Development and use of t cell adaptor therapeutics

Abstract

To provide a T cell adaptor therapeutic agent and use thereof.SOLUTION: The T-cell adaptor is a T-cell bispecific antibody comprising a first binding domain that binds to BCMA on the target cell surface and a second binding domain that binds to CD3 on the T-cell surface. The bispecific antibody is well tolerated, can effectively block the binding of BCMA to its receptor APRIL, and has no adverse effects such as weight loss.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to the development and use of T cell adaptor therapeutics, particularly those that bind to BCMA and CD3. The present invention relates to bispecific antibodies and uses thereof. [Background technology]

[0002] B-cell maturation antigen (BCMA) is a member of the tumor necrosis factor receptor superfamily and is widely expressed on the surface of myeloma cells. In normal tissues, it is expressed only on plasma cells, but also on naive B cells, CD34+ hematopoietic stem cells, and BCMA is not expressed in the cytoplasm of the B cell and other normal tissues. BCMA-deficient mice exhibit normal B cell development and proliferation. In samples from multiple myeloma patients, the ligand APRIL (A Pro BCMA binds to the BCMA and then binds to the bone marrow. It can inhibit tumor cell apoptosis while stimulating the growth and proliferation of myeloma cells. The expression level on malignant plasma cells is significantly higher than that on normal plasma cells. BCMA has become an important immunotherapeutic target.

[0003] Multiple myeloma is the second most common hematologic malignancy and occurs most frequently in middle-aged and elderly people. The accumulation of malignant proliferating plasma cells in the patient's bone marrow leads to bone marrow failure and immune dysfunction. This can lead to multiple osteolytic lesions, hypercalcemia, anemia, and kidney damage. In recent years, new drugs such as proteasome inhibitors, immunomodulators, and targeted antibodies have Although it significantly extends patient survival, nearly all patients still relapse. Novel therapeutic methods, such as CAR-T cells (chimeric antigen receptor T cells), The preparation process for HIV-1 receptor T-cells is complicated, the treatment costs are high, and recurrence is common. The rate is relatively high. ADCs (anticoagulants, anticoagulants) are available at low clinical doses due to the safety of the conjugated toxins. Dose-limited. Previously developed BCMA bispecific antibodies have shown excellent clinical results. Although it showed a good therapeutic effect, side effects also increased with increasing dose, and a strong cytokine stagnation occurred. This can lead to life-threatening neurological syndrome in patients. Therefore, new, safer and more effective drugs need to be developed.

[0004] T cell bispecific antibodies (also called T cell adaptors) bind to target cells at one end. It recognizes cell surface antigens (antigen arms) and binds to the T cell CD3 receptor (CD3 arms) at the other end. This is an antibody molecule that targets the CD3ε chain. Uses the body to aggregate CD3 on T cells in a manner similar to TCR / peptide / HLA This activates T cells and kills tumors. It targets BCMA expressed on myeloma cells and the CD3ε chain (CD3e) present on T cells. It is a T cell bispecific (TCB) antibody that targets BCMA and CD3. Mechanism of action of BCMA×CD3 bispecific molecules It simultaneously binds to BCMA+ myeloma cells and CD3+ T cells, promoting T cell activation and T cell proliferation. This includes causing cell-mediated cell killing.

[0005] During a normal immune response, TCRs react with low affinity (approximately 1-100 μM) to infection or sudden The exogenous peptide binds to the human leukocyte antigen (HLA) complex on the mutated cells and is activated by CD3. It transmits activation signals into the nucleus, and then activates transcription factors and their downstream proteins (cytokines, glutamate, etc.). This activates the expression of proteins such as ribozymes and perforins, and enhances the signal strength generated by the TCR complex. The degree of signaling determines the fate of the T cell. The ε, γ, δ, and ζ subunits of the signaling complex , which associate with each other to form CD3ε-γ heterodimers, CD3ε-δ heterodimers, and CD3ζ Many of the early CD3 bispecific antibodies developed were OKT 3, which are based on a small number of mouse-derived antibodies, such as L2K, UCHT1, and TR66; Relatively high affinity. Clinical studies have shown that CD3 antibodies with too high an affinity can cause T-cell This leads to excessive activation of the cells, releasing a large number of cytokines and causing cytokine storm syndrome. The high affinity of the antibody leads to the concentration of the bispecific antibody in secondary lymphoid organs. The ability of the antibody Fc portion to bind to Fcγ receptors increases drug safety. Another factor that influences immune responses is Fcγ receptors, which are expressed on a wide variety of normal tissues. Therefore, bispecific antibodies bind to Fcγ receptors on the cell membrane via the Fc domain and then bind to the other end of the Fc domain. The combined CD3 receptors are cross-linked and activated by aggregation of Fcγ receptors, resulting in severe off-target reactions. These drugs, for example, have been shown to cause adverse drug reactions, such as steroid steroids, which are often associated with steroid toxicity. ab) The Fc segment binds to Fcγ receptors expressed on hepatic Kupffer cells, resulting in rapid It induces rapid cytokine release. It is a human IgG2 subunit with weak binding ability to Fcγ receptors. IgG1 or IgG2 subtype, or further amino acid substitutions at the corresponding sites in CH2. For example, Armour et al. Substitution of (EU sequence number) with the corresponding sequence of IgG2 reduces binding to Fcγ receptors (Armour KL, et al, Recombinant human IgG molecules lacking Fcgamma receptor I bin ding and monocyte triggering activities, Eur J Immunol.1999 Aug;29(8):2613-24), Ne introduced the mutations Ser228Pro and Leu235Glu into IgG4. This stabilizes the IgG4 structure and reduces binding to Fcγ receptors (New man R,et al,Modification of the Fc regio n of a primatized IgG antibody to human CD4 retains its ability to modulate CD4 receptors but does not deplete CD4(+) T cells in chimpanzees.Clin Immunol.2001 F eb;98(2):164-74), Idusogie et al. 2. By substituting Pro329 or Pro331 with Ala, the complement of IgG1 is improved. We found that it can reduce binding to C1q (Idusogie EE, et al,Mapping of the C1q binding site on ri tuxan,a chimeric antibody with a human I gG1 Fc. J Immunol.2000 Apr 15;164(8):417 8-84). Summary of the Invention

