EA044652B1 - BCMA BINDING ANTIBODIES AND THEIR APPLICATIONS - Google Patents

Description

1. List of sequences

The invention contains a sequence listing that has been filed electronically in ASCII format and is thus incorporated herein by reference in its entirety. The ASCII copy in question, created on February 13, 2019, is called 298068-00272_SL.txt and is 26248 bytes in size.

2. Field of technology

The present invention relates to isolated antibodies or fragments thereof that bind to B-cell maturation antigen (BCMA), polynucleotides encoding the antibodies or fragments, host cells producing the antibodies or fragments, and methods of using or treating using the antibodies or fragments .

3. State of the art

BCMA (B cell maturation antigen; also referred to as TNFRSF17 or CD269) is a transmembrane protein belonging to the TNF receptor superfamily. BCMA is a B cell marker that is important for B cell development and homeostasis through its interaction with its ligands BAFF (B cell activating factor of the TNF family; also called TALL-1 or TNFSF13B) and APRIL (proliferation-inducing ligand).

BCMA expression is known to be restricted to B cell lineages, and the protein is primarily present on plasma cells and plasmablasts, and to some extent on memory B cells, but is virtually absent on peripheral and naïve B cells. Together with its family members, TACI (transmembrane activator and cyclophilin ligand-interacting) protein and BAFF-R (B-cell activating factor receptor of the TNF family), BCMA regulates various aspects of humoral immunity, B-cell development and homeostasis.

BCMA is also expressed on multiple myeloma (MM) cells. BCMA appears to support the growth and survival of multiple myeloma (MM) cells. MM cell lines and freshly isolated MM cells typically express BCMA and TACI proteins on their cell surface and have variable expression of BAFF-R protein on their cell surface. Multiple myeloma is the second most common hematologic malignancy, accounting for 2% of all cancer deaths. MM is a heterogeneous disease and is caused primarily by chromosomal translocations, including t(11;14), t(4;14), t(8;14), del(13), and del(17). Patients with MM may have a variety of disease-related symptoms due to bone marrow infiltration, bone destruction, renal failure, immunodeficiency, and the psychological burden associated with a cancer diagnosis.

Current treatments used to treat multiple myeloma usually do not cure the patient. Stem cell transplantation may not be suitable for many patients due to advanced age, other serious illnesses, or other physical limitations. Chemotherapy only partially controls multiple myeloma and rarely leads to complete remission. Thus, there is a need for new, innovative treatments for multiple myeloma and other plasma cell or B cell diseases or disorders.

4. Essence of the invention

In a first aspect, provided herein is an antibody that binds to B Cell Maturation Antigen (BCMA), or a BCMA-binding fragment thereof, comprising heavy chain CDR1, CDR2, or CDR3 sequences: SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9, respectively; SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively; SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15, respectively; SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively; SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO: 23, respectively; SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID NO: 26, respectively; SEQ ID NO: 29, SEQ ID NO: 30 and SEQ ID NO: 31, respectively; SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34, respectively; SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37, respectively; SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively; SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45, respectively; SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, respectively; SEQ ID NO: 51, SEQ ID NO: 52 and SEQ ID NO: 53, respectively; SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, respectively; or SEQ ID NO: 57, SEQ ID NO: 58 and SEQ ID NO: 59, respectively. In a specific embodiment, the antibody further comprises the light chain CDR1, CDR2, or CDR3 sequences of SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4, respectively. In another specific embodiment, the antibody further comprises the light chain CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively.

In a specific embodiment, provided herein is an antibody that binds to BCMA, or a BCMA-binding fragment thereof, comprising a light chain comprising the CDR1, CDR2, or CDR3 sequences of SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4 , respectively, and the heavy chain CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9, respectively; SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively; or SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15, respectively.

- 1 044652

In another specific embodiment, provided herein is an antibody that binds to BCMA, or a BCMA-binding fragment thereof, comprising a light chain comprising the sequences CDR1, CDR2 or CDR3 of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively, and the heavy chain CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively; SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO: 23, respectively; or SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID NO: 26, respectively.

In another specific embodiment, provided herein is an antibody that binds to BCMA, or a BCMA-binding fragment thereof, comprising a light chain comprising the sequences CDR1, CDR2 or CDR3 of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively, and the heavy chain CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 29, SEQ ID NO: 30 and SEQ ID NO: 31, respectively; SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34, respectively; or SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37, respectively.

In another specific embodiment, provided herein is an antibody that binds to BCMA, or a BCMA-binding fragment thereof, comprising a light chain comprising the sequences CDR1, CDR2 or CDR3 of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively, and the heavy chain CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively; SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45, respectively; or SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, respectively.

In another specific embodiment, provided herein is an antibody that binds to BCMA, or a BCMA-binding fragment thereof, comprising a light chain comprising the sequences CDR1, CDR2 or CDR3 of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively, and the heavy chain CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 51, SEQ ID NO: 52 and SEQ ID NO: 53, respectively; SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, respectively; or SEQ ID NO: 57, SEQ ID NO: 58 and SEQ ID NO: 59, respectively.

In another more specific embodiment of any of the above antibodies or BCMA-binding fragments thereof, the antibody or BCMA-binding fragment thereof comprises at least 80%, at least 85%, at least 90% of the light chain variable domain amino acid sequence. at least 95%, at least 96% identical, at least not 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO: 1, and the amino acid sequence of the heavy chain variable domain is at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, at least 99, or 100% identical to SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 28, SEQ ID NO: 39 or SEQ ID NO: 50.

In a more specific embodiment of any of the above antibodies or BCMA binding fragments thereof, the antibody or BCMA binding fragment thereof comprises a light chain variable domain amino acid sequence of SEQ ID NO: 1 or a light chain variable domain amino acid sequence of at least 80, 85, at least 90, at least 95, at least 96, at least 97, at least 98, or at least 99% identical to SEQ ID NO: 1. In another more specific embodiment implementation of any of the above antibodies or BCMA-binding fragments thereof, this antibody or BCMA-binding fragment thereof contains the amino acid sequence of the heavy chain variable domain of SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 28, SEQ ID NO: 39 or SEQ ID NO: 50, or the amino acid sequence of the heavy chain variable domain of at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, according to at least 98% or at least 99% identical to SEQ ID NO: 6, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 39, or SEQ ID NO: 50. In another more specific embodiment, any of of the above antibodies or BCMA-binding fragments thereof, the antibody or BCMA-binding fragment thereof contains the amino acid sequence of the light chain variable domain at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, at least 99, or 100% identical to SEQ ID NO: 1, and the amino acid sequence of the heavy chain variable domain at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, at least 99 or 100% identical to SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 28, SEQ ID NO: 39 or SEQ ID NO: 50.

In a more specific embodiment of any of the above antibodies or BCMA binding fragments thereof, the antibody or BCMA binding fragment thereof comprises a light chain variable domain amino acid sequence that comprises the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 , and is at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98 or at least 99 % identical to SEQ ID NO: 1. In other more specific embodiments, any of the above antibodies or BCMA-binding antibodies thereof

- 2 044652 fragments of an antibody or a BCMA-binding fragment thereof contains the amino acid sequence of the heavy chain variable domain of SEQ ID NO: 7-9, 10-12 or 13-15 and is at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, or at least 99% identical to SEQ ID NO: 6; the antibody or BCMA-binding fragment thereof contains the amino acid sequence of the heavy chain variable domain of SEQ ID NO: 18-20, 21-23 or 24-26 and is at least 85, at least 90, at least 95, according to at least 96, at least 97, at least 98, or at least 99% identical to SEQ ID NO: 17; the antibody or BCMA binding fragment thereof contains the amino acid sequence of the heavy chain variable domain of SEQ ID NO: 29-31, 32-34 or 35-37 and is at least 80, at least 85, at least 90 at least 95, at least 96, at least 97, at least 98, or at least 99% identical to SEQ ID NO: 28; the antibody or BCMA binding fragment thereof contains the amino acid sequence of the heavy chain variable domain of SEQ ID NO: 40-42, 43-45 or 46-48 and is at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, or at least 99% identical to SEQ ID NO: 39; the antibody or BCMA binding fragment thereof contains the amino acid sequence of the heavy chain variable domain of SEQ ID NO: 51-53, 54-56 or 57-59 and is at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, or at least 99% identical to SEQ ID NO: 50.

In specific embodiments of any of the anti-BCMA antibodies or BCMA-binding fragments thereof provided herein, the antibody is a monoclonal antibody, a chimeric antibody, a diantibody, a Fab fragment, a Fab' fragment, or an F(ab')=fragent, Fv, bispecific antibody, bispecific Fab ₂ , bispecific (mab) ₂ , humanized antibody, engineered human antibody, bispecific T cell activator, bispecific NK cell activator, single chain antibody (eg, single chain Fv fragment or scFv), Triomab, IgG variant ridge and valley (kih) with common light chain, CrossMAb platform antibodies, ortho-Fab IgG, DVD-Ig, 2 in 1-IgG, IgG-scFv, sdFv2-Fc, bi-nanobody, TandAb, reprogrammable platform antibody targeting and dual affinity (DART platform), DART-Fc, scFvHSA-scFv (where HSA is human serum albumin) or dock-and-lock antibody (DNL)-Fab3. In another specific embodiment of any of the anti-BCMA antibodies or BCMA-binding fragments thereof provided herein, the antibody or fragment is an antibody-drug conjugate.

In another aspect, provided herein is a polypeptide that comprises the heavy chain CDR1, CDR2, or CDR3 sequences of SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9, respectively; SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively; SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15, respectively; SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively; SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO: 23, respectively; SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID NO: 26, respectively; SEQ ID NO: 29, SEQ ID NO: 30 and SEQ ID NO: 31, respectively; SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34, respectively; SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37, respectively; SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively; SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45, respectively; SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, respectively; SEQ ID NO: 51, SEQ ID NO: 52 and SEQ ID NO: 53, respectively; SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, respectively; or SEQ ID NO: 57, SEQ ID NO: 58 and SEQ ID NO: 59, respectively. In another embodiment, provided herein is a polypeptide that contains the light chain sequences CDR1, CDR2, and CDR3 of SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4, respectively. In a specific embodiment of any of the polypeptides provided herein, the polypeptide further comprises the light chain CDR1, CDR2, or CDR3 sequences of SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4, respectively. In a specific embodiment of any of the polypeptides provided herein, the polypeptide further comprises the light chain sequences CDR1, CDR2, or CDR3 of SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4, respectively. Further provided herein is a polypeptide that contains a light chain variable sequence of SEQ ID NO: 1. Further provided herein is a polypeptide that contains a heavy chain variable sequence of SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 28, SEQ ID NO: 39 or SEQ ID NO: 50.

In another aspect, provided herein is a composition comprising any of the antibodies, binding fragments thereof, or polypeptides provided herein. In a specific embodiment, the composition is a pharmaceutical composition. In other specific embodiments, the composition is formulated for intravenous, intraarterial, intramuscular, intradermal, subcutaneous, intradural, intrathecal, or intraperitoneal delivery.

In another aspect, provided herein are polynucleotides encoding any of

- 3 044652 antibodies, antibody fragments or polypeptides proposed herein. In one embodiment, provided herein is a polynucleotide encoding an anti-BCMA antibody, a BCMA-binding antibody fragment, or a polypeptide comprising the heavy chain CDR1, CDR2, or CDR3 sequences of SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9. respectively; SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively; SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15, respectively; SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively; SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO: 23, respectively; SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID NO: 26, respectively; SEQ ID NO: 29, SEQ ID NO: 30 and SEQ ID NO: 31, respectively; SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34, respectively; SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37, respectively; SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively; SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45, respectively; SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, respectively; SEQ ID NO: 51, SEQ ID NO: 52 and SEQ ID NO: 53, respectively; SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, respectively; or SEQ ID NO: 57, SEQ ID NO: 58 and SEQ ID NO: 59, respectively. In a more specific embodiment of any of the polynucleotides provided herein, the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 5, SEQ ID NO: 16, SEQ ID NO: 27, SEQ ID NO: 38, SEQ ID NO: 49, or SEQ ID NO: 60. In another embodiment, provided herein is a polynucleotide encoding an anti-BCMA antibody, a fragment of a BCMA binding antibody, or a polypeptide comprising the light chain CDR1, CDR2, or CDR3 sequences of SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO : 4, respectively. In another embodiment, provided herein is a polynucleotide that encodes an anti-BCMA antibody, a BCMA-binding antibody fragment, or a polypeptide comprising a light chain variable sequence of SEQ ID NO: 1. In another embodiment, provided herein is a polynucleotide that encodes an anti-BCMA antibody , a BCMA-binding antibody fragment or polypeptide comprising a heavy chain variable sequence of SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 28, SEQ ID NO: 39, or SEQ ID NO: 50. In a particular embodiment herein The document provides a polynucleotide that encodes an anti-BCMA antibody, a BCMA-binding antibody fragment or a polypeptide containing a light chain variable sequence of SEQ ID NO: 1 and a heavy chain variable sequence of SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 28, SEQ ID NO: 39 or SEQ ID NO: 50.

In another aspect, provided herein are polynucleotide vectors comprising the polynucleotides provided herein. In a specific embodiment, the vector is an expression vector. In another specific embodiment, the vector is a retroviral vector or a lentiviral vector.

In another aspect, provided herein is a cell comprising any of the polynucleotides provided herein or expressing any of the anti-BCMA antibodies, BCMA-binding fragments thereof, or polypeptides provided herein. In a specific embodiment, the cell contains any of the vectors provided herein.

In another aspect, provided herein is a method for producing a polypeptide, comprising causing, in any of the cells provided herein, expression of a polynucleotide provided herein thereby producing said polypeptide; and isolating said polypeptide. In addition, provided herein is a method of producing an anti-BCMA antibody or a BCMA-binding fragment thereof, comprising inducing in any of the cells provided herein expression of a polynucleotide provided herein, thereby producing said antibody or fragment thereof; and isolating said antibody or fragment thereof.

In another aspect, provided herein is a method of depleting BCMA-expressing cells in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of any of the anti-BCMA antibodies, a BCMA-binding fragment thereof, or polypeptides provided herein. Further provided herein is a method of depleting BCMA expressing cells in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of any of the anti-BCMA antibodies, a BCMA binding fragment thereof, or polypeptides provided herein. Further provided herein is a method of treating a disease caused by BCMA-expressing cells, such as BCMA-expressing plasma cells, in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of an anti-BCMA antibody, a BCMA-binding fragment thereof, or polypeptides provided herein. in this document. Further provided herein is a method of treating a B cell disease associated with BCMA expression in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of any of the anti-BCMA antibodies, a BCMA binding fragment thereof, or polypeptides provided in this document. In specific embodiments, the B cell-related disease is multiple myeloma, plasmacytoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, follicular lymphoma

- 4 044652 foma, small cell lymphoma with non-split nucleus, endemic Burkitt lymphoma, sporadic Burkitt lymphoma, marginal zone lymphoma, extra-nodal lymphoma of mucosal associated lymphoid tissue, nodular monocytoid B-cell lymphoma, splenic lymphoma, mantle cell lymphoma, large cell lymphoma, diffuse mixed cell lymphoma, diffuse large B-cell lymphoma (DLBCL), indolent lymphoma, lymphoplasmacytic B-cell lymphoma, immunoblastic lymphoma, primary mediastinal B-cell lymphoma, pulmonary angiocentric B-cell lymphoma, small lymphocytic lymphoma, chronic lymphocytic leukemia, B-cell proliferator of uncertain malignant potential, lymphomatoid granulomatosis, post-transplant lymphoproliferative disease, immunoregulatory disease, rheumatoid arthritis, myasthenia gravis, idiopathic thrombocytopenic purpura, antiphospholipid syndrome, Chagas disease, Graves' disease, Wegener's granulomatosis, polyarteritis nodosa, Sjögre's syndrome on, vulgar pemphigus, scleroderma, multiple sclerosis, antiphospholipid syndrome, ANCA-associated vasculitis, Goodpasture's disease, Kawasaki disease, autoimmune hemolytic anemia, systemic lupus erythematosus, rapidly progressive glomerulonephritis, heavy chain disease, primary or immune cell-associated amyloidosis, cutaneous lupus erythematosus, monoclonal gammopathy of undetermined significance or glioblastoma.

In a further aspect, provided herein are chimeric antigen receptors (CARs) that comprise any of the heavy or light chain CDR sequences provided herein or any of the heavy or light chain variable sequences provided herein. In one embodiment, the CAR contains the specified heavy chain CDR1, CDR2 and CDR3 sequences and/or the specified light chain CDR1, CDR2 and CDR3 sequences in the extracellular BCMA-binding domain. In another embodiment, the CAR contains the specified light chain sequence and/or the specified heavy chain sequence in the extracellular BCMA-binding domain. In another embodiment, the CAR comprises or further comprises one or more of: a transmembrane domain, a primary signaling domain, and/or a co-stimulatory domain. Also provided herein are cells, such as immune cells, that express any of the CARs provided herein. In more specific embodiments, the cells are T cells, natural killer cells (NK cells), or natural killer T cells (NKT cells). Further provided herein is a method of depleting BCMA expressing cells, such as plasma cells, from a patient in need thereof, comprising administering to the patient a therapeutically effective amount of anti-BCMA CAR expressing cells as provided herein. A method of treating a B cell disease associated with BCMA expression in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of cells according to any of the embodiments provided herein. In a specific embodiment of any of the methods comprising a cell expressing an anti-BCMA CAR as provided herein, the B cell-related disease is multiple myeloma, plasmacytoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, follicular lymphoma, small cell nuclear fusion lymphoma , endemic Burkitt's lymphoma, sporadic Burkitt's lymphoma, marginal zone lymphoma, extra-nodal lymphoma of mucous membrane-associated lymphoid tissue, nodular monocytoid B cell lymphoma, splenic lymphoma, mantle cell lymphoma, large cell lymphoma, diffuse mixed cell lymphoma, diffuse large cell B -cell lymphoma (DLBCL), indolent lymphoma, lymphoplasmic B-cell lymphoma, immunoblastic lymphoma, primary mediastinal B-cell lymphoma, pulmonary angiocentric B-cell lymphoma, small lymphocytic lymphoma, chronic lymphocytic leukemia, proliferating B-cells of uncertain malignant potential, lymphomatoid granulomatosis, post-transplant lymphoproliferative disease, immunoregulatory disease, rheumatoid arthritis, myasthenia gravis, idiopathic thrombocytopenic purpura, antiphospholipid syndrome, Chagas disease, Graves' disease, Wegener's granulomatosis, polyarteritis nodosa, Sjögren's syndrome, pemphigus vulgaris, scleroderma, multiple sclerosis, antiphospholipid ny syndrome, ANCA-associated vasculitis , Goodpasture's disease, Kawasaki disease, autoimmune hemolytic anemia, systemic lupus erythematosus, rapidly progressive glomerulonephritis, heavy chain disease, primary or immune cell-associated amyloidosis, cutaneous lupus erythematosus, monoclonal gammopathy of undetermined significance, or glioblastoma.

