ability to specifically bind to the specified antigen. The antigen-binding functions of an antibody can be performed by fragments of an intact antibody. Examples of binding fragments included in the term "antigen-binding fragment" of an antibody include Fab-; Fab'-;F(ab')_2-; an Fd fragment consisting of the VH and CH1 domains; an Fv fragment consisting of the VL and VH domains of one arm of an antibody; a fragment of a single-domain antibody (dAb) (Ward et al., Nature 341:544-546, 1989) and an isolated complementarity determining region (CDR).

A "bispecific antibody" or "dual-specificity antibody" is a hybrid antibody with two different antigen-binding sites. The two antigen-binding sites of a bispecific antibody bind to two different epitopes, which may be located on the same or different target proteins.

A "bispecific anti-B-cell maturation antigen antibody" or "bispecific anti-BCMA antibody" is a bispecific antibody that specifically binds to BCMA and another antigen.

A "heterodimer," "heterodimeric protein," "heterodimeric complex," or "heteromultimeric polypeptide" is a molecule comprising a first polypeptide and a second polypeptide, wherein the second polypeptide differs in amino acid sequence from the first polypeptide by at least one amino acid residue.

An antibody, bispecific antibody, or polypeptide that "preferentially binds" or "specifically binds" (terms used interchangeably herein) to a target (e.g., a BCMA protein) is a term well known in the art, and methods for determining such specific or preferential binding are also well known in the art. A molecule is said to exhibit "specific binding" or "preferential binding" if it reacts or associates more frequently, more rapidly, with greater duration, and/or with greater affinity with a particular cell or substance than with alternative cells or substances. An antibody or bispecific antibody "specifically binds" or "preferentially binds" to a target if it binds with higher affinity, avidity, more readily, and/or with greater duration than it binds to other substances. For example, an antibody that specifically or preferentially binds to a BCMA epitope is one that binds to that epitope with greater affinity, avidity, greater facilitation, and/or longer duration than it binds to other BCMA epitopes or BCMA epitopes. It should also be understood when reading this definition that, for example, an antibody (or residue or epitope) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. Therefore, the terms "specific binding" or "preferential binding" do not necessarily require (although they may include) exclusive binding. Typically, but not necessarily, reference to "binding" means preferential binding.

The term "variable region" of an antibody refers to the variable region of the light chain of an antibody or the variable region of the heavy chain of an antibody, alone or in combination. As known in the art, the variable regions of the heavy and light chains each consist of four framework regions (FRs) connected by three complementarity-determining regions (CDRs), also known as hypervariable regions. The CDRs in each chain are held together in close proximity by the FRs and, together with the CDRs from the other chain, participate in forming the antigen-binding site of the antibody. There are at least two ways to define CDRs: (1) an approach based on interspecies sequence variability (i.e., Kabat et al., Sequences of Proteins of Immunological Interest, (5th ed., 1991, National Institutes of Health, Bethesda MD)); and (2) an approach based on crystallographic studies of antigen-antibody complexes (Al-lazikani et al., 1997, J. Molec. Biol. 273:927-948). As used herein, the term "CDR" may refer to CDRs determined using either approach or a combination of both approaches.

The "CDRs" of a variable domain are the amino acid residues in the variable region identified according to the definitions of Kabat, Chothia, the combination of Kabat and Chothia methods, AbM, the Contact system, and/or conformational definitions, or any method for defining CDRs well known in the art. Antibody CDRs can be identified as the hypervariable regions originally defined by Kabat et al. See, e.g., Kabat et al., 1992, Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, NIH, Washington D.C. CDR positions can also be identified as the structural loops originally described by Chothia et al. See, for example, Chothia et al., Nature 342:877-883, 1989. Other approaches to identifying CDRs include the "AbM definition," which is a compromise between Kabat and Chothia and is obtained using Oxford Molecular's AbM (now Accelrys®) antibody modeling software, or the "Contact definition" of CDRs based on observed antigen contacts, described in MacCallum et al., J. Mol. Biol., 262:732-745, 1996. In another approach, referred to herein as "conformational definition" of CDRs, CDR positions can be identified as residues that make an enthalpic contribution to antigen binding. See, for example, Makabe et al., Journal of Biological Chemistry, 283:1156-1166, 2008. Other definitions of CDR boundaries may not strictly follow one of the above approaches, but will still overlap with at least a portion of the Kabat CDRs, although they may be shortened or lengthened in light of the prediction or experimental findings that particular residues, or groups of residues, or even entire CDRs do not significantly affect antigen binding. As used herein, the term "CDR" may refer to CDRs defined by any approach known in the art, including combinations of approaches. The methods used herein may use CDRs defined by any of these approaches. In any specified aspect comprising more than one CDR, the CDR may be defined by any of the Kabat, Chothia, extended, AbM, Contact, and/or conformational definitions.

The terms "isolated antibody" and "isolated antibody fragment" refer to the state of purification and, in this context, mean that the named molecule is substantially free of other biological molecules, such as nucleic acids, proteins, lipids, carbohydrates, or other material, such as cellular debris and growth media. Generally, the term "isolated" is not intended to denote the complete absence of such material or the absence of water, buffers, or salts, unless they are present in amounts that substantially interfere with the experimental or therapeutic use of the binding compound as described herein.

As used herein, the term "monoclonal antibody," or "mAb," or "Mab," refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprising the population are identical in amino acid sequence, except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically include multiple different antibodies having different amino acid sequences in their variable domains, particularly the CDRs, which are often specific for different epitopes. The modifier "monoclonal" indicates the nature of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring the production of the antibody by any particular method. For example, monoclonal antibodies for use in the present invention can be produced by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or by recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567). "Monoclonal antibodies" can also be isolated from antibody phage libraries by methods described, e.g., in Clackson et al. (1991) Nature 352:624-628 and Marks et al. (1991) J. Mol. Biol. 222:581-597. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

The term "chimeric antibody" refers to an antibody in which a portion of the heavy and/or light chain is identical to or homologous with the corresponding sequences in an antibody derived from a particular biological species (e.g., human) or belonging to a particular class or subclass of antibodies, or, while the remainder of the chains is identical to or homologous with the corresponding sequences in an antibody derived from another biological species (e.g., mouse) or belonging to a different class or subclass of antibodies, as well as fragments of such antibodies, provided that they exhibit the desired biological activity.

The term "human antibody" refers to an antibody containing only human immunoglobulin protein sequences. A human antibody may contain mouse carbohydrate chains when produced in a mouse, mouse cell, or mouse cell-derived hybridoma. Similarly, the term "mouse antibody" or "rat antibody" refers to an antibody containing only mouse or rat immunoglobulin sequences, respectively.

The term "humanized antibody" refers to forms of antibodies that contain sequences from non-human (e.g., murine) antibodies, as well as human antibodies. Such antibodies contain a minimal sequence derived from non-human immunoglobulin. Generally, a humanized antibody will contain substantially all of at least one, and typically two, variable domains in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin, and all or substantially all of the FR regions are regions from a human immunoglobulin sequence. A humanized antibody will optionally also contain at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. The prefix "hum," "hu," or "h" is added to antibody clone designations, as necessary, to distinguish humanized antibodies from the parental rodent antibodies. Humanized forms of rodent antibodies will generally contain the same CDR sequences of the parent rodent antibodies, although some amino acid substitutions may be included to improve affinity, enhance the stability of the humanized antibody, or for other reasons.

The terms "malignant neoplasm" or "malignant" refer to or describe a physiological condition in mammals typically characterized by unregulated cell growth. The terms "malignant neoplasm" or "malignant tissue" may include tumor. Non-limiting examples of malignant neoplasms include carcinoma, lymphoma, leukemia, myeloma, blastoma, and sarcoma. Malignant neoplasms may include a malignant neoplasm and/or tumor-associated disease, including B-cell malignancies and/or tumor-associated diseases, including, but not limited to, multiple myeloma, malignant plasma cell neoplasia, lymphoma, Hodgkin's lymphoma, nodular Hodgkin's lymphoma with lymphocyte predominance, Kahler disease and myelomatosis, plasma cell leukemia, bone and extramedullary plasmacytoma with multiple myeloma, solitary bone and extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering myeloma, light chain amyloidosis, osteosclerotic myeloma, B-cell prolymphocytic leukemia, hairy cell leukemia, B-cell Non-Hodgkin's lymphoma (NHL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL), chronic myeloid leukemia (CML), follicular lymphoma, Burkitt's lymphoma, marginal zone cell lymphoma, mantle cell lymphoma, large cell lymphoma, precursor B-cell lymphoblastic lymphoma, myeloid leukemia, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma, mucosal lymphoid tissue lymphoma, small lymphocytic lymphoma, primary mediastinal (thymic) large B-cell lymphoma, lymphoplasmacytic lymphoma, marginal zone B-cell lymphoma, splenic marginal zone cell lymphoma, intravascular large B-cell lymphoma, Primary effusion lymphoma, Hodgkin's disease, T-cell/histiocytic large B-cell lymphoma, Primary central nervous system lymphoma, Primary cutaneous diffuse large B-cell lymphoma (leg type), EBV-positive diffuse large B-cell lymphoma of the elderly, Inflammation-associated diffuse large B-cell lymphoma, ALK-positive large B-cell lymphoma, Plasmoblastic lymphoma, Large B-cell lymphoma arising in HHV8-associated multicentric Castleman disease, Undifferentiated B-cell lymphoma intermediate in features between diffuse large B-cell lymphoma and Burkitt's lymphoma, Undifferentiated B-cell lymphoma intermediate in features between diffuse large B-cell lymphoma and classical Hodgkin's lymphoma and other B-cell lymphomas. Examples of malignant neoplasms and tumor-associated diseases are additionally presented in this description.

A "chemotherapeutic agent" is a chemical compound that can be used in the treatment of a malignant neoplasm and/or tumor-associated disease. Classes of chemotherapeutic agents include, but are not limited to: alkylating agents, antimetabolites, kinase inhibitors, plant spindle alkaloids, cytotoxic/antineoplastic antibiotics, topoisomerase inhibitors, photosensitizers, antiestrogens and selective estrogen receptor modulators (SERMs), antiprogesterones, estrogen receptor downregulators (ERDs), estrogen receptor antagonists, luteinizing factor releasing factor agonists, antiandrogens, aromatase inhibitors, EGFR inhibitors, VEGF inhibitors, and antisense oligonucleotides that inhibit the expression of genes involved in abnormal cell proliferation or tumor growth. Chemotherapeutic agents are further described herein.

For the purposes of this invention, the term "chemotherapy" refers to a chemotherapeutic agent as defined above, or a combination of two, three, or four chemotherapeutic agents for the treatment of a malignant neoplasm and/or tumor-associated disease. If chemotherapy consists of more than one chemotherapeutic agent, the chemotherapeutic agents may be administered to the patient on the same day or on different days within a single treatment cycle.

As used in the invention and claims, the term "consists essentially of" and variations thereof such as "consist essentially of" or "consisting essentially of" means the inclusion of any listed elements or group of elements and the optional inclusion of other elements of a similar or different nature than the listed elements that do not significantly alter the basic or novel properties of a particular dosage regimen, method, or composition.

The term "multiple myeloma therapy" refers to a drug, or a combination of two or more drugs, (1) approved by the United States Food and Drug Administration (USFDA) or the European Medicines Agency for the treatment of multiple myeloma or (2) in or having been in clinical trials in the United States or Europe for the treatment of multiple myeloma.

The term "conventional therapy for multiple myeloma" refers to a therapy for multiple myeloma approved by the USFDA or the European Medicines Agency, which may be a single drug, a combination therapy of two or more drugs.

For the purposes of this invention, the terms "IMiD drug," "imid drug," or "immunomodulatory agent" are used interchangeably and refer to a drug that a practitioner treating multiple myeloma considers to be an IMiD drug or immunomodulatory agent in the context of treating multiple myeloma. Non-limiting examples of IMiD drugs or immunomodulatory agents include thalidomide, lenalidomide, and pomalidomide.

The term "BCMA-targeted ADC therapy" refers to a therapy for multiple myeloma that includes an antibody-drug conjugate, where the antibody binds to the B-cell maturation antigen (BCMA). Non-limiting examples of BCMA-targeted ADCs include belantamab mafodotin-blmf, approved by the US FDA and marketed under the trade name BLENREP.

The terms "BCMA-targeted CAR-T cell therapy" or "anti-BCMA CAR-T cell" are used interchangeably herein and refer to a therapy for multiple myeloma involving a chimeric antigen receptor (CAR) T cell, wherein the CAR recognizes the B-cell maturation antigen (BCMA). Non-limiting examples of "BCMA-targeted CAR-T therapy" or "anti-BCMA CAR-T cell therapy" include idecabtagene vicleicel (ide-cel or bb2121) and JNJ-4528, also known as LCAR-B38M.

The term "BCMA-targeted therapy" refers to multiple myeloma therapies whose active ingredient includes a component that binds to a B-cell maturation antigen. BCMA-targeted therapies include BCMA-targeted ADCs, BCMA-targeted CAR-T therapies, and multiple myeloma therapies that include bispecific anti-BCMA antibodies.

The term "newly diagnosed multiple myeloma" refers to multiple myeloma where the patient (individual) has not yet undergone any treatment to diagnose multiple myeloma.

The term "homology" refers to the sequence similarity between two polypeptide sequences when they are optimally aligned. If a position in both of the two sequences being compared is occupied by the same amino acid monomer subunit, for example, if a position in the light chain CDR of two different Abs is occupied by alanine, then the two Abs are homologous at that position. Percent homology is the number of homologous positions shared by the two sequences divided by the total number of positions compared, ×100. For example, if 8 out of 10 positions in two sequences are the same or homologous in an optimal sequence alignment, then the two sequences are 80% homologous. Typically, comparisons are performed by aligning two sequences to obtain the maximum percentage of homology. For example, comparisons can be performed using the BLAST algorithm, where the algorithm parameters are selected to obtain the greatest match between corresponding sequences over the full length of the corresponding reference sequences.

The following references refer to BLAST algorithms frequently used for sequence analysis: BLAST ALGORITHMS: Altschul, S. F., et al., (1990) J. Mol. Biol. 215:403–410; Gish, W., et al., (1993) Nature Genet. 3:266–272; Madden, T. L., et al., (1996) Meth. Enzymol. 266:131–141; Altschul, S. F., et al., (1997) Nucleic Acids Res. 25:3389–3402; Zhang, J., et al., (1997) Genome Res. 7:649–656; Wootton, J. C., et al., (1993) Comput. Chem. 17:149-163; Hancock, J.M. et al., (1994) Comput. Appl. Biosci. 10:67-70; ALIGNMENT SCORING SYSTEMS: Dayhoff, M.O., et al., "A model of evolutionary change in proteins." in Atlas of Protein Sequence and Structure, (1978) vol. 5, suppl. 3. M.O. Dayhoff (ed.), pp. 345-352, Natl. Biomed. Res. Found., Washington, DC; Schwartz, R.M., et al., "Matrices for detecting distant relationships." in Atlas of Protein Sequence and Structure, (1978) vol. 5, suppl. 3. M. O. Dayhoff (ed.), pp. 353–358, Natl. Biomed. Res. Found., Washington, DC; Altschul, S. F., (1991) J. Mol. Biol. 219:555–565; States, D. J., et al., (1991) Methods 3:66–70; Henikoff, S., et al., (1992) Proc. Natl. Acad. Sci. USA 89:10915–10919; Altschul, S. F., et al., (1993) J. Mol. Evol. 36:290–300; STATISTICAL ALIGNMENT ANALYSIS: Karlin, S., et al., (1990) Proc. Natl. Acad. Sci. USA 87:2264-2268; Karlin, S., et al., (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877; Dembo, A., et al., (1994) Ann. Prob. 22:2022-2039; and Altschul, S.F. "Evaluating the statistical significance of multiple distinct local alignments." in Theoretical and Computational Methods in Genome Research (S. Suhai, ed.), (1997) pp. 1-14, Plenum, New York.

The term "patient" or "individual" refers to any living organism suffering from or predisposed to a condition that can be prevented or treated by administering a therapeutic agent or composition or combination as described herein, such as a malignant neoplasm and/or tumor-associated disease, and includes humans and animals. The terms "patients" and "individuals" include, but are not limited to, mammals (e.g., mice, monkeys, horses, cattle, pigs, dogs, cats, etc.), and preferably represent humans.

The term "sustained response" means a sustained therapeutic effect after cessation of treatment with the therapeutic agent or combination therapy described herein. In some aspects, a sustained response has a duration that is at least the same as the duration of treatment or at least 1.5, 2.0, 2.5, or 3 times longer than the duration of treatment.

As used herein, the term "administration" refers to the delivery of a therapeutic agent to an individual using any of the various delivery methods and systems known to those skilled in the art. Examples of routes of administration include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes, such as injection or infusion. As used herein, the phrase "parenteral administration" means routes of administration other than enteral and topical administration, typically by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrastermal injection and infusion, as well as in vivo electroporation. The therapeutic agent may be administered non-parenterally or orally. Other non-parenteral routes include topical, epidermal, or mucosal administration, such as intranasal, vaginal, rectal, sublingual, or topical. The administration may also be carried out, for example, once, multiple times and/or over one or more extended periods of time.

As used herein, "treating" or "treating" a malignant neoplasm and/or tumor-associated disease means subjecting an individual or patient with or diagnosed with a malignant neoplasm to the combination treatment of the present invention to achieve at least one beneficial therapeutic effect, such as, for example, a reduced number of malignant cells, a reduced tumor size, a reduced rate of infiltration of peripheral organs by malignant cells, or a reduced rate of metastasis or tumor growth, reversal, alleviation, inhibition of progression, or prevention of the disorder or condition to which such term refers, or one or more symptoms of such disorder or condition. As used herein, the term "treating," unless otherwise indicated, refers to the act of treating as "treating" is defined immediately above. The term "treating" also includes adjuvant and neoadjuvant treatment of an individual. For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: a decrease in the proliferation (or destruction) of a neoplastic or malignant cell; inhibition of metastasis or neoplastic cells; reduction in tumor size; remission of a malignancy; reduction in symptoms resulting from a malignancy; improvement in the quality of life of individuals suffering from a malignancy; reduction in the dose of other drugs required to treat a malignancy; delay in the progression of a malignancy; cure of a malignancy; overcoming one or more resistance mechanisms of a malignancy and/or prolonging the survival of patients with a malignancy. Positive therapeutic effects in a malignancy can be measured in a number of ways (see, e.g., W. A. Weber, J. Nucl. Med. 50:1S-10S (2009)). In some aspects, the treatment achieved with the combination of the invention is any of a partial response (PR), a complete response (CR), an overall response (OR), an objective response rate (ORR), a progression-free survival (PFS), a PFS without radiographic evidence of progression, a disease-free survival (DFS), and an overall survival (OS). PFS, also referred to as "time to tumor progression," refers to the length of time during and after treatment when the malignancy does not grow and includes the amount of time patients experience a CR or PR, as well as the amount of time patients experience stable disease (SD). The term "DFS" refers to the length of time during and after treatment when the patient is disease-free. The term "OS" refers to a prolongation of life expectancy compared to treatment-naive or untreated individuals or patients. In some aspects, the response to the combination of the invention is any of PR, CR, PFS, DFS, ORR, OR, or OS, as assessed using the Objective Response Evaluation Criteria in Solid Tumors (RECIST 1.1) (Eisenhauer et al., EA et al., Eur. J Cancer 45:228-247 (2009)). In some aspects, the anti-myeloma activity can be assessed using the overall response rate (ORR), time to response (TTR), complete response rate (CRR), duration of response (DOR), duration of complete response (DoCR), duration of stable disease (DOSD), progression-free survival (PFS), overall survival (OS) using the International Multiple Myeloma Study Group (IMWG) criteria. The treatment regimen for the combination therapy described herein is effective for treating a patient with a malignant neoplasm and may vary depending on factors such as the disease state, age and body weight of the patient, and the ability of the therapy to induce an antitumor response in an individual. Although an aspect of any of the aspects of the invention may not be effective in achieving a positive therapeutic effect in every individual, it should be so for a statistically significant number of individuals, determined using statistical analysis known in the art, such as, as non-limiting examples, the Cox log-rank test, the Cochran-Mantel-Haenszel log-rank test, the Student t-test, the Chi-square test, the Mann-Whitney U-test, the Kruskal-Wallis test (H-test), the Jonckheere-Terpstra test, and the Wilcoxon test. The term "treatment" also includes in vitro and ex vivo treatment, such as of a cell, with a reagent, diagnostic agent, binding compound, or another cell.

For the purposes of this disclosure, the term "pharmaceutical product" refers to a medicinal product containing an active pharmaceutical ingredient and regulated by the US FDA, EMA, or other regulatory authorities in other markets. A pharmaceutical product may be an investigational medicinal product or a finished dosage form already approved by a regulatory authority.

The terms "treatment regimen," "dosing protocol," and "administration regimen" are used interchangeably to refer to the dose and timing of administration of each therapeutic agent in the combination of the invention.

For the purposes of the invention, an "effective dose" or "effective amount" of a drug, compound, or pharmaceutical composition is an amount sufficient to produce any one or more beneficial or desired results. For prophylactic use, beneficial or desired results include the elimination or reduction of the risk, reduction in the severity, or delay in the onset of a disease, including the biochemical, histological, and/or behavioral symptoms of the disease, its complications, and intermediate pathological phenotypes present during the development of the disease. For therapeutic use, beneficial or desired results include clinical results such as a decrease in the incidence or improvement of one or more symptoms of various diseases or conditions (such as, for example, a malignancy), a reduction in the dose of other drugs required to treat the disease, an enhancement of the effect of another drug, and/or a delay in the progression of the disease. An effective dose can be administered in one or more administrations. For the purposes of the present invention, an effective dose of a drug, compound, or pharmaceutical composition is an amount sufficient to effect prophylactic or therapeutic treatment, directly or indirectly. As should be understood in a clinical context, an effective dose of a drug, compound, or pharmaceutical composition may or may not be achieved in combination with another drug, compound, or pharmaceutical composition. Thus, an "effective dose" can be considered in the context of administering one or more therapeutic agents, and an individual agent can be considered to be administered in an effective amount if the desired result can or is achieved in combination with one or more other agents.

As used herein, the term "dosing" refers to both a "dose amount," such as 1 mg, 20 mg, and a "frequency of administration," such as once daily (QD), once weekly (Q1W or QW), once every two weeks (Q2W), once every three weeks (Q3W), and once every four weeks (Q4W). Dosing may also include the route of administration of the drug, such as, for example, subcutaneous (SC), intravenous (IV), oral (PO), if indicated. Similarly, each of the terms "priming administration," "first therapeutic administration," "second therapeutic administration," etc., refers to a dose amount and a frequency of administration of such a dose and, optionally, also includes a route of administration, if indicated. In some embodiments, dosing includes one dose amount and one frequency of administration. In some embodiments, dosing includes more than one dose amount and/or more than one frequency of administration.

As used herein, "dose level", unless otherwise noted, when used to describe a dose amount of elranatamab (also known as PF06863135), refers to one of the following dose amounts: 4 mg, 8 mg, 12 mg, 16 mg, 20 mg, 24 mg, 32 mg, 44 mg, 76 mg, 116 mg, and 152 mg, where each of 8 mg, 12 mg, 16 mg, 20 mg, 24 mg, 32 mg, 44 mg, 76 mg, 116 mg, and 152 mg is one dose level higher than 4 mg, 8 mg, 12 mg, 16 mg, 24 mg, 32 mg, 44 mg, 76 mg, and 116 mg, respectively.