[0006] To solve the problems present in the prior art, the present disclosure provides a method for producing a BCMA protein with high affinity. Further, the present disclosure provides a novel BCMA antibody having the following structure: The present invention provides a novel BCMA-CD3 κλ bispecific antibody produced by the body. -CD3κλ bispecific antibody adopts full-length IgG structure, and has been optimized for charge introduction and affinity. This allows the κλ bispecific antibody to preferentially localize to BCMA+ tumor tissue, allowing it to be active at low concentrations. It can recruit activated T cells and effectively kill target cells, and in the absence of target cells In this case, T cells are not activated, and the κλ bispecific antibody binds FcγRI and FcγRIIA. It does not bind to receptors such as FcγRIIIA and FcγRIIIA, reducing the risk of producing a cytokine storm. In an immune reconstitution mouse model, this novel BCMA-C The D3κλ bispecific antibody can effectively inhibit tumor growth and has a therapeutic effect superior to that of the same type of antibody. It has been proven that this is the case.

[0007] In one aspect, the disclosure provides an antibody, or antigen-binding portion thereof, that binds to BCMA. do.

[0008] In one aspect, the present disclosure provides a bispecific antibody or antigen-binding portion thereof.

[0009] In one aspect, the present disclosure provides a method for encoding the bispecific antibody or antigen-binding portion thereof. , providing nucleic acids.

[0010] In one aspect, the present disclosure provides a vector comprising the nucleic acid.

[0011] In one aspect, the present disclosure provides a cell comprising the nucleic acid or vector.

[0012] According to the antibody or antigen-binding portion thereof, the antibody or antigen-binding portion thereof may be humanized. This is what was done.

[0013] In one aspect, the present disclosure provides the antibody or antigen-binding portion thereof, or a nucleic acid encoding the same. and a pharmaceutically acceptable carrier.

[0014] In one aspect, the present disclosure provides an antibody or antigen-binding portion thereof covalently linked to a therapeutic moiety. The present invention provides antibody-drug conjugates, including bispecific or multispecific molecules.

[0015] In one aspect, the present disclosure provides a therapeutically effective amount of the antibody or antigen-binding fragment thereof in a mammal. BCMA, comprising administering a nucleic acid, vector, cell, and / or pharmaceutical composition Methods for treating associated conditions are provided.

[0016] In one aspect, the present disclosure provides a drug or a method for treating a BCMA-associated disease in a mammal. The antibody or antigen-binding fragment thereof, nucleic acid, vector, cell and / or Uses of the pharmaceutical compositions are provided.

[0017] The antibodies of the present disclosure can be used to detect BCMA protein, diagnose, treat, or treat BCMA-associated diseases. It is used for several applications, including prophylaxis. [Brief explanation of the drawings]

[0018] [Figure 1] FIG. 1 shows the results of SDS-PAGE of human and cynomolgus monkey BCMA-mFc bivalent recombinant protein and BCMA-hFckih monovalent recombinant protein. [Figure 2]Figure 1 shows that human and cynomolgus monkey BCMA stably transfected cells were detected by flow cytometry, and both human BCMA stably transfected cells (CHO-hBCMA-2C2) and cynomolgus monkey BCMA stably transfected cells (CHO-cynoBCMA-1A2) express high levels of BCMA. [Figure 3] FIG. 1 shows the results of SDS-PAGE of CD3εγ-Fc recombinant protein. [Figure 4] FIG. 1 shows binding of humanized CD3 antibodies to CD3εγ-Fc recombinant protein. [Figure 5] FIG. 1 shows binding of humanized CD3 antibodies to Jurkat cells. [Figure 6] FIG. 1 shows binding of BCMA×CD3κλ bispecific antibodies to BCMA stably transfected cells. [Figure 7] FIG. 1 shows high affinity binding of BCMA×CD3κλ bispecific antibodies to BCMA+ tumor cells. [Figure 8] FIG. 1 shows binding of BCMA×CD3κλ bispecific antibodies to Jurkat cells. [Figure 9] FIG. 1 shows binding of BCMA×CD3κλ bispecific antibodies to human peripheral blood T cells. [Figure 10] FIG. 10 shows killing of NCI-H929 cells and activation of T cells by BCMA×CD3κλ bispecific antibodies, with FIG. 10A showing killing of NCI-H929 cells and FIG. 10B showing activation of T cells. [Figure 11] 11A and 11B show the killing of RPMI-8226 cells and the activation of T cells by BCMA×CD3κλ bispecific antibodies, where FIG. 11A shows the killing of RPMI-8226 cells and FIG. 11B shows the activation of T cells. [Figure 12] FIG. 1 shows activation of T cell signaling pathways by BCMA×CD3κλ bispecific antibodies. [Figure 13] FIG. 1 shows the activation of peripheral blood T cells by BCMA×CD3κλ bispecific antibodies. [Figure 14] FIG. 1 shows binding of BCMA×CD3κλ bispecific antibodies to activating FcγR receptors. [Figure 15] FIG. 1 shows blocking of BCMA binding to its receptor APRIL by BCMA×CD3κλ bispecific antibodies. [Figure 16] FIG. 1 shows the suppressive effect of the BCMA×CD3κλ bispecific antibody on a subcutaneous NCI-H929 tumor-implanted model in immunodeficient mice. DETAILED DESCRIPTION OF THE INVENTION

[0019] I. Definition In this disclosure, unless otherwise specified, scientific and technical terms used herein are understood to be of ordinary skill in the art. As used herein, the terms protein and nucleic acid have the meanings commonly understood by those skilled in the art. Terminology and laboratory procedures related to chemistry, molecular biology, cell and tissue culture, microbiology, and immunology The operating procedures are terms and routine procedures widely used in the corresponding fields. For better understanding, definitions and explanations of relevant terms are provided below.