5. Shapes

Fig. 1 is a schematic illustration of avid (1A) and non-avid (1B) binding.

In fig. 2 are histograms showing that human IgGs, 320262, 319966, 320111, 319883, and 320199, bind C6 cells expressing human BCMA.

In fig. 3 shows histograms demonstrating that human IgG, 320262, 319966,

- 5 044652

320111, 319883, and 320199 do not bind Hek293 cells expressing the TNF receptor family member, TNFRSF12A.

In fig. 4 shows histograms confirming that human IgG, 320262, 319966, 320111, 319883 and 320199 bind human multiple myeloma cells NCIH929, which endogenously express BCMA.

In fig. Figure 5 shows dose-response curves for the binding of human IgG, 320262, 319966, 320111, 319883 and 320199, to BCMA-positive human multiple myeloma cells U266B1 and KMS12BM. In fig. 5 also shows histograms depicting the expression profiles of TNF receptor family members, BCMA, TACI, and BAFF, in OPM2 and NUDHL1 cells using commercially available antibodies. Finally, the figure shows dose-response curves for the binding of antibodies 320111 and 320262 to OPM2 cells (BCMA⁺/TACI7BAFFR^-), but not with NUDHL1 cells (BCMA /TACI+/BAFFR+).

6. Detailed description

Provided herein are polypeptides, for example, BCMA-binding polypeptides that contain specific CDR1, CDR2, and CDR3 sequences, and BCMA-binding antibodies, antibody fragments, and polypeptides that contain such CDR1, CDR2, and CDR3 sequences. Also provided herein are methods of using such antibodies, antibody fragments and polypeptides for therapeutic purposes, for example, for treating subjects suffering from a plasma cell or B cell disease or disorder.

As used herein, CDR means complementarity determining region, which represents certain sequences of the heavy or light chains of an antibody that mediate binding between the antibody and the antigen or epitope to which the antibody is directed; amino acid residues of an antibody that (usually three or four short regions of extreme sequence variability) are found in the V region domain of an immunoglobulin that form the antigen-binding site and are the major determinants of antigen specificity. The CDR sequence may be defined or numbered, for example, according to the Kabat system (see Kabat, et al., 1983. Sequence of Proteins of Immunological Interest. National Institutes of Health, Bethesda, Maryland); according to the Chothia system (see Chothia &, Lesk, “Canonical Structures for the Hypervariable Regions of Immunoglobulins,”

I J. Mol. Biol. 196, 901-917 (1987)); or according to the IMGT system (see Lefranc et al., “IMGT Unique Numbering for Immunoglobulin and Cell Receptor Variable Domains and Ig superfamily V-like domains,” Dev. Comp. Immunol. 27, 55-77 (2003)). See also Abhinandan & Martin, “Analysis and Improvements to Kabat and Structurally Correct Numbering of Antibody Variable Domains,” Mol. Immunol. 45:3832 (2008).

6.1. Polypeptides.

Provided herein is a polypeptide that contains specific heavy chain CDR1, CDR2 and CDR3 sequences. In specific embodiments of the invention, such polypeptides are capable of binding BCMA, either alone or in combination with a polypeptide containing specific light chain CDR1, CDR2 and CDR3 sequences.

In a first embodiment, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 7, 8 and 9, respectively. In addition, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 10, 11 and 12, respectively. Further provided herein is a polypeptide that contains CDR1, CDR2 and/or CDR3 sequences of SEQ ID NOs: 13, 14 and 15, respectively. In a particular embodiment of any of these embodiments, the polypeptide comprises a heavy chain variable region sequence of SEQ ID NO: 2. In a particular embodiment of any of these embodiments, the polypeptide is a Fab fragment that contains a heavy chain variable region sequence of SEQ ID NO: 6. B In another specific embodiment of any of these embodiments, the polypeptide is an F(ab')2 fragment that contains the heavy chain variable region sequence of SEQ ID NO: 6.

In another embodiment, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 18, 19 and 20, respectively. Further provided herein is a polypeptide that contains CDR1, CDR2 and/or CDR3 sequences of SEQ ID NOs: 21, 22 and 23, respectively. Further provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 24, 25 and 26, respectively. In a particular embodiment of any of these embodiments, the polypeptide comprises a heavy chain variable region sequence of SEQ ID NO: 17. In a particular embodiment of any of these embodiments, the polypeptide is a Fab fragment that contains a heavy chain variable region sequence of SEQ ID NO: 17. In a different

- 6 044652 in a specific embodiment of any of these embodiments, the polypeptide is an F(ab') ₂ fragment that contains the heavy chain variable region sequence of SEQ ID NO: 17.

In another embodiment, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 29, 30 and 31, respectively. In addition, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 32, 33 and 34, respectively. Further provided herein is a polypeptide that contains CDR1, CDR2 and/or CDR3 sequences of SEQ ID NOs: 35, 36 and 37, respectively. In a particular embodiment of any of these embodiments, the polypeptide comprises a heavy chain variable region sequence of SEQ ID NO: 28. In a particular embodiment of any of these embodiments, the polypeptide is a Fab fragment that contains a heavy chain variable region sequence of SEQ ID NO: 28. In another specific embodiment of any of these embodiments, the polypeptide is an F(ab') ₂ fragment that contains the heavy chain variable region sequence of SEQ ID NO:28.

In another embodiment, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 40, 41 and 42, respectively. In addition, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 43, 44 and 45, respectively. In addition, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 46, 47 and 48, respectively. In a particular embodiment of any of these embodiments, the polypeptide comprises a heavy chain variable region sequence of SEQ ID NO: 39. In a particular embodiment of any of these embodiments, the polypeptide is a Fab fragment that contains a heavy chain variable region sequence of SEQ ID NO: 39. B In another specific embodiment of any of these embodiments, the polypeptide is an F(ab')2 fragment that contains the heavy chain variable region sequence of SEQ ID NO: 39.

In another embodiment, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 51, 52 and 53, respectively. In addition, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 54, 55 and 56, respectively. In addition, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 57, 58 and 59, respectively. In a particular embodiment of any of these embodiments, the polypeptide comprises a heavy chain variable region sequence of SEQ ID NO: 50. In a particular embodiment of any of these embodiments, the polypeptide is a Fab fragment that contains a heavy chain variable region sequence of SEQ ID NO: 50. B In another specific embodiment of any of these embodiments, the polypeptide is an F(ab') ₂ fragment that contains the heavy chain variable region sequence of SEQ ID NO: 50.

Also provided herein is an antibody that contains specific light chain CDR1, CDR2 and CDR3 sequences. In specific embodiments, such antibodies are capable of binding BCMA either alone or in combination with the heavy chain CDR1, CDR2 and CDR3 sequences. In a first embodiment, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 2, 3 and 4, respectively. In a specific embodiment of any of these embodiments, the antibodies comprise a light chain variable region sequence of SEQ ID NO: 1. In a particular embodiment of any of these embodiments, the antibodies are a Fab fragment that contains a light chain variable region sequence of SEQ ID NO: 1 In another specific embodiment, the antibody is an F(ab') ₂ fragment that contains the light chain variable region sequence of SEQ ID NO: 1.

Provided herein is an antibody that contains specific heavy chain CDR1, CDR2 and CDR3 sequences. In specific embodiments of the invention, such polypeptides are capable of binding BCMA, either alone or in combination with a polypeptide containing specific light chain CDR1, CDR2 and CDR3 sequences.

In a first embodiment, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 7, 8 and 9, respectively. In addition, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 10, 11 and 12, respectively. In addition, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 13, 14 and 15, respectively. In a specific embodiment of any of these embodiments, the antibody comprises a heavy chain variable region sequence of SEQ ID NO: 2. In a particular embodiment of any of these embodiments, the antibody is a Fab fragment that contains a heavy chain variable region sequence of SEQ ID NO: 6 In another specific embodiment, the antibody is an F(ab') ₂ fragment that contains

- 7 044652 heavy chain variable region sequence SEQ ID NO: 6.

In another embodiment, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 18, 19 and 20, respectively. In addition, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 21, 22 and 23, respectively. In addition, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 24, 25 and 26, respectively. In a specific embodiment of any of these embodiments, the antibody comprises a heavy chain variable region sequence of SEQ ID NO: 17. In a particular embodiment of any of these embodiments, the antibody is a Fab fragment that contains a heavy chain variable region sequence of SEQ ID NO: 17 In another specific embodiment of any of these embodiments, the antibody is an F(ab') ₂ fragment that contains the heavy chain variable region sequence of SEQ ID NO: 17.

In another embodiment, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 29, 30 and 31, respectively. In addition, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 32, 33 and 34, respectively. In addition, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 35, 36 and 37, respectively. In a specific embodiment of any of these embodiments, the antibody comprises a heavy chain variable region sequence of SEQ ID NO: 28. In a particular embodiment of any of these embodiments, the antibody is a Fab fragment that contains a heavy chain variable region sequence of SEQ ID NO: 28 In another specific embodiment of any of these embodiments, the antibody is an F(ab')2 fragment that contains the heavy chain variable region sequence of SEQ ID NO: 28.

In another embodiment, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 40, 41 and 42, respectively. In addition, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 43, 44 and 45, respectively. In addition, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 46, 47 and 48, respectively. In a particular embodiment of any of these embodiments, the antibody comprises a heavy chain variable region sequence of SEQ ID NO: 39. In a particular embodiment of any of these embodiments, the antibody is a Fab fragment that contains a heavy chain variable region sequence of SEQ ID NO: 39 In another specific embodiment of any of these embodiments, the antibody is an F(ab')2 fragment that contains the heavy chain variable region sequence of SEQ ID NO: 39.

In another embodiment, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 51, 52 and 53, respectively. In addition, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 54, 55 and 56, respectively. In addition, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 57, 58 and 59, respectively. In a specific embodiment of any of these embodiments, the antibody comprises a heavy chain variable region sequence of SEQ ID NO: 50. In a particular embodiment of any of these embodiments, the antibody is a Fab fragment that contains a heavy chain variable region sequence of SEQ ID NO: 50 In another specific embodiment of any of these embodiments, the antibody is an F(ab') ₂ fragment that contains the heavy chain variable region sequence of SEQ ID NO: 50.

Also provided herein is a polypeptide that contains specific light chain CDR1, CDR2 and CDR3 sequences. In specific embodiments, such polypeptides are capable of binding BCMA either alone or in combination with a polypeptide containing specific CDR1, CDR2 and CDR3 sequences. In a first embodiment, provided herein is a polypeptide that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 2, 3 and 4, respectively. In a particular embodiment of any of these embodiments, the polypeptide comprises a light chain variable region sequence of SEQ ID NO: 1. In a particular embodiment of any of these embodiments, the polypeptide is a Fab fragment that contains a light chain variable region sequence of SEQ ID NO: 1. In another specific embodiment of any of these embodiments, the polypeptide is an F(ab') ₂ fragment that contains the light chain variable region sequence of SEQ ID NO: 1.

Also provided herein is an antibody that contains specific sequences

- 8 044652 activity of CDR1, CDR2 and CDR3 of the light chain. In a first embodiment, provided herein is an antibody that contains the sequences CDR1, CDR2 and/or CDR3 of SEQ ID NO: 2, 3 and 4, respectively. In a specific embodiment of any of these embodiments, the antibody comprises a light chain variable region sequence of SEQ ID NO: 1. In a particular embodiment of any of these embodiments, the antibody is a Fab fragment that contains a light chain variable region sequence of SEQ ID NO: 1. B In another specific embodiment of any of these embodiments, the antibody is an F(ab') ₂ fragment that contains the light chain variable region sequence of SEQ ID NO: 1.

In more specific embodiments of any of the foregoing antibodies or BCMA-binding fragments thereof, the antibody or BCMA-binding fragment thereof comprises at least 80%, at least 85%, at least 90% of the light chain variable domain amino acid sequence. at least 95, at least 96, at least 97, at least 98, at least 99, or 100% identical to SEQ ID NO: 1, and the amino acid sequence of the heavy chain variable domain at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, at least 99, or 100% identical to SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 28, SEQ ID NO: 39 or SEQ ID NO: 50.

In a more specific embodiment of any of the above antibodies or BCMA binding fragments thereof, the antibody or BCMA binding fragment thereof comprises a light chain variable domain amino acid sequence of SEQ ID NO: 1, or a light chain variable domain amino acid sequence of at least 80 by at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, or at least 99% identical to SEQ ID NO: 1. In other more specific In embodiments of any of the above antibodies or BCMA-binding fragments thereof, the antibody or BCMA-binding fragment thereof comprises the heavy chain variable domain amino acid sequence of SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 28, SEQ ID NO: 39, or SEQ ID NO: 50, or the amino acid sequence of the heavy chain variable domain of at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, according to at least 98% or at least 99% identical to SEQ ID NO: 6, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 39, or SEQ ID NO: 50. In another more specific embodiment, any of of the above antibodies or BCMA-binding fragments thereof, the antibody or BCMA-binding fragment thereof contains the amino acid sequence of the light chain variable domain at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, at least 99, or 100% identical to SEQ ID NO: 1, and the amino acid sequence of the heavy chain variable domain at least 80, at least 85, according to at least 90, at least 95, at least 96, at least 97, at least 98, at least 99 or 100% identical to SEQ ID NO: 6, SEQ ID NO: 17 , SEQ ID NO: 28, SEQ ID NO: 39, or SEQ ID NO: 50.

In a more specific embodiment of any of the above antibodies or BCMA binding fragments thereof, the antibody or BCMA binding fragment thereof comprises a light chain variable domain amino acid sequence that comprises the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 and is at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98 or at least 99% identical to SEQ ID NO: 1. In other more specific embodiments of any of the above antibodies or BCMA binding fragments thereof, the antibody or BCMA binding fragment thereof comprises the amino acid sequence of the heavy chain variable domain SEQ ID NO: 7-9, 10-12 or 13- 15 and are at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98 or at least 99 % identical to SEQ ID NO: 6; the antibody or BCMA-binding fragment thereof contains the amino acid sequence of the heavy chain variable domain of SEQ ID NO: 18-20, 21-23 or 24-26 and is at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, or at least 99% identical to SEQ ID NO: 17; the antibody or BCMA-binding fragment thereof contains the amino acid sequence of the heavy chain variable domain of SEQ ID NO: 29-31, 32-34 or 35-37 and is at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, or at least 99% identical to SEQ ID NO: 28; the antibody or BCMA-binding fragment thereof contains the amino acid sequence of the heavy chain variable domain of SEQ ID NO: 40-42, 43-45 or 46-48 and is at least 80, at least 85, at least 90, at least 95, at least 96, according to

- 9 044652 at least 97, at least 98 or at least 99% identical to SEQ ID NO: 39; the antibody or BCMA-binding fragment thereof contains the amino acid sequence of the heavy chain variable domain of SEQ ID NO: 51-53, 54-56 or 57-59 and is at least 80%, at least 85%, at least 90 , at least 95, at least 96, at least 97, at least 98, or at least 99% identical to SEQ ID NO: 50.

6.2. Antibodies.

Also provided herein are antibodies that are or contain CDR sequences or heavy or light chain variable sequences.

The term antibody, as used herein, means any natural or engineered configuration of an antigen-binding polypeptide comprising one, two or three light chain CDRs and one, two or three heavy chain CDRs, wherein the polypeptide is capable of binding to an antigen.

In some embodiments, the antibody of the invention is an antibody with or without an Fc portion. In some embodiments, the antibodies provided herein are IgG antibodies, such as IgG1, IgG2, IgG3, or IgG4 antibodies. In some embodiments, the antibodies are IgA antibodies, IgE antibodies, IgD antibodies, or IgM antibodies. In some embodiments, the antibodies are monoclonal antibodies, such as IgG monoclonal antibodies.

In some embodiments, any antibody provided herein contains an Fc variant for the Fc region of a wild-type human IgG, wherein the antibody has reduced affinity for human FcyRIIIA and/or FcyRIIA and/or FcyRI compared to an antibody containing Fc- wild type IgG region.

In some embodiments, the antibodies are artificially produced fully human antibodies, such as mouse or rat antibodies, genetically modified to produce human antibodies, such as OMNIRAT or OMNIMOUSE (Ligand Pharmaceuticals). Such antibodies do not occur naturally, but have the framework and constant regions of human antibodies. Because human BCMA is a self-antigen, the human body does not produce antibodies to human BCMA. Human anti-BCMA antibodies may be antibodies having variable and constant regions substantially corresponding to human germline immunoglobulin sequences known in the art, including, for example, those described by Kabat et al. The human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or as a result of somatic mutation in vivo), for example, in the CDR, and in particular in CDR3. Human anti-BCMA antibodies provided herein may have at least one, two, three, four, five or more positions replaced by an amino acid residue that is not encoded by a human germline immunoglobulin sequence.