For the purposes of this invention, "relevant regulatory labeling of a pharmaceutical product" means the current US Prescribing Information (USPI) from the US Food and Drug Administration (FDA), the current Summary of Product Characteristics (SMPC) from the European Medicines Agency (EMA) for a pharmaceutical product, or similar pharmaceutical product labeling from regulatory authorities in other markets. In some embodiments, the term "relevant regulatory labeling of a pharmaceutical product" in a US patent or patent application refers to the current USPI of the pharmaceutical product, and in a patent or patent application of a European country accepting EMA marketing authorizations for the pharmaceutical product, the current SMPC of the pharmaceutical product, and similarly in other jurisdictions.

For the purposes of this invention, the term "subject response" refers to the clinical response of an individual treated with a pharmaceutical product containing elranatamab (PF006863135), either as monotherapy or in combination with a second therapeutic product for primary treatment. The term "subject response" includes one or more aspects of clinical efficacy, such as complete response, partial response, and duration of response. The term "subject response" may also include additional aspects, such as toxicity and adverse events.

For the purposes of the invention, the term "IMWG response" refers to the clinical response of a patient (individual) to a pharmaceutical product for the treatment of multiple myeloma, where the response, such as a complete response or a partial response, is defined in accordance with the most current definition of the International Multiple Myeloma Study Group.

As used herein, the terms "cycle" and "week" when used in the context of describing a method for treating a cancer, including its use, dose, or administration schedule, refer to a duration of time. A cycle is 21 days or 28 days, unless otherwise specified, when an individual is treated with a therapeutic agent, a pharmaceutical product thereof, such as elranatamab (PF06863135), or a pharmaceutical product thereof, as monotherapy or in combination with a second therapeutic agent. The term "week 1" refers to the first week when an individual is treated with the method or any of the doses or administration schedules within it, unless otherwise specified. Week 2 begins immediately after the end of week 1, week 3 begins immediately after the end of week 2, etc. Cycle 1 begins on the first day of week 1, the first day of week 2, or the first day of week three, unless otherwise specified. Unless otherwise specified, cycle 2 begins immediately after the end of cycle 1, cycle 3 begins immediately after the end of cycle 2, etc.

For the purposes of the invention, the term "eligible for stem cell transplantation" refers to a patient diagnosed with multiple myeloma who is not eligible for stem cell transplantation as a treatment for multiple myeloma.

The term "tumor," when used in reference to an individual diagnosed with or suspected of having a malignancy, refers to a malignant or potentially malignant neoplasm or tissue mass of any size and includes primary tumors and secondary neoplasms. A solid tumor is an abnormal growth or mass of tissue, typically without a cyst or fluid area. Different types of solid tumors are named by the type of cells that form them. Examples of solid tumors include sarcomas, carcinomas, and lymphomas. Leukemias (malignant neoplasms of the blood) generally do not form solid tumors (National Cancer Institute, Dictionary of Cancer Terms). Multiple myeloma is a malignancy of plasma cells.

The term "tumor burden," also referred to as "tumor mass," refers to the total amount of tumor material distributed throughout the body. The term "tumor burden" refers to the total number of malignant cells or the total size of tumors throughout the body, including lymph nodes and bone marrow. Tumor burden can be determined using various methods known in the art, such as by measuring the size of tumors after removal from an individual, such as with calipers, or while still in the body using imaging techniques such as ultrasound, bone scan, computed tomography (CT), or magnetic resonance imaging (MRI).

The term "tumor size" refers to the overall size of the tumor, which can be measured as its length and width. Tumor size can be determined using various methods known in the art, such as measuring tumor size after removal from the individual, such as with calipers, or while still in the body using imaging techniques such as bone scan, ultrasound, CT, or MRI.

The term "immunotherapy" refers to the treatment of an individual by a method that involves inducing, enhancing, suppressing, or otherwise modifying the immune response.

For the purposes of this invention, the term "effector immune cell" or "effector cell" refers to a cell within the natural repertoire of cells of the human immune system that can be activated to influence the viability of a target cell. Target cell viability may include survival, proliferation, and/or the cell's ability to interact with other cells.

The term "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" refers to a component that can be included in the compositions provided herein and will not cause significant toxicological adverse effects in an individual.

The terms "protein", "polypeptide" and "peptide" are used interchangeably herein and refer to any peptide-linked chain of amino acids, regardless of length and co-translational or post-translational modification.

As used herein, the terms "substantially" or "essentially" mean "almost entirely," such as 95% or more of the stated amount.

The term "substantially homologous" or "substantially identical" means that a specified sequence, such as a mutant sequence, differs from a reference sequence by one or more substitutions, deletions, or additions, the combined effect of which does not result in an undesirable functional divergence between the reference and specified sequences. For the purposes of the present invention, a sequence having greater than 95 percent homology (identity), equivalent biological activity (although not necessarily equivalent strength of biological activity), and equivalent expression characteristics with respect to the specified sequence is considered substantially homologous (identical). For the purposes of determining homology, truncation of the mature sequence is to be disregarded.

The terms "synergy" or "synergistic" are used to indicate that the result of a combination of two or more compounds, components, or targeted agents is greater than the sum of each agent combined. The terms "synergy" or "synergistic" also mean that an improvement in the condition of the disease or disorder being treated occurs when two or more compounds, components, or targeted agents are used, whereas each compound, component, or targeted agent acts individually. This improvement in the condition of the disease or disorder being treated constitutes a "synergistic effect." A "synergistic amount" is the amount of a combination of two compounds, components, or targeted agents that results in a synergistic effect as the term "synergistic" is defined herein. The definition of a synergistic interaction between one or two components, the optimal range for effect, and the absolute ranges of doses of each component for effect can be ultimately determined by administering the components in different ranges of mass/mass ratios and doses to patients in need of treatment. However, observation of synergy in in vitro models or in vivo models can be predictive of the effect in humans and other species, and in vitro models or in vivo models, as described herein, exist for measuring synergistic effects, and the results of such studies can also be used to predict the effective dose and ranges of plasma concentration ratios and absolute doses and plasma concentrations required for humans and other species using pharmacokinetic/pharmacodynamic methods.

Throughout the invention, the designation "PF-06863135" is used interchangeably with the term "elranatamab." PF06863135 is a bispecific antibody against BCMA×CD3. PF-06863135 is described, for example, in U.S. Patent No. 9,969,809. Selected sequences of PF-06863135 are provided in Table 15.

Unless otherwise defined, all technical and scientific terms used in this specification have the meaning commonly understood by one of ordinary skill in the art to which the invention pertains. In case of conflict, this specification, including definitions, will control. Throughout this specification and the claims, the term "comprise" or its variations, such as "comprises" or "comprising," shall be understood to imply the inclusion of the stated integer or group of integers, but not the exclusion of any other integer or group of integers. Unless the context otherwise indicates, singular terms shall include plurals, and plural terms shall include the singular.

This description provides examples of methods and materials, although methods and materials similar or equivalent to those described herein may also be used in practicing or testing the invention. The materials, methods, and examples are illustrative only and are not intended to be limiting.

II. Methods, application and medicinal products

The present invention relates to methods and compositions for treating a malignant neoplasm and/or a tumor-associated disease in an individual, comprising a combination treatment comprising at least a first therapeutic agent and a second therapeutic agent.

BCMA-specific therapeutic agents

In some aspects, the therapeutic agent may be a BCMA-specific therapeutic agent. In another aspect, the BCMA-specific therapeutic agent may be a polyspecific anti-BCMA antibody (e.g., bispecific and trispecific), an anti-BCMA antibody-drug conjugate, or a chimeric antigen receptor (CAR)-modified T cell therapeutic agent against BCMA. B-cell maturation antigen (BCMA, also known as TNFRSF17 and CD269) is a candidate for bispecific antibody-based immunotherapy. BCMA expression is upregulated during the maturation of B cells into plasmablasts and plasma cells, but is not expressed on naive B cells, hematopoietic stem cells, or normal tissues such as the heart, lungs, kidneys, or tonsils. In multiple myeloma, BCMA expression has been identified at every stage of the disease and in patients with different cytogenetic risks. Furthermore, BCMA expression was unaffected by autologous stem cell transplantation (ASCT) or chemotherapy. In vivo studies have shown that bispecific anti-BCMA antibodies induce T-cell activation, reduce tumor burden, and prolong survival.

Non-limiting examples of polyspecific anti-BCMA antibodies that can be used in the combination treatment methods of the present invention include AMG 420 (anti-BCMA×CD3 bispecific T cell engager, BiTE®, Amgen), AMG 701 (anti-BCMA×CD3 BiTE®, Amgen), CC-93269 (anti-BCMA×CD3 bispecific antibody, Celgene), JNJ-64007957 (Janseen), PF-06863135 (anti-BCMA×CD3 bispecific antibody, Pfizer Inc.), TNB-383B (TeneoBio/AbbVie), REGN5458 (anti-BCMA×CD3 bispecific antibody, Regeneron), AFM26 (anti-BCMA×CD16 tetravalent bispecific antibody, Affimed GmbH), HPN217 (trispecific antibody BCMA×ALB×CD3, Harpoon Therapeutics).

In some aspects, the BCMA-specific therapeutic agent is a bispecific antibody against the BCMA molecule. Bispecific anti-BCMA antibodies are monoclonal antibodies that have binding specificity for at least two different antigens (e.g., BCMA and CD3).

In some aspects, the bispecific anti-BCMA antibody comprises a first variable domain of the antibody and a second variable domain of the antibody, wherein the first variable domain of the antibody specifically binds to CD3, and wherein the second variable domain of the antibody specifically binds to BCMA.

In some aspects, the therapeutic agent in the combination therapy of the present invention is a bispecific anti-BCMA antibody. In some aspects, the bispecific anti-BCMA antibody may have any of the features or characteristics of any of the anti-BCMA bispecific antibodies described in WO2016166629, which is hereby incorporated by reference for all purposes.

In some aspects, the first variable domain of the antibody specifically binds to CD3. Information on CD3 is provided, for example, in UniProtKB #P07766. In some aspects, the first variable domain of the antibody comprises three CDRs of a heavy chain variable region (VH) comprising the amino acid sequence set forth in SEQ ID NO: 1 and/or three CDRs of a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO: 9. In some aspects, VH comprises VH CDR1 comprising the sequence set forth in SEQ ID NO: 2, 3 or 4, VH CDR2 comprising the sequence set forth in SEQ ID NO: 5 or 6, VH CDR3 comprising the sequence set forth in SEQ ID NO: 7, and/or VL comprises VL CDR1 comprising the sequence set forth in SEQ ID NO: 10, VL CDR2 comprising the sequence set forth in SEQ ID NO: 11, VL CDR3 comprising the sequence set forth in SEQ ID NO: 12. In some aspects, VH comprises the sequence set forth in SEQ ID NO: 1 and/or VL comprises the sequence set forth in SEQ ID NO: 9. In some aspects, the first antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 8 and/or a light chain comprising the amino acid sequence set forth in SEQ ID NO: 13.

In some aspects, the second variable domain of the antibody specifically binds to BCMA. Information on BCMA is provided, for example, in UniProtKB ID # Q02223. In some aspects, the second variable domain of the antibody comprises three CDRs of a heavy chain variable region (VH) comprising the amino acid sequence set forth in SEQ ID NO: 14 and/or three CDRs of a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO: 22. In some aspects, VH comprises a VH CDR1 comprising the sequence set forth in SEQ ID NO: 15, 16, or 17, a VH CDR2 comprising the sequence set forth in SEQ ID NO: 18 or 19, a VH CDR3 comprising the sequence set forth in SEQ ID NO: 20, and/or VL comprises a VL CDR1 comprising the sequence set forth in SEQ ID NO: 23, a VL CDR2 comprising the sequence set forth in SEQ ID NO: 24, a VL CDR3 comprising the sequence set forth in SEQ ID NO: 25. In some aspects, VH comprises the sequence set forth in SEQ ID NO: 14 and/or VL comprises the sequence set forth in SEQ ID NO: 22. In some aspects, the second antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 21 and/or a light chain comprising the amino acid sequence set forth in SEQ ID NO: 26.

In some aspects, the anti-BCMA bispecific antibody is PF-06863135, also known as elranatamab. The anti-BCMA bispecific antibody used in the examples provided herein was PF-06863135 unless otherwise noted. PF-06863135 is a heterodimeric humanized full-length bispecific antibody consisting of one B-cell maturation antigen (BCMA)-binding arm and one cluster of differentiation (CD3)-binding arm paired using hinge domain mutation technology. It utilizes a modified crystallizable fragment (Fc) of human IgG2Δa. PF-06863135 is described, for example, in U.S. Patent No. 9,969,809, which is hereby incorporated herein for all purposes. The sequences of PF-06863135 are shown in Table 19.

An effective amount of a BCMA-specific therapeutic agent can be administered in accordance with the dosages provided herein.

Anti-PD-1 and Anti-PD-L1 Therapies

In some aspects, a therapeutic agent for use in the combination therapy of the present invention may be an anti-PD-1 or anti-PD-L1 antibody. Programmed death receptor 1 (PD-1) and PD-1 ligands 1 and 2 (PD-L1 and PD-L2, respectively) play key roles in regulating the immune system. Expressed on activated T cells, PD-1 is activated by PD-L1 (also known as B7-H1) and PD-L2 expressed by stromal cells, tumor cells, or both, initiating T cell death and localized immunosuppression (Dong et al., Nat Med 1999; 5:1365-69; Freeman et al. J Exp Med 2000; 192:1027-34), potentially providing an immunotolerant environment for tumor development and growth. Conversely, inhibition of this interaction can enhance local T cell responses and mediate antitumor activity in non-clinical animal models (Iwai Y, et al. Proc Natl Acad Sci USA 2002; 99:12293-97).

Non-limiting examples of anti-PD-1 and anti-PD-L1 antibodies that can be used in the combination treatment of the present invention include atezolizumab (TECENTRIQ®, MPDL3280A, Roche Holding AG), durvalumab (IMFINZI®, AstraZeneca PLC), nivolumab (OPDIVO®, ONO-4538, BMS-936558, MDX1106, Bristol-Myers Squibb Company), pembrolizumab (KEYTRUDA®, MK-3475, lambrolizumab, Merck & Co., Inc.), BCD-100 (BIOCAD Biopharmaceutical Company), tislelizumab (BGB-A317, BeiGene Ltd./Celgene Corporation), genolimzumab (CBT-501, CBT Pharmaceuticals), CBT-502 (CBT Pharmaceuticals), GLS-010 (Harbin Gloria Pharmaceuticals Co., Ltd.), sintilimab (IBI308, Innovent Biologics, Inc.), WBP3155 (CStone Pharmaceuticals Co., Ltd.), AMP-224 (GlaxoSmithKline plc), BI 754091 (Boehringer Ingelheim GmbH), BMS-936559 (Bristol-Myers Squibb Company), CA-170 (Aurigene Discovery Technologies), FAZ053 (Novartis AG), spartalizumab (PDR001, Novartis AG), LY3300054 (Eli Lilly & Company), MEDI0680 (AstraZeneca PLC), PDR001 (Novartis AG), sasanlimab (PF-06801591, Pfizer Inc.), cemiplimab (LIBTAYO®, REGN2810, Regeneron Pharmaceuticals, Inc.), camrelizumab (SHR-1210, Incyte Corporation), TSR-042 (Tesaro, Inc.), AGEN2034 (Agenus Inc.), CX-072 (CytomX Therapeutics, Inc.), JNJ-63723283 (Johnson & Johnson), MGD013 (MacroGenics, Inc.), AN-2005 (Adlai Nortye), ANA011 (AnaptysBio, Inc.), ANB011 (AnaptysBio, Inc.), AUNP-12 (Pierre Fabre Medicament S.A.), BBI-801 (Sumitomo Dainippon Pharma Co., Ltd.), BION-004 (Aduro Biotech), CA-327 (Aurigene Discovery Technologies), CK-301 (Fortress Biotech, Inc.), ENUM 244C8 (Enumeral Biomedical Holdings, Inc.), FPT155 (Five Prime Therapeutics, Inc.), FS118 (F-star Alpha Ltd.), hAb21 (Stainwei Biotech, Inc.), J43 (Transgene S.A.), JTX-4014 (Jounce Therapeutics, Inc.), KD033 (Kadmon Holdings, Inc.), KY-1003 (Kymab Ltd.), MCLA-134 (Merus B.V.), MCLA-145 (Merus B.V.), PRS-332 (Pieris AG), SHR-1316 (Atridia Pty Ltd.), STI-A1010 (Sorrento Therapeutics, Inc.), STI-A1014 (Sorrento Therapeutics, Inc.), STI-A1110 (Les Laboratoires Servier), and XmAb20717 (Xencor, Inc.).

In some aspects, the therapeutic agent in the combination treatment of the present invention is an anti-PD-1 antibody. In some aspects, the anti-PD-1 antibody may have any of the features or characteristics of any of the antibodies presented in WO2016/092419, which is hereby incorporated by reference for all purposes.

In some aspects, an anti-PD-1 antibody comprises three CDRs of a heavy chain variable region (VH) comprising the amino acid sequence set forth in SEQ ID NO: 27 and/or three CDRs of a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO: 31. In some aspects, VH comprises a VH CDR1 comprising the sequence set forth in SEQ ID NO: 28, a VH CDR2 comprising the sequence set forth in SEQ ID NO: 29, a VH CDR3 comprising the sequence set forth in SEQ ID NO: 30, and/or VL comprises a VL CDR1 comprising the sequence set forth in SEQ ID NO: 32, a VL CDR2 comprising the sequence set forth in SEQ ID NO: 33, a VL CDR3 comprising the sequence set forth in SEQ ID NO: 34. In some aspects, VH comprises the sequence set forth in SEQ ID NO: 27 and/or VL comprises the sequence set forth in SEQ ID NO: 31.

In some aspects, the anti-PD-1 antibody is sasanlimab (PF-06801591). Sasanlimab is a humanized immunoglobulin G4 (IgG4) monoclonal antibody (mAb) that binds to the PD-1 receptor. By blocking its interaction with PD-L1 and PD-L2, PD-pathway-mediated inhibition of the immune response is abolished, resulting in an anti-tumor immune response. Clinical anti-tumor activity with sasanlimab was observed in a panel of anti-PD1 antibody-sensitive solid tumor types, including non-small cell lung cancer and urothelial carcinoma. Sasanlimab is described, for example, in U.S. Patent No. 10,155,037, which is hereby incorporated by reference for all purposes. The anti-PD-1 antibody used in the examples provided herein was a therapeutic humanized anti-human PD-1 antibody (hIgG2a-D265A) of in-house production unless otherwise indicated.

An effective amount of an anti-PD-1 antibody or an anti-PD-L1 antibody may be administered in accordance with the dosages provided herein.

Gamma-secretase inhibitor-based therapeutic agents

In some aspects, a therapeutic agent for use in the combination therapy of the present invention may be a gamma-secretase inhibitor (GSI). The terms "gamma-secretase inhibitor," "γ-secretase inhibitor," and "GSI" are used interchangeably herein to refer to compounds (including pharmaceutically acceptable salts, solvates, and prodrugs thereof) or other agents that inhibit or reduce the biological activity of gamma-secretase. Membrane-bound BCMA is actively cleaved from the tumor cell surface by the protease activity of gamma-secretase and can undergo gamma-secretase-mediated shedding. This may reduce the target density of BCMA-specific therapeutics on tumor cells and lead to the release of a soluble BCMA fragment (sBCMA) capable of antagonizing BCMA-specific therapeutics. By inhibiting gamma-secretase, membrane-bound BCMA can be preserved, increasing target density while reducing sBCMA levels. Thus, administration of GSIs may enhance the activity of BCMA-specific therapeutics.

Non-limiting examples of small molecule GSIs that can be used in the combination therapy of the present invention include the dipeptide class of GSIs, the sulfonamide class of GSIs, the transition state mimetic class of GSIs, the benzocaprolactam class of GSIs, and other GSIs known in the art. For example, the GSI can be selected from MK-0752 (Merck & Co., Inc.), MRK-003 (Merck & Co., Inc.), nirogacestat (PF-03084014, SpringWorks Therapeutics), RO4929097 (Roche), semagacestat (LY450139, Eli Lilly & Company), BMS-906024 (Bristol-Myers Squibb Company), and DAPT or pharmaceutically acceptable salts thereof. Additional examples of GSI include 1-(S)-endo-N-(1,3,3)-trimethylbicyclo[2.2.1]hept-2-yl)-4-fluorophenylsulfonamide, WPE-III-31C, (S)-3-[N'-(3,5-difluorophenyl-alpha-hydroxyacetyl)-L-alanyl]amino-2,3-dihydro-1-methyl-5-phenyl-1-H-1,4-benzodiazepin-2-one, and (N)-[(S)-2-hydroxy-3-methyl-butyryl]-1-(L-alanyl)-(S)-1-amino-3-methyl-4,5,6,7-tetrahydro-2H-3-benzazepin-2-one. See De Kloe & De Strooper (2014). Small Molecules That Inhibit Notch Signaling., In Bellen & Yamamoto (Eds.), Notch Signaling: Methods and Protocols, Methods in Mol. Biol., vol 1 187 (pp 311-322). New York, NY: Springer-Science+Business Media.

In some aspects, the therapeutic agent in the combination treatment of the present invention is a GSI. In some aspects, the GSI may have any of the features or characteristics of any of the GSIs disclosed in WO2005/092864, hereby incorporated by reference for all purposes. In some aspects, the GSI is nirogacestat (PF-03084014, SpringWorks Therapeutics) or a pharmaceutically acceptable salt thereof. Nirogacestat is an orally available, selective, small molecule GSI having the structure:

Nirogacestat is described, for example, in U.S. Patent No. 7,342,118, U.S. Patent No. 7,795,447, and U.S. Patent No. 7,951,958, which are hereby incorporated by reference for all purposes. Unless otherwise noted, the GSI used in the examples presented herein was nirogacestat.

An effective amount of GSI can be administered according to the dosages described herein. In some aspects, GSI is administered at a dose sufficient to upregulate BCMA surface expression on tumor cells. In some aspects, GSI is administered at a dose sufficient to reduce BCMA shedding on tumor cells. In some aspects, GSI is administered at a dose sufficient to reduce sBCMA levels. In some aspects, GSI is administered at a dose sufficient to improve the activity of BCMA-specific therapeutics.