[0020] As used herein, the term "BCMA" refers to BCMA present in animals, preferably humans. Refers to the concept collectively referring to itself and any variants, isoforms, and paralogs thereof. can be done.

[0021] The term "human BCMA" refers to BCMA of human origin, preferably as registered in Genbank. The amino acid sequence may be, but is not limited to, that of Accession No. AB052772.1.

[0022] The terms "anti-BCMA antibody" and "antibody that binds to BCMA" refer to the use of such antibodies as diagnostic agents. and / or B with sufficient affinity so as to be useful in targeting BCMA as a therapeutic agent. In one embodiment, the anti-BCMA antibody is For example, unrelated non-BCMA proteins measured by radioimmunoassay (RIA) The binding of the anti-BCMA antibody to the target substance is approximately 10% of the binding of the antibody to BCMA. In some embodiments, antibodies that bind BCMA have a mAb concentration of ≦1 μM, ≦100 μM, or nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (For example, 10-8M or less, for example, 10-8M to 10-13M, for example, 10-9M to In some embodiments, the anti-BCMA antibody has a dissociation constant (Kd) of 10 M. The antibody binds to a conserved BCMA epitope among BCMA from different species.

[0023] Unless otherwise specified, "CD3" refers to primate (e.g., human) and non-human primate (e.g., human) CD3 antibodies. mammals such as cynomolgus monkeys and rodents (e.g., mice and rats) The term refers to any native CD3 from any vertebrate, including "full length" and unprocessed CD3. This includes CD3 derived from normal CD3 and any form of CD3 processed intracellularly. The term also covers naturally occurring variants of CD3, such as splice variants or allelic variants. In one embodiment, the CD3 is human CD3, particularly the ε subunit of human CD3. The amino acid sequence of human CD3ε is available from UniProt (www .uniprot.org) accession number P07766 (version 144), or NCBI (www.ncbi.nlm.nih.gov / )RefSeq NP_000724. 1. Cynomolgus monkey [Macaca fascicularis] CD3ε antigen The amino acid sequence is shown in NCBI GenBank no. BAB71849.1.

[0024] The term "cell surface" is used according to its ordinary meaning in the art, where , including the outside of the cell that is accessible by binding to proteins and other molecules.

[0025] As used herein, the terms "about" or "approximately" mean, unless otherwise indicated. Within ±10% of a given value or range. Where required to be an integer, the term Round up or down to the nearest integer within ±10% of the given value or range.

[0026] With respect to antibody chain polypeptide sequences, the term "substantially identical" refers to a sequence of a reference polypeptide 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% of the sequence or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more With respect to nucleic acid sequences, the term can be understood as antibody chains that exhibit sequence identity with the reference nucleic acid sequence. At least 60% above, 65% above, 70% above, 75% above, 80% above for the column, Over 85%, Over 90%, Over 95%, Over 96%, Over 97%, Over 98%, Over 99% A nucleotide sequence that has a higher sequence identity than the original sequence may be understood as a nucleotide sequence that has a higher sequence identity than the original sequence.

[0027] Sequence "homology" or "identity" has a meaning well known in the art and can be determined using the disclosed techniques. to calculate the percentage of sequence identity between two nucleic acid or polypeptide molecules or regions. along the entire length of a polynucleotide or polypeptide or in a region of the molecule. Sequence identity can be measured along the There are several ways to measure the homology of a gene, but the term "homology" is well known to those skilled in the art. (Carrillo, H. & Lipman, D., SIAM J Applied Ma th 48:1073(1988)).

[0028] "Substitutional" variants are those in which at least one amino acid residue in a native sequence has been removed and its corresponding amino acid residue has been removed. A variant is one in which a different amino acid is inserted at the same position in the molecule. A single substitution where only one amino acid is substituted, or two or more amino acid substitutions in the same molecule. Multiple substitutions may occur at consecutive positions. The amino acid may be replaced by multiple residues, and such variants include both substitutions and insertions. "Insertional" variants are variants in which one or more amino acids are substituted for an amino acid at a particular position in the native sequence. The term "directly adjacent" refers to a mutant inserted adjacent to an amino acid. This means that the linker is attached to an α-carboxyl or α-amino functional group. A deletion variant is a variant in which one or more amino acids in the native amino acid sequence have been removed. The variants are those lacking one or two amino acids in a particular region of the molecule.

[0029] With respect to the variable domain of an antibody, the term "variable" refers to the specific antibody directed against a particular target. Identification of related molecules that differ significantly in sequence among antibodies used for specific recognition and binding of However, the variability is not evenly distributed throughout the variable domains of antibodies. The variability is found in the complementarity-determining regions (CDRs) within all light- and heavy-chain variable domains. It consists of three segments called CDR1, CDR2, and CDR3) or hypervariable regions. The more highly conserved parts of the variable domains are concentrated in the framework. The variable domains of each naturally occurring heavy and light chain are called framework sequences (FR) regions. The main domains are four domains connected by three CDRs and mainly adopt a β-sheet configuration. The CDRs form a loop connecting the β-sheet structure, the loop comprising: In some cases, they form part of a β-sheet structure. The CDRs of each chain are usually bounded by the FR regions. The CDRs from the other chain form the target binding site (epitope) of the antibody. As used herein, the amino acid residues of immunoglobulins contribute to the formation of a specific peptide or determinant. Numbering follows the immunoglobulin amino acid residue numbering scheme of Kabat et al. unless otherwise noted. A CDR may have the ability to specifically bind to a related epitope. .