In some embodiments, the antibodies provided herein are monoclonal antibodies. As used herein, the term monoclonal antibody refers to an antibody derived from a population of essentially homogeneous antibodies, that is, the individual antibodies comprising the population are identical except for possible mutations and/or post-translational modifications (e.g., isomerization, amidation) that may be present in small quantities. This term can be used to refer to such a homogeneous population of antibodies. Monoclonal antibodies are highly specific, directed against a single antigenic site. Additionally, unlike polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on an antigen. In addition to their specificity, monoclonal antibodies have the advantage that they can be synthesized by hybridoma cultures without being contaminated by other immunoglobulins. The designation monoclonal refers to the nature of the antibody as being derived from a substantially homogeneous population of antibodies, and should not be construed as requiring the antibody to be produced by any particular method. For example, the anti-BCMA monoclonal antibodies provided herein can be produced by the hybridoma method first described by Kohler et al., Nature, 256:495 (1975), or can be produced by recombinant DNA methods (see, for example, US Pat. No. 4,816,567). . Monoclonal antibodies can also be isolated from phage antibody libraries using techniques such as those described in Clackson et al., Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991).

In some embodiments, the antibodies provided herein, such as monoclonal antibodies, are chimeric antibodies (immunoglobulins) in which a region of the heavy and/or light chain is identical or homologous to the corresponding sequences in antibodies derived from specific biological species, or relative to

- 10 044652 refers to a specific class or subclass of antibodies, while the rest of the chain(s) is identical or homologous to the corresponding sequences in antibodies obtained from another biological species or belonging to a different class or subclass of antibodies, and also represent fragments of such antibodies. Preferably, such chimeric antibodies exhibit the desired biological activity, such as binding to BCMA (US Pat. No. 4,816,567; Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). In some embodiments, chimeric antibodies provided herein include primatized antibodies comprising antigen binding variable domain sequences derived from non-human primates (eg, cynomolgus monkeys, monkeys, etc.) and human antibody constant region sequences.

In some embodiments, the BCMA-binding antibodies provided herein are monovalent. In other embodiments, the BCMA-binding antibodies provided herein are multivalent, such as divalent, trivalent, or tetravalent. In some embodiments, the BCMA-binding antibodies provided herein are monospecific. In other embodiments, the BCMA-binding antibodies provided herein are multispecific, such as bispecific, trispecific, etc. In some embodiments, the BCMA-binding antibodies provided herein, which are multivalent, bind two or more epitopes on BCMA.

In some other embodiments, antibodies, such as monoclonal antibodies, provided herein may be humanized antibodies that can be produced using recombinant DNA techniques known in the art. Various approaches to the production of chimeric antibodies have been described. See, for example, Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851 (1985); Takeda et al., Nature 314:452 (1985), Cabilly et al., US Pat. No. 4,816,567; Boss et al., US Patent No. 4816397; Tanaguchi et al., EP 0171496; EP 0173494, GB 2177096.

In other embodiments, the antibody provided herein is a bispecific antibody; that is, it targets a second antigen in addition to BCMA. A bispecific or bifunctional antibody or immunoglobulin is an artificial hybrid antibody or immunoglobulin having two different heavy/light chain pairs and two different binding sites. Bispecific antibodies can be produced by a variety of methods, including hybridoma fusion or the binding of Fab' fragments. See, for example, Songsivilai & Lachmann, Clin. Exp. Immunol. 79: 315-321 (1990). Numerous methods known to those skilled in the art are available for the production of antibodies or antigen-binding fragments thereof. For example, antibodies can be produced using recombinant DNA techniques (see, for example, US Pat. No. 4,816,567). In the recent past, a large number of recombinant bispecific antibody formats have been developed (see, for example, Kontermann RE, mAbs 4:2, (2012) 1-16). In specific embodiments, the antibody is a diabody, a bispecific antibody, a peak-and-groove bispecific antibody, a CrossMAB bispecific antibody, for example, where VL and VH or CH1 and CL of one or more Fab fragments are swapped, bispecific Fab ₂ and /or bispecific (mab) ₂ , etc. In a specific embodiment, the antiBCMA Fab includes CrossMAB technology. In another specific embodiment, said second antigen is CD19.

In other specific embodiments, the antibody is a bispecific agent that interacts with T cells. In a more specific embodiment, the second antigen targeted by the T cell activator is CD3. In other specific embodiments, the antibody is a bispecific agent that interacts with NK cells. In a more specific embodiment, the second antigen targeted by the NK cell activator is NKG2D.

In some embodiments, the bispecific antibody contains no more than two Fab fragments from an anti-BCMA antibody and no more than one Fab fragment from a second antibody, for example, no more than one Fab fragment from an anti-CD3 antibody, (2+1 design) , optionally, with no more than one Fc part.

In other embodiments, provided herein are fragments of BCMA-binding antibodies, such as single chain antibodies, Fab fragments, or F'(ab) ₂ fragments.

Also provided herein are BCMA-binding antibodies in other configurations, e.g., wherein said antibody is or includes Triomab, a common light chain IgG, CrossMAb, e.g., wherein VL and VH or CH1 and CL of one or more Fab fragments are swapped in places, ortho-Fab IgG, DVD-Ig, 2 in 1-IgG, IgG-scFv, sdFv2-Fc, bi-nanobody, TandAb, antibody on a target-reprogrammable dual-affinity platform (DART platform, for example from Creative Biolabs) , DART-Fc, scFv-HSA-scFv (where HSA is human serum albumin), dock-and-lock antibody (DNL)-Fab3, ImmTAC, DAF,

- 11 044652

HSA body, IgG-finomer and ART-Ig.

6.3. Antibody-drug conjugates.

In another aspect, in some embodiments, each or any of the antibodies or BCMA-binding moieties thereof may be conjugated to a drug, eg, a toxin or toxic moiety, protein, polysaccharide, or small molecule compound, to form an antibody-drug conjugate. In one embodiment, the drug is, for example, one or more of an anti-apoptotic agent, a mitotic inhibitor, an antitumor antibiotic, an immunomodulatory agent, a gene therapy nucleic acid, an alkylating agent, an anti-angiogenic agent, an antimetabolite, a boron agent, a chemoprotective agent, hormonal agent, antihormonal agent, corticosteroid, photoactive therapeutic agent, oligonucleotide, radionuclide agent, radiosensitizer, topoisomerase inhibitor and tyrosine kinase inhibitor. In some embodiments, the mitotic inhibitor is dolastatin, auristatin, a maytansinoid, and a plant alkaloid. In some embodiments, the drug is a dolastatin, auristatin, a maytansinoid, and a plant alkaloid. An example of an auristatin is monomethyl auristatin F (MMAF) or monomethyl auristatin E (MMAE). Examples of maytansinoids include, but are not limited to, DM1, DM2, DM3 and DM4. In some embodiments, the antitumor antibiotic is selected from the group consisting of actinomycin, anthracycline, calicheamicin, and duocarmycin. In some embodiments, actinomycin is a pyrrolobenzodiazepine (PBD).

In some embodiments in which the drug is a maytansinoid, the maytansinoid is a thiol-containing maytansinoid prepared, for example, in accordance with the methods disclosed in US Pat. No. 6,333,410. In a particular embodiment, the maytansinoid is DM-1 (N2-deacetyl -N2-(3-mercapto-1oxopropyl)maytansine). DM-1 is 3-10 times more cytotoxic than maytansine and can be converted into a prodrug by linking it via disulfide bond(s) to the anti-BCMA antibody or BCMA-binding fragment thereof provided herein. Some of these conjugates (sometimes called tumor-activated prodrugs (TAPs)) are not cytotoxic in the bloodstream because they are activated upon binding to target cells and are internalized into the cell, thereby releasing the drug compound. In other specific embodiments, maytansinoids contain a side chain that contains a sterically closed thiol bond, such as, but not limited to, the maytansinoids N2deacetyl-N2-(4-mercapto-1-oxopentyl)-maytansine, also referred to as DM3 and N2-deacetyl-N2 -(4methyl-4-mercapto-1-oxopentyl) maytansine, also called DM4. DM4 differs from DM1 and DM3 in that it carries methyl groups on its AC. This results in steric hindrance when DM4 is attached via a linker, particularly but not limited to a disulfide bond-containing linker, to a targeting agent such as nBT062. A wide variety of maytansinoids bearing a sterically closed thiol group (having one or two substituents, particularly alkyl substituents such as the methyl substituents in DM4) are disclosed in US Patent Publication No. 2004/0235840, which is incorporated herein by reference in its entirety. Other effector molecules may be used containing substituents (such as alkyl groups) at positions that provide steric closure when the effector is attached to the targeting agent via a linker. In some specific embodiments, this closure causes a chemical modification, such as alkylation of the free drug compound, to increase its overall stability, which allows the drug compound not only to cause cell death or permanent cell cycle arrest in BCMA-expressing tumor cells, but optionally also may target supporting cells that, for example, support or protect the tumor from drugs, in particular cells of the tumor stroma and tumor vasculature, and that do not normally express BCMA, to reduce or suppress their supportive or protective function. DNA alkylating agents can also be used as effector molecules and include, without limitation, CC-1065 analogs or derivatives. CC-1065 is a potent antitumor antibiotic isolated from cultures of Streptomyces zelensis and has been shown to be exceptionally cytotoxic in vitro (US Patent No. 4,169,888).

In some other specific embodiments, the drug is a taxane, for example a taxane that contains one or more thiol or disulfide groups. Taxanes inhibit tubulin depolymerization, which leads to an increase in the rate of microtubule assembly and cell death. Taxanes that can be used in antibody-drug conjugates with the anti-BCMA antibodies or BCMA-binding fragments thereof provided herein are, for example, disclosed in US Patent Nos. 6,436,931, 6,340,701, 6,706,708 and US Patent Publications No. 2004/0087649. 2004/0024049 and 2003/0004210. Other taxanes that may be used are disclosed, for example, in US Pat. Nos. 6,002,023, 5,998,656, 5,892,063, 5,763,477, 5,705,508, 5,703,247, 5,367,086, and 6,596,757.

- 12 044652

In some embodiments, the drug is pomalidomide (4-amino-2-[(3RS)-(2,6-dioxoniperidin-3-yl)-1H-isoindole-1,3(2H)-dione). In another specific embodiment of the invention, the drug is thalidomide ((RS)-2-(2,6-dioxopiperidin-3-yl)-1H-isoindole-1,3(2H)-dione). In another specific embodiment, the drug is lenalidomide (3-(4-amino-1oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione). In another specific embodiment, the drug is 3-[4-(4-morpholin-4-ylmethylbenzyloxy)-1oxo-1,3-dihydro-isoindole-2-and 1]piperidine-2,6-dione. In another specific embodiment of the invention, the cereblon binding compound used in accordance with the methods described herein is 3-(4-((4-((4-(2,4-difluorophenyl)piperazin-1yl)methyl) benzyl)oxy)-1-oxoisoindolin-2-yl)piperidin-2,6-dione. See, for example, the disclosure of these compounds in US Patent Application Publication No. 2014/0162282, which is incorporated herein by reference in its entirety.

Compounds for the methods provided herein include, but are not limited to, immunomodulatory compounds (Celgene Corporation), a group of compounds that may be useful in the treatment of several types of human diseases, including some types of cancer.

As used herein (unless otherwise indicated), the term immunomodulatory compound may encompass certain small organic molecules that inhibit LPS-induced monocyte production of TNF-α, IL-1B, 1L-12, IL-6, MIP-1a, MCP-1, GM-CSF, GCSF and COX-2. These compounds can be produced synthetically or can be purchased commercially.

In some embodiments, the drug includes tetrasubstituted 2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolines described in US Pat. No. 5,798,368; 1-oxo- and 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)isoindolines (e.g. 4-methyl thalidomide derivatives), substituted 2-(2,6-dioxopiperidin-3-yl)phthalimides and substituted 2 -(2,6-dioxopiperidin-3yl)-1-oxoisoindoles, including, but not limited to, those disclosed in US patent No. 5635517, 6281230, 6316471, 6403613, 6476052 and 6555554; publication no. WO 02/059106). The entire contents of each of the patents and patent applications set forth herein are incorporated by reference.

The various compounds described herein contain one or more chiral centers and may exist as racemic mixtures of enantiomers or mixtures of diastereomers. Thus, the present document also provides for the use of stereomerically pure forms of such compounds, as well as the use of mixtures of these forms. For example, mixtures containing equal or unequal amounts of enantiomers of certain immunomodulatory compounds can be used. These isomers can be synthesized asymmetrically or separated using standard techniques such as chiral columns or chiral resolving agents. See, for example, Jacques, J., et al., Enantiomers, Racemates and Resolutions (Wiley-Interscience, New York, 1981); Wilen, S. H., et al., Tetrahedron 33:2725 (1977); Eliel, E. L., Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, S. H., Tables of Resolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind., 1972).

The immunomodulatory compounds provided herein include, but are not limited to, 1-oxo-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)isoindolines substituted with an amino group on the benzene ring as described in the patent US No. 5635517, which is incorporated herein by reference.

These compounds have structure I:

wherein one of X and Y is C=O, the other of X and Y is C=O or CH ₂ , and R ² is hydrogen or lower alkyl, particularly methyl. Specific immunomodulatory compounds include, but are not limited to:

- 13 044652

and their optically pure isomers.

The compounds can be prepared by standard synthetic methods (see, for example, US Pat. No. 5,635,517, incorporated herein by reference). Connections are also available from Celgene Corporation, Warren, NJ.

Other drugs used in the compositions described herein belong to the class of substituted 2-(2,6-dioxopiperidin-3-yl)phthalimides and substituted 2-(2,6dioxopiperidin-3-yl)-1-oxoisoindoles, such as disclosed in US Patent Nos. 6,281,230, 6,316,471, 6,335,349 and 6,476,052, and International Patent Application No. PCT/US97/13375 (International Publication No. WO 98/03502), each of which is incorporated herein by reference. Typical compounds have the formula:

R ⁴ in which: one of X and Y is C=O, and the other of X and Y is C=O or CH ₂ ; (i) each of R ¹ , R ² , R ³ and R ⁴ , independently of the others, is halogen, C1-4 alkyl or C1-4 alkoxy, or (ii) one of ^R1 , R ² , R ³ and R ⁴ are -NHR ⁵ and the remainder of R ¹ , R ² , R ³ and R ⁴ are hydrogen; R ⁵ represents hydrogen or alkyl with 1-8 carbon atoms; R ⁶ represents hydrogen, C1-8 alkyl, benzyl or halogen; provided that R ⁶ is not hydrogen if X and Y are C=O, and (i) each of R ¹ , R ² , R ³ and R ⁴ is fluorine, or (ii) one of R ¹ , R ² , R ³ or R ⁴ is an amino group.

Compounds representing this class have the formulas:

where R ¹ represents hydrogen or methyl. In a separate embodiment, provided herein is the use of enantiomerically pure forms (e.g., optically pure (R) or

- 14 044652 (S) enantiomers) of these compounds.

Other specific immunomodulatory compounds are the tetrasubstituted 2(2,6-dioxopiperidin-3-yl)-1-oxoisoindolines described in US Pat. No. 5,798,368, which is incorporated herein by reference. Typical compounds have the formula:

EL ^:

TO

O

R ¹ where each of R ¹ , R ² , R ³ and R ⁴ , independently of the others, represents a halogen, alkyl with 1-4 carbon atoms or an alkoxy group with 1-4 carbon atoms.

Other specific immunomodulatory compounds are 1-oxo- and 1,3-dioxo-2(2,6-dioxopiperidin-3-yl)isoindolines, disclosed in US Pat. No. 6,403,613, which is incorporated herein by reference. Typical compounds have the formula:

in which Y represents oxygen or _H2 , the first of ^R1 and ^R2 represents a halogen, alkyl, alkoxy group, alkylamino group, dialkylamino group, cyano group or carbamoyl group, the second of ^R1 and ^R2 , regardless of the first, represents hydrogen, a halogen, alkyl, alkoxy, alkylamino, dialkylamino, cyano or carbamoyl group, and

R ³ represents hydrogen, alkyl or benzyl.

Specific examples of compounds have the formula:

an alkoxy group containing from 1 to 4 carbon atoms, a dialkylamino group in which each alkyl has from 1 to 4 carbon atoms, a cyano group or a carbamoyl group; the second of R ¹ and R ² , regardless of the first, is hydrogen, halogen, alkyl containing from 1 to 4 carbon atoms, an alkoxy group containing from 1 to 4 carbon atoms, an alkylamino group in which the alkyl contains from 1 to 4 carbon atoms , a dialkylamino group in which each alkyl contains from 1 to 4 carbon atoms, a cyano group or a carbamoyl group; and R ³ represents hydrogen, alkyl containing from 1 to 4 carbon atoms, or benzyl. Specific examples include, but are not limited to, 1-oxo-2-(2,6dioxopiperidin-3-yl)-4-methylisoindoline.

where: the first of R ¹ and R ² is a halogen, an alkyl containing from 1 to 4 carbon atoms, an alkoxy group containing from 1 to 4 carbon atoms, a dialkylamino group in which each alkyl contains from 1 to 4 carbon atoms, a cyano group or a carbamoyl group group; the second of R ¹ and R ² , regardless of the first, is hydrogen, halogen, alkyl containing from 1 to 4 carbon atoms, an alkoxy group containing from 1 to 4 carbon atoms, an alkylamino group in which the alkyl contains from 1 to 4 carbon atoms , a dialkylamino group in which each alkyl contains from 1 to 4 carbon atoms, a cyano group or a carbamoyl group; and R ³ represents hydrogen, alkyl containing from 1 to 4 carbon atoms, or benzyl.