Therapeutic agents

In some aspects, a therapeutic agent for use in the combination treatment of the present invention may include one or more of a biotherapeutic agent, a chemotherapeutic agent, an immunomodulatory agent (e.g., thalidomide, lenalidomide, pomalidomide, iberdomide, and apremilast), a proteasome inhibitor (e.g., bortezomib, carfilzomib, and ixazomib), a corticosteroid (e.g., dexamethasone and prednisone), a histone deacetylase (HDAC) inhibitor (e.g., panobinostat), and a nuclear export inhibitor (e.g., selinexor). Additional therapeutic agents for use in the combination treatment of the present invention include tumor vaccines, immune cell therapy (e.g., CAR-T cell therapy), radiation therapy, a vaccine, cytokine therapy (e.g., immunostimulatory cytokines, including various signaling proteins that stimulate the immune response, such as interferons, interleukins, and hematopoietic growth factors), a targeted cytokine, an inhibitor of other immunosuppressive pathways, angiogenesis inhibitors, a T cell activator, a metabolic pathway inhibitor, an mTOR (mammalian target of rapamycin) inhibitor (e.g., rapamycin, rapamycin derivatives, sirolimus, temsirolimus, everolimus, and deforolimus), an adenosine pathway inhibitor, a gamma-secretase inhibitor (e.g., nirogacestat), a tyrosine kinase inhibitor, including, but not limited to, ^INLYTA® inhibitors ALK (anaplastic lymphoma kinase) (e.g., crizotinib, ceritinib, alectinib, and sunitinib), BRAF inhibitor (e.g., vemurafenia and dabrafenib), PI3K inhibitor, HPK1 inhibitor, epigenetic modifier, inhibitors or depletor of T _reg cells and/or myeloid-derived suppressor cells, JAK (Janus kinase) inhibitor (e.g., ruxolitinib and tofacitinib, varicitinib, filgotinib, gandotinib, lestaurtinib, momelotinib, pacritinib, and upadacitinib), STAT (signal transducers and activators of transcription) inhibitor (e.g., STAT1, STAT3, and STAT5 inhibitors such as fludarabine), inhibitor of cyclin-dependent kinases (CDKs) or other cell cycle factors, immunogenic agent (e.g., attenuated Malignant cells, tumor antigens, antigen-presenting cells such as dendritic cells complemented with antigen or nucleic acids of tumor origin, a MEK inhibitor (e.g., trametinib, cobimetinib, binimetinib, and selumetinib), a GLS1 inhibitor, a PARP inhibitor (e.g., talazoparib, olaparib, rucaparib, niraparib), an oncolytic virus, gene therapies including DNA, RNA delivered directly or via adeno-associated viruses (AAV) or nanoparticles, modulators of the innate immune response (e.g., TLR, KIR, NKG2A), an IDO (indoleamine pyrrole-2,3-dioxygenase) inhibitor, a PRR (pattern recognition receptor) agonist, and cells transfected with genes encoding immunostimulatory cytokines such as, but not limited to, GM-CSF).

In some aspects, therapeutic agents for use in the combination treatment of the present invention may comprise an antibody, including, but not limited to, an anti-CTLA-4 antibody, an anti-CD3 antibody, an anti-CD4 antibody, an anti-CD8 antibody, an anti-4-1BB antibody, an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-TIM3 antibody, an anti-LAG3 antibody, an anti-TIGIT antibody, an anti-OX40 antibody, an anti-IL-7R-alpha (CD127) antibody, an anti-IL-8 antibody, an anti-IL-15 antibody, an anti-HVEM antibody, an anti-BTLA antibody, an anti-CD38 antibody, an anti-CD40 antibody, an anti-CD40L antibody, an anti-CD47 antibody, an anti-CSF1R antibody, an anti-CSF1 antibody, an anti-IL-7R antibody, an anti-MARCO antibody, an anti-CXCR4 antibody, an anti-VEGF antibody, an anti- VEGFR1, anti-VEGFR2, anti-TNFR1, anti-TNFR2, bispecific anti-CD3, anti-CD19, anti-CD20, anti-Her2, anti-EGFR, anti-ICOS, anti-CD22, anti-CD52, anti-CCR4, anti-CCR8, anti-CD200R, anti-VISG4, anti-CCR2, anti-LILRb2, anti-CXCR4, anti-CD206, anti-CD163, anti-KLRG1, anti-FLT3, anti-B7-H4, anti-B7-H3, anti-KLRG1, anti-BTN1A1, anti-BCMA, anti-SLAMF7, anti-avb8, anti-CD80 or anti-GITR antibody.

In some aspects, other exemplary therapeutic agents for use in the combination treatment of the present invention may be directed or targeted to 5T4; A33; folate receptor alpha 1 (e.g., mirvetuximab soravtansine); Alk-1; BCMA (e.g., see WO2016166629 et al. cited herein); BTN1A1 (see, e.g., WO2018222689); CA19-9; CA-125 (e.g., abagovomab); carbonic anhydrase IX; CCR2; CCR4 (e.g., mogamulizumab); CCR5 (e.g., leronlimab); CCR8; CD3 [e.g., blinatumomab (anti-CD3/CD19 bispecific antibody), PF-06671008 (anti-CD3/P-cadherin bispecific antibody), PF-06863135 (anti-CD3/BCMA bispecific antibody)]; CD19 (e.g., blinatumomab, MOR208); CD20 (e.g., ibritumomab tiuxetan, obinutuzumab, ofatumumab, rituximab, ublituximab); CD22 (inotuzumab ozogamicin, moxetumomab pasudotox); CD25; CD28; CD30 (e.g., brentuximab vedotin); CD33 (e.g., gemtuzumab ozogamicin); CD38 (eg, daratumumab, daratumumab and hyaluronidase, and isatuximab), CD40; CD-40L; CD44v6; CD47 (eg, Hu5F9-G4, CC-90002, SRF231, B6H12); CD52 (eg, alemtuzumab); CD56; CD63; CD79 (eg, polatuzumab vedotin); CD80; CD86; CD123; CD276/B7-H3 (eg, omburtamab); CDH17; CEA; ClhCG; CTLA-4 (eg, ipilimumab, tremelimumab), CXCR4; desmoglein 4; DLL3 (eg, rovalpituzumab tesirin); DLL4; E-cadherin; EDA; EDB; EFNA4; EGFR (e.g., cetuximab, depatuxizumab mafodotin, necitumumab, panitumumab); EGFRvIII; Endosialin; EpCAM (e.g., oportuzumab monatox); FAP; fetal acetylcholine receptor; FLT3 (see, e.g., WO2018/220584); 4-1BB (CD137) [e.g., utomilumab/PF-05082566 (see WO2012/032433) or urelumab/BMS-663513], GD2 (e.g., dinutuximab, 3F8); GD3; GITR (e.g., TRX518); GloboH; GM1; GM2; HER2/neu [e.g., margetuximab, pertuzumab, trastuzumab; ado-trastuzumab emtansine, trastuzumab duocarmazine, PF-06804103 (see U.S. Patent No. 8,828,401)]; HER3; HER4; ICOS; IL-10; ITG-AvB6; LAG-3 (e.g., relatlimab, IMP701); Lewis antigen Y; LG; Ly-6; M-CSF [e.g., PD-0360324 (see U.S. Patent No. 7,326,414)]; (membrane-bound) IgE; MCSP; mesothelin; MIS receptor type II; MUC1; MUC2; MUC3; MUC4; MUC5AC; MUC5B; MUC7; MUC16; Notch1; Notch3; nectin-4 (e.g., enfortumab vedotin); OX40 [e.g., PF-04518600 (see U.S. Patent No. 7,960,515)]; P-cadherin [e.g., PF-06671008 (see WO2016/001810)]; PCDHB2; PD-1 [e.g., BCD-100, camrelizumab, cemiplimab, genolimzumab (CBT-501), MEDI0680, nivolumab, pembrolizumab, sasanlimab (PF-06801591, see WO2016/092419), sintilimab, spartalizumab, STI-A1110, tislelizumab, TSR-042, and others described herein]; PD-L1 (e.g., atezolizumab, durvalumab, BMS-936559 (MDX-1105), LY3300054, and others described herein); PDGFRA (e.g., olaratumab); plasma cell antigen; poly-SA; PSCA; PSMA; PTK7 [e.g., PF-06647020 (see U.S. Patent No. 9,409,995)]; Ror1; SAS; SLAMF7 (e.g., elotuzumab); SHH; SIRPa (e.g., ED9, Effi-DEM); STEAP; sTn; TGF-beta; TIGIT; TIM-3; TMPRSS3; TNF-alpha precursor; TROP-2 (e.g., sacituzumab govitecan); TSPAN8; VEGF (e.g., bevacizumab, brolucizumab); VEGFR1 (eg, ranibizumab); VEGFR2 (eg, ramucirumab, ranibizumab) and Wue-1.

In some aspects, a therapeutic agent for use in the combination therapy of the present invention may be a therapeutic antibody in any suitable format. For example, therapeutic antibodies may have any format as described elsewhere herein. In some aspects, a therapeutic antibody may be a naked antibody. In some aspects, a therapeutic antibody may be linked to a drug/agent (also known as an "antibody-drug conjugate" (ADC)). Drugs or agents that can be linked to an antibody in ADC format may include, for example, cytotoxic agents, immunomodulatory agents, imaging agents, therapeutic proteins, biopolymers, or oligonucleotides. Examples of cytotoxic agents that can be included in an ADC include anthracycline, auristatin, dolastatin, combretastatin, duocarmycin, pyrrolobenzodiazepine dimer, indolinobenzodiazepine dimer, enediyne, geldanamycin, maytansine, puromycin, taxane, vinca alkaloid, camptothecin, tubulysin, hemiasterlin, spliceostatin, pladienolide and their stereoisomers, isomers, analogs or derivatives.

In some aspects, a therapeutic antibody against a specific antigen can be included in a multispecific antibody (e.g., a bispecific or trispecific antibody). Bispecific antibodies are monoclonal antibodies that have binding specificity for at least two different antigens. In some aspects, the bispecific antibody comprises a first variable domain of the antibody and a second variable domain of the antibody, wherein the first variable domain of the antibody can recruit the activity of a human effector immune cell by specifically binding to an effector antigen localized on the human effector immune cell, and wherein the second variable domain of the antibody can specifically bind to a target antigen, as described herein. In some aspects, the antibody has an IgG1, IgG2, IgG3, or IgG4 isotype. In some aspects, the antibody comprises an immunologically inert Fc region. In some aspects, the antibody is a human antibody or a humanized antibody.

A human effector immune cell may be any of a variety of effector immune cells known in the art. For example, an effector immune cell may be a member of the human lymphoid lineage, including, but not limited to, a T cell (e.g., a cytotoxic T cell), a B cell, and a natural killer (NK) cell. An effector immune cell may also be, but not limited to, a member of the human myeloid lineage, including, but not limited to, a monocyte, a neutrophil granulocyte, and a dendritic cell. Such effector immune cells may have a cytotoxic or apoptotic effect on a target cell or another desired effect regarding activation through binding to an effector antigen.

An effector antigen is an antigen (e.g., a protein or polypeptide) expressed on a human effector immune cell. Non-limiting examples of effector antigens that can be bound by a heterodimeric protein (e.g., a heterodimeric antibody or a bispecific antibody) include human CD3 (or CD3 (cluster of differentiation) complex), CD16, NKG2D, NKp46, CD2, CD28, CD25, CD64, and CD89. A target antigen is typically expressed on a target cell in a diseased state (e.g., a malignant cell). Examples of target antigens for use in bispecific antibodies are provided herein.

In some aspects, the bispecific antibody provided herein binds to two different target antigens on a single target cell (e.g., two different antigens on a single tumor cell). Such antibodies may be advantageous, for example, to achieve increased specificity for a target cell of interest (e.g., a tumor cell expressing two specific tumor-associated antigens of interest). For example, in some aspects, the bispecific antibody provided herein comprises a first variable domain of the antibody and a second variable domain of the antibody, wherein the first variable domain of the antibody can specifically bind to a first target antigen, as provided herein, and the second variable domain of the antibody can specifically bind to a second target antigen, as provided herein.

In some aspects, a therapeutic agent for use in the combination therapy of the present invention may include immunomodulatory agents, including thalidomide, lenalidomide, pomalidomide, iberdomide, and apremilast, which can stimulate an immune response in an individual. Additional immunomodulatory agents include pattern recognition receptor (PRR) agonists, immunostimulatory cytokines, immune cell therapy, and tumor vaccines.

Pattern recognition receptors (PRRs) are receptors expressed by cells of the immune system that recognize various molecules associated with pathogens and/or cell injury or death. PRRs are involved in the innate and adaptive immune responses. PRR agonists can be used to stimulate the immune response in an individual. There are many classes of PRR molecules, including Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs), C-type lectin receptors (CLRs), and stimulator of interferon genes (STING) protein.

Examples of TLR agonists provided herein include TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, and TLR9 agonists. Examples of RLR agonists that can be used in the treatment methods, medicaments, and uses of the present invention include, for example, short double-stranded RNA with an uncapped 5'-triphosphate (RIG-I agonist); poly-I:C (MDA-5 agonist), and BO-112 (MDA-A agonist). Examples of NLR agonists that can be used in the treatment methods, medicaments, and uses of the present invention include, for example, liposomal muramyl tripeptide/mifamurtide (NOD2 agonist). Examples of CLR agonists that can be used in the treatment methods, medicaments, and uses of the present invention include, for example, MD fraction (purified soluble beta-glucan extract from Grifola frondosa ) and Imprime PGG (beta-1,3/1,6-glucan PAMP obtained from yeast). Examples of STING agonists that can be used in the treatment methods, medicaments, and uses of the present invention include various immunostimulatory nucleic acids, such as synthetic double-stranded DNA, cyclic di-GMP, cyclic-GMP-AMP (cGAMP), synthetic cyclic dinucleotides (CDNs) such as MK-1454 and ADU-S100 (MIW815), and small molecules such as P0-424. Other PRRs include, for example, DNA-dependent activator of interferon-regulatory factors (DAI) and Absent in melanoma 2 (AIM2).

Immunostimulatory cytokines include, but are not limited to, various signaling proteins that stimulate the immune response, such as interferons, interleukins, and hematopoietic growth factors. In some aspects, non-limiting examples of immunostimulatory cytokines include GM-CSF, G-CSF, IFNγ, IFNα, IL-2 (e.g., denileukin diftitox), IL-6, IL-7, IL-10, IL-11, IL-12, IL-15, IL-18, IL-21, and TNFα. Immunostimulatory cytokines can be in any suitable format. In some aspects, an immunostimulatory cytokine can be a recombinant version of a wild-type cytokine. In some aspects, an immunostimulatory cytokine can be a mutein having one or more amino acid changes compared to the corresponding wild-type cytokine. In some aspects, an immunostimulatory cytokine can be incorporated into a chimeric protein comprising the cytokine and at least one other functional protein (e.g., an antibody). In some aspects, the immunostimulatory cytokine can be covalently linked to a drug/agent (e.g., any drug/agent described elsewhere herein as a possible component of an ADC). In some aspects, the cytokines are PEGylated.

Immune cell therapy involves treating patients with immune cells capable of targeting malignant cells. Examples of immune cell therapy include tumor-infiltrating lymphocytes (TILs) and chimeric antigen receptor T cells (CAR-T cells).

Tumor vaccines include various compositions containing tumor-associated antigens (or that can be used to generate a tumor-associated antigen in an individual) and, thus, can be used to initiate an immune response in an individual that is directed against tumor cells containing the tumor-associated antigen. Examples of materials that can be included in a tumor vaccine include attenuated malignant cells, tumor antigens, and antigen-presenting cells such as dendritic cells complemented with an antigen or tumor-derived nucleic acids encoding tumor-associated antigens. In some aspects, a tumor vaccine can be produced using a patient's own malignant cells. In some aspects, a tumor vaccine can be produced using biological material other than the patient's own malignant cells. Cancer vaccines include, for example, sipuleucel-T and talimogene laherparepvec (T-VEC).

The combination therapy provided herein may include one or more chemotherapeutic agents. Examples of chemotherapeutic agents include 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 altretamine, triethylenemelamine, triethylenephosphamide, triethylenethiophosphamide, and trimethylolomelamine; acetogenins (particularly bullatacin and bullatacinone); camptothecin (including the synthetic analog topotecan); bryostatin; callistatin; CC-1065 (including its synthetic analogs adozelesin, carzelesin, and bizelesin); Cryptophycins (especially cryptophycin 1 and cryptophycin 8); Dolastatin; Duocarmycin (including synthetic analogues, KW-2189 and CBI-TMI); Eleutherobin; Pancratistatin; Sarcodictin; Spongistatin; Nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichine, phenesterine, prednimustine, trofosfamide, uracil mustard; Nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as enediyne antibiotics (e.g., calicheamicin, particularly calicheamicin gamma II and calicheamicin phi II, see, e.g., Agnew, Chem. Intl. Ed. Engl., 33:183-186 (1994)); dynemicin, including dynemicin A; bisphosphonates such as clodronate; esperamicin; and neocarzinostatin chromophore and related chromoprotein chromophore-enediyne antibiotics), aclacinomycins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, cacinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, Porfiromycin, puromycin, quelamicin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; Anti-adrenal agents such as aminoglutethimide, mitotane, trilostane; Folic acid supplement such as folinic acid; FOLFOX including folinic acid, 5-FU and oxaliplatin; Aceglatone; Aldophosphamide glycoside; Aminolevulinic acid; Eniluracil; Amsacrine; Bestrabucil; Bisantrene; Edatraxate; Defofamine; Demecolcine; Diaziquone; Elfornithine; Elliptinium acetate; Epothilone; Etoglucide; Gallium nitrate; Hydroxyurea; Lentinan; Lonidamine; Maytansinoids such as maytansine and ansamitocins; Mitoguazone; Mitoxantrone; Mopidamol; Nitracrine; Pentostatin; Phenamet; Pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; razoxan; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2′,2″-trichlorotriethylamine; trichothecenes (eg, T-2 toxin, verraccurin A, roridin A, and anguidine); urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosin; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids such as paclitaxel and doxetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogues such as carboplatin; Cisplatin; Vinblastine; Platinum; Etoposide (VP-16); Ifosfamide; Mitoxantrone; Vincristine; Vinorelbine; Novantrone; Teniposide; Edatrexate; Daunomycin; Aminopterin; Xeloda; Ibandronate; CPT-11; Topoisomerase inhibitor RFS 2000; Difluoromethylornithine (DMFO); Retinoids such as retinoic acid; Capecitabine and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing.

In some aspects, a therapeutic agent for use in the combination treatment of the present invention may be an antihormonal agent that acts by regulating or inhibiting the action of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen, raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone and toremifene (fareston); aromatase inhibitors that inhibit the aromatase enzyme that regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazoles, aminoglutethimide, megestrol acetate, exemestane, formestane, fadrozole, vorozole, letrozole and anastrozole.

In some aspects, a therapeutic agent for use in the combination treatment of the present invention may be antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, fluridil, apalutamide, enzalutamide, cimetidine and goserelin; KRAS inhibitors; MCT4 inhibitors; MAT2a inhibitors; tyrosine kinases/vascular endothelial growth factor (VEGF) receptor inhibitors such as sunitinib, axitinib, sorafenib, tivozanib; alk/c-Met/ROS inhibitors such as crizotinib, lorlatinib; mTOR inhibitors such as temsirolimus, gedatolisib; src/abl inhibitors such as bosutinib; Cyclin-dependent kinase (CDK) inhibitors such as palbociclib, PF-06873600, abemaciclib, and ribociclib; erb inhibitors such as dacomitinib; PARP inhibitors such as talazoparib, olaparib, rucaparib, niraparib; SMO inhibitors such as glasdegib, PF-5274857; EGFR T790M inhibitors such as PF-06747775; EZH2 or other epigenetic modifier inhibitors such as PF-06821497; PRMT5 inhibitors such as PF-06939999; TGFRβr1 inhibitors such as PF-06952229; and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing.

Treatment

Each therapeutic agent in the combination treatment of the invention may be administered alone or in a medicament (also referred to herein as a pharmaceutical composition) comprising the therapeutic agent and one or more pharmaceutically acceptable carriers, excipients and diluents in accordance with standard pharmaceutical practice.

Each therapeutic agent in the combination therapy of the invention can be administered simultaneously (i.e., in a single medicinal agent), simultaneously (i.e., in separate medicinal agents administered immediately after each other in any order), or sequentially in any order. Sequential administration is particularly suitable when the therapeutic agents in the combination therapy are in different dosage forms (one agent is a tablet or capsule, and the other agent is a sterile liquid) and/or they are administered according to different administration schedules, for example, a chemotherapeutic agent that is administered at least daily and a therapeutic agent that is administered less frequently, for example, weekly, biweekly, or triweekly.

In some aspects, the therapeutic agent in the combination therapy may be administered using the same dosing regimen (dose, frequency, and duration of treatment) as would typically be used when the agent is used as monotherapy for the treatment of the same malignancy. In other aspects, the patient may be administered a lower total amount of at least one of the therapeutic agents in the combination therapy than when the agent is used as monotherapy, such as lower doses, less frequent dosing, and/or a shorter duration of treatment.

The therapeutic agents in the combination therapy of the invention can be administered by any suitable enteral route or parenteral route. The term "enteral route" of administration refers to administration via any part of the gastrointestinal tract. Examples of enteral routes include the oral, mucosal, buccal and rectal route or the intragastric route. The term "parenteral route" of administration refers to a route of administration other than the enteral route. Examples of parenteral routes of administration include intravenous, intramuscular, intradermal, intraperitoneal, intratumoral, intravesical, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, transtracheal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal, subcutaneous or topical administration. Therapeutic agents of the invention can be administered by any suitable route, such as orally, via a nasogastric tube, gastrostomy tube, injection, infusion, implantable infusion pump, or osmotic pump. The appropriate route and method of administration may vary depending on a number of factors, such as the specific therapeutic agent used, the desired rate of absorption, the specific formulation or dosage form used, the type or severity of the disorder being treated, the specific site of action, and the patient's condition. Examples of parenteral routes of administration also include intraosseous and intrapleural.

Oral administration of a solid dosage form of a therapeutic agent can be accomplished, for example, in discrete units such as hard or soft capsules, pills, cachets, lozenges, or tablets, each containing a predetermined amount of at least one therapeutic agent. In another aspect, oral administration can be in the form of a powder or granule. In another aspect, the oral dosage form is sublingual, such as, for example, a lozenge. In such solid dosage forms, the therapeutic agents are typically combined with one or more excipients. Such capsules or tablets may contain a controlled-release formulation. Capsules, tablets, and pills may also contain buffering agents or may be prepared using enteric coatings.

In another aspect, oral administration of the therapeutic agent can be carried out in a liquid dosage form. Liquid dosage forms for oral administration include, for example, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art (e.g., water). Such compositions may also contain excipients such as humectants, emulsifiers, suspending agents, flavoring agents (e.g., sweeteners), and/or aromatizing agents.

In some aspects, therapeutic agents are administered in parenteral dosage form. "Parenteral administration" includes, for example, subcutaneous injections, intravenous injections, intraperitoneal injections, intramuscular injections, intrasternal injections, and infusion. Injectable preparations (i.e., sterile injectable aqueous or oleaginous suspensions) can be formulated by known methods using suitable dispersing, wetting, and/or suspending agents, and include depot formulations.

In some aspects, therapeutic agents are administered in a topical dosage form. The term "topical administration" includes, for example, transdermal administration, such as via transdermal patches or iontophoresis devices, intraocular administration, or intranasal or inhalation administration. Compositions for topical administration also include, for example, topical gels, sprays, ointments, and creams. The topical formulation may include a compound that enhances the absorption or penetration of the active ingredient through the skin or other affected areas. If therapeutic agents are administered via a transdermal device, administration will be accomplished using a reservoir-type patch and a porous membrane or a solid matrix-type patch. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, powders, dressings, foams, films, skin patches, implantable capsules, implants, sponges, fibers, bandages, and microemulsions. Liposomes can also be used. Typical carriers include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethyleneglycol, and propylene glycol. Penetration enhancers can be included; see, for example, Finnin and Morgan, J. Pharm. Sci., 88 (10), 955–958 (1999).

Other carrier materials and routes of administration known in the pharmaceutical art may also be used with therapeutic agents. The above considerations regarding effective formulations and routes of administration are well known in the art and are described in standard textbooks. Compounding of drugs is described, for example, in Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1975; Liberman et al., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Kibbe et al., Eds., Handbook of Pharmaceutical Excipients (3rd sup. Ed.), American Pharmaceutical Association, Washington, 1999.