[0030] As used herein, an "antibody fragment" or "antigen-binding fragment" of an antibody is less than full-length. The variable region (e.g., one or more CDRs and / or one The antibody fragments may contain at least a portion of the full-length antibody (or multiple antibody binding sites) to enhance binding specificity and enhance the full-length antibody fragments. It refers to any portion of a full-length antibody that retains at least a portion of its specific binding ability. Thus, an antigen-binding fragment is an antigen-binding portion that binds to the same antigen as the antibody from which the antibody fragment is derived. Antibody fragments refer to antibody derivatives produced by enzyme-catalyzed treatment of full-length antibodies, including: as well as synthetically produced derivatives, e.g., recombinantly produced derivatives. , antibody fragments. Examples of antibody fragments include Fab, Fab', F(ab')2, single chain Fv (scFv), Fv, dsFv, diabodies, Fd, and Fd' fragments and modifications Other fragments include, but are not limited to, fragments of the lysosome. The antibody may comprise multiple chains linked by sulfide bonds and / or peptide linkers. Fragments generally contain more than or about 50 amino acids, typically 200 The antigen-binding fragment contains at least 200 amino acids or about 200 amino acids. When inserted into a gene (e.g., by replacing the corresponding region), it is immunospecific (i.e., A Ka of at least 10 to 10 M or a Ka of at least about 10 to 10 M The term "functional fragment" or "antibody fragment" includes any antibody fragment that provides an antibody that binds to an antigen (as shown). "CMA antibody analogs" refer to antibodies that inhibit the ability of the receptor to bind to a ligand or initiate signal transduction. As used herein, a fragment or analogue that inhibits or substantially reduces the ability of a gene to initiate a gene expression pathway. The functional fragments referred to herein generally have the same meaning as "antibody fragments" and, in relation to antibodies, inhibit or substantially reduce the ability of a compound to bind to a ligand or initiate signal transduction An "Fv" fragment may refer to a fragment that binds to the antibody, such as Fv, Fab, or F(ab')2. It is formed by the non-covalent association of one heavy chain variable domain and one light chain variable domain. In this configuration, the VH-VL dimer is the same as that of an intact antibody. Similarly, the three CDRs of each variable domain interact to form target binding sites on the surface of the VH-VL dimer. The six CDRs confer target binding specificity to the intact antibody. However, a single variable domain (or a polypeptide containing only three target-specific CDRs) Even half of the Fv) can still function to recognize and bind to the target.

[0031] As used herein, "bispecific antibodies" The term BsAb (antibody) refers to an antibody that can specifically bind to two different antigenic determinants. Generally, bispecific antibodies and / or antigen-binding molecules are It contains two antigen-binding sites, each with specificity for a different antigenic determinant. In some embodiments, the bispecific antibody and / or antigen-binding molecule binds two antigenic determinants. capable of simultaneously binding to two antigenic determinants, especially those expressed on two different cells .

[0032] As used herein, "monoclonal antibody" refers to a population of identical antibodies; This means that each individual antibody molecule in a population of monoclonal antibodies is identical to every other antibody molecule. This property is the opposite of that of polyclonal populations of antibodies, which contain antibodies with multiple distinct sequences. Monoclonal antibodies can be prepared by a number of well-known methods (Smith, et al. (2004) J. Clin. Pathol. 57, 912-917, and Nelson et al., J Clin Pathol (2000), 53, 111 -117) For example, monoclonal antibodies can be prepared by immortalizing B cells, e.g. For example, by fusing with myeloma cells to produce hybridoma cell lines, or by using viruses such as EBV. They can be prepared by infection of B cells with a virus. Recombinant techniques can be used to generate antibodies that encode The gene is produced by transforming a host cell with a plasmid carrying an artificial sequence of nucleotides that Antibodies can also be prepared in vitro from clonal populations of host cells.

[0033] As used herein, the term "hybridoma" or "hybridoma cell" refers to a Produced by the fusion of antibody-producing lymphocytes with non-antibody-producing cancer cells (usually myeloma or As known to those skilled in the art, hybrids are referred to as cells or cell lines (e.g., cells that are lymphoma cells). The cells grow and produce specific monoclonal antibodies, providing a continuous supply. Methods for producing hybridomas are known in the art. When the term "hybridoma cell" is mentioned, it refers to a subclonal hybridoma. This also includes the cells and progeny thereof.

[0034] As used herein, a full-length antibody refers to an antibody that comprises two full-length heavy chains (e.g., VH-CH1-C H2-CH3 or VH-CH1-CH2-CH3-CH4) and two full-length light chains (VL- CL) and hinge region, e.g., naturally occurring antibodies secreted by antibody-secreting B cells. and synthetically produced antibodies with the same domains.

[0035] The term "chimeric antibody" refers to an antibody in which the variable region sequences are derived from one species and the constant region sequences are derived from another. Antibodies derived from a species, e.g., whose variable region sequences are derived from a mouse antibody and whose constant region sequences are derived from a human antibody, Refers to antibodies derived from the body.