Other specific compounds provided herein have the formula:

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and their pharmaceutically acceptable salts, solvates and stereoisomers, where:

R ¹ represents: hydrogen; halogen; -(CH ₂ ) _p OH; (C ₁ -C ₆ )alkyl, optionally substituted with one or more halogens; (C1- _C6 )alkoxy group, optionally substituted with one or more halogens; or

-(CH2) _n NHR ^a , where R ^a is:

hydrogen;

(C1- _C6 )alkyl, optionally substituted with one or more halogens;

-(CH ₂ ) _n -(6-10 membered aryl);

-C(O)-(CH ₂ ) _n -(6-10 membered aryl) or -C(O)-(CH ₂ ) _n -(6-10 membered heteroaryl), where the aryl or heteroaryl is optionally substituted by one or several of: halogen; -SCF3; (C1- _C6 )alkyl, itself optionally substituted with one or more halogens; or a (C1- _C6 )alkoxy group, itself optionally substituted with one or more halogens;

-C(O)-(C ₁ -C ₈ )alkyl, where the alkyl is optionally substituted with one or more halogens;

-C(O)-(CH ₂ ) _n -(C ₃ -C ₁₀ -cycloalkyl);

-C(O)-(CH2) _n -NR ^b R ^c where R ^b and R ^c are each independently: hydrogen;

(C1- _C6 )alkyl, optionally substituted with one or more halogens;

(C1- _C6 )alkoxy group, optionally substituted with one or more halogens; or

6-10 membered aryl, optionally substituted with one or more of the following groups: gene halo;

(C1- _C6 )alkyl, itself optionally substituted with one or more halogens; or a (C1- _C6 )alkoxy group, itself optionally substituted with one or more halogens;

-C(O)-(CH2)n-O-(C1-C6)alkyl; or

-C(O)-(CH2)n-O-(CH2)n-(6-10 membered aryl);

R ² represents: hydrogen; -(CH ₂ ) _p OH; phenyl, -O-(C1- _C6 )alkyl; or (C1- _C6 )alkyl, optionally substituted with one or more halogens;

R ³ represents hydrogen; or (C1- _C6 )alkyl, optionally substituted with one or more halogens; and n has the value 0, 1 or 2.

Specific examples include, but are not limited to, 3-(5-amino-2-methyl-4-oxo-4Hquinazolin-3-yl)piperidin-2,6-dione (Compound A), which has the following structure:

or an enantiomer thereof, or a mixture of enantiomers thereof, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

Compound A can be prepared in accordance with the methods described in the Examples section presented herein or as described in US Pat. No. 7,635,700, the disclosure of which is incorporated herein by reference in its entirety. The compound may also be synthesized by other methods apparent to those skilled in the art based on the ideas presented herein. In some embodiments, Compound A is in the crystalline form described in US Provisional Patent Application No. 61/451,806, filed March 11, 2011, which is incorporated herein by reference in its entirety. In some embodiments, the hydrochloride salt of Compound A is used in the methods provided herein. Methods for treating, preventing and/or controlling cancer and other diseases using Compound A are described in US Pat. No. 8,802,685, which is incorporated herein by reference in its entirety.

Other specific compounds provided herein have the formula:

or is a pharmaceutically acceptable salt, solvate or stereoisomer thereof, wherein:

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X represents C=O or CH2;

R ¹ represents Y-R3;

R ² represents H or (C1-C ₆ )alkyl;

Y is a 6-10 membered aryl, heteroaryl or heterocycle, each of which may be optionally substituted with one or more halogens; or communication;

R ³ represents:

- (CH ₂ ) _n -aryl, -O-(CH ₂ ) _n -aryl or -(CH ₂ ) _n -O-aryl, where aryl is optionally substituted by one or more of: (C ₁ -C ₆ )alkyl, itself optionally substituted with one or more halogens; (C1- _C6 )alkoxy group, itself substituted by one or more halogens; oxo groups; amino groups; carboxyl group; cyano groups; hydroxyl group; halogen; deuterium; 6-10 membered aryl or heteroaryl, optionally substituted with one or more (C ₁ -C ₆ )alkyl, (C1C ₆ )alkoxy groups or halogens; -CONH2; or -COO-(C ₁ -C ₆ )alkyl, where the alkyl may be optionally substituted by one or more halogens;

- (CH ₂ ) _n -heterocycle, -O-(CH ₂ ) _n -heterocycle or -(CH ₂ ) _n -O-heterocycle, where the heterocycle is optionally substituted by one or more of: (C1-C ₆ )alkyl, optionally itself substituted with one or more halogens; (C1- _C6 )alkoxy group, itself substituted by one or more halogens; oxo groups; amino groups; carboxyl group; cyano groups; hydroxyl group; halogen; deuterium; 6-10 membered aryl or heteroaryl, optionally substituted with one or more (C ₁ C ₆ ) alkyls, (C 1 - C ₆ ) alkoxy groups or halogens; -CONH2; or -COO-(C1-C ₆ )alkyl, where the alkyl may be optionally substituted with one or more halogens; or

- (CH ₂ ) _n -heteroaryl, -O-(CH ₂ ) _n -heteroaryl or -(CH ₂ ) _n -O-heteroaryl, where the heteroaryl is optionally substituted by one or more of: (C1-C ₆ )alkyl, optionally itself substituted with one or more halogens; (C1- _C6 )alkoxy group, itself substituted by one or more halogens; oxo groups; amino groups; carboxyl group; cyano groups; hydroxyl group; halogen; deuterium; 6-10 membered aryl or heteroaryl, optionally substituted with one or more ( _C1C6 )alkyl, (C1- _C6 )alkoxy groups or halogens; -CONH2; or -COO-(C1-C ₆ )alkyl, where the alkyl may be optionally substituted with one or more halogens; and n is 0, 1, 2 or 3.

Specific examples include, but are not limited to, 3-(4-((4(morpholinomethyl)benzyl)oxy)-1-oxoisoindolin-2-yl)piperidin-2,-6-dione. In one embodiment, the compound is the (S) stereoisomer of 3-(4-((4(morpholinomethyl)benzyl)oxy)-1-oxoisoindolin-2-yl)piperidin-2,-6-dione (Compound B), e.g. , for use in the methods described herein. Racemic 3-(4-((4(morpholinomethyl)benzyl)oxy)-1-oxoisoindolin-2-yl)piperidin-2,6-dione and methods for its preparation are described in US Patent Publication No. 2011/0196150, which is incorporated in its entirety. this document by reference. Compound B has the following structure:

All described compounds can either be purchased from companies or prepared in accordance with methods described in patents or patent publications disclosed herein. In addition, optically pure compounds can be synthesized asymmetrically or separated using known resolving agents or chiral columns, as well as other standard methods of synthetic organic chemistry. Additional information regarding immunomodulatory compounds, their preparation and use can be found, for example, in US Patent Application Publications Nos. US 20060188475, US 20060205787 and US 20070049618, each of which is incorporated herein by reference in its entirety.

If there is a discrepancy between the structure depicted and the name given to that structure, the structure in the image is given greater weight. In addition, if the stereochemistry of a structure or portion of a structure is not indicated, for example, by thick or dotted lines, then the structure or portion of a structure should be interpreted as encompassing all of its stereoisomers.

6.4. Chimeric antigen receptors.

In some embodiments, any of the anti-BCMA antibodies or

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The BCMA-binding sites are included in the chimeric antigen receptor, for example, as a BCMA targeting domain or part of a BCMA targeting domain. In specific embodiments, the CAR comprises a fragment of a BCMA binding antibody provided herein, for example, a single chain Fv fragment or a Fab fragment.

In some embodiments, chimeric antigen receptors (CARs) are artificial membrane-bound proteins that direct T cells or natural killer (NK) cells to an antigen, such as BCMA, and stimulate the T cells or NK cells to kill the cell presenting the antigen. , for example BCMA. See, for example, Eshhar, US Pat. No. 7,741,465. At a minimum, the CARs provided herein contain an extracellular domain that binds BCMA; a transmembrane domain and an intracellular (cytoplasmic) signaling domain that transmits the primary activation signal to the immune cell. Under all other conditions, when CAR is expressed on the surface of, for example, a T lymphocyte, and the extracellular domain of CAR binds to BCMA, the intracellular signaling domain signals the T lymphocyte to activate and/or proliferate, and if BCMA is present on the cell surface, to kill the BCMA-expressing cell.

Because T cells require at least two signals, a primary activation signal and a co-stimulatory signal, to be fully activated, typically CARs also contain a co-stimulatory domain such that binding of the extracellular domain to BCMA on the cell surface results in transmission of both the primary activation signal and the co-stimulatory signal .

In some embodiments, the intracellular domain of a CAR is or includes an intracellular domain or motif of a protein that is expressed by T lymphocytes and triggers the activation and/or proliferation of said T lymphocytes. Such a domain or motif is capable of transmitting the primary antigen-binding signal, which is necessary for T-lymphocyte activation in response to antigen binding to the extracellular part of the CAR. Typically this domain or motif contains or is an ITAM (immunoreceptor tyrosine activation motif). ITAM-containing polypeptides suitable for CAR include, for example, the CD3 zeta chain (CD3Z) or ITAM-containing regions thereof. In a specific embodiment, the intracellular domain is a CD3Z intracellular signaling domain. In other specific embodiments, the intracellular domain is derived from a lymphocyte receptor chain, a TCR/CD3 complex protein, an Fc receptor subunit, or an IL-2 receptor subunit. In some embodiments, the primary signaling domain is or comprises a signaling domain from TCRZ, FcRy, FcRe, CD3y, CD3δ, CD3ε, CD5, CD22, CD79a, CD79b, CD278, FceRI, DAP10, DAP12, or CD66d.

In some embodiments, the CAR further comprises one or more co-stimulatory domains or motifs, for example, as part of the intracellular domain of the polypeptide. The one or more costimulatory domains or motifs may be or comprise one or more CD27 costimulatory polypeptide sequences, CD28 costimulatory polypeptide sequences, OX40 costimulatory polypeptide sequences (CD134), 4-1BB costimulatory polypeptide sequences (CD137), or inducible T cell costimulatory polypeptide sequences (CD137). ICOS) polypeptide, or other co-stimulatory domain or motif. In some other embodiments, the co-stimulatory domain is or includes a functional signaling domain derived from one or more MHC class I molecules, TNF receptor, immunoglobulin-like protein, cytokine receptor, integrin, signaling lymphocyte activation molecule (SLAM protein), NK-activating protein. cellular receptor, BTLA, Toll ligand receptor,

CD2, CD7, CD27, CD30, CD40, CDS, ICAM-1, LFA-1(CD1 la/CD18), B7-NZ, CDS, ICAM-1,

ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD 19, CD4, CD8aa, CD8p, IL2Rp, IL2Ry, IL7Ra, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CDlld, ITGAE, CD103, ITGAL, CDlla, LFA-1, ITGAM, CDllb, ITGAX, CDllc, ITGB1, CD29, ITGB2, CD18, LFA1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96, CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69 , SLAMF6 (NTB-A, Lyl08), SLAM (SLAMF1, CD 150, IPO-3), BLAME (SLAMF8), SELPLG (CD 162), LTBR, LAT, GADS, SLP-76 or PAG/Cbp.

The transmembrane region can be any transmembrane region that can be included in a functional CAR, typically a transmembrane region from a CD4 or CD8 molecule. In some embodiments, the transmembrane domain of the CAR is derived from an immune system protein that typically conveys an inhibitory signal to such immune system cells, for example, the transmembrane domain from CTLA4 (cytotoxic T lymphocyte antigen 4 or cytotoxic T lymphocyte associated protein 4) or PD- 1 (programmed death protein-1).

- 18 044652

In some embodiments, any of the T lymphocytes provided herein that contain multiple cell death polypeptides contain a transmembrane domain from CTLA4 or PD-1. In a specific embodiment of the invention, a T lymphocyte expressing said polypeptide or any of such polypeptides described herein is activated or stimulated to proliferate when said polypeptide binds to said antigen. In a specific embodiment of the invention, the polypeptide, when expressed on the surface of a T lymphocyte, directs the T lymphocyte to destroy a cell expressing the specified antigen.

In specific embodiments of any of the polypeptides herein in which the transmembrane domain of the polypeptide is derived from CTLA4, the transmembrane domain of CTLA4 is derived from mammalian, eg, human, primate, or rodent CTLA4, eg, murine CTLA4. Preferably, the transmembrane domain does not contain amino acids from the intracellular domain, extracellular domain, or intracellular or extracellular domain of CTLA4 or PD-1. Specific non-limiting examples of CTLA4 or PD-1 transmembrane domain sequences are provided below.

In a specific embodiment, the CTLA4 transmembrane domain is a polypeptide sequence encoded by exon 3 of the human CTLA4 gene. In another specific embodiment, the transmembrane domain of CTLA4 is or contains the amino acid sequence PEPCPDSDFLLWILAAVSSGLFFYSFLLTAVSLSKM (in three-letter code

Pro-Glu-ProCys-Pro-Asp-Ser-Asp-Phe-Leu-Leu-Trp-Ile-Leu-Ala-Ala-Val-Ser-Ser-Gly-Leu-Phe-Phe-TyrS er-Phe-Leu -Leu-Thr-Al a-Vai - S er-Leu-S er-Ly s-Met) (SEQ ID NO: 61). In another specific embodiment, the CTLA4 transmembrane domain is or comprises the polypeptide sequence encoded by nucleotides 610-722 of GenBank accession number NM_005214.4. In another specific embodiment, the transmembrane domain of CTLA4 is or includes the amino acid sequence PDSDFLLWILAAVSSGLFFYSFLLTAVSL (in three-letter code

Pro-Asp-Ser-Asp-PheLeu-Leu-Trp-Ile-Leu-Ala-Ala-Val-Ser-Ser-Gly-Leu-Phe-Phe-Tyr-Ser-Phe-Leu-Leu-Thr-AlaVal- Ser-Leu) (SEQ ID NO: 62). In another specific embodiment, the CTLA4 transmembrane domain is or comprises the polypeptide sequence encoded by nucleotides 636-699 of GenBank accession number NM_005214.4. In another specific embodiment, the transmembrane domain of CTLA4 is or contains the amino acid sequence FLLWILAAVSSGLFFYSFLLTAV (in three-letter code

Phe-Leu-LeuTrp-Ile-Leu-Ala-Ala-Val-Ser-Ser-Gly-Leu-Phe-Phe-Tyr-Ser-Phe-Leu-Leu-Thr-Ala-Val) (SEQ ID NO: 63 ). See, for example, the protein reference with accession number ENSP00000303939.3 in the Ensembl database. In another specific embodiment, the transmembrane domain of CTLA4 is or includes the polypeptide sequence FLLWILAAVSSGLFFYSFLLT (in three letter code

Phe-Leu-LeuTrp-Il e-Leu-Ala-Ala-Val-Ser-Ser-Gly-Leu-Phe-Phe-Tyr-Ser-Phe-Leu-Leu-Thr) (SEQ ID NO: 64), cm ., for example, the sequence with registration number UNIPROT P16410.

In another specific embodiment, the transmembrane domain of CTLA4 is or contains the polypeptide sequence FLLWILVAVSLGLFFYSFLVSAVSLS (in three-letter code

Phe-Leu-Leu-Trp-Ile-Leu-ValAla-Val-Ser-Leu-Gly-Leu-Phe-Phe-Tyr-Ser-Phe-Leu-Val-Ser-Ala-Val-Ser-Leu-Ser) (SEQ ID NO: 65). See, for example, Shin et al., Blood 119:5678-5687 (2012). In another specific embodiment, the transmembrane domain of PD-1 is or contains the amino acid sequence TLVVGVVGGLLGSLVLLVWVLAVICSRAA (in three letter code

Thr-Leu-Val-Val-Gly-Val-Val-Gly-Gly-Leu-Leu-Gly-Ser-Leu-Val-LeuLeu-Val-Trp-Val-Leu-Ala-Val-Ile-Cys-Ser- Arg-Ala-Ala) (SEQ ID NO: 66). See Finger et al., Gene 197 (1-2): 177-187 (1997). In another specific embodiment, the transmembrane domain of PD-1 is or contains the amino acid sequence VGVVGGLLGSLVLLVWVLAVI (in three-letter code

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Val-Gly-ValVal-Gly-Gly-Leu-Leu-Gly-Ser-Leu-Val-Leu-Leu-Val-Trp-Val-Leu-Ala-Val-Ue) (SEQ ID NO: 67). See, for example, sequence registration number UNIPROT Q15116. In another specific embodiment, the transmembrane domain of PD-1 is or contains the amino acid sequence FQTLVVGVVGGLLGSLVLLVWVLAVI (in three letter code

Phe-Glu-Thr-Leu-Val-ValGly-Val-Val-Gly-Gly-Leu-Leu-Gly-Ser-Leu-Val-Leu-Leu-Val-Trp-Val-Leu-Ala-Val-Ile) (SEQ ID NO: 68). See, for example, sequence GenBank accession number NM_005018.2.

In some embodiments, the nucleotide sequence encoding one of the transmembrane polypeptides disclosed herein comprises a nucleotide sequence that encodes any of the amino acid sequences disclosed in SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, or SEQ ID NO: 68. In another specific embodiment, the PD-1 transmembrane domain is or contains at least 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 consecutive amino acids disclosed in SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, or SEQ ID NO: 68. In some embodiments, a nucleotide sequence that encodes one of the polypeptides disclosed herein includes a nucleotide sequence that encodes at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 consecutive amino acids disclosed in SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, or SEQ ID NO: 68. When constructing a polypeptide, such as a CAR, in some embodiments, human sequences may be combined with human sequences. animals. For example, a polypeptide, such as CAR, containing the amino acid sequences of the extracellular and intracellular domains of a human protein may include a transmembrane domain from a protein of a non-human species; for example, may contain the mouse CTLA4 transmembrane domain or the mouse PD-1 transmembrane domain. In a more specific embodiment, the invention may include a mouse CTLA4 transmembrane domain or a mouse PD-1 transmembrane domain. In a more specific embodiment, a polypeptide, such as a CAR, contains human amino acid sequences for extracellular and intracellular domains and contains a transmembrane domain having or consisting of the amino acid sequence of SEQ ID NO: 65.

6.5. Pharmaceutical compositions.

In another aspect, provided herein are pharmaceutical compositions comprising any of the anti-BCMA antibodies or BCMA-binding fragments thereof. In some embodiments, the pharmaceutical compositions contain a pharmaceutically acceptable carrier.