The selection of a dosing regimen (also referred to herein as a dosing schedule) for the combination therapy of the invention may depend on several factors, including the turnover rate of the substance in the serum or tissue, the level of symptoms, the immunogenicity of the substance, and the availability of the target cells, tissue, or organ in the body of the individual being treated. Preferably, the dosing regimen maximizes the amount of each therapeutic agent administered to the patient consistent with an acceptable level of side effects. Thus, the dose amount and frequency of administration of each therapeutic agent or chemotherapeutic agent in the combination depends, in part, on the particular therapeutic agent, the severity of the malignancy being treated, and the characteristics of the patient. Guidelines for selecting appropriate doses of antibodies, cytokines, and small molecules are available. See, for example, Wawrzynczak (1996) Antibody Therapy, Bios Scientific Pub. Ltd, Oxfordshire, UK; Kresina (ed.) (1991) Monoclonal Antibodies, Cytokines and Arthritis, Marcel Dekker, New York, NY; Bach (ed.) (1993) Monoclonal Antibodies and Peptide Therapy in Autoimmune Diseases, Marcel Dekker, New York, NY; Baert et al. (2003) New Engl. J. Med. 348:601-608; Milgrom et al. (1999) New Engl. J. Med. 341:1966-1973; Slamon et al. (2001) New Engl. J. Med. 344:783-792; Beniaminovitz et al. (2000) New Engl. J. Med. 342:613-619; Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J. Med. 343:1594-1602; Physicians' Desk Reference 2003 (Physicians' Desk Reference, 57th Ed); Medical Economics Company; ISBN: 1563634457; 57th edition (November 2002). The determination of the appropriate dosage regimen will be made by the clinician, for example, using parameters or factors known or suspected in the art to influence treatment or to be prognostic of treatment, and will depend on, for example, the patient's medical history (e.g., previous therapy), the type and stage of the malignancy being treated, and biomarkers of response to one or more of the therapeutic agents in combination therapy.

In some aspects, the therapeutic agents in the combination treatment of the invention can be administered to an individual at a dose of about 0.01 μg/kg, 0.02 μg/kg, 0.03 μg/kg, 0.04 μg/kg, 0.05 μg/kg, 0.06 μg/kg, 0.07 μg/kg, 0.08 μg/kg, 0.09 μg/kg, 0.1 μg/kg, 0.2 μg/kg, 0.3 μg/kg, 0.4 μg/kg, 0.5 μg/kg, 0.6 μg/kg, 0.7 μg/kg, 0.8 μg/kg, 0.9 μg/kg, 1 μg/kg, 2 μg/kg, 3 μg/kg, 4 µg/kg, 5 µg/kg, 6 µg/kg, 7 µg/kg, 8 µg/kg, 9 µg/kg, 10 µg/kg, 15 µg/kg, 20 µg/kg, 25 µg/kg, 30 µg/kg, 35 µg/kg, 40 µg/kg, 45 µg/kg, 50 µg/kg, 60 µg/kg, 70 µg/kg, 80 µg/kg, 90 µg/kg, 100 µg/kg, 110 µg/kg, 120 µg/kg, 130 µg/kg, 140 µg/kg, 150 µg/kg, 200 µg/kg, 250 µg/kg, 300 mcg/kg, 400 mcg/kg, 500 mcg/kg, 600 mcg/kg, 700 mcg/kg, 800 mcg/kg, 900 mcg/kg, 1000 mcg/kg, 1200 mcg/kg, 1400 mcg/kg or more.

In some aspects, the therapeutic agents in the combination treatment of the invention can be administered to an individual at a dose of from about 1 mg/kg to about 1000 mg/kg, from about 2 mg/kg to about 900 mg/kg, from about 3 mg/kg to about 800 mg/kg, from about 4 mg/kg to about 700 mg/kg, from about 5 mg/kg to about 600 mg/kg, from about 6 mg/kg to about 550 mg/kg, from about 7 mg/kg to about 500 mg/kg, from about 8 mg/kg to about 450 mg/kg, from about 9 mg/kg to about 400 mg/kg, from about 5 mg/kg to about 200 mg/kg, from about 2 mg/kg to about 150 mg/kg, from about 5 mg/kg to about 100 mg/kg, from about 10 mg/kg to about 100 mg/kg or from about 10 mg/kg to about 60 mg/kg.

In some aspects, the therapeutic agents in the combination treatment of the invention can be administered to the individual at a dose of at least 0.05 μg/kg, 0.2 μg/kg, 0.5 μg/kg, 1 μg/kg, 10 μg/kg, 100 μg/kg, 0.2 mg/kg, 1.0 mg/kg, 2.0 mg/kg, 3.0 mg/kg, 5.0 mg/kg, 10 mg/kg, 25 mg/kg, 50 mg/kg body weight, or more. See, e.g., Yang et al. (2003) New Engl. J. Med. 349:427-434; Herold et al. (2002) New Engl. J. Med. 346:1692-1698; Liu et al. (1999) J. Neurol. Neurosurg. Psych. 67:451-456; Portielji et al. (20003) Cancer Immunol. Immunother. 52:133-144.

In some aspects, a patient may be administered a fixed dose of a therapeutic agent of about or at least about 0.05 mcg, 0.2 mcg, 0.5 mcg, 1 mcg, 10 mcg, 100 mcg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 75 mg, 80 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 350 mg, 700 mg, 750 mg, 800 mg, 900 mg, 1000 mg, or 1500 mg or more. The fixed dose can be administered at intervals, such as daily, every other day, three times a week, or once a week, fortnightly, three weeks, monthly, every 2 months, every 3 months, every 4 months, etc.

In the case of oral administration, therapeutic agents (e.g., typically small molecule chemotherapeutic agents) can be provided in the form of tablets in the dose of the therapeutic agent provided herein.

In some aspects, the therapeutic agents in the combination treatment of the invention can be administered at least daily, once daily, twice daily, three times daily, four times daily, every two days, every three days, once weekly, every two weeks, every three weeks, every four weeks, every 30 days, every five weeks, every six weeks, once monthly, every two months, every three months, or every four months in an oral, IV, or SC dose.

The treatment methods provided herein can be used for as long as the clinician caring for the patient deems the treatment method to be effective. Non-limiting parameters indicative of a treatment method being effective include any one or more of the following: a reduction in tumor size (in terms of mass and/or volume); a reduction in the number of individual tumor colonies; tumor elimination; and progression-free survival. A change in tumor size can be determined by any suitable method, such as imaging. Various diagnostic imaging methods well known in the art can be used, such as computed tomography (CT), dual-source CT, positron emission tomography, ultrasound, computed axial tomography, and MRI. In some aspects, the combination treatment of the invention is used to treat a tumor large enough to be detected by palpation or by imaging methods well known in the art, such as MRI, ultrasound, or computed axial tomography.

Examples of time periods associated with a course of therapy include approximately one week; approximately two weeks; approximately three weeks; approximately four weeks; approximately five weeks; approximately six weeks; approximately seven weeks; approximately eight weeks; approximately nine weeks; approximately ten weeks; approximately eleven weeks; approximately twelve weeks; approximately thirteen weeks; approximately fourteen weeks; approximately fifteen weeks; approximately sixteen weeks; approximately seventeen weeks; approximately eighteen weeks; approximately nineteen weeks; approximately twenty weeks; approximately twenty-one weeks; approximately twenty-two weeks; approximately twenty-three weeks; approximately twenty-four weeks; approximately seven months; approximately eight months; approximately nine months; approximately ten months; approximately eleven months; approximately twelve months; approximately thirteen months; approximately fourteen months; approximately fifteen months; approximately sixteen months; approximately seventeen months; approximately eighteen months; approximately nineteen months; approximately twenty months; approximately twenty-one months; approximately twenty-two months; approximately twenty-three months; approximately twenty-four months; approximately thirty months; approximately three years; approximately four years; and approximately five years.

The combinations and methods of the invention can be used to treat a patient suffering from any condition that can be treated or prevented by the methods provided herein, such as a malignant neoplasm and/or a tumor-associated disease.

In some aspects, the condition is a malignancy, including, but not limited to, carcinoma, lymphoma, leukemia, myeloma, blastoma, and sarcoma. In some aspects, the malignancy may include tumor-associated diseases including a B-cell malignancy and/or tumor-associated disease including, but not limited to, multiple myeloma, malignant plasma cell neoplasia, lymphoma, Hodgkin's lymphoma, nodular Hodgkin's lymphoma with lymphocyte predominance, Kahler's disease and myelomatosis, plasma cell leukemia, plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering myeloma, light chain amyloidosis, osteosclerotic myeloma, B-cell prolymphocytic leukemia, hairy cell leukemia, B-cell non-Hodgkin's lymphoma (NHL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL), Chronic myeloid leukemia (CML), follicular lymphoma, Burkitt's lymphoma, marginal zone cell lymphoma, mantle cell lymphoma, large cell lymphoma, precursor B-cell lymphoblastic lymphoma, myeloid leukemia, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma, mucosal lymphoid tissue lymphoma, small lymphocytic lymphoma, primary mediastinal (thymic) large B-cell lymphoma, lymphoplasmacytic lymphoma, marginal zone B-cell lymphoma, splenic marginal zone cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, Hodgkin's disease, T-cell/histiocytic large B-cell lymphoma, primary central lymphoma nervous system, primary cutaneous diffuse large B-cell lymphoma (foot type), EBV-positive diffuse large B-cell lymphoma of the elderly, inflammation-associated diffuse large B-cell lymphoma, ALK-positive large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma arising in HHV8-associated multifocal Castleman disease, undifferentiated B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and Burkitt's lymphoma, undifferentiated B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin's lymphoma, and other B-cell lymphomas.

In some aspects, the malignancy is gastric cancer, small bowel cancer, head and neck cancer (e.g., squamous cell carcinoma of the head and neck), thymus cancer, epithelial cancer, salivary gland cancer, liver cancer, biliary tract cancer, neuroendocrine tumors, gastric cancer, thyroid cancer, lung cancer (e.g., non-small cell lung cancer, small cell lung cancer), mesothelioma, ovarian cancer, breast cancer, prostate cancer, kidney cancer, esophageal cancer, pancreatic cancer, glioma, kidney cancer (e.g., renal cell carcinoma), bladder cancer, cervical cancer, uterine cancer, vulvar cancer, endometrial cancer, penile cancer, testicular cancer, anal cancer, choriocarcinoma, colon cancer, colorectal cancer, oral cancer, skin cancer, Merkel cell carcinoma, glioblastoma, brain tumor, malignant neoplasm of bone tissue, ocular malignancy, melanoma, or malignancy with high microsatellite instability (MSI-H).

The combination treatment of the invention can be used before or after surgery to remove the tumor and before, during or after radiation therapy.

In some aspects, the combination therapy of the invention is administered to a patient who has not previously been treated with a therapeutic or chemotherapeutic agent, i.e., previously treated. In other aspects, the combination therapy is administered to a patient who has failed to achieve a sustained response to prior therapy with a therapeutic or chemotherapeutic agent, i.e., previously treated. In some aspects, the individual has previously undergone therapy for the treatment of a tumor, and the tumor is relapsed or refractory.

The present invention relates to methods of combination therapy that have additive potency or additive therapeutic effect while reducing or avoiding unwanted or adverse effects. The invention also relates to synergistic combinations in which the therapeutic efficacy is greater than additive while unwanted or adverse effects are reduced or eliminated. In certain aspects, the methods and compositions provided herein enable the treatment or prevention of diseases and disorders wherein the treatment is improved by an enhanced antitumor response using smaller and/or less frequent doses of at least one therapeutic agent in the combination therapy for at least one of: i) reducing the incidence of unwanted or adverse effects caused by administering the therapeutic agents separately while at least maintaining the effectiveness of the treatment; ii) increasing patient compliance with the treatment regimen; and iii) improving the effectiveness of the antitumor treatment.

Sets

The therapeutic agents of the combination treatment methods of the present invention may be conveniently combined in the form of a kit suitable for co-administration of the compositions.

In one aspect, the kit comprises at least a first container, a second container, and a package insert. The first container contains at least one dose of a first therapeutic agent, and the second container contains at least one dose of a second therapeutic agent of the combination therapy. The package insert/labeling contains instructions for treating a patient for a cancer and/or tumor-associated disease using the therapeutic agents. The first and second containers may be the same or different shapes (e.g., vials, syringes, and bottles) and/or materials (e.g., plastic or glass). The kit may further contain other materials that can be used in administering the therapeutic agents, such as diluents, filters, IV bags and systems, needles, and syringes.

Clinical trials

An open-label, multi-dose, multicenter, Phase 1 dose-escalation, safety, pharmacokinetics (PK), and pharmacodynamic study of PF-06863135 is ongoing in adult patients with advanced multiple myeloma who have relapsed on or are refractory to standard therapy (NCT03269136). This is a two-part study to evaluate the safety and tolerability of escalating dose levels of PF-06863135 in Part 1 and to determine the recommended Phase 2 dose (RP2D) in Part 2. This Phase 1 study is described in Example 10. Two additional clinical studies of PF06863135 (elranatamab) monotherapy are described in Examples 11 and 12.

Further clinical evaluation of PF-06863135 may be performed in combination with any of the therapeutic agents described herein: PF-06863135 in combination with an anti-PD-1/PD-L1 antibody (e.g., sasanlimab/PF-06801591), PF-06863135 in combination with an immunomodulatory agent (e.g., thalidomide, lenalidomide, pomalidomide, iberdomide, and apremilast), PF-06863135 in combination with a gamma-secretase inhibitor (e.g., nirogacestat), PF-06863135 in combination with other treatment modalities, such as a biotherapeutic agent (e.g., the anti-CD38 antibodies daratumumab, daratumumab and hyaluronidase, and isatuximab, and the anti-SLAMF7 antibody elotuzumab), a chemotherapeutic agent (e.g., melphalan, vincristine, cyclophosphamide, etoposide, doxorubicin, liposomal doxorubicin, and bendamustine), a proteasome inhibitor (e.g., bortezomib, carfilzomib, and ixazomib), a corticosteroid (e.g., dexamethasone and prednisone), a histone deacetylase (HDAC) inhibitor (e.g., panobinostat), and a nuclear export inhibitor (e.g., selinexor). Examples 12-16 describe several planned clinical trials of the combination treatment PF06863135 (elranatamab).

General methods

Standard molecular biology techniques are described by Sambrook, Fritsch, and Maniatis (1982 & 1989 2nd Edition, 2001 3rd Edition) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Sambrook and Russell (2001) Molecular Cloning, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Wu (1993) Recombinant DNA, Vol. 217, Academic Press, San Diego, CA). Standard techniques are also described in Ausbel et al. (2001) Current Protocols in Molecular Biology, Vols. 1–4, John Wiley and Sons, Inc. New York, NY, which describes cloning in bacterial cells and DNA mutagenesis (Vol. 1), cloning in mammalian cells and yeast (Vol. 2), glycoconjugates and protein expression (Vol. 3), and bioinformatics (Vol. 4).

Protein purification methods, including immunoprecipitation, chromatography, electrophoresis, centrifugation, and crystallization, are described (Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 1, John Wiley and Sons, Inc., New York). Chemical analysis, chemical modification, post-translational modification, production of fusion proteins, and protein glycosylation are described (see, e.g., Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 2, John Wiley and Sons, Inc., New York; Ausubel, et al. (2001) Current Protocols in Molecular biology, Vol. 3, John Wiley and Sons, Inc., NY, NY, pp. 16, 0, 5-16, 22, 17; Sigma-Aldrich, Co. (2001) Products for Life Science Research, St. Louis, MO; pp. 45-89; Amersham Pharmacia Biotech (2001) BioDirectory, Piscataway, N.J., pp. 384-391). The production, purification, and fragmentation of polyclonal and monoclonal antibodies have been described (Coligan, et al. (2001) Current Protocols in Immunology, Vol. 1, John Wiley and Sons, Inc., New York; Harlow and Lane (1999) Using Antibodies, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Harlow and Lane, supra). Standard methods for characterizing ligand/receptor interactions are available (see, e.g., Coligan, et al. (2001) Current Protocols in Immunology, Vol. 4, John Wiley, Inc., New York).

Monoclonal, polyclonal, and humanized antibodies can be produced (see, e.g., Sheperd and Dean (eds.) (2000) Monoclonal Antibodies, Oxford Univ. Press, New York, NY; Kontermann and Dubel (eds.) (2001) Antibody Engineering, Springer-Verlag, New York; Harlow and Lane (1988) Antibodies A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 139–243; Carpenter et al. (2000) J. Immunol. 165:6205; He et al. (1998) J. Immunol. 160:1029; Tang et al. (1999) J. Biol. Chem. 274:27371–27378; Baca et al. (1997) J. Biol. Chem. 272:10678–10684; Chothia et al. (1989) Nature 342:877–883; Foote and Winter (1992) J. Mol. Biol. 224:487–499; U.S. Patent No. 6,329,511).

An alternative to humanization is the use of human antibody libraries displayed on phage or human antibody libraries in transgenic mice (Vaughan et al. (1996) Nature Biotechnol. 14:309–314; Barbas (1995) Nature Medicine 1:837–839; Mendez et al. (1997) Nature Genetics 15:146–156; Hoogenboom and Chames (2000) Immunol. Today 21:371–377; Barbas et al. (2001) Phage display: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; Kay et al. (1996) Phage display of Peptides and Proteins: A Laboratory Manual, Academic Press, San Diego, CA; de Bruin et al. (1999) Nature Biotechnol. 17:397–399).

Antigen purification is not necessary to produce antibodies. Animals can be immunized with cells bearing the antigen of interest. Splenocytes can then be isolated from the immunized animals, and the splenocytes can be fused with a myeloma cell line to produce a hybridoma (see, e.g., Meyaard et al. (1997) Immunity 7:283–290; Wright et al. (2000) Immunity 13:233–242; Preston et al., supra; Kaithamana et al. (1999) J. Immunol. 163:5157–5164).

Antibodies can be conjugated to, for example, small molecule drugs, enzymes, liposomes, polyethyleneglycol (PEG). Antibodies can be used for a therapeutic agent, a diagnostic agent, a kit, or other purposes, and include antibodies linked to, for example, dyes, radioactive isotopes, enzymes, or metals such as colloidal gold (see, for example, Le Doussal et al. (1991) J. Immunol. 146:169-175; Gibellini et al. (1998) J. Immunol. 160:3891-3898; Hsing and Bishop (1999) J. Immunol. 162:2804-2811; Everts et al. (2002) J. Immunol. 168:883-889).

Flow cytometry methods are available, including fluorescence-activated cell sorting (FACS) (see, e.g., Owens et al. (1994) Flow Cytometry Principles for Clinical Laboratory Practice, John Wiley and Sons, Hoboken, NJ; Givan (2001) Flow Cytometry, 2nd ed.; Wiley-Liss, Hoboken, NJ; Shapiro (2003) Practical Flow Cytometry, John Wiley and Sons, Hoboken, NJ). Fluorescent reagents suitable for modifying nucleic acids, including nucleic acid primers and probes, polypeptides, and antibodies, are available for use, for example, as diagnostic reagents (Molecular Probes (2003) Catalogue, Molecular Probes, Inc., Eugene, OR; Sigma-Aldrich (2003) Catalogue, St. Louis, MO).

Standard techniques for histology of the immune system have been described (see, e.g., Muller-Harmelink (ed.) (1986) Human Thymus: Histopathology and Pathology, Springer Verlag, New York, NY; Hiatt, et al. (2000) Color Atlas of Histology, Lippincott, Williams, and Wilkins, Phila, PA; Louis, et al. (2002) Basic Histology: Text and Atlas, McGraw-Hill, New York, NY).

Software packages and databases are available for determining, for example, antigen fragments, leader sequences, protein folding, functional domains, glycosylation sites, and sequence alignments (see, for example, GenBank, Vector NTI® Suite (Informax, Inc, Bethesda, MD); GCG Wisconsin Package (Accelrys, Inc., San Diego, CA); DeCypher® (TimeLogic Corp., Crystal Bay, Nevada); Menne, et al. (2000) Bioinformatics 16: 741–742; Menne, et al. (2000) Bioinformatics Applications Note 16:741–742; Wren, et al. (2002) Comput. Methods Programs Biomed. 68:177–181; von Heijne (1983) Eur. J. Biochem. 133:17–21; von Heijne (1986) Nucleic Acids Res. 14:4683-4690).

EXAMPLES

Example 1: Research in vitro induction of PD-1 on CD8 ⁺ T cells cocultured with MM.1S multiple myeloma cells treated with a bispecific antibody against BCMA×CD3

This example shows that treatment with a bispecific antibody against BCMA×CD3 induces PD-1 expression on CD8 ⁺ T cells.

CD3 ⁺ T cells from PBMC (Stem Cell Technologies) were negatively selected using the EasySep Human T Cell Enrichment Kit (Stem Cell Technologies). 10,000 target MM.1S multiple myeloma cells expressing luciferase (MM.1S-luc) were seeded with 50,000 CD3 ⁺ pan-T cells in a clear 96-well V-bottom plate. Cells were treated with 1 nM anti-BCMA×CD3 bispecific antibody, and PD-1 expression was analyzed at 3, 24, 48, and 72 hours after addition of the anti-BCMA×CD3 bispecific antibody. At selected time points, cells were harvested from wells, washed with PBS + 2% FBS, and stained with ZombieNIR viability stain (Biolegend) in PBS for 20 min at room temperature, followed by staining with anti-human CD8 and PD-1 antibodies (Biolegend). Samples were analyzed using FlowJo flow cytometry software. Dead cells were removed from the analysis by gating on the ZombieNIR-negative population. Samples were additionally gated on the CD8 ⁺ population. The percentage of PD-1 ⁺ cells is expressed as PD-1-positive cells in the CD8 ⁺ population. The results, summarized in Fig. 1 and Table 1, indicate that treatment of BCMA-expressing MM.1S multiple myeloma cells with the anti-BCMA×CD3 bispecific antibody induces PD-1 expression on CD8 ⁺ T cells.

Table 1. % PD-1 ⁺ cells after treatment with bispecific antibody against BCMA×CD3

Watch % PD-1 (from CD8 ⁺ T cells) [± standard deviation] Carrier Bispecific antibody against BCMA×CD3 3 17.1±0.6 12.8±0.6 24 10.3±0.4 18.9±1.2 48 29.8±3.0 69.9±3.1 72 13.6±0.5 69.9±3.1

Example 2: Research in vivo bispecific antibody against BCMA×CD3 in combination with an antibody against PD-1 in an orthotopic and subcutaneous MM.1S-PDL1 mouse model

This example shows the efficacy of a combination of an anti-BCMA×CD3 bispecific antibody with an anti-PD-1 antibody in (A.) MM.1S-Luc-PD-L1 and (B.) MM.1S-PD-L1 orthotopic multiple myeloma models compared to either the anti-BCMA×CD3 bispecific antibody or the anti-PD-1 antibody alone.

A. Orthotopic mouse model

Multiple myeloma MM.1S-Luc cells were engineered to express PD-L1 and designated MM.1S-Luc-PD-L1. MM.1S-Luc-PD-L1 cells were prepared as a single-cell suspension at 5× ¹⁰⁶ cells for intravenous (IV) inoculation into NSG mice.

Tumor growth was monitored by fluorescence imaging using intraperitoneal (IP) injection of luciferin in DPBS and visualized using a Perkin Elmer IVIS Spectrum camera system. Animals were injected IV with 2 × 10 ⁷ expanded human T cells 19 days after tumor cell inoculation. Two days after T cell inoculation, a single dose of anti-BCMA×CD3 bispecific antibody (10 μg/kg) was administered as a bolus IV injection. Anti-PD-1 antibody was administered twice weekly as a bolus IP injection at 5 mg/kg for a total of six injections.