[0036] "Humanized" antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. immunoglobulins, immunoglobulin chains, or fragments thereof (e.g., Fv, Fab, Fab') , F(ab')2 or other antigen-binding subsequence of an antibody), Preferably, a humanized antibody is an antibody prepared from human immunoglobulins (recipient). the recipient antibody, wherein residues in the complementarity determining regions (CDRs) of the recipient antibody are those that bind to the desired specificity of non-human species (donor antibodies) such as mouse, rat, or rabbit with the same sex, affinity, and potency. It is a human immunoglobulin with the CDR residues replaced.

[0037] In addition, in humanization, CDR1, CDR2 and / or CDR3 of VH and / or VL By mutating amino acid residues within the region, one or more binding properties of the antibody (e.g., the parent It is also possible to improve the compatibility of the clones, for example by PCR-mediated mutagenesis. Mutations can be introduced and their effect on antibody binding or other functional properties determined as described herein. The mutation can be evaluated by the in vitro or in vivo tests described in Such mutations may be amino acid substitutions, additions, or deletions. Furthermore, the number of mutations in a CDR is usually not more than one or two. Humanized antibodies further include antibodies containing one or two amino acid mutations within the CDRs.

[0038] As used herein, the term "CDR" refers to a complementarity determining region. The tarity-determining region of an antibody molecule Light chains are known to have three CDRs, also called hypervariable regions, It exists in the variable regions of each heavy and light chain of an antibody and has extremely high variability in the primary structure of CDR. As used herein, the CDRs of a heavy chain are those that are located at the amino-terminal ends of the amino acid sequence derived from the heavy chain. The CDRs of the light chain are represented by CDR1, CDR2, and CDR3 at the ends, and the CDRs of the light chain are represented by CDR1, CDR2, and CDR3 at the ends. The amino-terminal sequences are designated CDR1, CDR2, and CDR3. The positions are adjacent to each other in the tertiary structure and determine the specificity of antigen binding to the antibody.

[0039] As used herein, the term "epitope" refers to the region of an antigen to which the paratope of an antibody binds. Epitopes are usually sequences of amino acids or sugar side chains. It contains chemically active surface groups of molecules, usually with specific three-dimensional structural features and specific charge characteristics. Has.

[0040] As used herein, with respect to an antibody or antigen-binding fragment thereof, "specifically binds" or The term "immunospecifically binds" is used interchangeably herein and refers to a binding site between an antibody or an antigen. The binding fragment binds to its cognate antigen via non-covalent interactions between the antibody and the antibody-binding site of the antigen. The term "antigen" refers to the ability to form one or more non-covalent bonds. The antigen may be an isolated antigen. , may be present on tumor cells. Usually, they immunospecifically bind (or specifically bind) to an antigen. ) antibodies have an affinity constant Ka(1) of about 1 x 107 M-1 or about 1 x 108 M-1 or greater. binds to the antigen with a dissociation constant (Kd) of 1×10 M or less The affinity constant can be determined by standard kinetic methods of antibody reactions, e.g., immunoassays, surface plasmon resonance assays, etc. SPR(Rich and Myszka(2000)Curr.Opin.Bi otechnol 11:54, Englebienne (1998) Analyst. 123:1599), isothermal titration calorimetry (ITC) or other kinetic phase analysis methods known in the art. An exemplary S for calculating the binding affinity of an antibody can be measured by interaction assays. (See U.S. Patent No. 7,229,619, which describes PR and ITC methods.) Devices and methods for real-time detection and monitoring of blood pressure are known and commercially available. Available for purchase (BiaCore 2000, Biacore AB, Upsala, SW eden and GE Healthcare Life Sciences;Mal (See mqvist (2000) Biochem. Soc. Trans. 27:335) .

[0041] As used herein, the terms "polynucleotide" and "nucleic acid molecule" refer to two or more refers to an oligomer or polymer containing nucleotides or nucleotide derivatives linked together Deoxyribonucleic acid (DNA) and ribonucleic acid, which are usually linked by phosphodiester bonds, As used herein, the term "nucleic acid molecule" includes DNA molecules, RNA molecules, and the like. Nucleic acid molecules may be single-stranded or double-stranded, and may be It may also be DNA.

[0042] As used herein, an isolated nucleic acid molecule refers to an isolated nucleic acid molecule that is isolated from the natural source of the nucleic acid molecule. A nucleic acid molecule that is isolated from other nucleic acid molecules. For example, an "isolated" nucleic acid molecule is a cDNA molecule. The acid molecule, when prepared by recombinant techniques, does not substantially alter other cellular material or culture medium. free of, or if chemically synthesized, substantially free of, chemical precursors or other chemical components Exemplary isolated nucleic acid molecules provided herein are not intended to be limiting unless otherwise specified. The present invention includes isolated nucleic acid molecules encoding the recombinant fragments.

[0043] As used herein, "operably" refers to a nucleic acid sequence, region, element, or domain. By "linked" is meant that the nucleic acid regions are functionally related to each other. For example, The promoter encodes a polypeptide so that it can regulate or mediate transcription of the nucleic acid. It can be operably linked to a nucleic acid.