In some embodiments, the anti-BCMA antibody formulations or BCMA binding fragments thereof provided herein are formulated for storage and use by combining the purified binding agent of the present invention with a pharmaceutically acceptable carrier (eg, carrier or excipient). Suitable pharmaceutically acceptable carriers include, but are not limited to, non-toxic buffers such as phosphate, citrate and other organic acids; salts such as sodium chloride; antioxidants including ascorbic acid and methionine; preservatives such as octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkylparabens such as methyl or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol and mcresol; low molecular weight polypeptides (eg, less than about 10 amino acid residues); proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; carbohydrates such as monosaccharides, disaccharides, glucose, mannose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes such as Zn protein complexes; and nonionic surfactants such as TWEEN or polyethylene glycol (PEG). (See, for example, Remington: The Science and Practice of Pharmacy, ^22nd Edition, 2012, Pharmaceutical Press, London).

The pharmaceutical compositions provided herein can be formulated for administration by any route for local or systemic treatment. The pharmaceutical compositions provided herein may be formulated for topical use, for example, in the form of epidermal or transdermal patches, ointments, lotions, creams or gels; or in the form of drops, suppositories, sprays, liquids and powders; for pulmonary administration by inhalation or insufflation

- 20 044652 tion, including using a nebulizer, or intratracheal or intranasal administration; or for parenteral administration, including intravenous, intraarterial, intratumoral, subcutaneous, intraperitoneal, intramuscular (eg, injection or infusion), or intracranial (eg, intrathecal or intraventricular) administration.

The therapeutic formulation of anti-BCMA antibodies or BCMA-binding fragments thereof provided herein may be in a unit dosage form. Such formulations include tablets, pills, capsules, powders, granules, solutions or suspensions in water or non-aqueous media, or suppositories, or plastic blood bags or similar devices suitable, for example, for single intravenous or intra-arterial administration.

In some embodiments, anti-BCMA antibodies or BCMA-binding fragments thereof provided herein are contained in microcapsules. Such microcapsules are prepared, for example, by coacervation or interfacial polymerization methods, for example, hydroxymethylcellulose or gelatin microcapsules and polymethylmethacylate microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions, as described in Remington: The Science and Practice of Pharmacy, ^22nd Edition, 2012, Pharmaceutical Press, London.

In some embodiments, the pharmaceutical compositions provided herein include one or more anti-BCMA antibodies or BCMA-binding fragments thereof provided herein in combination with liposomes. Methods for preparing liposomes are known to those skilled in the art. For example, some liposomes can be prepared by reverse phase evaporation with a lipid composition containing phosphatidylcholine, cholesterol and PEG-modified phosphatidylethanolamine (PEG-PE). Liposomes can be extruded through filters of specific pore sizes to produce liposomes of the desired diameter.

6.6. Polynucleotides and methods for producing polypeptides and antibodies.

In another aspect, provided herein are polynucleotides, eg, polynucleotide sequences that encode any of the anti-BCMA antibodies or BCMA-binding fragments thereof, including heavy or light chains, and/or CDR sequences for each chain. In some embodiments, provided herein are polynucleotides comprising polynucleotides that encode a polypeptide (or fragment of a polypeptide) that specifically binds BCMA. The term polynucleotides encoding a polypeptide includes a polynucleotide that includes only coding sequences for the polypeptide, as well as a polynucleotide that includes additional coding and/or non-coding sequences. For example, in some embodiments, the invention provides a polynucleotide comprising a polynucleotide sequence that encodes an anti-human BCMA antibody or encodes a fragment of such an antibody (eg, a fragment containing a BCMA binding site). The polynucleotides of the invention may be in the form of RNA or in the form of DNA. In specific embodiments, the DNA may be, for example, cDNA, genomic DNA, and synthetic DNA; and may be double-stranded or single-stranded, and, in single-stranded form, may be a coding strand or a non-coding (antisense) strand.

In specific embodiments, the polynucleotides provided herein encode an antibody light chain variable region comprising, consisting essentially of, or consisting of the nucleotide sequence of SEQ ID NO: 5.

In other specific embodiments, the polynucleotides provided herein encode an antibody heavy chain variable region comprising, consisting essentially of, or consisting of the nucleotide sequence of SEQ ID NO: 16, SEQ ID NO: 27, SEQ ID NO: 38, SEQ ID NO: 49 or SEQ ID NO: 60.

In other specific embodiments, the polynucleotides provided herein encode an antibody light chain variable region comprising, consisting essentially of, or consisting of the nucleotide sequence of SEQ ID NO: 5, and encode an antibody heavy chain variable region comprising, consisting of substantially of or consisting of the nucleotide sequence of SEQ ID NO: 16. In other specific embodiments, the polynucleotides provided herein encode an antibody light chain variable region comprising, consisting primarily of or consisting of the nucleotide sequence of SEQ ID NO: 5, and encode an antibody heavy chain variable region comprising, consisting essentially of, or consisting of the nucleotide sequence of SEQ ID NO: 27. In other specific embodiments, the polynucleotides provided herein encode an antibody light chain variable region containing, consisting essentially of of or consisting of the nucleotide sequence of SEQ ID NO: 5, and encode an antibody heavy chain variable region comprising, consisting essentially of, or consisting of the nucleotide sequence of SEQ ID NO: 38. In other specific embodiments, the polynucleotides provided herein are encodes an antibody light chain variable region comprising, consisting essentially of, or consisting of the nucleotide sequence of SEQ ID

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NO: 5, and encode an antibody heavy chain variable region comprising, consisting essentially of, or consisting of the nucleotide sequence of SEQ ID NO: 49. In other specific embodiments, the polynucleotides provided herein encode an antibody light chain variable region comprising consisting essentially of or consisting of the nucleotide sequence of SEQ ID NO: 5, and encodes an antibody heavy chain variable region comprising, consisting essentially of or consisting of the nucleotide sequence of SEQ ID NO: 60.

Any of the polynucleotides provided herein may be contained in and/or expressed from a polynucleotide vector. In some embodiments, the vector is contained in a host cell. Such a host cell, when transformed or transfected with a vector provided herein, expresses or is capable of expressing polynucleotide sequences encoding the anti-BCMA antibodies provided herein. In some embodiments, the polynucleotides provided herein are operably linked to one or more regulatory sequences in a vector to allow expression of a BCMA-binding antibody.

The term vector as used herein means a nucleic acid molecule used as a carrier for the transfer of genetic material into a cell and includes, without limitation, plasmids, viral genomes (including replication-incompetent viral genomes and viral genomes in multiple segments), cosmids and artificial chromosomes. In general, the constructed vectors contain an origin of replication, a region with multiple cloning sites, and a selectable marker. The vector itself is typically a nucleotide sequence, typically a DNA sequence, that contains an insert (a transgene, e.g., any of the anti-BCMA antibody coding sequences described herein) and a longer sequence that serves as the backbone of the vector. Vectors may include additional functions beyond transgene insertion and scaffold: promoter, genetic marker, antibiotic resistance, reporter gene, guide sequence, protein purification tag. Vectors, called expression vectors, provide expression of a transgene in a target cell and typically have regulatory sequences, such as a promoter sequence, that direct expression of the transgene. Incorporation of a vector into a target cell is usually called transformation for bacterial cells and transfection for eukaryotic cells; Also, the insertion of a viral vector into a mammalian cell is called transduction.

As used herein, the term host cell refers to a cell into which a polynucleotide encoding a BCMA-binding antibody provided herein is introduced through transformation, transfection, or the like. The term refers not only to the specific cells that are transfected, transformed or transduced, but also to the descendants of such cells. Since certain modifications may arise in subsequent generations due to mutations or environmental influences, such progeny may not be identical to the parent cells, but are still within the scope of the term as used herein.

As used herein, the term expression includes any step involved in the production of a binding molecule of the invention, including, but not limited to, transcription (for example, from a polynucleotide provided herein or a polynucleotide encoding a BCMA-binding antibody or a BCMA-binding fragment thereof provided herein), and, in some embodiments, post-transcriptional modification, translation, post-translational modification and secretion of, for example, a BCMA binding antibody or a BCMA binding fragment thereof provided herein.

The term regulatory sequences refers to nucleotide sequences necessary for the expression of an operably linked coding sequence in a particular host organism. Regulatory sequences that are suitable for prokaryotes, for example, include a promoter, optionally an operator sequence and a ribosome binding site. It is known that eukaryotic cells are used, and the vector may contain: promoters, polyadenylation signals and enhancers.

As used herein, a nucleic acid or polynucleotide is operably linked when it is in operative relationship with another nucleotide sequence. For example, the nucleotide sequence of a promoter is operably linked to a nucleotide sequence encoding a BCMA-binding antibody or a BCMA-binding fragment thereof, if the promoter drives expression of the BCMA-binding antibody or a BCMA-binding fragment thereof; or, as another example, the nucleotide sequence for a pre-sequence or secretory leader is operably linked to a nucleic acid encoding a polypeptide if expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects transcription of the sequence; or the ribosome binding site is operably linked to the coding sequence if it is

- 22 044652 is placed so as to facilitate broadcasting. Generally, operably linked means that the linked nucleic acid sequences are contiguous, and in the case of a secretory leader, contiguous and in the same reading frame. However, enhancers do not have to be contiguous. Linking can be accomplished, for example, by ligation at convenient restriction sites. If such sites do not exist, synthetic oligonucleotide adapters or linkers are used in accordance with standard practice.

The terms host cell, target cell or recipient cell include any individual cell or culture of cells that can be or have been recipients for vectors or for the incorporation of exogenous nucleic acid molecules, polynucleotides and/or proteins. These terms also include the progeny of a single cell, and these progeny need not be completely identical (in morphology, genomic or total DNA) to the original parent cell due to natural, accidental or intentional mutation. The cells may be prokaryotic or eukaryotic and include, but are not limited to, bacterial cells, yeast cells, animal cells and mammalian cells such as mouse, rat, macaque or human. Suitable host cells include prokaryotic and eukaryotic host cells, including yeast, fungi, insect cells and mammalian cells.

Antibodies or fragments thereof can be produced by bacteria. Once expressed, the binding molecule of the invention is preferably recovered from the E. coli cell pellet in a soluble fraction and can be purified, for example, by affinity chromatography and/or size exclusion chromatography. Final purification can be carried out similarly to the purification process for antibodies expressed, for example, in CHO cells.

In addition to prokaryotes, suitable hosts for cloning or expression of the binding molecule of the invention include eukaryotic microorganisms such as filamentous fungi or yeast. Saccharomyces cerevisiae, or common baker's yeast, is the most commonly used lower eukaryotic microorganism host. However, a number of other genera, species and strains are commonly available and suitable for use in the present invention, such as Schizosaccharomyces pombe, Kluyveromyces host cells such as, for example, K. lactis, K. fragilis (ATCC 12424), K. bulgaricus (ATCC 16045), K. wickeramii (ATCC 24178), K. waltii (ATCC 56500), K. drosophilarum (ATCC 36906), K. thermotolerans and K. marxianus; Yarrowia (EP 402 226); Pichia pastoris (EP 183 070); Candida; Trichoderma reesia (EP 244 234); Neurospora crassa; Schwanniomyces such as Schwanniomyces occidentalism and filamentous fungi such as, for example, Neurospora, Penicillium, Tolypocladium, and Aspergillus host cells such as A nidulans and A. niger.

Suitable host cells for expressing the glycosylated binding molecule of the invention, preferably antibody-derived binding molecules, are from multicellular organisms. Examples of invertebrate cells include plant and insect cells. Numerous strains and variants of baculoviruses and their corresponding permissive insect host cells have been identified, such as Spodoptera frugiperda (fall armyworm caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruit fly) and Bombyx mori. A variety of viral strains for transfection are publicly available, for example Autographa californica NPV variant L-1 and Bombyx mori NPV strain Bm-5, and such viruses can be used as the virus herein according to the present invention, in particular for transfecting Spodoptera frugiperda cells .

Plant cell cultures from cotton, corn, potato, soybean, petunia, tomato, Arabidopsis or tobacco can also be used as hosts. Cloning and expression vectors useful for producing proteins in plant cell culture are known to those skilled in the art. See, for example, Hiatt et al., Nature (1989) 342: 76-78, Owen et al. (1992) Bio/Technology 10: 790794, Artsaenko et al. (1995) The Plant J. 8: 745-750 and Fecker et al. (1996) Plant Mol. Biol. 32: 979-986.

The propagation of vertebrate cells in culture (tissue culture) and the expression of proteins in them has become a routine procedure. Examples of suitable mammalian host cell lines include monkey kidney line CV1 transformed with SV40 (C0S-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese Hamster Ovary Cells/-DHFR (CHO, Urlaub et al., Proc. Natl. Acad. Sci. USA 77: 4216 (1980)); mouse Sertoli cells (TM4, Mather, Biol. Reprod. 23: 243-251 (1980)); monkey kidney cells (CV1, ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); gray rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, 1413 8065); mouse mammary tumor cells (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; and a human hepatoma cell line (Hep G2).

Using recombinant methods, BCMA-binding antibodies or fragments thereof provided herein can be produced intracellularly, in the periplasmic

- 23 044652 space or directly secreted into the environment. If the binding molecule is produced intracellularly, in a first step the debris particles, whether host cells or lysed fragments, are removed, for example by centrifugation or ultrafiltration. Carter et al., Bio/Technology 10: 163-167 (1992) describe a procedure for isolating antibodies that are secreted into the periplasmic space of E. coli. Briefly, the cell pellet is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonyl fluoride (PMSF) for approximately 30 min. Remaining cells can be removed by centrifugation. When the antibody is secreted into the medium, the supernatants of such expression systems are typically first concentrated using a commercially available protein concentration filter, such as an Amicon or Millipore Pellicon ultrafiltration device. A protease inhibitor such as PMSF may be included at any of the above steps to inhibit proteolysis, and antibiotics may be included to prevent the growth of ingested microorganisms.

Anti-BCMA antibodies or BCMA-binding fragments thereof obtained from host cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis and affinity chromatography, with affinity chromatography being the preferred purification technique. The carrier to which the affinity ligand is attached is most often agarose, but other carriers are available. Mechanically stable supports such as controlled pore glass or poly(styrene divinyl)benzene provide higher flow rates and shorter treatment times than can be achieved with agarose. If the binding molecule of the present invention contains a CH3 domain, ABXMresin Bakerbond (JT Baker, Phillipsburg, NJ) can be used for purification. Depending on the antibody to be isolated, other protein purification methods may be used, such as ion exchange column fractionation, ethanol precipitation, reverse phase HPLC, silica chromatography, heparin-SEPHAROSE!!! chromatography, anion or cation exchange resin chromatography (such as a polyaspartic acid column), chromatofocusing, SDS-PAGE and ammonium sulfate precipitation according to standard procedures.

In another aspect, methods are provided for producing the binding molecules of the present invention, wherein the methods comprise culturing a host cell as defined herein under conditions allowing expression of the binding molecule, and isolating the resulting binding molecule from the culture.

The term culture refers to the maintenance, differentiation, growth, proliferation and/or propagation of cells in vitro under suitable environmental conditions.

6.7. Methods of application.

In another aspect, provided herein are methods of using anti-BCMA antibodies or BCMA-binding fragments thereof, such as methods of treatment using such antibodies or antibody fragments.

In some embodiments, provided herein is a method of treating a subject suffering from a BCMA-related disease or disorder, comprising administering to the subject a therapeutically effective amount of one or more anti-BCMA antibodies or BCMA-binding fragments thereof. Also provided herein is a method of treating a subject suffering from a plasma cell-related disease or disorder, comprising administering to the subject a therapeutically effective amount of one or more anti-BCMA antibodies or BCMA-binding fragments thereof. Also provided herein is a method of treating a subject suffering from a B cell-related disease or disorder, comprising administering to the subject a therapeutically effective amount of one or more anti-BCMA antibodies or BCMA-binding fragments thereof. In specific embodiments, the BCMA-related disease is macroglobulinemia, amyloidosis, Waldenström's macroglobulinemia, solitary plasmacytoma of bone, extramedullary plasmacytoma, osteosclerotic myeloma, heavy chain diseases, monoclonal gammopathy of undetermined significance, and smoldering multiple myeloma.

Also provided herein is a method of inhibiting the growth of multiple myeloma cells, comprising contacting the multiple myeloma cells with one or more anti-BCMA antibodies or BCMA-binding fragments thereof provided herein. Further provided herein is a method of reducing the growth rate of multiple myeloma cells comprising contacting the multiple myeloma cells with one or more anti-BCMA antibodies or BCMA-binding fragments thereof provided herein.

In any of the methods, for example, methods of treatment proposed herein, the antibody or BCMA-binding fragment thereof is a naked antibody or fragment, that is, the antibody or fragment is not modified to include a toxic fragment. In any of the methods, such as the treatments provided herein, the antibody or a BCMA binding fragment thereof is part of an antibody-drug conjugate (ADC), e.g.

- 24 044652

ADC described in section 5.3 above.

In any of the treatment methods provided herein, anti-BCMA antibodies or BCMA-binding fragments thereof are used or administered in a therapeutically effective amount. In various embodiments, the therapeutically effective amount is from 1x105 to 5x105, from 5x105 to 1x10 ⁶ , from 1x10 ⁶ to 5x10 ⁶ , from 5x10 ⁶ to 1x10 ⁷ , from 1x10 ⁷ to 5x10 ⁷ , from 5x10 ⁷ to 1x10 ⁸ , from 1x10 ⁸ up to 5x10 ⁸ , from 5x10 ⁸ to 1x10 ⁹ nanomoles of the indicated antibodies or fragments. In various other embodiments of the invention, the therapeutically effective amount is from 1x10 ⁵ to 5x105, from 5x105 to 1x10 ⁶ , from 1x10 ⁶ to 5x10 ⁶ , from 5x10 ⁶ to 1x10 ⁷ , from 1x10 ⁷ to 5x10 ⁷ , from 5x10 ⁷ to 1x10 ⁸ , from 1x10 ⁸ to 5x10 ⁸ , from 5x10 ⁸ to 1x10 ⁹ nanomoles of the indicated antibodies or fragments.