Tumor growth was monitored by image measurements collected twice weekly. Mice were imaged using a Perkin Elmer IVIS Spectrum camera system with automatic parameter detection and a maximum imaging time of 3 minutes. Living Image software was used for data collection. Regions of interest (ROIs) were drawn around the entire mouse body, excluding the tail whenever possible. Background flux measured through the anesthesia channel was subtracted from each ROI. Tumor measurements are expressed as total flux, expressed as photons/sec. The study was terminated at day 40 after tumor inoculation. The results, summarized in Fig. 2A and Table 2, indicate that treatment with the anti-BCMA×CD3 bispecific antibody and anti-PD-1 antibody is more effective than treatment with the bispecific antibody or antibody alone.

Table 2. Tumor measurements expressed as total flux (photons/sec) after treatment

Days after tumor cell inoculation Bispecific negative control antibody + anti-PD-1 antibody (5 mg/kg) Bispecific antibody against BCMA×CD3 (10 μg/kg + antibody against PD-1 (5 mg/kg) Bispecific antibody against BCMA×CD3 (10 μg/kg) Lack of processing 15 4.01×10 ⁷
± 2.25×10 ⁷ 1.31133334×10 ⁷
± 3.1997153×10 ⁶ 1.51333334×10 ⁷
± 2.325463×10 ⁶ 7.2×10 ⁷
± 5×10 ⁷ 18 5.535×10 ⁷
± 1.735×10 ⁷ 6.54333334×10 ⁷
± 1.36827548×10 ⁷ 8.34333334×10 ⁷
± 2.40911003×10 ⁷ 1.68×10 ⁸
± 1.54×10 ⁸ 20 1.855×10 ⁸
± 4.75×10 ⁷ 1.586666667×10 ⁸
± 1.92382721×10 ⁷ 1.788×10 ⁸
± 6.89374596×10 ⁷ 3.76×10 ⁸
± 1.31×10 ⁸ 25 1.052×10 ⁹
± 2.08×10 ⁸ 3.4466667×10 ⁶
± 2.8719195×10 ⁶ 2.2729×10 ⁷
± 1.99389279×10 ⁷ 2.455×10 ⁹
± 9.55×10 ⁸ 28 1.73×10 ⁹
± 1.3×10 ⁸ 5,792×10 ⁶
± 4.8055591×10 ⁶ 7.08166667×10 ⁷
± 6.7101553×10 ⁷ 5.555×10 ⁹
± 2.455×10 ⁹ 32 1.73×10 ⁹
± 1.3×10 ⁸ 5,792×10 ⁶
± 4.8055591×10 ⁶ 7.07566667×10 ⁷
± 6.71310595×10 ⁷ 5.545×10 ⁹
± 2.455×10 ⁹ 35 1.93×10 ⁹
± 7.8×10 ⁸ 7.373334×10 ⁵
± 4.980711×10 ⁵ 8.712333334×10 ⁸
± 8.444262754×10 ⁸ 1.8295×10 ¹⁰
± 1.3305×10 ¹⁰ 39 2.825×10 ⁹
± 1.745×10 ⁹ 4.654667×10 ⁵
± 3.153991×10 ⁵ 6.775666667×10 ⁸
± 6.363327336×10 ⁸ 2,446×10 ¹⁰
± 1.454×10 ¹⁰

B. Subcutaneous mouse model

MM.1S multiple myeloma cells were engineered to express PD-L1 and designated MM.1S-PD-L1. Preactivated and expanded T cells (20 × 10 ⁶ ) were administered on day 19 after subcutaneous (SC) inoculation with MM.1S-PDL1 tumor cells. Anti-BCMA×CD3 bispecific antibody (0.3 or 1 mg/kg) or negative control bispecific antibody (1 mg/kg) were administered IV on day 21 on a Q7Dx3 schedule. Anti-PD-1 mAb was administered at 5 mg/kg intraperitoneally (IP) twice weekly starting on day 21. Tumor measurements were recorded 2 to 3 times weekly using a digital caliper. N (at baseline) represents 5–12 animals per group. Results summarized in Fig. 2B and Table 3 indicate that treatment with the bispecific anti-BCMA×CD3 antibody and anti-PD-1 antibody is more effective than treatment with the bispecific antibody or treatment with the antibody alone.

Table 3. Post-treatment tumor measurements (tumor volume ± SEM (mm ³ )).

Days after tumor cell inoculation Lack of processing Bispecific negative control antibody Bispecific negative control antibody + anti-PD-1 antibody (5 mg/kg) Bispecific antibody against BCMA×CD30 (3 mg/kg) Bispecific antibody against BCMA×CD3 (1 mg/kg) Separately + antibody against PD-1 (5 mg/kg) Separately + antibody against PD-1 (5 mg/kg) 14 39.6
± 6.9 34.1
± 5.3 30.7
±4.6 34.4
± 9.5 33.6
± 5.3 26.4
± 7.5 33.6
± 5.3 18 80.3
± 16.9 71.5
± 13.7 64.5
± 10 91.6
± 18 74.5
± 12.2 92
± 17.1 74.5
± 12.2 20 108.7
± 27 121.4
± 27.3 114.7
± 21 122.5
± 20.7 110.5
± 18.6 130.7
± 24.6 110.5
± 18.6 25 407.8
± 87.4 401.7
± 30.5 401.3
± 42.9 403
± 59 504.4
± 67.4 601.1
± 70 504.4
± 67.4 28 699.4
± 126.2 761.6
± 97.1 694.7
± 69.5 403.3
± 67.6 341
± 86.9 482.9
± 32.6 341
± 86.9 32 1379.8
±308.6 1789.8
± 233.4 1238.6
± 146.2 603.6
± 151.6 164.9±108.9 410.5
± 82.7 164.9
± 108.9 35 2125.2
± 61.6 2562.5
± 81.8 1861.3
± 238.4 835.1
± 77.2 18.5
±9.9 627.9
± 111.8 18.5
± 9.9 39 4017.3
± 103 4012.4
± 106.2 2792.6
± 4.7 1569.3±151.8 2.9
± 2.9 1436.8±276.4 2.9
± 2.9

Example 3: Research in vitro for detection of BCMA expression on the cell surface in multiple myeloma cell lines treated with gamma-secretase inhibitors (GSI)

This example shows upregulation of BCMA cell surface expression in multiple myeloma cell lines treated with GSI.

Multiple myeloma cells (MM.1S, OPM2, H929, Molp8, RPMI8226) were seeded in a 96-well U-bottom plate at 40,000 cells/well. Cells were incubated for 24 h in the presence of GSI diluted in RPMI (0.1% DMSO). The following GSI concentrations were tested: 1000 nM, 500 nM, 100 nM, 50 nM, 25 nM, 10 nM, 5 nM, 2.5 nM, 1 nM, 0.1 nM, 0.01 nM. After 24 h, cells were harvested and washed with PBS + 2% FBS, followed by staining with ZombieNIR viability stain (Biolegend) at a 1:500 dilution in PBS at room temperature for 20 min. Cells were then washed with PBS + 2% FBS and stained with PE-labeled anti-BCMA antibody (Biolegend) diluted in PBS + 2% FBS for 30 min at 4°C. Cells were processed with a BD flow cytometer and analyzed using FlowJo flow cytometry software. Dead cells were removed from the analysis by gating on the ZombieNIR-negative population. The mean fluorescence intensity (MFI) of BCMA was plotted against the concentration of GSI to determine the _EC50 .

The results summarized in Figs. 3A–3E and Table 4 indicate that GSI treatment positively regulates cell surface BCMA expression in the multiple myeloma cell lines MM.1S, OPM2, H929, Molp8, and RPMI8226, respectively.

Table 4. Mean fluorescence intensity ± standard deviation

GSI [nM] MFI BCMA (-PE) MM.1S OPM2 H929 Molp8 RPMI8226 1000 35780±21.2 14747.5±965.2 21156±338 6401±550.1 14362±705.7 500 35780±4279.4 15910.5±307.6 20285.5±542.4 5640.5±758.7 13464.5±624.4 100 37582.5±1033.1 14364±403.1 19455.5±563.6 5412±383.3 13416±452.5 50 38111±2506 10863±19.8 17424.5±48.8 5120±533.2 13190.5±555.1 25 37026.5±370 6575.5±115.3 18445.5±309 3691±323.9 12617±762.3 10 31248±335.2 3190.5±88.4 15670±1516 2183.5±81.3 11181.5±193 5 28832±1926.2 2788.5±177.5 13226±1531.6 1761.5±74.2 10589.5±351.4 2.5 21412.5±614.5 2358.5±136.5 10344.5±1617.2 1401±107.5 10226.5±277.9 1 14396±6222.5 2185±15.6 8615.5±1656.8 1288.5±16.3 10483.5±536.7 0.1 15775.5±316.1 2079.5±82.7 5305.5±290.6 1242.5±137.9 10099±455.4 0.01 15589.5±230 2093.5±71.4 4824.5±183.1 1191.5±188.8 10028±29.7 0 15362 2069.5±23.3 4908±138.6 1231±79.2 10142±448.3

Example 4: Research in vitro for the detection of cell surface BCMA expression as a function of time in GSI-treated multiple myeloma cell lines

This example shows that treatment of multiple myeloma cell lines with GSI increases cell surface BCMA expression in a time-dependent manner, and that surface BCMA levels return to baseline levels after removal of GSI from the cultures.

Multiple myeloma cells (MM.1S, OPM2, H929, Molp8, RPMI8226) were seeded in 6-well plates at 800,000 cells/2 ml/well with GSI diluted at a concentration of 1 μM in RPMI medium (with 0.1% DMSO). Cells were harvested for assessment of cell surface BCMA expression at baseline and then at 3 hours, 6 hours, and 24 hours after GSI addition. After 24 hours of incubation with GSI, cells were washed twice with PBS and reseeded in new 6-well plates. Cells were additionally harvested for staining at 3 hours, 6 hours, and 24 hours after GSI washout. At the indicated time points, samples were stained with ZombieNIR viability stain (Biolegend) for 20 minutes at room temperature at a 1:500 dilution in PBS, washed with PBS + 2% FBS, and further stained with PE-labeled anti-BCMA antibody diluted in PBS + 2% FBS for 30 minutes at 4°C. Samples were processed using a BD flow cytometer and analyzed using FlowJo software. Dead cells were removed from the analysis by gating on the ZombieNIR-negative population. The BCMA MFI was plotted as a histogram.

The results summarized in Figs. 4A-4E and Table 5 indicate that GSI upregulates BCMA expression on the surface of MM.1S, OPM2, H929, Molp8, and RPMI8226 cells, respectively, in a time-dependent manner, and that upregulated BCMA surface expression is not maintained after GSI is removed from the cultures.

Table 5. Mean fluorescence intensity ± standard deviation

Time point MFI BCMA (-PE) MM.1S OPM2 H929 Molp8 RPMI8226 Beginner level 7614 3127 2387 2384 2991 3 hours after GSI treatment 13690 9257 3653 2461 4437 6 hours after GSI treatment 14920 9854 3423 3098 4527 24 hours after GSI treatment 25602 20592 8245 5307 6340 3 hours after washing 33509 21621 15422 7523 6557 5 hours after washing 23906 11050 9313 5050 4437 24 hours after washing 11170 2624 8423 2960 2688

Example 5: Research in vitro to determine soluble BCMA (sBCMA) levels in GSI-treated multiple myeloma cell lines

This example shows reduced sBCMA shedding in multiple myeloma cell lines treated with GSI.

Multiple myeloma cells (MM.1S, OPM2, H929, Molp8, RPMI8226) were seeded in a 96-well U-bottom plate at 40,000 cells/well. Cells were incubated for 24 h in the presence of GSI diluted in RPMI medium (0.1% DMSO). The following GSI concentrations were tested: 1000 nM, 500 nM, 100 nM, 50 nM, 25 nM, 10 nM, 5 nM, 2.5 nM, 1 nM, 0.1 nM, 0.01 nM. After 24 h, cell culture medium was collected and the sBCMA concentration in the supernatants was measured using the human BCMA/TNFRSF17 DuoSet ELISA kit (R&D Systems) according to the manufacturer's instructions.

The results summarized in Figs. 5A-5E and Table 6 indicate that GSI treatment blocks sBCMA shedding in the multiple myeloma cell lines MM.1S, OPM2, H929, Molp8, and RPMI8226, respectively.

Table 6. Mean fluorescence intensity ± standard deviation

GSI [nM] BCMA MFI (-PE) MM.1S OPM2 H929 Molp8 RPMI8226 1000 350±494.9 0 625.1±28.7 51.6±19.2 0±0 500 151.8±214.7 23.8±33.7 716.5±40.2 87.3±32.2 0±0 100 21.3±30.2 1238.6±70.3 1110.6±46.4 431.8±28.2 64.1±28.3 50 279.9±32.6 3055.7±267.3 1460.7±55.6 1044.3±161.3 249.5±99.2 25 630.5±33.2 4487.9±76.6 2336.4±10.3 3021.7±244.1 836.5±339.2 10 1661.5±114.7 6488.6±254.6 4901.7±9.3 5958.9±461.5 1926.2±569.7 5 2453.6±36 7617.3±624.4 8438.2±1019.6 5989.3±566.5 2525.6±267.3 2.5 3988.3±251.6 7726.5±64.5 12874.8±1086.7 6765.5±1147.3 2398±289.5 1 5143.1±296.4 8014±815.1 19516.4±615.8 6072.9±1772.4 2647.5±158.6 0.1 5797.5±228.2 8872.2±24.9 27414.8±4090.6 6363.4±628.8 3333.8±171.3 0.01 3794.2±471.1 6043.3±41.7 24084.8±3954.7 6887.3±879.1 4710.8±1361.5 0 4276.9±277.1 5948.9±288.1 24399.1±658.2 6859.8±541.7 3333.8±440.4

Example 6: Bispecific antibody against BCMA×CD3 in combination with GSI in multiple myeloma

This example shows that treatment with the BCMA×CD3 bispecific antibody in combination with GSI demonstrates enhanced killing of multiple myeloma cells cultured with human T cells compared to the BCMA×CD3 bispecific antibody alone.

CD3 ⁺ T cells from PBMC (Stem Cell Technologies) were negatively selected using the EasySep Human T Cell Enrichment Kit (Stem Cell Technologies). Luciferase-expressing multiple myeloma cells (MM.1S-luc, OPM2-luc, H929-luc, Molp8-luc, RPMI8226-luc) were treated with 1 μM GSI 10,000. After 24 h, cells were seeded with 50,000 CD3 ⁺ pan-T cells in a clear 96-well V-bottom plate. Cells were further treated with a range of concentrations of the anti-BCMA×CD3 bispecific antibody with or without 1 μM GSI. After 60 hours of treatment, luciferase activity in treated cells was analyzed using the NeoLite reagent kit (Perkin Elmer) and processed on a VictorX multi-mode plate reader (Perkin Elmer). Cell viability was calculated by dividing the luciferase activity of treated cells by the luciferase activity of the untreated control (without the addition of the bispecific anti-BCMA×CD3 antibody).

The results summarized in Figs. 6A-6E and Tables 7-8 indicate that GSI treatment enhances BCMA×CD3 bispecific antibody (“BCMA×CD3” in Tables 7 and 8)-mediated cell killing in multiple myeloma cell lines (MM.1S (21-fold), OPM2 (21-fold), H929, Molp8, RPMI8226 (24-fold), respectively) when co-cultured with human T cells.

Table 7. Mean fluorescence intensity ± standard deviation

BCMA×CD3 [nM] Viability of MM.1S cells (% of control) H929 cell viability (% control) Molp8 cell viability (% control) BCMA×CD3 BCMA×CD3
+GSI BCMA×CD3 BCMA×CD3
+GSI BCMA×CD3 BCMA×CD3
+GSI 4 1.3±0.8 1.7±2.1 8.6±3 6±1 5.2±0.3 7.3±0.3 0.8 2.4±0.5 2±1.5 11.3±1.5 6.8±2.3 6.1±0.7 7.6±0.2 0.16 1.7±0.8 2±0.3 23.2±7.2 11.2±4.1 8±2 7.6±4.3 0.032 2.5±0.3 2.1±0.2 98.6±1.2 47.2±2.5 66.1±4.9 40.7±34.1 0.0064 108±4.5 7.2±4.1 81.2±3 127.5±9.5 55.2±1.3 94±9.1 0.00128 87.7±4.4 56.6±0.6 77.8±3.3 186.9±6.5 66.7±0.3 149±6.3 0.00026 78±0.9 116.2±2.6 79±0.8 134.9±6.8 58.5±0.6 119.4±4.4 0.00005 79.8±2.9 101.5±15.3 81.1±0.2 121.5±0.9 59.1±4.6 112.3±0.6 0.00001 80.6±3.9 95.6±5.7 80.2±0.3 116.1±3.3 64±11.6 106.7±3.1

Table 8. Mean fluorescence intensity ± standard deviation

BCMA×CD3
[nM] OPM2 cell viability (% control) Viability of RPMI8226 cells (% control) BCMA×CD3 BCMA×CD3 +GSI BCMA×CD3 BCMA×CD3 +GSI 50 3.3±0.3 2.1±0.9 19.4±5 7.3±2 10 2±0.2 1.9±0.2 19.9±1.1 6.3±0.2 2 2.8±0.8 1.9±0.5 25.7±5.5 10.9±2.3 0.4 3.1±0.8 1.9±0.3 28.7±13.6 8.4±1.5 0.08 43.6±6.4 1.7±0.3 54.2±16.9 9.8±1.5 0.016 68.4±2.4 4±1.5 113.4±1.3 8.8±2.7 0.0032 84.3±3.1 35.3±2.5 130.5±19.5 22.7±0.9 0.00064 83.8±3.4 61.3±2.1 113.2±10.6 58±6.3

Example 7: Research in vitro to determine cell surface expression of BCMA in lymphoma cell lines treated with GSI

This example shows upregulation of BCMA expression on the surface of lymphoma cells treated with GSI.

Lymphoma cells (Raji cell line) were seeded in a 96-well U-bottom plate at 40,000 cells/well. Cells were incubated for 24 hours in the presence of GSI diluted in RPMI medium (0.1% DMSO). The following GSI concentrations were tested: 1000 nM, 500 nM, 100 nM, 50 nM, 25 nM, 10 nM, 5 nM, 2.5 nM, 1 nM, 0.1 nM, 0.01 nM. After 24 hours, cells were harvested and washed with PBS + 2% FBS, followed by staining with ZombieNIR viability stain (Biolegend) at a 1 in 500 dilution in PBS at room temperature for 20 minutes. Cells were then washed with PBS + 2% FBS and stained with PE-labeled anti-BCMA antibody (Biolegend) diluted in PBS + 2% FBS for 30 min at 4°C. Cells were processed using a BD flow cytometer and analyzed using FlowJo flow cytometry software. Dead cells were removed from the analysis by gating on the ZombieNIR-negative population. BCMA MFI was plotted against GSI concentration to determine the _EC50 .

The results summarized in Fig. 7 and Table 9 indicate that GSI treatment positively regulates BCMA expression on the surface of Raji lymphoma cells.

Table 9. Mean fluorescence intensity ± standard deviation

GSI [nM] MFI BCMA 1000.00 534.5±67.2 500.00 568.5±50.3 100.00 498.5±20.6 50.00 535±65.1 25.00 532±42.5 10.00 515.5±47.4 5.00 484±43.9 2.50 469±31.2 1.00 398±34 0.10 147.5±20.6 0.01 142.5±0.8 0.0 141±4.3

Example 8: Research in vitro to determine cell surface BCMA expression over time in GSI-treated lymphoma cell lines

This example shows that treatment of lymphoma cell lines with GSI increases cell surface BCMA expression in a time-dependent manner, and that surface BCMA levels return to baseline levels after GSI removal from the culture.

Lymphoma cells (Raji) were seeded in 6-well plates at 800,000 cells/2 ml/well with GSI diluted at 1 μM in RPMI medium (with 0.1% DMSO). Cells were harvested for assessment of cell surface BCMA expression at baseline and then at 3 hours, 6 hours, and 24 hours after GSI addition. After 24 hours of incubation with GSI, cells were washed twice with PBS and reseeded in new 6-well plates. Cells were further harvested for staining at 3 hours, 6 hours, and 24 hours after GSI washout. At the indicated time points, samples were stained with ZombieNIR viability stain (Biolegend) for 20 minutes at room temperature at a 1:500 dilution in PBS, washed with PBS + 2% FBS, and further stained with PE-labeled anti-BCMA antibody diluted in PBS + 2% FBS for 30 minutes at 4°C. Samples were processed using a BD flow cytometer and analyzed using FlowJo software. Dead cells were removed from the analysis by gating on the ZombieNIR-negative population. The mean fluorescence intensity (MFI) of BCMA was plotted as a histogram.

The results summarized in Fig. 7B and Table 10 indicate that GSI upregulates BCAM expression on the surface of Raji cells in a time-dependent manner and that upregulated BCMA surface expression is not maintained after GSI removal from cultures.

Table 10. Mean fluorescence intensity ± standard deviation

Time point MFI BCMA (-PE) Raji Beginner level 647 3 hours after GSI treatment 699 6 hours after GSI treatment 735 24 hours after GSI treatment 1506 3 hours after washing 1619 5 hours after washing 1822 24 hours after washing 795

Example 9A: Bispecific antibody against BCMA×CD3 in combination with GSI in lymphoma cells

This example shows that treatment with a BCMA×CD3 bispecific antibody in combination with GSI demonstrates enhanced cell killing in BCMA-poor lymphoma cells cultured with human T cells compared to the BCMA×CD3 bispecific antibody alone.

CD3 ⁺ T cells from PBMCs (Stem Cell Technologies) were negatively selected using the EasySep Human T Cell Enrichment Kit (Stem Cell Technologies). 10,000 target luciferase-expressing lymphoma cells (Raji-luc) were treated with 1 μM GSI. After 24 h, the cells were seeded with 50,000 CD3 ⁺ pan-T cells in a clear 96-well V-bottom plate. Cells were further treated with a range of concentrations of the anti-BCMA×CD3 bispecific antibody with or without 1 μM GSI. Sixty hours after treatment, luciferase activity in treated cells was analyzed using the NeoLite reagent kit (Perkin Elmer) and processed on a VictorX multi-mode plate reader (Perkin Elmer). Cell viability was calculated by dividing the luciferase activity of treated cells by the luciferase activity of untreated control cells (without addition of the bispecific anti-BCMA×CD3 antibody).

The results summarized in Fig. 8 and Table 11A indicate that GSI treatment enhances BCMA×CD3 bispecific antibody-mediated cell killing in a lymphoma cell line (Raji) when co-cultured with human T cells.

Table 11A . Cell viability ± standard deviation

BCMA×CD3
[nM] Raji cell viability (% control) BCMA×CD3 BCMA×CD3 +GSI 50 80±13.2 41.5±9.3 10 82.1±9.7 46.4±14.1 2 71±9.7 45.3±4.3 0.4 88.1±18.4 61.9±3 0.08 89.3±5.7 56.3±0.3 0.016 105.8±10.2 78.2±9.1 0.0032 107±4.5 97.8±8.2 0.00064 94±3.7 92.4±10.8

Example 9B: Gamma-secretase inhibitor action enhances cytotoxic effect in vitro bispecific antibody against BCMA×CD3 PF06863135 (elranatamab) against multiple myeloma cells in a coculture assay

This example demonstrates the combined beneficial effect of treatment of multiple myeloma cells with GSI and the anti-BCMA×CD3 bispecific antibody PF06863135 (elranatamab) compared to the anti-BCMA×CD3 antibody alone in in vitro cytotoxic T lymphocyte (CTL) coculture assays.