[0044] "Conservative sequence modifications" of the sequences in the sequence listing herein, i.e., nucleotide Nucleotides that do not eliminate binding of an antibody containing an amino acid sequence to an antigen. Conservative nucleotide and amino acid sequence modifications are also provided. This includes nucleotide and amino acid substitutions, and additions and deletions of nucleotides and amino acids. by standard techniques well known in the art (e.g., site-directed mutagenesis and PCR-mediated mutagenesis). Modifications may be introduced into the sequence listings provided herein. Conservative sequence modifications are Similar side chains include amino acid substitutions, in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having side chains of the formula: The family consists of amino acids with basic side chains (e.g., lysine, arginine, and histidine). ), amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar Amino acids with side chains (e.g., glycine, asparagine, glutamine, serine, threonine) amino acids with non-polar side chains (e.g., tryptophan, tyrosine, cysteine, tryptophan); , alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine amino acids with β-branched side chains (e.g., threonine, valine, isoleucine) and Amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, Histidine). Therefore, predicted non-essential amino acid residues in anti-BCMA antibodies are Preferably, the amino acid residue is replaced with another amino acid residue from the same side chain family. Methods for identifying conservative substitutions of nucleotides and amino acids that do not disrupt the sequence are well known in the art. (e.g., Brummell et al., Biochem. 32:1180-1 187(1993);Kobayashi et al.Protein Eng.12( 10):879-884(1999);Burkset al,Proc.Natl.A cad.Sci.USA 94:412-417(1997)).

[0045] Alternatively, in another embodiment, anti-BCMA antibodies can be generated, e.g., by saturation mutagenesis. Random mutations can be introduced along all or part of the antibody coding sequence. The resulting modified anti-BCMA antibodies are then screened for improved binding activity. This can be done.

[0046] As used herein, "expression" refers to the production of a polypeptide by transcription and translation of a polynucleotide. The expression level of a polypeptide can be determined, for example, by the amount of polypeptide produced by a host cell. using any method known in the art, including methods for determining the amount of polypeptide produced. Such methods can be used to assess the activity of polypeptides in cell lysates by ELISA. quantification of ATP, gel electrophoresis followed by Coomassie blue staining, Lowry protein assay, and and Bradford protein assay. .

[0047] As used herein, a "host cell" refers to a cell that receives, maintains, replicates, and expresses a vector. The host cell is a cell used to amplify the vector encoded by the vector. When the host cell divides, the vector is transformed into a polypeptide. The nucleic acid contained in the target is replicated, and the nucleic acid is amplified. Suitable host cells include CHO cells, various COS cells, and the like. cells, HeLa cells, HEK cells, e.g., HEK293 cells, but are not limited to these. do not have.

[0048] As used herein, a "vector" is a replicable nucleic acid that can be used to infect a suitable host. When transformed into a cell, the vector can express one or more heterologous proteins. The vector can be used to express the polypeptide or a fragment thereof, typically by restriction digestion and ligation. The vector includes a vector into which a nucleic acid encoding a polypeptide can be introduced. Further included are vectors containing the nucleic acid. The vectors may be used for amplification of the nucleic acid or for encoding the nucleic acid. The nucleic acid encoding the polypeptide is introduced into a host cell for expression / display of the polypeptide. Vectors are usually episomal but can also be used to deliver genes or A part of the chromosome may be designed to be integrated into the genome. Vectors that are artificial chromosomes, such as mammalian artificial chromosomes, are also contemplated. The selection and use of such a filter is well known to those skilled in the art.

[0049] As used herein, a vector is a "viral vector" or "viral vector." Viral vectors can also be used to deliver exogenous genes (vehicles or shuttles). engineered gene operably linked to an exogenous gene for transfer into a cell (as a It's a virus.

[0050] As used herein, an "expression vector" refers to a vector capable of expressing DNA. The DNA may contain, for example, a promoter region, a control that affects the expression of the DNA fragment. Such additional fragments may be operably linked to promoter and terminator sequences. - sequences, and optionally one or more origins of replication, one or more selectable markers The expression vehicle may contain a sequence, an enhancer, a polyadenylation signal, etc. may be derived from plasmid or viral DNA, or contain elements of both Thus, expression vehicles can be recombinant DNA or RNA constructs, such as plasmids, Refers to a phage, recombinant virus, or other vector that, when introduced into a suitable host cell, The expression vectors result in expression of the cloned DNA. Suitable expression vectors are well known to those skilled in the art. expression vectors that are replicable in eukaryotic and / or prokaryotic cells, and episomal These include expression vectors that remain intact or that integrate into the host cell genome.

[0051] As used herein, "treating" an individual with a disease or symptoms of a disease means treating said individual This means that the symptoms of a disease are partially or totally alleviated or remain unchanged after treatment. Thus, treatment includes prophylaxis, treatment and / or cure. Prophylaxis includes the prevention and treatment of potential diseases. Treatment refers to the prevention of the worsening of symptoms or progression of the disease. or antigen-binding fragments thereof, and any pharmaceutical use of the compositions provided herein. Further includes:

[0052] As used herein, "therapeutic benefit" refers to the benefit obtained by treating an individual with a disease. A change in the symptoms of a disease or disease state, usually an improvement or amelioration of the disease or disease state, or is a cure for a disease or disease state.

[0053] As used herein, a "therapeutically effective amount" or "therapeutically effective dose" refers to a therapeutically effective amount of a compound that is effective to treat a patient after administration to a subject. A drug, compound, substance, or composition containing a compound, at least sufficient to achieve the effect It therefore refers to the amount of a substance that prevents, cures, improves, suppresses or partially cures the symptoms of a disease or condition. This is the amount necessary to suppress the

[0054] As used herein, a "prophylactically effective amount" or "prophylactically effective dose" refers to a dose that, upon administration to a subject, e.g., For example, preventing or delaying the onset or recurrence of a disease or condition, reducing the likelihood of the onset or recurrence of a disease or condition, a substance, compound, material, or composition containing a compound that has a desired preventative effect, such as reducing the risk of The full prophylactically effective dose does not need to be achieved by administering a single dose. A prophylactically effective amount may be achieved only when a series of doses is administered. Thus, a prophylactically effective amount may be achieved only when a series of doses is administered. Several doses may be administered.