Anti-BCMA antibodies or BCMA-binding fragments may be administered to a patient who requires it at any dosing schedule deemed appropriate by the treating physicians or clinicians, for example, once every 1, 2, 3, 4, 5, 6 or 7 days or once every 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 weeks. Antibodies against BCMA or BCMA-binding fragments thereof can be administered for a specified period of time, for example, 1, 2, 3 or 4 weeks, followed by rest without administration of the antibody or antibody fragment. This insertion-rest cycle can be repeated 2, 3, 4 or 5 times.

In some embodiments, the patient being administered anti-BCMA antibodies or BCMA-binding fragments thereof has previously received 1, 2, 3, or more lines of therapy, wherein the previous lines of therapy may include administration of one or more of the following drugs: lenalidomide (Revlimid) , pomalidomide (Pomalista), thalidomide (Thalomida), bortezomib (Velkada), dexamethasone, cyclophosphamide, doxorubicin (Adriamycin, Rubexa), carfilzomib (Kripolis), ixazomib (Ninlaro), cisplatin (Platinol), doxorubicin (Adriamycin), etoposide (Etopofos ), anti-CD38 antibodies such as daratumumab (Darzalex); panobinostat or elotuzumab (Emplicity). In specific embodiments of the invention, such patients received therapy with bortezomib, lenalidomide and dexamethasone (RVD); bortezomib, cyclophosphamide and dexamethasone (BCD); bortezomib, doxorubicin and dexamethasone; carfilzomib, lenalidomide and dexamethasone (CRD); ixazomib, lenalidomide and dexamethasone; bortezomib and dexamethasone; bortezomib, thalidomide and dexamethasone; lenalidomide and dexamethasone; dexamethasone, thalidomide, cisplatin, doxorubicin, cyclophosphamide, etoposide and bortezomib (VTDRACE); lenalidomide and dexamethasone in low doses; bortezomib, cyclophosphamide and dexamethasone; carfilzomib and dexamethasone; lenalidomide only; bortezomib only; daratumumab only; bortezomib, lenalidomide and dexamethasone; daratumumab, bortezomib and dexamethasone; daratumumab, lenalidomide and dexamethasone; elotuzumab, lenalidomide and dexamethasone; elotuzumab, lenalidomide and dexamethasone; bendamustine, bortezomib and dexamethasone; bendamustine, lenalidomide and dexamethasone; pomalidomide and dexamethasone; pomalidomide, bortezomib and dexamethasone; pomalidomide, carfilzomib and dexamethasone; bortezomib and liposomal doxorubicin; cyclophosphamide, lenalidomide and dexamethasone; elotuzumab, bortezomib and dexamethasone; ixazomib and dexamethasone; panobinostat, bortezomib and dexamethasone; panobinostat and carfilzomib; or pomalidomide, cyclophosphamide and dexamethasone.

Administration of antibodies against BCMA or BCMA-binding fragments thereof to a patient in need thereof may be accompanied by one or more additional treatments. For some types of cancer, such as multiple myeloma, one or more additional therapeutic regimens may include one or more of the following drugs: lenalidomide, pomalidomide, thalidomide, bortezomib, dexamethasone, cyclophosphamide, doxorubicin, carfilzomib, iaxizomib, cisplatin, doxorubicin, etoposide, an anti-CD38 antibody such as daratumumab, panobinostat and/or elotuzumab, alone or in one of the combinations listed above, or in any other combination.

In some embodiments, one or more anti-BCMA antibodies provided herein are administered to a patient in need thereof in combination with one or more compounds described in section 5.3 above, for example, in combination with lenalidomide or pomalidomide.

In some other embodiments of the invention, one or more anti-BCMA antibodies provided herein are administered to a patient in need thereof in combination with one or more of the following compounds.

Examples of compounds include, but are not limited to, N-{[2-(2,6-dioxo(3-nineridyl)-1,3dioxoisoindolin-4-yl]methyl}cyclopropyl-carboxamide; 3-[2-(2,6 -dioxopiperidin-3-yl)-1,3-dioxo2,3-dihydro-1H-isoindol-4-ylmethyl]-1,1-dimethylurea; (-)-3 -(3,4-dimethoxyphenyl)-3-( 1 -oxo-1,3-dihydroisoindol-2-yl)propionamide; (+)-3-(3,4-dimethoxyphenyl)-3 -(1-oxo-1,3-dihydroisoindol-2yl)propionamide; (-)-{ 2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione}; (+)-{2-[1-(3-ethoxy-4-methoxyphenyl) -2-methylsulfonylethyl]-4acetylaminoisoindoline-1,3-dione}; difluoromethoxy-SelCID; 1-phthalimido-1-(3,4-diethoxyphenyl)ethane; 3- 25 044652 (3,4-dimethoxyphenyl)-3-(3 ,5-dimethoxyphenyl)acrylonitrile; 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline; 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline ; 4-amino-2-(3-methyl2,6-dioxopiperidin-3-yl)isoindole-1,3-dione; 3-(3-acetoamidophthalimido)-3-(3-ethoxy-4methoxyphenyl)-N-hydroxypropionamide; 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4-methylisoindoline; cyclopropyl-N-{2-[(1S)-1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethyl]-3-oxoisoindolin-4yl}carboxamide; substituted 2-(3-hydroxy-2,6-dioxopiperidin-5-yl)isoindoline; N-[2-(2,6dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1 H-isoindol-5-ylmethyl]-4-trifluoromethoxybenzamide; (S)-4-chloro-N-((2-(3-methyl-2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)methyl)benzamide; pyridin-2-carboxylic acid[2-[(3S)-3-methyl-2,6-diokCOpiperidin-3-yl]-1,3-dioxo-2,3-dihydro-1Nisoindol-5-ylmethyl]amide; (S)-N-((2-(3-methyl-2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5yl)methyl)-4-(trifluoromethyl)benzamide; 3-(2,5-dimethyl-4-oxo-4H-quinazolin-3-yl)piperidin-2,6-dione, etc.

In some embodiments, one or more anti-BCMA antibodies provided herein are administered to a patient in need thereof in combination with one or more cyano- and carboxy-substituted styrene derivatives, such as those disclosed in US Pat. No. 5,929,117 ; 1-oxo-2-(2,6-dioxo-3-fluoropiperidin-3-yl)isoindolines and 1,3-dioxo-2(2,6-dioxo-3-fluoropiperidin-3-yl)isoindolines, such as those described in US patent Nos. 5874448 and 5955476; 1-oxo- and 1,3-dioxoisoindolines substituted at the 4th or 5th position of the indoline ring (for example, 4-(4-amino-1,3-dioxoisoindolin-2-yl)-4-carbamoylbutanoic acid), described in US Pat. No. 6,380,239; isoindolin-1-one and isoindolin-1,3-dione substituted at the 2-position with 2,6-dioxo-3-hydroxypiperidin-5-yl (for example, 2-(2,6-dioxo-3-hydroxy-5fluoropiperidine -5-yl)-4-aminoisoindolin-1-one), described in US patent No. 6458810; representatives of the class of non-polypeptide cyclic amides disclosed in US patent Nos. 5698579 and 5877200; and isoindoleimide compounds such as those described in US Patent Publication No. 2003/0045552, US Patent Publication No. 2003/0096841 and International Application No. PCT/US01/50401 (International Publication No. WO 02/059106). US Patent Publication No. 2006/0205787 describes compositions of 4amino-2-(3-methyl-2,6-dioxopiperidin-3-yl)isoindole-1,3-dione. US Patent Publication No. 2007/0049618 describes isoindoleimide compounds.

In some embodiments, a patient in need thereof is administered one or more of the anti-BCMA antibodies provided herein in combination with one or more members of the isoindoleimide class disclosed in U.S. Patent No. 7,091,353, U.S. Patent Publication No. 2003/0045552, and International Application No. PCT/US01/50401 (International Publication No. WO 02/059106), each of which is incorporated herein by reference. Typical compounds have formula II:

O

and their pharmaceutically acceptable salts, hydrates, solvates, clathrates, enantiomers, diastereomers, racemates and mixtures of stereoisomers, where: one of X and Y is C=O and the other is CH ₂ or C=O; R ¹ represents H, (C1-C8)alkyl, (C3-C7)cycloalkyl, (C2-C8)alkenyl, (C2C8)kynyl, benzyl, aryl, (C0-C4)alkyl-(C1-C6)heterocycloalkyl, (C0-C4)alkyl-(C2-C5)heteroaryl, C(O)R ³ , C(S)R ³ , C(O)OR ⁴ , (C1-C ₈ )alkyl-N(R ⁶ )2, (C1-C8)alkyl-OR ⁵ , (C1-C ₈ )alkyl-C(O)ORS, C(O)NHR ³ , C(S)NHR ³ , C(O)NR ³ R ³ ', C( S)NR ³ R ³ ' or (C1-C8)αlkyl-O(CO)R ⁵ ; R ² represents H, F, benzyl, (CrC8)alkyl, (C2-C8)alkenyl or (C2-C8)alkynyl; R ³ and R ³ ' are independently (C1- _C8 )αalkyl, ( _C3C7 ) _cycloalkyl , ( _C2 - _C8 )alkenyl, ( _C2 - _C5 )alkynyl, benzyl, aryl, ( _C0 -C ₄ )alkyl-(C1-C ₆ )heterocycloalkyl, (C ₀ -C ₄ )alkyl-(C ₂ -C ₅ )heteroaryl, (C ₀ -C ₈ )alkyl-N(R ⁶ )2, (C1 -C8)alkyl-OR ⁵ , (C1-C ₈ )alkyl-C(O)OR ⁵ , (C1C8)αalkyl-O(CO)R ⁵ or C(O)OR ⁵ ; R ⁴ is (C1- _C8 )alkyl, ( _C2 - _C8 )alkenyl, ( _C2 - _C8 )kynyl, (C1 _{- C4} )alkyl- ^OR5 , benzyl, aryl, (C0- C4)alkyl-(C1-C6)heterocycloalkyl or (C0-C4)alkyl-(C2C5)heteroaryl; _R ⁵ is (C1- _C8 )alkyl, ( _C2 - _C8 )alkenyl, ( _C2 - _C8 )alkynyl, benzyl, aryl, or ( _C2C5 )heteroaryl; in all cases, R ⁶ is independently H, (C ₁ -C ₈ )αalkyl, (C ₂ -C ₈ )alkenyl, (C ₂ C ₈ )alkynyl, benzyl, aryl, (C ₂ -C ₅ )heteroaryl or ( C ₀ -C ₈ )alkyl-C(O)O-R ⁵ , or R ⁶ groups can combine to form a heterocycloalkyl group;

n has the value 0 or 1; and * represents a chiral carbon center.

In specific compounds of Formula II, when n is 0, then R ¹ is (C ₃ -C ₇ )cycloalkyl, (C ₂ -C ₈ )alkenyl, (C ₂ -C ₈ )alkynyl, benzyl, aryl, ( C ₀ -C ₄ )alkyl-(C ₁ -C ₆ )heterocycloalkyl, (C0-C4)alkyl-(C2-C5)heteroaryl, C(O)R ³ , C(O)OR ⁴ , (C1-C8) alkyl-N(R ⁶ )2, (C1-C8)alkyl-OR ⁵ , (C1-C8)alkylC(O)OR ⁵ , C(S)NHR ³ or (C1-C8)alkyl-O(CO)R ⁵ ;

R ² represents H or (C1-C8)alkyl; and R ³ represents (C1-C8)alkyl, (C ₃

- 26 044652

C7)cycloalkyl, ( _C2 - _C8 )alkenyl, ( _C2 - _C8 )alkynyl, benzyl, aryl, ( _C0 - _C4 )alkyl-(C1- _C6 )heterocycloalkyl, (Co-C4)alkyl- (C2-C5)heteroaryl, (C5-C8)alkyl-N(R ⁶ )2; (Co-C8)alkyl-NH-C(O)O-R ⁵ ; (C1-C8)alkyl-OR ⁵ , (Cr C8)alkyl-C(O)OR ⁵ , (C1-C ₈ )alkyl-O(CO)R ⁵ or C(O)OR ⁵ ; and other variables have the same definitions.

In other specific compounds of Formula II, R ² represents H or (C1-C ₄ )alkyl.

In other specific compounds of Formula II, R ¹ is (C ₁ -C ₈ )αalkyl or benzyl.

In other specific compounds of formula II, R ¹ represents H, (C ₁ -C ₈ )alkyl, benzyl, w^ch ₂ -$

CH2OCH3, CH2OCH3, CH2CH2OCH3 or ⁰

In another embodiment of the compounds of Formula II, R ¹ is

where Q is O or S, and in each case R7 is independently H, ( _C1C8 )alkyl, (C3-C7)cycloalkyl, ( _C2 - _C8 )alkenyl, ( _C2 - _C8 )alkynyl, benzyl , aryl, halogen, (C ₀ -C ₄ )alkyl-(C1C ₆ )heterocycloalkyl, (C ₀ -C ₄ )alkyl-(C2-C ₅ )heteroaryl, (C ₀ -C ₈ )alkyl-N(R ⁶ )2, (C1-C8)alkyl- ^OR5 , (C1C8)alkyl-C(O) ^OR5 , (C1 _{- C8} )alkyl-O(CO) ^R5 or C(O) ^OR5 , or neighboring R7 can be combined to form a bicyclic alkyl or aryl ring.

In other specific compounds of Formula II, R ¹ is C(O)R ₃ .

In other specific compounds of formula II, R ³ is (C ₀ -C ₄ )alkyl-(C ₂ C ₅ )heteroaryl, (C ₁ -C ₈ )alkyl, aryl or (C ₀ -C ₄ )alkyl-OR ⁵ .

In other specific compounds of Formula II, heteroaryl is pyridyl, furyl or thienyl.

In other specific compounds of Formula II, R ¹ is C(O)OR ⁴ .

In other specific compounds of Formula II, the H in C(O)NHC(O) may be replaced by ( _C1C4 )alkyl, aryl or benzyl.

Additional examples of compounds in this class include, but are not limited to: [2-(2,6dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-ylmethyl]amide; (2(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-ylmethyl)carbamic acid tert-butyl ester; 4(aminomethyl)-2-(2,6-dioxo(3-piperidyl))isoindoline-1,3-dione; N-(2-(2,6-dioxopiperidin-3-yl)-1,3dioxo-2,3-dihydro-1H-isoindol-4-ylmethyl)acetamide; N-{(2-(2,6-dioxo(3-piperidyl)-1,3dioxoisoindolin-4-yl)methyl}cyclopropylcarboxamide; 2-chloro-N-{(2-(2,6-dioxo(3-piperide yl))-1,3-dioxoisoindolin-4-yl)methyl}acetamide; N-(2-(2,6-dioxo(3-ninerid yl))-1,3-dioxoisoindolin-4-yl)3-pyridylcarboxamide; 3 -{1-oxo-4-(benzylamino)isoindolin-2-yl}piperidin-2,6-dione; 2-(2,6dioxo(3-piperidyl))-4-(benzylamino)isoindoline-1,3-dione ; N-{(2-(2,6-dioxo(3-piperidyl))-1,3dioxoisoindolin-4-yl)methyl}propanamide; N-{(2-(2,6-dioxo(3-niperidyl)) -1,3-dioxoisoindolin-4yl)methyl}-3-pyridylcarboxamide; N-{(2-(2,6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4yl)methyl}heptanamide; N-{ (2-(2,6-dioxo(3-πpiperidyl))-1,3-dioxoisoindolin-4-yl)methyl}-2furylcarboxamide; (N-(2-(2,6-dioxo(3-pineridyl)))- 1,3-dioxoisoindolin-4yl)carbamoyl}methyl acetate; N-(2-(2,6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl)pentanamide; N(2-(2,6 -dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl)-2-thienylcarboxamide; N-{[2-(2,6-dioxo(3piperidyl))-1,3-dioxoisoindolin-4-yl ]methyl}(butylamidenet)carboxamide; N-{[2-(2,6-dioxo(3piperidyl))-1,3-dioxoisoindolin-4-yl]methyl}(octylamino)carboxamide; and N-{[2-(2,6-dioxo(3piperidyl))-1,3-dioxoisoind olin-4-yl]methyl}(benzylamino)carboxamide.

In some other embodiments of the invention, one or more anti-BCMA antibodies provided herein are administered to a patient in need thereof in combination with one or more members of the isoindolimide class disclosed in US Patent Application Publication No. US 2002/0045643, International Publication No. WO 98/54170 and US Patent No. 6395754, each of which is incorporated herein by reference. Typical compounds have formula III:

O R

-Y /—

JL JZ )=° id and their pharmaceutically acceptable salts, hydrates, solvates, clathrates, enantiomers, diastereomers, racemates and mixtures of stereoisomers, where: one of X and Y represents C=O and the other represents

- 27 044652 is CH2 or C=O;

R is H or CH2OCOR';

(i) each of R ¹ , R ² , R ³ or R ⁴ , independently of the others, is halogen, a C 1-4 alkyl or a C 1-4 alkoxy group, or (ii) one of R ¹ , R ² , R ³ or R ⁴ is nitro or -NHR5 and the remainder of R ¹ , R ² , R ³ or R ⁴ is hydrogen; R ⁵ is hydrogen or C1-8 alkyl; ^R6 is hydrogen, C1-8 alkyl, benzo, chlorine or fluorine;

R' is R ⁷ -CHR ¹⁰ -N(R ⁸ R ⁹ );

R ⁷ represents m-phenylene or p-phenylene or -(C _n H _2n )-, in which n is from 0 to 4; each of ^R8 and ^R9 , independently of each other, represents hydrogen or alkyl containing from 1 to 8 carbon atoms, or ^R8 and ^R9 taken together represent tetramethylene, pentamethylene, hexamethylene or -CH2CH2X1CH2CH2-, in in which X1 represents -O-, -S- or -NH-; R ¹⁰ represents hydrogen, alkyl with 8 carbon atoms or phenyl; and * represents a chiral carbon center.