Luciferase-expressing multiple myeloma cell lines (H929-Luc, Molp8-Luc, OPM2-Luc, and RPMI8226-Luc) were cultured at 37°C and 5% _CO2 with 1 mM GSI for 24 h or left untreated. Myeloma cells were then harvested and transferred to 96-well U-bottom plates at 10,000 cells/well along with 50,000 CD3 ⁺ T cells/well enriched from human PBMC using a pan-T cell negative selection kit (Miltenyi Biotec). Media with or without 1 mM GSI containing serial dilutions of the BCMA×CD3 bispecific antibody PF06863135 were additionally added to the wells before incubating the plates at 37°C and 5% _CO2 for 72 h. At the end of the incubation period, Bright-Glo substrate (Promega) was added to the wells and luminescence was measured using a SpectraMax plate reader. Percent cell viability was calculated by using the luminescence signal value for each test well, dividing it by the average signal from the control wells without antibody treatment, and then multiplying by 100. Additionally, _EC50 values were calculated by fitting a four-parameter dose-response curve to the cell viability data against the antibody dose concentration using GraphPad Prism. Table 11B shows that GSI treatment enhances anti-BCMA×CD3 antibody-mediated killing of multiple myeloma cells (H929, Molp8, OPM2, and RPMI8226) treated in co-cultures with human T cells.

Table 11B. Anti-BCMA×CD3 bispecific antibody PF06863135 mediated killing of multiple myeloma cells

BCMA×CD3 BCMA×CD3+GSI Cell line EC ₅₀ (nM) SD N EC ₅₀ (nM) SD N EC ₅₀ fold change using GSI H929-Luc 0.1668 0.1126 6 0.0055 0.0042 6 30.3 times Molp8-Luc 0.1085 0.0456 6 0.0202 0.0177 6 5.4 times OPM2-Luc 0.0528 0.0122 6 0.0033 0.0008 6 15.9 times RPMI8226-Luc 0.0092 0.0065 6 0.0017 0.0008 6 5.5 times

Example 10: A first-in-human Phase 1 clinical trial of the BCMA×CD3 bispecific antibody elranatamab (PF-06863135).

This example demonstrates an ongoing, open-label, multicenter, Phase 1 clinical trial of PF-06863135 (a bispecific antibody against BCMA×CD3) as monotherapy and in combination with sasanlimab, lenalidomide, or pomalidomide in adult patients with advanced multiple myeloma that has relapsed or is refractory to standard therapy. The trial is registered on ClinicalTrials.gov under identifier NCT03269136 and was first published in August 2017. This example describes the results for Part 1 and additional arms of the study.

The study groups and initial designs of the administration regimens are summarized in the table. 12. In each study arm, drug treatment will continue until disease progression, patient withdrawal (withdrawal of consent), or unacceptable toxicity occurs.

Table 12. Treatment with PF-06863135 in a first-in-human clinical trial

Study groups Medicines Dose levels and administration schedule Part 1 Monotherapy PF06863135 IV: 0.1, 0.3, 1, 3, 10, 30, and 50 (mcg/kg) Q1W;
SC: 80, 130, 215, 360, 600 and 1000 (mcg/kg), Q1W;
Selected doses from those listed above for Q2W Part 1.1 Monotherapy PF-06863135 Doses in Part 1 and a priming dose on Cycle Zero Day 1. Then set to a priming of 600 (mcg/kg) followed by 1000 (mcg/kg) Q1W; or a priming of 600 (mcg/kg) followed by 1000 (mcg/kg) Q2W Part 1B PF-06863135 and sasanlimab PF-06863135: TBD
Sasanlimab: 300 mg Q4W, SC Part 1C PF-06863135 and lenalidomide PF-06863135: TBD
Lenalidomide: 25 mg orally daily on days 1–21 without dexamethasone
Then modified to 15 mg QD on days 1-21 Part 1D PF-06863135 and pomalidomide PF-06863135: TBD
Pomalidomide: 4 mg orally daily on days 1–21 without dexamethasone Part 2A Monotherapy PF-06863125 RP2D, to be determined Part 2B PF-06863135 and sasanlimab PF-06863135: TBD
Sasanlimab: 300 mg Q4W, SC Part 2C PF-06863135 and lenalidomide PF-06863135: TBD
Lenalidomide: 25 mg orally daily on days 1–21 (without dexamethasone) Part 2D PF-06863135 and pomalidomide PF-06863135: TBD
Pomalidomide: 4 mg orally daily on days 1-21 (without dexamethasone)

The RP2D dose was then determined based on the clinical outcome of Part 1 and a maintenance dose of 76 mg Q1W SC was chosen with a single priming dose of 44 mg SC administered one week before the first maintenance dose.

In Part 1 of the combination dose-finding study, it was decided that subjects would receive fixed doses of PF06863135, with maintenance doses beginning one week after the priming dose and one dose level below the RP2D of monotherapy, and the dose would be escalated to the RP2D or decreased to the RP2D within 2 dose levels. Table 12A shows potential fixed dose levels in the study of PF06863135 in combination with a second agent. For Part 1C, the starting dose of lenalidomide was modified to 15 mg orally QD on days 1–21 of a 28-day cycle beginning 7 days after the priming dose of PF06863135.

Table 12A. Potential fixed dose levels for combination studies

Dose level Priming dose (mg) Maintenance dose (mg) PR2D minus 2 24 32 RP2D minus 1 32 44 RP2D dose 44 76

Part 1 of the study is a dose escalation cohort of PF-06813135 monotherapy at dose levels of 0.1, 0.3, 1, 3, 10, 30, and 50 mcg/kg Q1W via intravenous (IV) administration and at dose levels of 80, 130, 215, 360, 600, and 1000 mcg/kg Q1W via subcutaneous (SC) administration. After reaching the maximum tolerated dose (MTD)/maximum administered dose (MAD), patients can be treated at a dose level selected from the dose levels described above in this paragraph and below the MTD/MAD for IV and SC Q2W administration to further support the recommended phase 2 dose (RP2D) decision. For the study, the dose-limiting toxicity observation period was defined as 21 days for Q1W administration and 28 days for Q2W administration. The treatment cycle, also referred to as a "cycle," for Q1W administration will be 3 weeks, and for Q2W administration it will be 4 weeks.

Clinical outcome of Part 1 of the study . As of April 15, 2020, a total of 23 patients were enrolled in Part 1 of the study and treated with PF-06863135 administered intravenously (IV) at doses of 0.1 (N=2), 0.3 (N=3), 1 (N=2), 3 (N=3), 10 (N=2), 30 (N=5), and 50 (N=6) mcg/kg. As of August 21, 2020, a total of 30 patients were enrolled in Part 1 of the study and treated with PF-06863135 administered subcutaneously (SC) at doses of 80 (N=6), 130 (N=4), 215 (N=4), 360 (N=4), 600 (N=6), and 1000 (N=6) mcg/kg. Safety and efficacy data are available for 23 patients on IV administration and 30 patients on SC administration according to IMWG (International Myeloma Study Group) criteria.

Of the patients in cohorts IV, 2 patients (1 patient in the 30 mcg/kg cohort and 1 patient in the 50 mcg/kg cohort) experienced dose-limiting toxicities (DLTs) of grade 3 febrile neutropenia and grade 1 QT prolongation on ECG. No patients in the SC cohorts experienced DLTs. Cytokine release syndrome (CRS) was the most common adverse event reported. In cohorts IV, CRS was observed in 1 (50.0%), 4 (80.0%), and 6 (100.0%) patients in the 10, 30, and 50 mcg/kg cohorts. Of all IV-treated patients, 6 (26.1%) experienced maximal grade 1 CRS, while 5 (21.7%) experienced maximal grade 2 CRS. CRS began within the first 2 days of administration in each of the 11 patients with CRS. CRS also occurred in 3 patients at 50 mcg/kg: after the second dose in 1 patient, after the second and third doses in 1 patient, and after the third and fourth doses in 1 patient.

In the SC cohorts, CRS was observed in 3 (50.0%), 2 (50.0%), 3 (75.0%), 3 (75.0%), 6 (100%), and 6 (100%) patients in the 80, 130, 215, 360, 600, and 1000 mcg/kg groups, respectively. Of all SC-treated patients, 18 (60.0%) experienced maximal Grade 1 CRS, while 5 (16.7%) experienced maximal Grade 2 CRS. CRS primarily began within the first 2 days of administration. Table 13 provides additional details of CRS in the SC cohorts.

Table 13. Cytokine release syndrome (CRS) in the SC cohorts of Part 1 and Part 1.1 of the study

80 mcg/kg
Q1W
N=6 130 mcg/kg
Q1W
N=4 215 mcg/kg
Q1W
N=4 360 mcg/kg
Q1W
N=4 600 mcg/kg
Q1W
N=6 1000 mcg/kg
Q1W
N=6 Priming 600 (mcg/kg), 1000 (mcg/kg) Q1W or Q2W
N=20 Patients with CRS 3 (50%) 2 (50%) 3 (75%) 3 (75%) 6 (100%) 6 (100%) 20
(100%) Degree 1 2 2 3 2 4 5 11 Degree 2 1 0 0 1 2 1 9 Average duration Day 1 3 days 3 days 1.7 days 1.7 days 4 days 2 days

In cohorts IV, 2 patients achieved a minimal response at 3 mcg/kg and 50 mcg/kg IV, and 1 patient achieved a complete response at 50 mcg/kg IV. Ten individuals in cohorts IV (0.3-50 mcg/kg) achieved the best response—stable disease.

In the SC cohorts, the efficacy results are summarized in Table 14 below.

Table 14. Patient response in the SC cohorts of Part 1 and Part 1.1 of the study

215 mcg/kg
N=4 360 mcg/kg
N=4 600 mcg/kg
N=6 1000 mcg/kg
N=6 Priming 600 (mcg/kg), 1000 (mcg/kg) Q1W
N=7 Priming 600 (mcg/kg), 1000 (mcg/kg) Q2W
N=13 OR 3/4 (75%) 3/4 (75%) 4/6 (67%) 5/6 (83%) 4/7 (57%) 7/13 (54%) sCR 2 1 2 CR 1 2 VGPR 2 2 3 1 4 PR 1 1 1 3 SD 1 1 1 PD 1 1 1 3

These results indicate that at the highest dose levels of 600 and 1000 mcg/kg SC, clinical efficacy was observed in the majority of patients and toxicity was tolerable and manageable, with less severe CRS occurring in SC-treated patients despite a higher overall dose exposure in SC-treated compared to IV-treated patients.

Part 1.1 of the study is an alternative maintenance dose escalation arm for PF-06863135 monotherapy. If excess toxicity occurs or the maximum tolerated dose (MTD)/maximum administered dose (MAD) is reached at a dose level earlier than described for Part 1 of the study above, a priming dose will be administered one week prior to the Day 1 Cycle 1 dose (maintenance dose) at that dose level and for all subsequent dose levels in the dose escalation phase that can be initiated for Part 1.1. The priming dose will be administered at a lower dose level than the maintenance dose.

Clinical outcome of Part 1.1 of the study. As of February 4, 2021, a total of 20 patients were enrolled in Part 1.1 of the study, with a cohort of 7 patients receiving a 600 mcg/kg priming dose followed by 1000 mcg/kg Q1W and a cohort of 13 patients receiving a 600 mcg/kg priming dose followed by 1000 mcg/kg Q2W. CRS in these two cohorts is shown in Table 13. The inclusion of the priming dose reduced the mean CRS duration by 50% from 4 days to 2 days. The frequency of administration in Part 1.1 of the study (Q1W vs. Q2W) did not affect CRS. Patient responses in Part 1.1 of the study are shown in Table 14.

Part 2A of the study represents the expansion cohort of the established dose of monotherapy PF-06863135. Based on the clinical data from the monotherapy dose escalation phase, IV or SC administration, including a priming and maintenance dose and administration Q1W or Q2W, will be chosen for Part 2A of the study. Specifically, SC administration at a dose level of 215, 360, 600, or 1000 mcg/kg Q1W or Q2W without a priming dose, or SC administration at a maintenance dose level of 215, 360, 600, or 1000 mcg/kg Q1W or Q2W with a priming dose on day 1 of cycle zero at a dose level lower than the maintenance dose, appears promising as the RP2D for the Phase 2A study.

Preliminary pharmacokinetic (PK) analysis showed that body weight is not a clinically relevant factor for PF-06863135 exposure. Therefore, a fixed dose is appropriate for administration of PF-06863135. Given the encouraging efficacy and safety data obtained in Part I of the study, a promising RP2D for Part 2A of the study may be a fixed dose equivalent to 1000 mcg/kg (i.e., 76 mg) of PF-06863135 administered Q1W or Q2W. A fixed dose equivalent to 600 mcg/kg (i.e., 44 mg) will likely be used as a priming dose on cycle zero, day 1. The initial dose of 44 mg serves as a priming dose and is intended to reduce CRS symptoms at the 76 mg dose. Given the results of Part 1 of the study, CRS primarily occurs after the priming dose. Then, 44 mg (priming) and 76 mg (maintenance) were chosen as the RP2D monotherapy dose. The patient will receive a single priming dose of 44 mg SC PF06863135 followed by a maintenance dose of 76 mg Q1W SC or 76 mg Q2W, starting 7 days after the single priming dose.

Part 1B and Part 2B of the study are combination treatments with PF-06863135 and sasanlimab, an anti-PD-1 antibody. The treatment cycle is 28 days. Sasanlimab will be administered at a dose of 300 mg SC Q4W, beginning on Day 1 of Cycle 1. PF-06863135 will be administered SC or IV at the selected dose Q1W or Q2W, beginning on Day 1 of Cycle 1, with or without a priming dose one week before Day 1 of Cycle 1.

In Part 1B, the dose of PF-06863135 will be determined based on the results of Part 1 and Part 1.1 of the study, starting at the RP2D described for Part 2A of the study mentioned above, or at the MTD/MAD minus one dose level, whichever is lower. If the combination treatment regimen is not well tolerated, PF-06863135 will be tapered to a lower dose level to select the dose level for Part 2B.

In Part 2B, PF06863135 will be administered at a dose level based on the results of Part 1B.

Part 1C and Part 2C of the study are combination treatment with PF-06863135 and lenalidomide. The treatment cycle is 28 days. Lenalidomide will be administered daily on days 1–21 at a dose of 25 mg orally (PO) without dexamethasone, beginning on day 1 of cycle 1. PF-06863135 will be administered SC or IV at the selected dose Q1W or Q2W, beginning on day 1 of cycle 1, with or without a priming dose one week before day 1 of cycle 1.

In Part 1C, the dose of PF-06863135 will be determined based on the results of Part 1 and Part 1.1 of the study, and the initial plan was to start with the RP2D described for Part 2A of the study mentioned above or with the MTD/MAD, whichever is lower. If the combination treatment regimen is not well tolerated, PF-06863135 will be reduced to a lower dose level to select the dose level for Part 2C. The decision was then made to start with a dose level of PF06863135 one level lower than the RP2D of monotherapy, as shown in Table 12A. The starting dose of lenalidomide was modified to 15 mg QD orally on days 1-21 of a 28-day cycle, beginning 7 days after the priming dose of PF06863135.

In Part 2C, PF-06863135 will be administered at a dose level based on the results of Part 1C.

Part 1D and Part 2D of the study are combination treatment with PF06863135 and pomalidomide. The treatment cycle is 28 days. Pomalidomide will be administered daily on days 1-21 at a dose of 4 mg PO without dexamethasone, beginning on day 1 of cycle 1. PF-06863135 will be administered SC or IV at the selected dose Q1W or Q2W, beginning on day 1 of cycle 1, with or without a priming dose one week before day 1 of cycle 1.

In Part 1D, the dose of PF-06863135 will be determined based on the results of Part 1 and Part 1.1 of the study, starting at the RP2D described for Part 2A of the study mentioned above, or at the MTD/MAD, whichever is lower. If the combination treatment regimen is not well tolerated, PF-06863135 will be reduced to a lower dose level to select the dose level for Part 2D. The decision was then made to start at a dose level of PF06863135 one level below the RP2D of monotherapy, as shown in Table 12A.

In Part 2D, PF-06863135 will be administered at a dose level based on the results of Part 1D.

Patient Inclusion Criteria . For all study arms, the patient inclusion criteria presented herein include that patients must have progressed on or are intolerant to conventional therapies known to provide clinical benefit in multiple myeloma, including a proteasome inhibitor, immunomodulatory imid (ImiD) drugs, and anti-CD38 mAbs, approved and available, in combination or as monotherapy, and that patients must not be candidates for treatment regimens known to provide clinical benefit in relapsed or refractory multiple myeloma, as determined by the investigator.

The primary and secondary objectives of the study include (1) assessing the preliminary clinical efficacy of PF-06863135 in RP2D, (2) further characterizing safety and tolerability, (3) assessing the PK of PF-06863135 in RP2D, (4) assessing the immunogenicity of PF-06863135, (5) characterizing the effect of PF-06863135 on systemic soluble immune factors, with each of (1)-(5) pertaining to PF-06863135 as monotherapy and in combination with sasanlimab, lenalidomide, or pomalidomide.

Example 11. A phase 2 clinical trial of PF-06863135 bispecific anti-BCMA×CD3 monotherapy in a participant with multiple myeloma refractory to at least one proteasome inhibitor, one IMiD, and one anti-CD38 monoclonal antibody.

This study is an open-label, multicenter, non-randomized, phase 2 study to evaluate the efficacy and safety of PF-06863135 in participants with refractory/relapsed multiple myeloma (RRMM) refractory to at least one proteasome inhibitor (PI), one IMiD, and one anti-CD38 mAb. To determine the effects of prior BCMA-targeted therapy on response to PF-06863135 monotherapy, this study will include 2 independent and parallel cohorts, one with participants naive to BCMA-targeted therapy (Cohort A; approximately 90 participants) and one with participants exposed to prior BCMA-targeted ADC or BCMA-targeted CAR-T cell therapy, approved or investigational (Cohort B; approximately 60 participants). The primary objective for each independent cohort will be to determine the efficacy (i.e., ORR) of PF-06863135, assessed by independent, blinded, centralized review (BICR) determined by the International Myeloma Study Group (IMWG). The study design is shown in Table 15 below.

Table 15. Study treatment from the phase 2 clinical trial of BCMA×CD3 bispecific antibody monotherapy PF-06863135

Cohorts Number of patients and previous therapy Intervention/Introduction A N=90; RRMM. Refractory to all three of IMiDs, PI, and anti-CD38 antibody; no prior BCMA-targeted therapy PF-06863135: priming dose of 44 mg SC at C1D1; then maintenance dose of 76 mg SC Q1W starting at C1D8 and optionally switching to 76 mg SC Q2W after 6 cycles (each cycle is 28 days) B N=60; RRMM. Refractory to all three of IMiDs, PI, and anti-CD38 antibody; prior BCMA-targeted ADC or CAR-T cell therapy, but no prior BCMA-targeted bispecific Ab therapy.

Administration : A participant in each cohort will receive an initial dose of 44 mg PF-06863135 via subcutaneous injection (SC) on Cycle 1 Day 1 (C1D1). Each treatment cycle is 28 days. The initial 44 mg dose serves as a priming dose and is expected to reduce CRS symptoms primarily expected after the initial dose. The priming dose will then be modified to 12 mg PF06863135 administered on C1D1, followed by 32 mg PF06863135 administered on C1D4. The PF-06863135 dose will be escalated to 76 mg SC Q1W beginning on Cycle 1 Day 8 provided the participant meets all three of the following criteria:

(1) ANC ≥1.0×10 ⁹ /L;

(2) Platelets ≥25×10 ⁹ /L; and

(3) Recovery of treatment-related non-hematological toxicities to baseline severity or grade ≤1 (or, at the discretion of the investigator, grade ≤2 if not considered a risk to the safety of the participant).

If a participant does not meet these criteria on Cycle 1 Day 8, the start of 76 mg dosing should be delayed until these criteria are met. If a participant has been on Q1W for at least 6 cycles and achieved an IMWG response of PR or better, with responses maintained for at least 2 months, the dosing interval should be changed from Q1W to Q2W, as a lower dose intensity may be adequate to maintain response given the reduced disease burden in these participants. However, the participant may remain on the Q1W schedule based on the investigator's medical judgment and after consultation with the study sponsor. After changing to the Q2W interval, the dosing interval may be returned to Q1W based on the investigator's medical judgment.

For each study cohort, treatment with PF-06863135 will continue until disease progression, patient withdrawal (withdrawal of consent), or unacceptable toxicity. The study will be concluded when all participants discontinue study treatment and undergo overall survival (OS) follow-up for at least 2 years.

Primary endpoints : determination of overall response rate (ORR) by independent, blinded central review (BICR) according to the International Myeloma Study Group (IMWG)

Secondary endpoints : (1) duration of response (DOR) by BICR and investigator assessment by IMWG; (2) cumulative complete response rate (CCRR) by BICR and investigator assessment by IMWG; (3) ORR assessed by investigator assessment by IMWG; (4) duration of cumulative complete response (DOCCR) by BICR and investigator assessment by IMWG; (5) progression-free survival (PFS) by BICR and investigator assessment by IMWG; (6) overall survival (OS); (7) time to response (TTR) by BICR and investigator assessment by IMWG; (8) minimal residual disease (MRD) negativity rate (central laboratory assessment) by IMWG; (9) AEs and laboratory abnormalities classified according to NCI Common Terminology Criteria for Adverse Events (CTCAE) v5.0; (10) severity of CRS and immune cell-associated neurotoxicity syndrome (ICANS) assessed according to the American Society for Transplantation and Cellular Therapy (ASTCT) criteria. (11) concentrations before and after administration of PF-06863135 and (12) ADA and NAb against PF-06863135

Example 12. An open-label, multicenter, phase 1/2 study to evaluate two-stage escalated priming doses and longer dosing intervals of elranatamab monotherapy (PF-06863135) in participants with relapsed/refractory multiple myeloma

The objective of this study is to evaluate grade 2 or greater CRS when elranatamab is administered using a 2-prime escalation schedule with a premedication agent. Additionally, this study will evaluate the safety, tolerability, PK, and preliminary anti-myeloma activity of elranatamab at doses greater than 76 mg with varying dosing intervals (QW, Q2W, and Q4W) in participants with relapsed/refractory multiple myeloma (RRMM). A full 76 mg QW schedule of elranatamab for 6 cycles followed by Q2W (part 2) will also be evaluated. Cycle 1 begins on the day of the participant's first priming dose.

All doses of elranatamab will be administered subcutaneously (SC).

In the first cycle (C1) of elranatamab treatment, the following treatment regimen will be evaluated for all study participants:

C1D1: premedication drug + elranatamab 12 mg; C1D4: premedication drug + elranatamab 32 mg; C1D8: premedication drug + elranatamab 76 mg; C1D15 and C1D22: elranatamab 76 mg.

Premedication medications are required approximately 60 minutes before the priming doses (in C1D1 and C1D4) and the first full dose of elranatamab (C1D8). The premedication medications used are acetaminophen 650 mg (or paracetamol 500 mg), diphenhydramine 25 mg orally or IV, and dexamethasone 20 mg (or equivalent) orally or IV.

In Cycle 2 and beyond, the following will be assessed:

Part 1A . In the dose level 1 cohort, participants will receive 116 mg Q2W during C2-C6, optionally switching to 116 mg Q4W for participants with an IMWG response of PR or better for at least 2 cycles on Q2W. If dose level 1 is tolerated, in the dose level 2 cohort, participants will receive 152 mg Q2W during C2-C6, optionally switching to 152 mg Q4W for participants with an IMWG response of PR or better for at least 2 cycles on Q2W. For dose level 1 and dose level 2, if after switching to the Q4W interval, a participant began to experience increasing disease burden not yet classified as PD by IMWG criteria, the dosing interval should be returned to Q2W at the same dose level (e.g., from 152 mg Q4W to 152 mg Q2W).