[0055] As used herein, the term "patient" refers to a mammal, such as a human.

[0056] II. Specific Embodiments

[0057] In one embodiment, the present disclosure provides a method for detecting a BCMA antigen comprising: (a) detecting a first antibody that binds to a BCMA antigen on the surface of a target cell; a first antigen-binding portion or antigen-binding fragment thereof, the first antigen-binding portion comprising a first heavy chain and a first and the first antigen-binding portion comprises a first binding domain that binds to the first antigen. , the first binding domain is selected from the amino acid sequences SEQ ID NO: 12, 13, 14, 26, 27, 28, 36, 37, 43, 44, 50, 55, 56, 57, 65, 66, 71, 7 2, 73, 147, 186 or any variant thereof, and / or Amino acid sequences SEQ ID NO: 17, 18, 19, 22, 23, 31, 32, 33, 40, 47, 60, 61, 62, 76, 77, 78, 83, 84, 87, 88, 89, 9 a first antigen-binding portion comprising a light chain CDR selected from 2, 144, or any variant thereof; an antigen-binding fragment, and (b) a second antigen-binding portion or antigen-binding fragment thereof that binds to the CD3 antigen on the surface of T cells; wherein the second antigen-binding portion comprises a second heavy chain and a second light chain, and the second antigen-binding portion is a second antigen-binding moiety comprising a second binding domain that binds to a second antigen, or The present invention provides bispecific antibodies or antigen-binding portions thereof, including fragments thereof.

[0058] In the antibody or antigen-binding portion thereof according to the above embodiment, the first binding domain comprises the amino acid Selected from the sequences SEQ ID NO: 12, 26, 55, 65, 71 or any variant thereof Heavy chain CDR1, amino acid sequences SEQ ID NOs: 13, 27, 36, 43, 50, a heavy chain CDR2 selected from 56, 66, 72, 147, 186, or any variant thereof; The amino acid sequences SEQ ID NO: 14, 28, 37, 44, 57, 73 or any variant thereof and / or the first binding domain comprises a heavy chain CDR3 selected from the amino acid sequence Column SEQ ID NO: 17, 22, 31, 47, 60, 76, 83, 87, 92, 14 4 or any variant thereof, the amino acid sequence of which is selected from SEQ ID NO: 18, 23, 32, 40, 61, 77, 84, 88 or any variant thereof Chain CDR2, amino acid sequence SEQ ID NO: 19, 33, 62, 78, 89 or It comprises a light chain CDR3 selected from any variant.

[0059] According to any of the above-described embodiments of the antibody or antigen-binding portion thereof, the first binding domain The heavy chain CDR1, CDR2 and CDR3 sequences are the amino acid sequences SEQ ID NO:12, 13, 14. First heavy chain CDR1, CDR2 and CDR3 sequences, amino acid sequence SEQ ID NO: The first heavy chain CDR1, CDR2 and CDR3 sequences of D NO:12, 186 and 14, The first heavy chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 26, 27, 28 DR3 sequence, first heavy chain CDR1 of amino acid sequence SEQ ID NO: 12, 36, 37 , CDR2 and CDR3 sequences, amino acid sequences SEQ ID NO: 12, 43, 44 Heavy chain CDR1, CDR2 and CDR3 sequences of 1, amino acid sequence SEQ ID NO:12 , 50, 14, the first heavy chain CDR1, CDR2 and CDR3 sequences, amino acid sequence SEQ The first heavy chain CDR1, CDR2 and CDR3 sequences of ID NOs: 55, 56 and 57, The first heavy chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 65, 66, 14 DR3 sequence, first heavy chain CDR1 of amino acid sequence SEQ ID NO: 71, 72, 73 , CDR2 and CDR3 sequences, amino acid sequences of SEQ ID NOs: 71, 147, 73 a first heavy chain CDR1, CDR2, and CDR3 sequence selected from the first binding domain; The light chain CDR1, CDR2 and CDR3 sequences of SEQ ID NO:17 , 18, 19, the first light chain CDR1, CDR2 and CDR3 sequences, amino acid sequence SEQ The first light chain CDR1, CDR2 and CDR3 sequences of ID NOs: 22, 23 and 19, The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 31, 32, 33 DR3 sequence, first light chain CDR1 of amino acid sequence SEQ ID NO: 17, 40, 19 , CDR2 and CDR3 sequences, amino acid sequences SEQ ID NO: 47, 18, 19 1, light chain CDR1, CDR2 and CDR3 sequences of amino acid sequence SEQ ID NO:60 , 61, 62, the first light chain CDR1, CDR2 and CDR3 sequences, amino acid sequence SEQ The first light chain CDR1, CDR2 and CDR3 sequences of ID NOs: 76, 77 and 78, The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 83, 84, 19 DR3 sequence, first light chain CDR1 of amino acid sequence SEQ ID NO: 87, 88, 89 , CDR2 and CDR3 sequences, amino acid sequences SEQ ID NO: 92, 23, 19 Light chain CDR1, CDR2 and CDR3 sequences of 1, amino acid sequence SEQ ID NO:14 4, 77, 78.

[0060] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N O: The first heavy chain CDR1, CDR2 and CDR3 sequences of 12, 13 and 14, and The first light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 17, 18, 19 Contains the R3 sequence.

[0061] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N O: The first heavy chain CDR1, CDR2 and CDR3 sequences of 12, 13 and 14, and The first light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 22, 23, 19 Contains the R3 sequence.

[0062] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N O: The first heavy chain CDR1, CDR2 and CDR3 sequences of 26, 27 and 28, and amino acid sequence The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 31, 32, 33 Contains the R3 sequence.