Other typical compounds have the formula:

V o, o G R®

II Ί / ^Х У-N—С1Г-О —С —R7—С —N

[| H X / \ ' IH

I Σ )= ^{0 Rt}

R ⁴ where: one of X and Y is C=O, and the other of X and Y is C=O or CH2;

(i) each of R ¹ , R ² , R ³ or R ⁴ , independently of the others, is halogen, a C 1-4 alkyl or a C 1-4 alkoxy group, or (ii) one of R ¹ , R ² , R ³ and R ⁴ are -NHR5 and the remainder of R ¹ , R ² , R ³ or R ⁴ are hydrogen;

R ⁵ represents hydrogen or alkyl with 1-8 carbon atoms;

R ⁶ is hydrogen, C1-C8 alkyl, benzo, chlorine or fluorine;

R ⁷ represents m-phenylene or p-phenylene or -(CnH2n)-, in which n is from 0 to 4;

each of ^R8 and ^R9 , independently of each other, represents hydrogen or alkyl containing from 1 to 8 carbon atoms, or ^R8 and ^R9 taken together represent tetramethylene, pentamethylene, hexamethylene or -CH2CH2X1CH2CH2-, in in which X1 represents -O-, -S- or -NH-; And

R ¹⁰ represents hydrogen, alkyl up to 8 carbon atoms or phenyl.

Other typical compounds have the formula:

| O

X X >X

R ⁴ in which one of X and Y is C=O and the other of X and Y is C=O or CH2;

each of R ¹ , R ² , R ³ and R ⁴ , independently of the others, is a halogen, a C 1-4 alkyl or a C 1-4 alkoxy group, or (ii) one of R ¹ , R ² , R ³ and R ⁴ represents a nitro group or a protected amino group, and the remainder of R ¹ , R ² , R ³ and R ⁴ represent hydrogen; And

R ⁶ represents hydrogen, alkyl with 1-8 carbon atoms, benzo, chlorine or fluorine.

Other typical compounds have the formula:

X X /X x°

Ri X---/

R ^J in which: one of X and Y is C=O, and the other of X and Y is C=O or CH2;

(i) each of R ¹ , R ² , R ³ and R ⁴ , independently of the others, is halogen, C 1-4 alkyl or C 1-4 alkoxy, or (ii) one of

R ¹ , R ² , R ³ and R ⁴ are -NHR5 and the remainder of R ¹ , R ² , R ³ and R ⁴ are hydrogen;

R ⁵ is hydrogen, alkyl of 1-8 carbon atoms, or CO-R ⁷ -CH(R ¹⁰ )NR ⁸ R ⁹ , in which R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each as defined herein document; And

R ⁶ represents C1-C8 alkyl, benzo, chlorine or fluorine.

Specific examples of compounds have the formula

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one of X and Y is C=O, and the other of X and Y is C=O or CH2;

R ⁶ represents C1-C8 alkyl, benzyl, chlorine or fluorine;

R ⁷ represents m-phenylene, p-phenylene or -(C _p H _2p )-, in which n is from 0 to 4;

each of R ⁸ and R ⁹ , independently of each other, is hydrogen or alkyl containing from 1 to 8 carbon atoms, or R ⁸ and R ⁹ taken together are tetramethylene, pentamethylene, hexamethylene or -CH ₂ CH ₂ X ₁ CH ₂ CH ₂ -, in which X ₁ represents -O-, -S- or -NH-; And

R ¹⁰ represents hydrogen, alkyl with 1-8 carbon atoms or phenyl.

Other specific immunomodulatory compounds are 1-oxo-2-(2,6-dioxo-3fluoropiperidin-3-yl)isoindolines and 1,3-dioxo-2-(2,6-dioxo-3-fluoropiperidin-3-yl)isoindolines , such as those described in US Patent Nos. 5,874,448 and 5,955,476, each of which is incorporated herein by reference. Typical compounds have the formula:

represents hydrogen, halogen, alkyl of 1-4 carbon atoms, alkoxy group of 1-4 carbon atoms or an amino group.

Other specific immunomodulatory compounds disclosed herein are 1-oxo- and 1,3-dioxoisoindolines substituted at the 4th or 5th position of the indoline ring described in US Pat. No. 6,380,239 and US Pat. No. 7,244,759, which are both included incorporated into this document by reference. Typical compounds have the formula:

in which the carbon atom designated C* is a chiral center (when n is not zero and R ¹ is not the same as R ² ); one of X ¹ and X ² represents an amino group, a nitro group, alkyl having one to six carbon atoms, or NH-Z, and the other of X ¹ or X ² represents hydrogen; R ¹ and R ² are each, independently of the other, hydroxy or NH-Z; R ³ represents hydrogen, alkyl of one to six carbon atoms, halogen or haloalkyl; Z represents hydrogen, aryl, alkyl of one to six carbon atoms, formyl or acyl of one to six carbon atoms; and n is 0, 1 or 2; with the proviso that if X ¹ is an amino group and n is 1 or 2, then R ¹ and R ² are both non-hydroxy; and their salts.

Other typical compounds have the formula: in which the carbon atom, designated C*, represents a chiral center when n is not zero and R ¹ is not R ² ; one of X ¹ and X ² represents an amino group, a nitro group, alkyl having one to six carbon atoms, or NH-Z, and the other of X ¹ or X ² represents hydrogen; R ¹ and R ² are each, independently of the other, hydroxy or NH-Z; R ³ represents alkyl of one to six carbon atoms, halogen or hydrogen; Z represents hydrogen, aryl or alkyl or acyl of one to six carbon atoms; and n has the value 0, 1 or 2.

Specific examples include, but are not limited to, 2-(4-amino-1-oxo-1,3-dihydroisoindol-2-yl)-4-carbamoylbutyric acid and 4-(4-amino-1-oxo-1,3-dihydroisoindole -2-yl)4-carbamoylbutyric acid, which have the following structures, respectively, and their pharmaceutically acceptable salts, solvates, prodrugs and stereoisomers:

- 29 044652

in which the carbon atom designated C* represents a chiral center when n is not zero and R ¹ is not R ² ; one of X ¹ and X ² represents an amino group, a nitro group, alkyl having one to six carbon atoms, or NH-Z, and the other of X ¹ or X ² represents hydrogen; R ¹ and R ² are each, independently of the other, hydroxy or NH-Z; R ³ represents alkyl of one to six carbon atoms, halogen or hydrogen; Z represents hydrogen, aryl or alkyl or acyl of one to six carbon atoms; and n is 0, 1 or 2; and their salts.

Specific examples include, but are not limited to, 4-carbamoyl-4-{4-[(furan-2ylmethyl)amino]-1,3-dioxo-1,3-dihydroisoindol-2-yl}butyric acid, 4-carbamoyl- 2-{4-[(furan-2ylmethyl)amino]-1,3-dioxo-1,3-dihydroisoindol-2-yl}butyric acid, 2-{4-[(furan-2ylmethyl)amino]-1,3 -dioxo-1,3-dihydroisoindol-2-yl}-4-p-genylcarbamoyl-butyric acid and 2-{4[(furan-2-ylmethyl)amino]1,3-dioxo-1,3-dihydroisoindole-2 -yl}pentane acid, which have the following structures, respectively, and their pharmaceutically acceptable salts, solvates, prodrugs and stereoisomers:

- 30 044652 in which: one of X ¹ and X ² represents nitro or NH-Z, and the other of X ¹ or X ² represents hydrogen;

R ¹ and R ² are each, independently of the other, hydroxy or NH-Z;

R ³ represents alkyl of one to six carbon atoms, halogen or hydrogen;

Z represents hydrogen, phenyl, acyl of one to six carbon atoms or alkyl of one to six carbon atoms; and n is 0, 1 or 2; and if -COR ² and -(CH ₂ ) _n COR ¹ are different, the carbon atom designated C* represents the chiral center.

Other typical compounds have the formula:

in which: one of X ¹ and X ² is an alkyl containing from one to six carbon atoms;

R ¹ and R ² are each, independently of the other, hydroxy or NH-Z;

R ³ represents alkyl of one to six carbon atoms, halogen or hydrogen;

Z represents hydrogen, phenyl, acyl of one to six carbon atoms or alkyl of one to six carbon atoms; and n is 0, 1 or 2; and if -COR ² and -(CH ₂ ) _n COR1 are different, the carbon atom designated C* represents the chiral center.

Other specific immunomodulatory compounds are isoindolin-1-one and isoindolin-1,3-dione, substituted at the 2-position with 2,6-dioxo-3-hydroxypiperidin-5-yl, described in US Pat. No. 6,458,810, which is included in this document by reference.

Typical compounds have the formula:

in which: the carbon atoms designated * represent chiral centers; X represents -C(O)- or _-CH2- ;

R ¹ represents alkyl with 1-8 carbon atoms or -NHR ³ ;

R ² represents hydrogen, alkyl with 1-8 carbon atoms or halogen; And

R ³ represents hydrogen, alkyl of 1-8 carbon atoms, unsubstituted or substituted alkoxy of 1-8 carbon atoms, halogen, amino or alkylamino group of 1-4 carbon atoms, cycloalkyl of 3-18 carbon atoms, phenyl, unsubstituted or substituted by alkyl of 1-8 carbon atoms, alkoxy group of 1-8 carbon atoms, halogen, amino or alkylamino group of 1-4 carbon atoms, benzyl, unsubstituted or substituted by alkyl group of 1-8 carbon atoms, alkoxy group of 1-8 carbon atoms , halogen, amino or alkylamino group with 1-4 carbon atoms or -COR ⁴ in which R ⁴ represents hydrogen, alkyl with 1-8 carbon atoms, unsubstituted or substituted with alkoxy group with 1-8 carbon atoms, halogen, amino or alkylamino group with 1-4 carbon atoms, cycloalkyl with 3-18 carbon atoms, phenyl, unsubstituted or substituted with alkyl with 1-8 carbon atoms, alkoxy group with 1-8 carbon atoms, halogen, amino or alkylamino group with 1-4 carbon atoms, or benzyl, unsubstituted or substituted with alkyl of 1-8 carbon atoms, alkoxy group of 1-8 carbon atoms, halogen, amino or alkylamino group of 1-4 carbon atoms.

7. Examples

In the experiments described in examples 1-3, the following materials and methods were used.

Cell culture: Hek293 cells were grown in DMEM (ThermoFisher) supplemented with 10% fetal bovine serum (FBS). C6 cells were cultured in Ham's F12 nutrient mixture (ThermoFisher) supplemented with 10% FBS. NCIH929, U266B1, KMS12BM, OPM2 and NUDHL1 cells were cultured in RPMI-1640 medium supplemented with 10% FBS.

Plasmids: cDNA encoding full-length human BCMA (UniProtKB-Q02223; TNFRSF17) was cloned into the expression vector SF#10008 and transfected into C6 cells. The full-length cDNA encoding human TNFRSF12A (TWEAKR; NP_057723.1) was subcloned into the expression vector pCDNA3.1 (+), which was subsequently transfected into Hek293 cells. Transfectants were selected using puromycin or neomycin, and clonal populations were generated and analyzed by flow cytometry.

FACS Analysis: FACS antibodies used in this study include: 1) LS-C106982-100, a mouse monoclonal anti-human TNFRSF12A antibody conjugated to R-phycoerythrin

- 31 044652 (PE) (LifeSpan BioSciences, Inc.); 2) EM901, which was previously characterized as a BCMA-binding antibody (Mab42; international application number WO2017/021450); 3) human Fcy-specific IgG from Ebiosciences, labeled with PE; 4) human IgG isotype control; 5) anti-TACI antibody [1A1] (Abcam); 6) antibody to the BAFF receptor 8A7 (ThermoFisher); 7) anti-BCMA antibody 19F2 (BioLegend). Each cell type (75,000 cells/well) was incubated with primary antibodies in FACS buffer (PBS containing 0.2% BSA) for 25 min in a 96-well plate. Cells were washed once with FACS buffer and incubated with a 1:100 dilution of secondary antibody for 25 min. After washing once, cells were fixed for 10 min with 1% paraformaldehyde, washed with FACS buffer, and resuspended in 100 μl of FACS buffer. Labeled cells were analyzed using an Attune NxT flow cytometer, and data were processed using FlowJo 10.0.8rl (FLOWJO LLC) according to the manufacturer's instructions.

7.1. Example 1.

B cell maturation antigen (BCMA), also known as tumor necrosis factor receptor superfamily member 17 (TNFRSF17), is a protein that is found in many different types of cancer.

Preparation of anti-BCMA antibodies: Anti-human BCMA antibodies were obtained by immunizing animals with OmniFlic (Ligand Pharmaceuticals)x RAT5Lew, carried out at Aldevron Freiburg (Freiburg, Germany) with repeated use of a DNA vector containing BCMA-(aa 1-54) and recombinant extracellular human BCMA domain (aa 1-54), purchased from AcroBiosystem (Newark, DE, 19711). Lymph nodes from immunoreactive animals were collected, and total RNA was extracted from primary lymphocytes at Teneobio, Inc. (Palo Alto, California). IgG variable regions were amplified from each sample prior to paired-end next-generation sequencing followed by rank order analysis to quantify the expression level for each unique human IgG variable region.

Expression of anti-BCMA antibodies: The previously described five VH regions, along with a panel of putative BCMA-reactive VHs, were cotransfected with one common _VL in a standard IgG1/Carr antibody format. All antibody sequences were subcloned into the mammalian protein expression vector pTT5 (NRC-CNRC, Ottawa, ON, Canada). Antibodies were transiently transfected into Expi293 or ExpiCHO-S cells (Thermo Fisher Scientific, Waltham, MA) in 24-deep well plates (cat. no. P-DW-10ML-24-CS, Axygen, Tewksbury, MA) according to the manufacturer's protocols. For each antibody, a 1:1 DNA ratio for LC:HC was used for expression at 0.5 μg/ml DNA/ml expression medium, and cultures were shaken at 500 rpm using a 3 mm orbital shaking platform at 37°C at atmosphere 5% CO ₂ . Six to seven days after transfection, the media in the plates was clarified by centrifugation (3724 RCF, 4°C, 30 min), and the supernatants were transferred to new multiwell plates designed for titer determination using Protein A biosensors on an Octet Red 384 device (Pall ForteBio , Fremont, California) and screened for activity.

After identification of positive supernatants, potential anti-BCMA molecules were transiently transfected into ExpiCHO-S cells in a volume of 60 ml in 250 ml Erlenmeyer flasks (Cat. No: 431144, Corning, Tewksbury, MA) according to the manufacturer's instructions and shaken at 37°C and 120 rpm on a shaking platform with a 25 mm orbit. After one week of culture, supernatants were collected by centrifugation (3724 RCF, 4°C, 30 min) followed by filtration (Cat. No. 89220-720, VWR, Rednor, PA) and then purified.

Anti-BCMA Antibody Purification: Filtered supernatants were purified using 5 mL of Mab Select Sure Lx Resin (Cat. No. 29157185, GE Healthcare) on an ACT Pure Chromatography System (Cat. No. 29046694, GE Healthcare). These samples were eluted from the column using sodium citrate, pH 3.0, and neutralized to pH 5.5 with 3M Tris-HCl. The final samples were dialyzed into laboratory universal buffer (10 mM sodium acetate, pH 5.2 and 9 w/v% sucrose). Concentration and purity (aggregation assessment) were measured using a SEC column (cat. no. PL 1580-3301, Agilent, USA).

Surface Plasmon Resonance Binding Assay for Anti-BCMA Antibodies: The running buffer used in this assay was HBS-EP (150 mM NaCl, 10 mM Hepes pH 7.4, 3 mM EDTA, and 0.005% P20 surfactant) . For non-avid measurements, antibodies 320199 and 319883 were immobilized on a Protein A chip (GE Healthcare) at a concentration of 2 μg/ml, while recombinant Hu-BCMA (human BCMA) from Acro Biosystems (BCA-H522y) as well as Cyno -BCMA (cynomolgus BCMA) with the cleaved Fc was passed over them, starting at a maximum concentration of 50 nM followed by 3-fold dilutions to 0.6 nM (50, 16.66, 5.55, 1.85, 0.6 and 0 nM). Assays were performed at 25°C and 37°C and a flow rate of 10 μL.

Six anti-BCMA clones from Teniobio (320199, 319883, 319952, 320262, 319966, 320111) were tested for binding to Human-BCMA, Cyno-BCMA (cynomolgus), and Cyno-TACI (CAML transmembrane activator protein-interacting protein). cynomolgus) expressed in mouse Fc-tagged 293 cells using a Biacore-8K instrument (GE Healthcare). Were

- 32 044652 measurements of avidity (Fig. 1A) and non-avidity (Fig. 1B) were carried out.

To measure avidity, antigens were captured on surfaces capable of binding to murine antibodies at a concentration of 2 μg/ml, while antibodies were run at an initial maximum concentration of 50 nM followed by 3-fold dilutions to 0.6 nM (50, 16,66 , 5.55, 1.85, 0.6 and 0 nm). Assays were performed at 25°C and a flow rate of 10 μL/min for uptake, 30 μL/min for 3 min for association, and 5 min for dissociation. The surface was regenerated after each cycle with 10 mM glycine, pH 1.5, for 1 min. The running buffer used in this assay was HBS-EP (150 mM NaCl, 10 mM Hepes pH 7.4, 3 mM EDTA, and 0.005% surfactant P20).

Data analysis was performed using Biacore 8K Analysis software (GE Healthcare), considering data with duplicate controls and using a 1:1 Langmuir model for curve fitting. Anti-BCMA antibody from EngMab was used as a positive control, and anti-RSV1-IgG1 antibody was used as a negative control.

Immunohistochemistry: Single-color immunohistochemical (IHC) staining using DAB binding molecules in a human or rabbit IgG backbone with normal human and rabbit IgG (Abcam, ab172730) as isotype controls was performed on a Bond-Ill Autostainer (Leica Microsystems) with a Bond polymer refine detection kit (Leica, DS9800). Formalin-fixed, paraffin-embedded (FFPE) sections (4 μm) of the cell pellet were antigen unmasked using epitope retrieval solution 1 (Leica, AR9961) for 20 min at 100°C and blocked with peroxide blocking agent for 5 min. Sections were incubated with binding agents at appropriate dilutions for 15 min. For binding agents in the human IgG framework, a secondary rabbit anti-human IgG antibody (Abcam, ab2410, 1:100) was used for 8 min. This step was not required for the binding agents in the rabbit IgG scaffold. Sections were then incubated in polymer for 8 min, developed in DAB for 10 min, stained with hematoxylin for 5 min, and covered with a Sakura Finetek coverslip.