Part 1B begins after identification of a potential MTD/RP2D from Part 1A and will be an expansion of the cohort at the selected dose level.

Part 1C . This is started only if dose level 1 and dose level 2 in Part 1A are tolerated. In this case, during C2-C3, participants will receive 116 mg Q1W or 152 mg Q1W. During C4-C6, for participants with an IMWG response of PR or better at C2 and C3, 116 mg or 152 mg Q2W will be administered. During C7 and thereafter, for participants with an IMWG response of PR or better for at least two cycles on Q2W, 116 mg or 152 mg Q4W will be administered.

Part 2 : 76 mg Q1W will be administered at C2-C6. For participants with an IMWG response of PR or better for at least two cycles on Q1W, 76 mg Q2W will be administered at C7 and beyond. If, after switching to the Q2W interval, the participant begins to experience increasing disease burden not yet classified as PD according to the IMWG criteria, the dosing interval should be returned to 76 mg Q1W.

Example 13. An open-label, multicenter, randomized phase 3 study to evaluate the efficacy and safety of elranatumab (PF06863135) and daratumumab in participants with relapsed/refractory multiple myeloma (RRMM)

The objectives of Part 1 of this study are to evaluate the DLT, safety, and tolerability of elranatamab and daratumumab to select the RP3D combination. The objectives of Part 2 are to compare the efficacy of elranatamab (Arm A) and the combination of elranatamab and daratumumab (Arm B) with the control combination of daratumumab, pomalidomide, and dexamethasone (Arm C). The objectives of Part 1 of this study also include assessing the CRS rate of grade 2 or higher with elranatamab administered alone or in combination with two escalating priming doses along with a premedication agent. The study treatments are shown in Table 16. The cycle is 28 days.

Table 16. Study treatment with the combination of elranatamab and daratumumab

Group Medicines and administration Part 1, dose level minus 1 Elranatamab: priming 12 mg (C1D1) + 32 mg (C1D4), maintenance dose 44 mg QW (starting at C1D8), optional, switch to 44 mg Q2W after 6 cycles
Daratumumab: 1800 mg SC, QW with C1D15, Q2W with C3D15, Q4W with C8D1 Part 1, Dose Level 1 Elranatamab: priming 12 mg (C1D1) + 32 mg (C1D4), maintenance dose 76 mg QW (starting at C1D8), optional, switch to 76 mg Q2W after 6 cycles
Daratumumab: 1800 mg SC, QW with C1D15, Q2W with C3D15, Q4W with C8D1 Part 1, dose cohort expansion Elranatamab: selected dose level minus 1 or dose level 1
Daratumumab: 1800 mg SC, QW with C1D15, Q2W with C3D15, Q4W with C8D1 Part 2, Group A Elranatamab Part 2, Group B Elranatamab, TBD administration taking into account part 1
Daratumumab: 1800 mg SC, QW with C1D15, Q2W with C3D15, Q4W with C8D1 Part 2, Group C Daratumumab, 1800 mg SC, QW with C1D1, Q2W with C3 D1, Q4W with C7D1;
Pomalidomide: 4 mg PO QD on days 1-21 of each 28-day cycle;
Dexamethasone: 40 mg (20 mg for participants aged 75 years and older) PO QW

Elranatamab administration: In Part 1, dose level minus 1, after a participant has been on 44 mg QW until the end of cycle 6, 44 mg Q2W should be administered thereafter to participants with an IMWG response of PR or better that has been maintained for at least 2 cycles. Similarly, 76 mg QW replaces 76 mg Q2W in Part 1, dose level 1, and similarly, QW will replace Q2W in Arm A and Arm B in Part 2. Then, if an increase in disease burden (not classifiable as PD by IMWG criteria) is observed, the dosing interval should be returned to QW.

Daratumumab administration: A subcutaneous injection of 1800 mg Q1W, followed by Q2W, and then Q4W will be used according to the USPI administration schedule of the FDA-approved daratumumab and hyaluronidase-fihj product.

Premedication is required approximately 60 minutes before the priming dose (in C1D1 and C4D1) and the first full dose of elranatamab (C1D8). Premedication is also required 1-3 hours before each dose of daratumumab, except in Part 2 of Group C, where the dexamethasone component of the regimen should be administered before daratumumab and will serve as premedication. If elranatamab and daratumumab are to be administered on the same day, premedication should be administered only once on that day, before elranatamab and daratumumab. The premedication drug used is acetaminophen 650-1000 mg (or paracetamol 500 mg), diphenhydramine 25-50 mg orally or IV, or dexamethasone 20 mg (or equivalent) orally or IV.

Example 14. A randomized, 2-arm phase 3 study of elranatamab (PF-06863135) and lenalidomide versus lenalidomide in patients with newly diagnosed multiple myeloma (NDMM) who were minimal residual disease (MRD) positive after autologous stem cell transplantation (ASCT)

The objectives of this study include comparing the efficacy of combination therapy with elranatamab and lenalidomide (Arm A) with lenalidomide alone (Arm B) and determining the safety and tolerability of combination therapy with elranatamab and lenalidomide. Participants will be individuals with newly diagnosed multiple myeloma who are minimal residual disease (MRD)-positive following autologous stem cell transplantation. Table 16 below shows the planned dosing regimens for each arm of the study.

Table 16. Investigational combination treatment with elranatamab and lenalidomide versus lenalidomide alone in patients with NDMM who are MRD-positive after ASCT

Groups Medicines and administration A Elranatamab: 12 mg priming dose in C1D1; 32 mg priming dose in C1D4; full dose (to be determined) QW in C1D8 until the end of C6, optionally followed by Q2W, dose to be determined based on Example 16, Substudy B (Arm B1, B2, and B3). Elranatamab will be administered subcutaneously.
Lenalidomide: 10 mg PO. Cycle 1: daily from day 8 (or day 15) to day 28. Cycle 2 and thereafter: day 1 to day 28, may be increased to 15 mg PO daily after 3 cycles B Lenalidomide: 10 mg PO daily, from day 1 to day 28 of each 28-day cycle; may be increased to 15 mg PO daily after 3 cycles.

Premedication medications are required approximately 60 minutes before the priming dose (in C1D1 and C1D4) and the first full dose of elranatamab (C1D8). The premedication medications used are acetaminophen 650 (or paracetamol 500 mg), diphenhydramine 25 mg orally or IV, and dexamethasone 20 mg (or equivalent) orally or IV.

Example 15. A randomized, controlled, 2-arm phase 3 study of elranatamab (PF06863135) and lenalidomide versus control in patients with newly diagnosed multiple myeloma (NDMM) ineligible for stem cell transplantation

The objectives of this study include comparing the efficacy of combination therapy with elranatamab and lenalidomide (Arm A) with a lenalidomide control group and determining the safety and tolerability of elranatamab. Participants will be individuals with newly diagnosed multiple myeloma who are ineligible for stem cell transplantation. Table 17 below shows the planned administration schedules for each arm of the study.

Table 17. Investigational combination treatment with elranatamab and lenalidomide in patients with NDMM unsuitable for stem cell transplantation

Groups Medicines and administrations Experimental group Induction, elranatamab: TBD based on results from Example 16, arm B1, B2, and B3. Lenalidomide: 5-25 mg PO daily on 14-21 consecutive days in each cycle, cycle 1 starting on day 1 of week 3; dexamethasone: 10, 20, 40 mg PO daily on selected days (e.g., day 1, 8, 15, 22) in at least cycle 1 and cycle 2.
Maintenance, elranatamab: TBD based on results from Example 16, arm B1, B2, and B3; lenalidomide: 5-15 mg PO QD D1-D28 every cycle Control group Induction, lenalidomide, bortezomib, dexamethasone, standard dose and regimen
Maintenance, lenalidomide, standard dose and regimen

Example 16. An open-label, umbrella, phase 1b and 2 study of elranatamab (PF06863135) in combination with other anticancer agents in participants with relapsed/refractory multiple myeloma (RRMM)

The study objectives include assessing the safety and tolerability of elranatamab in combination with other anticancer therapies in participants with RRMM to determine the RP2D combination. Table 18 provides several examples of this combination therapy study design.

Table 18. Investigational combination therapy with elranatamab and other antitumor therapies in relapsed/refractory multiple myeloma (RRMM)

Groups Medicines and administration A1 Elranatamab: priming at 12 mg+32 mg at week 1 followed by full dose of 44 mg (or 76 mg) Q1W for weeks 23 or 24, with the possibility of tapering to 44 mg (or 76 mg) Q2W thereafter.
Sasanlimab; 300 mg Q4W A2 Elranatamab: priming at 12 mg+32 mg at week 1 followed by full dose of 44 mg (or 76 mg) Q1W for weeks 23 or 24, with the possibility of tapering to 44 mg (or 76 mg) Q2W thereafter.
Isatuximab: TBD B1 Elranatamab: priming at 12 mg+32 mg at week 1 followed by a full dose of 44 mg (or 76 mg) Q1W for 25 weeks with the possibility of tapering to 44 mg (or 76 mg) Q2W thereafter, 76
Lenalidomide: 5-25 mg PO QD, days 1-14 of a 21-day cycle or 1-21 of a 28-day cycle, cycle 1 starts on day 1 of week 3.
Dexamethasone: 10, 20, 40 mg orally daily on selected days (e.g., day 1, 8, 15, 22) in at least cycle 1 and cycle 2. B2 TBD results B1
Elranatamab: priming at 12 mg+32 mg at week 1 followed by a full dose of 44 mg (or 76 mg) Q1W for 25 weeks with the possibility of tapering to 44 mg (or 76 mg) Q2W thereafter, 76
Lenalidomide: 5-25 mg PO QD, days 1-14 of a 21-day cycle or 1-21 of a 28-day cycle, cycle 1 starts on day 1 of week 3.
Dexamethasone: 10, 20, 40 mg orally daily on selected days (e.g., day 1, 8, 15, 22) in at least cycle 1 and cycle 2. B3 Repeat administration for group B1 or group B2, but with expansion of the dose cohort

Sequences

Table 19 shows the sequences of the bispecific anti-BCMA×CD3 antibody PF-06863135 and the anti-PD-1 antibody sasanlimab and the corresponding SEQ ID NOs referenced herein. SEQ ID NOs: 1-13 are the sequences of the CD3 arm of PF-06863135, SEQ ID NOs: 14-26 are the sequences of the BCMA arm of PF-06863135. SEQ ID NOs: 27-34 are the sequences of the anti-PD-1 antibody sasanlimab.

Table 19. Sequences of PF-06863135 and sasanlimab

SEQ ID NO (descriptions) Sequences 1 (VH CD3-arm) EVQLVESGGG LVQPGGSLRL SCAASGFTFS DYYMTWVRQA PGKGLEWVAF IRNRARGYTS DHNPSVKGRF TISRDNAKNS LYLQMNSLRA EDTAVYYCAR DRPSYYVLDY WGQGTTVTVSS 2 (CDR1 VH CD3-arm) DYYMT 3 (CDR1 VH CD3-arm) GFTFSDY 4 (CDR1 VH CD3-arm) GFTFSDYYMT 5 (CDR2 VH CD3-arm) RNRARGYT 6 (CDR2 VH CD3-arm) FIRNRARGYTSDHNPSVKG 7 (CDR3 VH CD3-arm) DRPSYYVLDY 8 (full-length heavy chain CD3-arm) EVQLVESGGG LVQPGGSLRL SCAASGFTFS DYYMTWVRQA PGKGLEWVAF IRNRARGYTS DHNPSVKGRF TISRDNAKNS LYLQMNSLRA EDTAVYYCAR DRPSYYVLDY WGQGTTVTVS Sastkgpsvf plapcsrsts estaalgclv kdyfpepvtv swnsgaltsg vhtfpavlqs sglyslssvv tvpssnfgtq tytcnvdhkp sntkvdktve rkcrvrcprc pappvagpsv flfppkpkdt lmisrtpevt cvvvavshed pevqfnwyvd gvevhnaktk preeqfnstf rvvsvltvvh qdwlngkeyk ckvsnkglps siektisktk gqprepqvyt lppsreemtk nqvsltclvk gfypsdiave wesngqpenn ykttppmlds dgsfflysrl tvdksrwqqg nvfscsvmhe alhnhytqks lslspgk 9 (VL CD3-shoulder) DIVMTQSPDS LAVSLGERAT INCKSSQSLF NVRSRKNYLA WYQQKPGQPP KLLISWASTR ESGVPDRFSG SGSGTDFTLT ISSLQAEDVA VYYCKQSYDL FTFGSGTKLE IK 10 (CDR1 VL CD3-arm) KSSQSLFNVRSRKNYLA 11 (CDR2 VL CD3-arm) WASTRES 12 (CDR3 VL CD3-arm) KQSYDLFT 13 (full-length CD3-arm light chain) DIVMTQSPDS LAVSLGERAT INCKSSQSLF NVRSRKNYLA WYQQKPGQPP KLLISWASTR ESGVPDRFSG SGSGTDFTLT ISSLQAEDVA VYYCKQSYDL FTFGSGTKLE IKrtvaapsv fifppsdeql ksgtasvvcl lnnfypreak vqwkvdnalq sgnsqesvte qdskdstysl sstltlskad yekhkvyace vthqglsspv tksfnrgec 14 (VH BCMA-shoulder) EVQLLESGGG LVQPGGSLRL SCAASGFTFS SYPMSWVRQA PGKGLEWVSA IGGSGGSLPY ADIVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARYW PMDIWGQGTL VTVSS 15 (CDR1 VH BCMA-arm) GFTFSSY 16 (CDR1 VH BCMA-arm) SYPMS 17 (CDR1 VH BCMA-arm) GFTFSSYPMS 18 (CDR2 VH BCMA-arm) GGSGGS 19 (CDR2 VH BCMA-arm) AIGGSGGSLPYADIVKG 20 (CDR3 VH BCMA-arm) YWPMDI 21 (full-length BCMA-arm heavy chain) EVQLLESGGG LVQPGGSLRL SCAASGFTFS SYPMSWVRQA PGKGLEWVSA IGGSGGSLPY ADIVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARYW PMDIWGQGTL VTVSSastkg psvfplapcs rstsestaal gclvkdyfpe pvtvswnsga ltsgvhtfpa vlqssglysl ssvvtvpssn fgtqtytcnv dhkpsntkvd ktverkceve cpecpappva gpsvflfppk pkdtlmisrt pevtcvvvav shedpevqfn wyvdgvevhn aktkpreeqf nstfrvvsvl tvvhqdwlng keykckvsnk glpssiekti sktkgqprep qvytlppsre emtknqvslt cevkgfypsd iavewesngq pennykttpp mldsdgsffl yskltvdksr wqqgnvfscs vmhealhnhy tqkslslspg k 22 (VL BCMA-shoulder) EIVLTQSPGT LSLSPGERAT LSCRASQSVS SSYLAWYQQK PGQAPRLLMY DASIRATGIP DRFSGSGSGT DFTLTISRLE PEDFAVYYCQ QYQSWPLTFG QGTKVEIK 23 (CDR1 VL BCMA-arm) RASQSVSSSYLA 24 (CDR2 VL BCMA-arm) DASIRAT 25 (CDR3 VL BCMA-arm) QQYQSWPLT 26 (full-length BCMA-arm light chain) EIVLTQSPGT LSLSPGERAT LSCRASQSVS SSYLAWYQQK PGQAPRLLMY DASIRATGIP DRFSGSGSGT DFTLTISRLE PEDFAVYYCQ QYQSWPLTFG QGTKVEIKrt vaapsvfifp psdeqlksgt asvvcllnnf ypreakvqwk vdnalqsgns qesvteqdsk dstyslsstl tlskadyekh kvyacevthq glsspvtksf nrgec 27 (VH antibodies against PD-1) QVQLVQSGAE VKKPGASVKV SCKASGYTFT SYWINWVRQA PGQGLEWMGN IYPGSSLTNY NEKFKNRVTM TRDTSTSTVY MELSSLRSED TAVYYCARLS TGTFAYWGQG TLVTVSS 28 (CDR1 VH antibodies against PD-1) SYWIN 29 (CDR2 VH antibodies against PD-1) NIYPGSSLTNYNEKFKN 30 (CDR3 VH antibodies against PD-1) LSTGTFAY 31 (VL antibodies against PD-1) DIVMTQSPDS LAVSLGERAT INCKSSQSLW DSGNQKNFLT WYQQKPGQPP KLLIYWTSYR ESGVPDRFSG SGSGTDFTLT ISSLQAEDVA VYYCQNDYFY PHTFGGGTKV EIK 32 (CDR1 VL anti-PD-1 antibodies) KSSQSLWDSGNQKNFLT 33 (CDR2 VL anti-PD-1 antibodies) WTSYRES 34 (CDR3 VL anti-PD-1 antibodies) QNDYFYPHT

--->

SEQUENCE LIST

<110> PFIZER INC.

<120> METHODS, THERAPY AND APPLICATION FOR THE TREATMENT OF MALIGNANT

NEOPLASMS

<130> PC072602A

<150> US 63/024,016

<151> 2020-05-13

<150> US 63/078,211

<151> 2020-09-14

<150> US 63/106,302

<151> 2020-10-27

<150> US 63/185,357

<151> 2021-05-06

<160> 34

<170> PatentIn version 3.5

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Tyr Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

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Ala Phe Ile Arg Asn Arg Ala Arg Gly Tyr Thr Ser Asp His Asn Pro

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Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser

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Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr

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Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr Val Leu Asp Tyr Trp Gly

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Gln Gly Thr Thr Val Thr Val Ser Ser

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Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

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Tyr Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

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Ala Phe Ile Arg Asn Arg Ala Arg Gly Tyr Thr Ser Asp His Asn Pro

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Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser

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Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr

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Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr Val Leu Asp Tyr Trp Gly

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Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

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Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala

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Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val

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Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

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Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

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Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His

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Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Arg

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Val Arg Cys Pro Arg Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val

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Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

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Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu

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Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

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Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser

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Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

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Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

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Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

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Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

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Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

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Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser

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Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

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Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

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His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

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Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

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Arg Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

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Pro Pro Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val

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Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

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Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln

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Ser Tyr Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

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Arg Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

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Pro Pro Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val

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Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

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Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln

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Ser Tyr Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

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Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

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Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

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Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

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Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

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Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

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Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

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Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

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Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

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Pro Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

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Ser Ala Ile Gly Gly Ser Gly Gly Ser Leu Pro Tyr Ala Asp Ile Val

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Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

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Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

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Pro Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

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Ser Ala Ile Gly Gly Ser Gly Gly Ser Leu Pro Tyr Ala Asp Ile Val

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Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

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Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

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Ala Arg Tyr Trp Pro Met Asp Ile Trp Gly Gln Gly Thr Leu Val Thr

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Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro

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Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val

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Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

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Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

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Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly

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Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

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Val Asp Lys Thr Val Glu Arg Lys Cys Glu Val Glu Cys Pro Glu Cys

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Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

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Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

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Val Val Ala Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr

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Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

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Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His

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Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

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Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln

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Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met

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Thr Lys Asn Gln Val Ser Leu Thr Cys Glu Val Lys Gly Phe Tyr Pro

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Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

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Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu

385 390 395 400

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

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Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

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Lys Ser Leu Ser Leu Ser Pro Gly Lys

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Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

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Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

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Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Met Tyr Asp Ala Ser Ile Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gln Ser Trp Pro

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Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

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Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala

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Asp Ala Ser Ile Arg Ala Thr

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Gln Gln Tyr Gln Ser Trp Pro Leu Thr

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Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

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Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

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Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Met Tyr Asp Ala Ser Ile Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gln Ser Trp Pro

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Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

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Ser Phe Asn Arg Gly Glu Cys

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Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

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Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

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Trp Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

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Gly Asn Ile Tyr Pro Gly Ser Ser Leu Thr Asn Tyr Asn Glu Lys Phe

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Lys Asn Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

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Ala Arg Leu Ser Thr Gly Thr Phe Ala Tyr Trp Gly Gln Gly Thr Leu

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Val Thr Val Ser Ser

115

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Ser Tyr Trp Ile Asn

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Asn Ile Tyr Pro Gly Ser Ser Leu Thr Asn Tyr Asn Glu Lys Phe Lys

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Asn

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Leu Ser Thr Gly Thr Phe Ala Tyr

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Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Trp Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Thr Ser Tyr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Phe Tyr Pro His Thr Phe Gly Gly Gly Thr Lys Val Glu Ile

100 105 110

Lys

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Lys Ser Ser Gln Ser Leu Trp Asp Ser Gly Asn Gln Lys Asn Phe Leu

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Thr

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Trp Thr Ser Tyr Arg Glu Ser

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Gln Asn Asp Tyr Phe Tyr Pro His Thr

1 5

<---

Claims (142)

1. A method of treating multiple myeloma in an individual, comprising subjecting the individual to a combination treatment comprising administering a first therapeutic agent and a second therapeutic agent, wherein the first therapeutic agent is elranatamab and the second therapeutic agent is sasanlimab, lenalidomide or pomalidomide. 2. The method of claim 1, wherein the method comprises administering elranatamab to the individual according to the administration regimen: (a) 80, 130, 215, 360, 600, or 1000 mcg/kg Q1W subcutaneously (SC); (b) 80, 130, 215, 360, 600, or 1000 μg/kg Q2W SC; (c) 16 to 80 mg Q1W SC or Q2W SC; (d) 16 to 20, 40 to 44, or 76 to 80 mg Q1W SC; (e) 16 to 20, 40 to 44, or 76 to 80 mg Q2W SC; (f) 40 mg Q1W SC or Q2W SC; (g) 44 mg Q1W SC or Q2W SC; (h) 76 mg Q1W SC or Q2W SC; (i) 80 mg Q1W SC or Q2W SC; (j) a priming dose of 44 mg Q1W SC for 1-4 weeks or a priming dose of 32 mg Q1W SC for 1-4 weeks followed by a first treatment dose of 76 mg Q1W SC or Q2W SC; (k) priming with 40 mg Q1W SC for 1-4 weeks followed by the first treatment with 80 mg Q1W SC or Q2W SC; (l) a priming administration of 44 mg Q1W SC for 1-4 weeks or a priming administration of 32 mg Q1W SC for 1-4 weeks followed by a first treatment administration of 76 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47 or 48 weeks followed by a second treatment administration of 76 mg Q2W SC; (m) a priming dose of 40 mg Q1W SC for 1-4 weeks, followed by a first treatment dose of 80 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47 or 48 weeks, followed by a second treatment dose of 80 mg Q2W SC; (n) priming with 44 mg Q1W SC for 1 week followed by the first treatment dose of 76 mg Q1W SC or Q2W SC; (o) a priming dose of 32 mg Q1W SC for 1 week followed by a first treatment dose of 76 mg Q1W SC or Q2W SC; (p) priming administration of 40 mg Q1W SC for 1 week followed by the first treatment administration of 80 mg Q1W SC or Q2W SC; (q) a priming dose of 44 mg Q1W SC for 1 week followed by a first treatment dose of 76 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47 or 48 weeks followed by a second treatment dose of 76 mg Q2W SC; (r) a priming dose of 44 mg Q1W SC for 1 week followed by a first treatment dose of 76 mg Q1W SC for 23 weeks followed by a second treatment dose of 76 mg Q2W SC; (s) a priming dose of 44 mg Q1W SC for 1 week followed by a first treatment dose of 76 mg Q1W SC for 24 weeks followed by a second treatment dose of 76 mg Q2W; (t) a priming dose of 32 mg Q1W SC for 1 week followed by a first treatment dose of 76 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47 or 48 weeks followed by a second treatment dose of 76 mg Q2W SC; (u) a priming dose of 40 mg Q1W SC for 1 week followed by a first treatment dose of 80 mg Q1W SC for 2 to 20, 21, 22, 23, 24, 25 to 46, 47, or 48 weeks followed by a second treatment dose of 80 mg Q2W SC; or (v) a priming dose of 40 mg Q1W SC for 1 week followed by a first treatment dose of 80 mg Q1W for 23 or 24 weeks followed by a second treatment dose of 80 mg Q2W SC. 3. The method of claim 2, wherein the individual is administered elranatamab in a first therapeutic administration of 76 mg Q1W SC and after at least 23 weeks of such first therapeutic administration, the individual is administered elranatamab in a second therapeutic administration of 76 mg Q2W or continues to be administered elranatamab in the first therapeutic administration. 4. The method of claim 1, wherein the method comprises administering elranatamab to the individual, subcutaneously, a first therapeutic administration at 23, 24, or 25 weeks, followed by a second therapeutic administration, wherein (a) the first treatment administration is 4 mg Q1W and the second treatment administration is 4 mg Q2W; (b) the first treatment administration is 12 mg Q1W and the second treatment administration is 12 mg Q2W; (c) the first treatment administration is 24 mg Q1W and the second treatment administration is 24 mg Q2W; (d) the first treatment administration is 32 mg Q1W and the second treatment administration is 32 mg Q2W; (e) the first treatment administration is 44 mg Q1W and the second treatment administration is 44 mg Q2W; or (f) the first treatment administration is 76 mg Q1W and the second treatment administration is 76 mg Q2W. 5. The method of claim 4, wherein if the amount of the dose within the first therapeutic administration is 32 mg or more, the method further comprises administering elranatamab to the individual within a priming administration, and the priming administration is carried out for one week, the first dose within the first therapeutic administration is administered in the week immediately following the week in which the priming administration is carried out, and wherein (1) The priming dose is a single priming dose, and the single priming dose is 24 mg; (2) the priming administration includes a first priming dose of 4 mg and a second priming dose of 20 mg, and the two priming doses are administered on two different days and the first priming dose is administered before the second priming dose; (3) the priming administration includes a first priming dose of 8 mg and a second priming dose of 16 mg, and the two priming doses are administered on two different days and the first priming dose is administered before the second priming dose; (4) the priming administration includes a first priming dose of 12 mg and a second priming dose of 12 mg, and the two priming doses are administered on two different days and the first priming dose is administered before the second priming dose; (5) the priming administration includes a first priming dose of 8 mg and a second priming dose of 24 mg, and the two priming doses are administered on two different days and the first priming dose is administered before the second priming dose; or (6) The priming administration includes a first priming dose of 4 mg and a second priming dose of 28 mg, and the two priming doses are administered on two different days and the first priming dose is administered before the second priming dose. 6. The method of claim 1, wherein the method comprises administering to the individual elranatamab: (a) the first therapeutic administration of 32 mg to 76 mg Q1W SC, starting at week 1; or (b) a priming administration during week 1, and a first treatment administration beginning in week 2, wherein the priming administration is (i) a first reconciliation dose of 4 mg SC to 32 mg SC and a second reconciliation dose of 12 mg SC to 44 mg SC, wherein the first reconciliation dose and the second reconciliation dose are administered sequentially in week 1, or (ii) a single reconciliation dose of 24 mg to 44 mg SC, and wherein the first treatment administration is 32 mg to 76 mg Q1W SC or 32 mg to 152 mg Q2W SC, beginning in week 2, and wherein the dose amount within the first treatment administration is higher than the dose amount of each of the respective single priming dose, first priming dose, and second priming dose; where week 1, week 2 and any subsequent weeks refer to the first, second and any subsequent weeks that the individual is administered elranatamab, respectively, and elranatamab is administered to the individual in the form of a pharmaceutical preparation containing elranatamab. 7. The method of claim 6, wherein the individual is subjected to a priming administration, wherein the priming administration is a single priming dose of 24 mg SC, 32 mg SC, or 44 mg SC at week 1, or the priming administration is (i) a first priming dose of 12 mg SC and a second priming dose of 32 mg SC; (ii) a first priming dose of 4 mg SC and a second priming dose of 20 mg; (iii) a first priming dose of 8 mg and a second priming dose of 16 mg; (iv) a first priming dose of 12 mg and a second priming dose of 12 mg; or (v) a first priming dose of 8 mg and a second priming dose of 24 mg. 8. The method according to claim 6 or 7, wherein the first therapeutic administration is 32 mg Q1W SC or 32 mg Q2W SC, 44 mg Q1W SC or 44 mg Q2W SC. 9. The method of claim 8, wherein the individual is subjected to the first treatment administration until at least the end of cycle 1 or at least the end of cycle 6, wherein the cycle is 21 days or 28 days, cycle 1 begins on day 1 of week 1, day 1 of week 2, or day 1 of week 3, and cycle 1, cycle 2, and subsequent cycles refer to the first, second, and subsequent cycles that the individual is administered elranatamab, respectively. 10. The method of claim 9, further comprising administering to the individual elranatamab in a second therapeutic administration of 32 mg to 152 mg Q2W SC, 32 mg to 152 mg Q3W SC, or 32 mg to 152 mg Q4W SC, after which the individual is no longer in the period of the first therapeutic administration, wherein the second therapeutic administration is administered at a frequency less than the frequency of the corresponding first therapeutic administration, or the second therapeutic administration has a smaller dose amount than the first therapeutic administration. 11. The method of claim 9, wherein after subjecting the individual to the first treatment administration, at least until the end of cycle 6, the individual is subjected to a second treatment administration of elranatamab instead of the first treatment administration, or the individual may continue to be subjected to the first treatment administration, and wherein the second treatment administration is from 32 mg to 152 mg Q2W SC, from 32 mg to 152 mg Q3W SC, or from 32 mg to 152 mg Q4W SC, wherein the second treatment administration is administered at a frequency less than the frequency of the first treatment administration, or the second treatment administration has a smaller dose amount than the first treatment administration. 12. The method of claim 1, wherein the method comprises administering elranatamab to the individual in a first treatment administration, wherein the first treatment administration is (i) 76 mg Q1W SC, (ii) 76 mg Q2W SC, or (iii) 76 mg Q1W SC for three weeks followed by 116 mg Q1W SC, or (iv) 76 mg Q1W SC for three weeks followed by 152 mg Q1W SC. 13. The method of claim 12, wherein the individual is subjected to the first treatment administration until at least the end of cycle 1, at least the end of cycle 3, or at least the end of cycle 6, wherein the cycle is 21 days or 28 days, and cycle 1 begins on day 1 of week 1, day 1 of week 2, or day 1 of week 3, and cycle 1, cycle 2, and subsequent cycles refer to the first, second, and subsequent cycles that the individual is administered elranatamab, respectively. 14. The method of claim 13, further comprising administering to the individual elranatamab in a second therapeutic administration of 44 mg to 152 mg Q2W SC, 44 mg to 152 mg Q3W SC, or 44 mg to 152 mg Q4W SC, after which the individual is no longer in the period of the first therapeutic administration, wherein the second therapeutic administration is administered at a frequency less than the frequency of the first therapeutic administration, or the second therapeutic administration has a smaller dose amount than the first therapeutic administration. 15. The method of claim 13, wherein after subjecting the individual to the first therapeutic administration, at least until the end of cycle 6, the individual is subjected to a second therapeutic administration of 44 mg to 152 mg Q2W SC, 44 mg to 152 mg Q3W SC, or 44 mg to 152 mg Q4W SC instead of the first therapeutic administration, or the individual may continue to be subjected to the first therapeutic administration, wherein the second therapeutic administration is carried out at a frequency less than the frequency of the corresponding first therapeutic administration, or the second therapeutic administration has a smaller dose amount than the first therapeutic administration. 16. The method of claim 14 or 15, wherein the first therapeutic administration is 76 mg Q1W SC, and the second therapeutic administration is 44 mg Q2W SC, 76 mg Q2W SC, 116 mg Q2W SC, 152 mg Q2W SC, 44 mg Q3W SC, 76 mg Q3W SC, 116 mg Q3W SC, 152 mg Q3W SC, 44 mg Q4W SC, 76 mg Q4W SC, 116 mg Q4W SC, or 152 mg Q4W SC. 17. The method of any one of claims 14 or 15, wherein the first therapeutic administration is 76 mg Q2W SC, and the second therapeutic administration is 44 mg Q2W SC, 44 mg Q3W SC, 76 mg Q3W SC, 116 mg Q3W SC, 152 mg Q3W SC, 44 mg Q4W SC, 76 mg Q4W SC, 116 mg Q4W SC, or 152 mg Q4W SC. 18. The method of any one of claims 6-17, wherein the individual is administered elranatamab in a first treatment administration until the end of cycle 1 followed by a second treatment administration, wherein the cycle is 21 days or 28 days, cycle 1 begins on day 1 of week 1, or day 1 of week 2, or day 1 of week 3, and cycle 1, cycle 2 and subsequent cycles refer to the first, second and subsequent cycles that the individual is administered elranatamab, respectively. 19. The method of claim 18, wherein the second therapeutic administration is carried out at least until the end of cycle 6, and then the individual is subjected to a third therapeutic administration of 76 mg to 152 mg Q3W SC or 76 mg to 152 mg Q4W SC instead of the second therapeutic administration, or the individual continues to be subjected to the second therapeutic administration. 20. The method of claim 19, wherein the second therapeutic administration is administered before the end of cycle 6, the first dose in the third therapeutic administration is administered in cycle 7, and the third therapeutic administration is 116 mg Q4W SC or 152 mg Q4W SC. 21. The method of claim 19 or 20, wherein the first therapeutic administration is 76 mg Q1W SC, the second therapeutic administration is 116 mg Q2W SC, and the third therapeutic administration is 116 mg Q4W SC. 22. The method of claim 19 or 20, wherein the first therapeutic administration is 76 mg Q1W SC, the second therapeutic administration is 152 mg Q2W SC, and the third therapeutic administration is 152 mg Q4W SC. 23. The method of claim 1, wherein the method comprises administering elranatamab to the individual according to a dosing schedule as shown below, and wherein the dosing schedule is described by a week number, a dose amount, and a frequency of administration corresponding to each number of weeks: (a) Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-24 32; 44; 76; 116 or 152 Weekly 25 and beyond 32; 44; 76; 116 or 152 Weekly; once every two weeks; once every three weeks or once every four weeks (b) Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-25 32; 44; 76; 116 or 152 Weekly 26 and further 32; 44; 76; 116 or 152 Weekly; once every two weeks; once every three weeks or once every four weeks (c) Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-26 44; or 76 Weekly 27 and further 44; or 76 Weekly; once every two weeks; once every three weeks or once every four weeks (d) Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-24 32; 44; 76; 116 or 152 Once every two weeks 25 and beyond 32; 44; 76; 116 or 152 Once every two weeks; once every three weeks or once every four weeks (e) Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-25 32; 44; 76; 116 or 152 Once every two weeks 26 and further 32; 44; 76; 116 or 152 Once every two weeks; once every three weeks or once every four weeks or (f) Week number Dose amount (mg) Frequency of administration 1 44; 32; 12 and 32 or A and B Weekly 2-26 32; 44; 76; 116 or 152 Once every two weeks 27 and further 32; 44; 76; 116 or 152 Once every two weeks; once every three weeks or once every four weeks where if the dose amount is 12 mg and 32 mg during week 1, the dose amount of 12 mg is administered on one day, then the dose amount of 32 mg is administered on another day, where A and B are 4 (A) and 20 (B), 8 (A) and 16 (B), 12 (A) and 12 (B), or 8 (A) and 24 (B), and where if the dose amount is A mg and B mg during week 1, the dose amount of A mg is administered on one day, then the dose amount of B mg is administered on another day. 24. The method of claim 23, wherein the individual is administered elranatamab according to the administration regimen as shown below. (a) Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-24 76 Weekly 25 and beyond 76 Weekly; once every two weeks; once every three weeks or once every four weeks (b) Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-25 76 Weekly 26 and further 76 Weekly; once every two weeks; once every three weeks or once every four weeks (c) Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-26 76 Weekly 27 and further 76 Weekly; once every two weeks; once every three weeks or once every four weeks (d) Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-24 76 Once every two weeks 25 and beyond 76 Once every two weeks; once every three weeks or once every four weeks (e) Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-25 76 Once every two weeks 26 and further 76 Once every two weeks; once every three weeks or once every four weeks or (f) Week number Dose amount (mg) Frequency of administration 1 44 Weekly 2-26 76 Once every two weeks 27 and further 76 Once every two weeks; once every three weeks or once every four weeks. 25. The method of claim 24, wherein the individual is administered elranatamab according to the administration schedule (a), (b), or (c), and the frequency of administration during week 25 onward, week 26 onward, and week 27 onward in the administration schedule (a), (b), and (c), respectively, is (i) weekly, (ii) biweekly, or (iii) weekly or biweekly. 26. The method of claim 23, wherein the individual is administered elranatamab according to the administration regimen as shown below. (a) Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-24 76 Weekly 25 and beyond 76 Weekly; once every two weeks; once every three weeks or once every four weeks (b) Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-25 76 Weekly 26 and further 76 Weekly; once every two weeks; once every three weeks or once every four weeks (c) Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-26 76 Weekly 27 and further 76 Weekly; once every two weeks; once every three weeks or once every four weeks (d) Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-24 76 Once every two weeks 25 and beyond 76 Once every two weeks; once every three weeks or once every four weeks (e) Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-25 76 Once every two weeks 26 and further 76 Once every two weeks; once every three weeks or once every four weeks or (f) Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-26 76 Once every two weeks 27 and further 76 Once every two weeks; once every three weeks or once every four weeks. 27. The method of claim 26, wherein the individual is administered elranatamab according to the administration schedule (a), (b), or (c), and the frequency of administration during week 25 onward, week 26 onward, and week 27 onward in the administration schedule (a), (b), and (c), respectively, is (i) weekly, (ii) biweekly, or (iii) weekly or biweekly. 28. The method of claim 23, wherein the individual is administered elranatamab according to the administration regimen as shown below. (a) Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-24 44 Weekly 25 and beyond 44 Weekly; once every two weeks; once every three weeks or once every four weeks (b) Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-25 44 Weekly 26 and further 44 Weekly; once every two weeks; once every three weeks or once every four weeks (c) Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-26 44 Weekly 27 and further 44 Weekly; once every two weeks; once every three weeks or once every four weeks (d) Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-24 44 Once every two weeks 25 and beyond 44 Once every two weeks; once every three weeks or once every four weeks (e) Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-25 44 Once every two weeks 26 and further 44 Once every two weeks; once every three weeks or once every four weeks or (f) Week number Dose amount (mg) Frequency of administration 1 32; or 12 and 32 Weekly 2-26 44 Once every two weeks 27 and further 44 Once every two weeks; once every three weeks or once every four weeks. 29. The method of claim 28, wherein the individual is administered elranatamab according to the administration schedule (a), (b), or (c), and the frequency of administration during week 25 onward, week 26 onward, and week 27 onward in the administration schedule (a), (b), and (c), respectively, is (i) weekly, (ii) biweekly, or (iii) weekly or biweekly. 30. The method of any one of claims 23-29, wherein the amount of dose and the frequency of administration during week 1 are collectively referred to as a priming administration, and if the individual is administered only one dose of elranatamab as part of the priming administration, such single dose is referred to as a single remediation dose, if the individual is administered two doses of elranatamab sequentially during week 1, the two doses are referred to as the first remediation dose and the second remediation dose, respectively; the amount of dose and the frequency of administration during weeks 2-24, weeks 2-25 and weeks 2-26 in the respective administration schedules (a) and (d), (b) and (e) and (c) and (f), respectively, in each administration schedule are collectively referred to as the first therapeutic administration, the amount of dose and the frequency of administration during week 25 and onward, week 26 and onward and week 27 and onward in the respective administration doses (a) and (d), (b) and (e) and (c) and (f) in each administration schedule are collectively referred to as the second therapeutic administration. 31. The method of claim 30, wherein the individual is subjected to a second therapeutic administration of elranatamab for 6 to 18 cycles, then the individual is subjected to a third therapeutic administration of elranatamab subcutaneously, wherein the third therapeutic administration is 32 mg Q2W, 32 mg Q4W, 44 mg Q2W, 44 mg Q4W, 76 mg Q2W, 76 mg Q4W, 116 mg Q2W, 116 mg Q4W, 152 mg Q2W, or 152 mg Q4W, wherein the cycle is 21 days or 28 days, and cycle 1 begins on day 1 of week 1, day 1 of week 2, or day 1 of week 3. 32. The method of claim 23, wherein (i) the first therapeutic administration is 32 mg Q1W, the second therapeutic administration is 32 mg Q1W or 32 mg Q2W, and the third therapeutic administration is 32 mg Q2W or 32 mg Q4W, (ii) the first therapeutic administration is 32 mg Q1W, the second therapeutic administration is 32 mg Q2W, and the third therapeutic administration is 32 mg Q4W, (iii) the first therapeutic administration is 44 mg Q1W, the second therapeutic administration is 44 mg Q1W or 44 mg Q2W, and the third therapeutic administration is 44 mg Q2W or 44 mg Q4W; (iv) the first therapeutic administration is 44 mg Q1W, the second therapeutic administration is 44 mg Q2W, and the third therapeutic administration is 44 mg Q4W; (v) the first therapeutic administration is 76 mg Q1W, the second therapeutic administration is 76 mg Q1W or 76 mg Q2W, and the third therapeutic administration is 76 mg Q2W or 76 mg Q4W, (vi) the first therapeutic administration is 76 mg Q1W, the second therapeutic administration is 76 mg Q2W, and the third therapeutic administration is 76 mg Q4W, (vii) the first therapeutic administration is 116 mg Q1W, the second therapeutic administration is 116 mg Q1W or 116 mg Q2W, and the third therapeutic administration is 116 mg Q2W or 116 mg Q4W; (viii) the first therapeutic administration is 116 mg Q1W, the second therapeutic administration is 116 mg Q2W, and the third therapeutic administration is 116 mg Q4W, (ix) the first therapeutic administration is 152 mg Q1W, the second therapeutic administration is 152 mg Q1W or 152 mg Q2W, and the third therapeutic administration is 152 mg Q2W or 152 mg Q4W, or (x) the first therapeutic administration is 152 mg Q1W, the second therapeutic administration is 152 mg Q2W, and the third therapeutic administration is 152 mg Q4W. 33. The method of claim 1, wherein the method comprises administering elranatamab to the individual according to a dosing schedule as shown below, and wherein the dosing schedule is described by a week number, a dose amount, and a frequency of administration corresponding to each number of weeks: Week number Dose amount (mg) Frequency of administration 1 44; or 32; or 12 and 32; or A and B Weekly 2-4 from 44 to 152 Weekly 5-24 from 44 to 152 Weekly; or biweekly 25 and beyond from 44 to 152 Once every two weeks; once every three weeks or once every four weeks where if the dose amount is 12 mg and 32 mg during week 1, the dose amount of 12 mg is administered on one day, then the dose amount of 32 mg is administered on another day, where A and B are 4 (A) and 20 (B), 8 (A) and 16 (B), 12 (A) and 12 (B), or 8 (A) and 24 (B), and where the dose amount of A mg is administered on one day, then the dose amount of B mg is administered on another day. 34. The method of claim 33, wherein elranatamab is administered to the individual according to the following administration schedule: Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 76 Weekly 5-24 116 Once every two weeks 25 and beyond 116 Once every two weeks; once every three weeks or once every four weeks. 35. The method of claim 33, wherein elranatamab is administered to the individual according to the following administration schedule: Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 76 Weekly 5-24 152 Once every two weeks 25 and beyond 152 Once every two weeks; once every three weeks or once every four weeks. 36. The method of any one of claims 33-35, wherein the dose amount and the frequency of administration during week 1 are collectively referred to as a priming administration, and if the individual is administered only one dose of elranatamab as part of the priming administration, such single dose is referred to as a single remediation dose, if the individual is administered two doses of elranatamab sequentially during week 1, the two doses are referred to as a first remediation dose and a second remediation dose, respectively, the dose amount and the frequency of administration during weeks 2-4 are collectively referred to as a first therapeutic administration, the dose amount and the frequency of administration during weeks 5-24 are collectively referred to as a second therapeutic administration, and the dose amount and the frequency of administration during week 25 and thereafter are collectively referred to as a third therapeutic administration. 37. The method of claim 1, wherein the method comprises administering elranatamab to the individual according to a dosing schedule as shown below, and wherein the dosing schedule is described by a week number, a dose amount, and a frequency of administration corresponding to each number of weeks: Week number Dose amount (mg) Frequency of administration 1 44; or 32; or 12 and 32; or A and B Weekly 2-4 from 44 to 76 Weekly 5-12 from 44 to 152 Weekly 13-24 from 44 to 152 Weekly; or biweekly 25 and beyond from 44 to 152 Once every two weeks; once every three weeks or once every four weeks where if the dose amount is 12 mg and 32 mg during week 1, the dose amount of 12 mg is administered on one day, then the dose amount of 32 mg is administered on another day, where A and B are 4 (A) and 20 (B), 8 (A) and 16 (B), 12 (A) and 12 (B), or 8 (A) and 24 (B), and where the dose amount of A mg is administered on one day, then the dose amount of B mg is administered on another day. 38. The method of claim 37, wherein elranatamab is administered to the individual according to the following administration schedule: Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 76 Weekly 5-12 116 Weekly 13-24 116 Weekly; or biweekly 25 and beyond 116 Once every two weeks; once every three weeks or once every four weeks. 39. The method of claim 37, wherein elranatamab is administered to the individual according to the following administration schedule: Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 76 Weekly 2-12 152 Weekly 13-24 152 Weekly or biweekly 25 and beyond 152 Once every two weeks or once every four weeks. 40. The method of claim 37, wherein elranatamab is administered to the individual according to the following administration schedule: Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 76 Weekly 5-12 76 Weekly 13-24 76 Weekly; or biweekly 25 and beyond 76 Once every two weeks; once every three weeks or once every four weeks. 41. The method of claim 37, wherein elranatamab is administered to the individual according to the following administration schedule Week number Dose amount (mg) Frequency of administration 1 12 and 32 Weekly 2-4 44 Weekly 5-12 44 Weekly 13-24 44 Weekly or biweekly 25 and beyond 44 Once every two weeks; once every three weeks or once every four weeks. 42. The method according to any one of claims 37-41, wherein the amount of dose and the frequency of administration during week 1 are collectively referred to as a priming administration, and if the individual is administered only one dose of elranatamab as part of the priming administration, such a single dose is referred to as a single remediation dose, if the individual is administered two doses of elranatamab sequentially during week 1, the two doses are referred to as a first remediation dose and a second remediation dose, respectively, the amount of dose and the frequency of administration during weeks 2-4 and the amount of dose and the frequency of administration during weeks 5-12 are collectively referred to as a first therapeutic administration, the amount of dose and the frequency of administration during weeks 13-24 are collectively referred to as a second therapeutic administration, and the amount of dose and the frequency of administration during week 25 and thereafter are collectively referred to as a third therapeutic administration. 43. The method according to any one of claims 2-42, further comprising administering daratumumab to the individual. 44. The method of any one of claims 2-42, further comprising administering to the individual isatuximab. 45. The method of any one of paragraphs 2-42, further comprising administering to the individual at least one dose of a premedication drug on the day the individual is administered a single priming dose, a first priming dose, a second priming dose, or a first dose of a first treatment administration of elranatamab, wherein the premedication drug is acetaminophen, diphenhydramine, or dexamethasone. 46. The method according to any one of paragraphs 2-42, further comprising subjecting the individual to radiation therapy.