7.2. Anti-BCMA antibody binding studies. Example 2.

This example demonstrates that human IgGs, 320262, 319966, 320111, 319883 and 320199, recognize C6 cells expressing human BCMA protein. C6 cells stably transfected with human BCMA or mock-transfected C6 cells were incubated with 0.4 μg/ml EM901, 320262, 319966, 320111, 319883, or 320199. The reactivity of each antibody was monitored by FACS. EM901, an antibody previously shown to bind to BCMA (patent number WO 2017021450), bound C6-BCMA transfectants well, confirming BCMA expression by these cells. Compared to untransfected C6 cells (left histogram (blue) curve in each panel), histograms corresponding to numbers 320262, 319966, 320111, 319883, and 320199 (right histogram (red) curve in each panel) showed significant shifts to the right (Figure 2 ). These results indicate that 320262, 319966, 320111, 319883 and 320199 react with human BCMA expressed on the cell surface. Geometric means quantifying this binding are included (Figure 2).

7.3. Anti-BCMA antibody binding studies. Example 3.

This example shows that 320262, 319966, 320111, 319883 and 320199 do not bind Hek293 cells expressing full-length human TNFRSF12A/TWEAKR, a member of the BCMA-related TNF receptor family. Hek293 cells stably transfected with human TNFRSF12A were incubated with 0.4 μg/ml human IgG isotype control, 320262, 319966, 320111, 319883, or 320199. The reactivity of each antibody was monitored by FACS. Hek293TNFRSF12A cells stained with anti-TNFRSF12A antibody indicate expression of TNFRSF12A. Relative to the isotype control (blue histogram), no rightward shifts were detected in the histograms corresponding to numbers 320262, 319966, 320111, 319883, and 320199 (shown in red) (Fig. 3). These results indicate that 320262, 319966, 320111, 319883, and 320199 did not exhibit significant binding activity toward human TNFRSF12A. Geometric means to quantify this binding are shown (Figure 3).

7.4. Anti-BCMA antibody binding studies. Example 4.

This example demonstrates that 320262, 319966, 320111, 319883 and 320199 recognize NCIH929 multiple myeloma cells that endogenously express human BCMA. NCIH929 cells were incubated with 1.0 μg/ml human IgG isotype control, EM901, 320262, 319966, 320111, 319883, or 320199. The reactivity of each antibody was monitored by FACS. The BCMA-recognizing antibody EM901 binds strongly to NCIH929 cells. Relative to the isotype control (left histogram (blue) curve in each panel), histograms corresponding to numbers 320262, 319966, 320111, 319883, and 320199 (right histogram (red) curve in each panel) showed significant shifts to the right (Fig. 4). These results indicate that 320262, 319966, 320111, 319883, and 320199 interacted with NCIH929 cells. Geometric means to quantify this binding are shown (Figure 4).

7.5 Anti-BCMA antibody binding studies. Example 5.

- 33 044652

This example demonstrates that 320262, 319966, 320111, 319883 and 320199 recognize BCMA+, U266B1 and KMS12BM multiple myeloma cells incubated with 0.016-10 μg/ml human IgG isotype control, EM901, Figure 5. The reactivity of each antibody was monitored by FACS. EM901, 320262, 319966, 320111, 319883, and 320199 bound U266B1 and KMS12BM relative to the isotype control in a dose-dependent manner (Fig. 5). In addition, 320262 and 320111 recognized OPM2 multiple myeloma cells that express BCMA, but not TACI or BAFF receptor, other members of the BCMA-related TNF receptor family (dose-response curve and histogram, Fig. 5). In contrast, 320262 and 320111 do not bind NUDHL1 cells, which are BCMA'/TACI ⁺ /BAFFR ⁺ (dose response curve and histogram, Fig. 5). These results indicate that 320262, 319966, 320111, 319883 and 320199 exhibit BCMA-dependent, dose-dependent binding to cells. Geometric means to quantify this binding are shown (Figure 5).

SEQ ID NO. DESCRIPTION SUBSEQUENCE 1 Generic VL light chain EIVMTQSPATLSVSPGERATLLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSG SGTEFTLTISSLQSEDFAVYYCQQYNNWPWTFGQGTKVEIK 2 cLC CDR1 (according to Kabat, Chotiya, IMGT) RASQSVSSNLA 3 cLC CDR2 (according to Kabat, Chotiya, IMGT) GASTRAT 4 cLC CDR3 (according to Kabat, Chotiya, IMGT) QQYNNWPWT 5 cLC GAAATTGTGATGACTCAGTCGCCCGCCACCCTGTCCGTGTCTCCGGGAGAGCGGGCCACTCTGAGCTGT CGCGCGTCACAGTCGGTGTCCTCCAACCTCGCCTGGTACCAGCAGAAGCCTGGACAGGCCCCAAGACTG CTGATCTACGGCGCCTCCACCCGGGCCACCGGGATTCCTGCCCGGTTCTCCGGCTCCGGTTCCGGCACTG AGTTCACCCTGACCATCAGCTCACTGC AGTCCGAGGACTTCGCCGTGTACTACTGCCAGCAGTACAACA A CTGGCCGTGGACCTTTGGCCAAGGAACCAAGGTCGAAATCAAG 6 320111 VH QLPLQESGPGLVKPSETLSLTCTVSGGSIRSSSYYWGWIRQPPGKGLEWIGTIYYSGSTYYNPSLKS RVTISVDTSKNQLSLKLSSVTAADTAVYYCARPYYDSSGYYYYWGQGTLVTVSS 7 320111 VH SSSYYWG CDR1 according to Kabat 8 320111 VH CDR2 by Kabat TIYYSGTYYNPSLKS 9 320111 VH CDR3 by Kabat PYYDSSGYYYY 10 320111 VH CDR1 by Chotiya GGSIRSSSY AND 320111 VH CDR2 according to Chotiya YYSGS 12 320111 VH CDR3 according to Chotiya PYYDSSGYYYY 13 320111 VH CDR1 no IMGT GGSIRSSSYYWG 14 320111 VH CDR2 no IMGT TIYYSGSTYYNPSLKS 15 320111 VH CDR3 no IMGT PYYDSSGYYYY 16 320111 VH CAGCTGCCGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCACT GTTTCTGGTGGCTCCATCAGGAGTAGTAGTTACTACTGGGGCTGGATCCGGCAGCCCCCAGGGAAGGGG CTGGAGTGGATTGGGACTATCTATTATAGTGGGAGCACCTACTACAACCCGTCCCTCAAGAGTCGAGTC ACCATATCCGTAGACACGTCCAAGAACCAGCTCTCCCTGAAGCTGAGCTCTGTGACCGCCGCAGACACG

- 34 044652

GCTGTGTATTACTGTGCGAGACCTTACTATGATAGTAGTGGTTATTACTACTACTGGGGCCAGGGCACC TGGTCACCGTCTCCTCA 17 320199 VH QLQLQESGPGLVKPSETLSLTCTVSGGSISSSNSYWGWIRQSPGRGLEWIGRIYYSGITHYNPSLKSRVTISVD TSKNQFSLKLSSVTAADTAVYYCASPYKWNDGNFFGWGQGTLVTVSS 18 320199 VH CDR1 according to Kabat SSNSYWG 19 320199 VH CDR2 by Kabat RIYYSGITHYNPSLKS 20 320199 VH CDR3 by Kabat PYKWNDGNFG 21 320199 VH CDR1 by Chotiya GGSISSSNS 22 320199 VH CDR2 according to Chotiya YYSGI 23 320199 VH CDR3 according to Chotiya PYKWNDGNFG 24 320199 VH CDR1 no IMGT GGSISSSNSYWG 25 320199 VH CDR2 no IMGT RIYYSGITHYNPSLKS 26 320199 VH CDR3 no IMGT PYKWNDGNFG 27 320199 VH CAGCTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCACT GTCTCTGGTGGCTCCATCAGCAGTAGTAATTCCTACTGGGGCTGGATCCGCCAGTCCCCAGGGAGGGGG CTGGAGTGGATTGGGAGGATCTATTATAGTGGGATCACCCACTACAACCCGTCCCTCAAGAGTCGAGTC ACCATATCCGTAGACACGTCCAAGAACCAGTTCT CCCTGAAGCTGAGCTCTGTGACCGCCGCAGACACG GCTGTGTATTACTGTGCGAGTCCGTATAAGTGGAACGACGGGAATTTTTTTGGTTGGGGCCAGGGCACC CTGGTCACCGTCTCCTCA 28 320262 VH QLQLQESGPGLVKPSETLSLTCTVSGGSISNSNYYWGWIRQPPGKGLEWIGNIYYSGRTY YTPSLKSRVTISEDTSKNQFSLKVRSVTVADTGVYYCARPDYYGSGTIAWGQGTLVTVSS 29 320262 VH CDR1 according to Kabat NSNYYWG thirty 320262 VH CDR2 by Kabat NIYYSGRTYYTPSLKS 31 320262 VH CDR3 by Kabat PDYYGSGTIA 32 320262 VH CDR1 by Chotiya GGSISNSNY 33 320262 VH CDR2 by Chotiya YYSGR 34 320262 VH CDR3 by Chotiya PDYYGSGTIA 35 320262 VH GGSISNSNYYWG

- 35 044652

CDR1 no IMGT 36 320262 VH CDR2 no IMGT NIYYSGRTYYTPSLKS 37 320262 VH CDR3 no IMGT PDYYGSGTIA 38 320262 VH CAGCTGCAGCTACAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCACT GTCTCTGGTGGCTCCATCAGCAATAGTAATTATTACTGGGGCTGGATCCGCCAGCCCCCAGGAAAGGGG CTGGAGTGGATTGGGAATATCTATTATAGTGGGAGAACCTATTACACCCCGTCCCTCAAGAGTCGCGTC ACCATATCCGAAGACACGTCCAAGAACCAGTTCTCCCTGAAGGTGAGGTCTGTGACCGTCGCAGACAC GGGTGTGTATTACTGTGCGAGACCGGATTACTATGGTTCGGGGACTATCGCGTGGGGCCAGGGCACCCT GGTCACCGTCTCCTCA 39 319883 VH QLQLQESGPGLVKPSDTLSLTCTVSGGSISSSNSYWGWIRQSPGRGLEWIGRIYYSGITH YNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASPYKWNDGNFFGWGQGTLVTVSS 40 319883 VH CDR1 according to Kabat SSNSYWG 41 319883 VH CDR2 by Kabat RIYYSGITHYNPSLKS 42 319883 VH CDR3 by Kabat PYKWNDGNFG 43 319883 VH CDR1 by Chotiya GGSISSSNSY 44 319883 VH CDR2 by Chotiya YYSGI 45 319883 VH CDR3 by Chotiya PYKWNDGNFG 46 319883 VH CDR1 no IMGT GGSISSSNSYWG 47 319883 VH CDR2 no IMGT RIYYSGITHYNPSLKS 48 319883 VH CDR3 no IMGT PYKWNDGNFG 49 319883 VH CAGCTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGACACCCTGTCCCTCACCTGCACT GTCTCTGGTGGCTCCATCAGCAGTAGTAATTCCTACTGGGGCTGGATCCGCCAGTCCCCAGGGAGGGGG CTGGAGTGGATTGGGAGGATCTATTATAGTGGGATCACCCACTACAACCCGTCCCTCAAGAGTCGAGTC ACCATATCCGTAGACACGTCCAAGAACCAGTTCT CCCTGAAGCTGAGCTCTGTGACTGCCGCGGACACG GCTGTGTATTACTGTGCGAGTCCGTATAAGTGGAACGACGGGAATTTTTTTGGTTGGGGCCAGGGCACC CTGGTCACCGTCTCCTCA 50 319966 VH QLQLQESGPGLVKPSETLSLTCTVSGSSISRSNYYWGWIRQPPGKGLEWIGTFYYSGTTY YNPSLKSRVTISEDTSKKQLSLNLRSVTAADTAVYYCARPSGYTTSWGQGTLVTVSS 51 319966 VH CDR1 according to Kabat RSNYYWG 52 319966 VH CDR2 by Kabat TFYYSGTTYYNPSLKS 53 319966 VH CDR3 by Kabat PSGYTTS 54 319966 VH GSSISRSNY

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Claims (14)

CDR1 by Chotiya 55 319966 VH CDR2 by Chotiya YYSGT 56 319966 VH CDR3 by Chotiya PSGYTTS 57 319966 VH CDR1 no IMGT GSSISRSNYYWG 58 319966 VH CDR2 no IMGT TFYYSGTTYYNPSLKS 59 319966 VH CDR3 no IMGT PSGYTTS 60 319966 VH CAGCTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCACT GTCTCTGGAAGCTCCATCAGCAGGAGTAATTACTACTGGGGCTGGATCCGCCAGCCCCCAGGGAAGGGT CTGGAGTGGATTGGGACTTTCTATTATAGTGGGACCACCTACTACAACCCGTCCCTCAAGAGTCGAGTC A CCATATCCGAAGACACGTCCAAGAAACAGTTATCCCTGAACCTGAGGTCTGTGACCGCCGCAGACACGG CTGTGTATTACTGTGCGAGACCTTCCGGATATACCACCAGCTGGGGCCAGGGCACCCTGGTCACCGTCT C CTCA CLAIM 1. An antibody that binds to B-cell maturation antigen (BCMA), or a BCMA-binding antibody fragment containing: (A) heavy chain CDR1, CDR2 and CDR3 sequences: a) SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9, respectively; b) SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively; c) SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15, respectively; d) SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively; e) SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO: 23, respectively; f) SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID NO: 26, respectively; g) SEQ ID NO: 29, SEQ ID NO: 30 and SEQ ID NO: 31, respectively; h) SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34, respectively; i) SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37, respectively; j) SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively; k) SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45, respectively; l) SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, respectively; m) SEQ ID NO: 51, SEQ ID NO: 52 and SEQ ID NO: 53, respectively; n) SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, respectively; or o) SEQ ID NO: 57, SEQ ID NO: 58 and SEQ ID NO: 59, respectively; and (B) the light chain CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively. 2. An antibody or a BCMA-binding fragment thereof according to claim 1, comprising a light chain containing CDR1, CDR2 or CDR3 with the sequences SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively, and a heavy chain, containing CDR1, CDR2 and CDR3 with the sequences: a) SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9, respectively; b) SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively; or c) SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15, respectively. 3. The antibody or BCMA-binding fragment thereof according to claim 1, containing a light chain containing CDR1, CDR2 or CDR3 with the sequences SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively, and a heavy chain, containing CDR1, CDR2 and CDR3 with the sequences: a) SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively; b) SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO: 23, respectively; or c) SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID NO: 26, respectively. 4. An antibody or a BCMA-binding fragment thereof according to claim 1, comprising a light chain containing CDR1, CDR2 or CDR3 with the sequences SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively, and a heavy chain, containing the sequences CDR1, CDR2 and CDR3: a) SEQ ID NO: 29, SEQ ID NO: 30 and SEQ ID NO: 31, respectively; b) SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34, respectively; or - 37 044652 c) SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37, respectively. 5. An antibody or a BCMA-binding fragment thereof according to claim 1, comprising a light chain containing CDR1, CDR2 or CDR3 with the sequences SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively, and a heavy chain, containing CDR1, CDR2 and CDR3 with the sequences: a) SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively; b) SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45, respectively; or c) SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, respectively. 6. An antibody or a BCMA-binding fragment thereof according to claim 1, comprising a light chain containing CDR1, CDR2 or CDR3 with the sequences SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively, and a heavy chain, containing CDR1, CDR2 and CDR3 with the sequences: a) SEQ ID NO: 51, SEQ ID NO: 52 and SEQ ID NO: 53, respectively; b) SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, respectively; or c) SEQ ID NO: 57, SEQ ID NO: 58 and SEQ ID NO: 59, respectively. 7. The antibody or binding fragment thereof according to claim 1, which contains the light chain variable domain of SEQ ID NO: 1. 8. The antibody or binding fragment thereof of claim 1, which comprises the heavy chain variable domain of SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 28, SEQ ID NO: 39, or SEQ ID NO: 50. 9. The antibody of claim 1, wherein said antibody is Triomab, knob-in-hole (kih) common light chain IgG, CrossMAb, ortho-Fab IgG, DVD-Ig, 2 in 1-IgG, IgG-scFv, scFv ₂ -Fc, bi-nanobody, TandAb, DART, DART-Fc, scFv-HSA-scFv mniDNL-Fab3. 10. The antibody or BCMA binding fragment thereof according to claim 1, wherein the binding fragment is a single chain variable fragment (scFv), a Fab fragment or an F(ab') ₂ fragment. 11. Polypeptide that contains: (A) Heavy chain CDR1, CDR2 or CDR3 with sequences: a) SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9, respectively; b) SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively; c) SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15, respectively; d) SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively; e) SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO: 23, respectively; f) SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID NO: 26, respectively; g) SEQ ID NO: 29, SEQ ID NO: 30 and SEQ ID NO: 31, respectively; h) SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34, respectively; i) SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37, respectively; j) SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively; k) SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45, respectively; l) SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, respectively; m) SEQ ID NO: 51, SEQ ID NO: 52 and SEQ ID NO: 53, respectively; n) SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, respectively; or o) SEQ ID NO: 57, SEQ ID NO: 58 and SEQ ID NO: 59, respectively; and (B) light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively; and where the heavy chain and light chain form an antigen binding domain that binds to BCMA. 12. The polypeptide according to claim 11, which contains a light chain variable domain with the sequence SEQ ID NO: 1. 13. The polypeptide according to claim 11, which contains a heavy chain variable domain with the sequence SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 28, SEQ ID NO: 39 or SEQ ID NO: 50. 14. A method of depleting BCMA-expressing cells in a patient, comprising administering to the patient a therapeutically effective amount of an antibody or a BCMA-binding fragment or polypeptide thereof according to claim 1. -