[0063] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain CDR1, CDR2 and CDR3 sequences of O:12, 36, 37, and amino acid sequence The first light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 17, 40, 19 Contains the R3 sequence.

[0064] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain CDR1, CDR2 and CDR3 sequences of O:12, 43, 44, and amino acid sequence The first light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 47, 18, 19 Contains the R3 sequence.

[0065] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N O: The first heavy chain CDR1, CDR2 and CDR3 sequences of 12, 50, 14, and amino acid sequence The first light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 17, 40, 19 Contains the R3 sequence.

[0066] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain CDR1, CDR2 and CDR3 sequences of O:55, 56, 57, and amino acid sequence The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 60, 61, 62 Contains the R3 sequence.

[0067] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N O: The first heavy chain CDR1, CDR2 and CDR3 sequences of 12, 13 and 14, and The first light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 17, 40, 19 Contains the R3 sequence.

[0068] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain CDR1, CDR2 and CDR3 sequences of O:65, 66, 14, and amino acid sequence The first light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 17, 40, 19 Contains the R3 sequence.

[0069] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain CDR1, CDR2 and CDR3 sequences of O:71, 72, 73, and amino acid sequence The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 76, 77, 78 Contains the R3 sequence.

[0070] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N O: The first heavy chain CDR1, CDR2 and CDR3 sequences of 12, 13 and 14, and The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 83, 84, 19 Contains the R3 sequence.

[0071] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N O: The first heavy chain CDR1, CDR2 and CDR3 sequences of 12, 13 and 14, and The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 87, 88, 89 Contains the R3 sequence.

[0072] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N O: The first heavy chain CDR1, CDR2 and CDR3 sequences of 12, 13 and 14, and The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 92, 23, 19 Contains the R3 sequence.

[0073] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain CDR1, CDR2 and CDR3 sequences of O:71, 147, 73, and The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 144, 77, 78 Contains the CDR3 sequence. In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain CDR1, CDR2 and CDR3 sequences of O:12, 186, 14, and The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 17, 40, 19 Contains the DR3 sequence. In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain CDR1, CDR2 and CDR3 sequences of O:12, 186, 14, and The first light chain CDR1, CDR2 and CDR3 of the amino acid sequence SEQ ID NO: 22, 23, 19 Contains the DR3 sequence.

[0074] According to any of the above-described embodiments of the antibody or antigen-binding portion thereof, the first binding domain comprises , amino acid sequences SEQ ID NO: 10, 24, 34, 41, 48, 53, 63, 69 , 79, 145, 150, 154, 158, 162, 166, 170, 187 or any of the above a first heavy chain variable region selected from any variant thereof and / or an amino acid sequence SEQ ID NO: NO:15, 20, 29, 38, 45, 51, 58, 67, 74, 81, 85, 90, 1 42, 148, 152, 156, 160, 164, 168, 188 or any variant thereof The first light chain variable region is selected from: In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:187 or any variant thereof, and the amino acid sequence SEQ. The first light chain variable region of D NO:188 or any variant thereof.

[0075] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:10 or any variant thereof, and the amino acid sequence SEQ ID The first light chain variable region of NO:15 or any variant thereof.

[0076] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:10 or any variant thereof, and the amino acid sequence SEQ ID The first light chain variable region of NO:20 or any variant thereof.

[0077] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:24 or any variant thereof and the amino acid sequence SEQ ID The first light chain variable region of NO:29 or any variant thereof.

[0078] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:34 or any variant thereof and the amino acid sequence SEQ ID NO: The first light chain variable region of NO:38 or any variant thereof.

[0079] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:41 or any variant thereof and the amino acid sequence SEQ ID NO: The first light chain variable region of NO:45 or any variant thereof.

[0080] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:48 or any variant thereof and the amino acid sequence SEQ ID NO: The first light chain variable region of NO:51 or any variant thereof.

[0081] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:53 or any variant thereof and the amino acid sequence SEQ ID The first light chain variable region of NO:58 or any variant thereof.

[0082] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:10 or any variant thereof, and the amino acid sequence SEQ ID The first light chain variable region of NO:38 or any variant thereof.

[0083] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:63 or any variant thereof and the amino acid sequence SEQ ID The first light chain variable region of NO:67 or any variant thereof.

[0084] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:69 or any variant thereof and the amino acid sequence SEQ ID The first light chain variable region of NO:74 or any variant thereof.

[0085] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:79 or any variant thereof and the amino acid sequence SEQ ID The first light chain variable region of NO:81 or any variant thereof.

[0086] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:10 or any variant thereof, and the amino acid sequence SEQ ID The first light chain variable region of NO:85 or any variant thereof.

[0087] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:10 or any variant thereof, and the amino acid sequence SEQ ID The first light chain variable region of NO:90 or any variant thereof.

[0088] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:145 or any variant thereof, and the amino acid sequence SEQ. The first light chain variable region of D NO:142 or any variant thereof.

[0089] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:150 or any variant thereof, and the amino acid sequence SEQ. The first light chain variable region of D NO:148 or any variant thereof.

[0090] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:154 or any variant thereof, and the amino acid sequence SEQ. The first light chain variable region of D NO:152 or any variant thereof.

[0091] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:158 or any variant thereof, and the amino acid sequence SEQ. D NO:156 or any variant thereof.

[0092] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:162 or any variant thereof, and the amino acid sequence SEQ. The first light chain variable region of D NO:160 or any variant thereof.

[0093] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:166 or any variant thereof, and the amino acid sequence SEQ. The first light chain variable region of D NO:164 or any variant thereof.

[0094] In some embodiments, the first binding domain has the amino acid sequence SEQ ID N The first heavy chain variable region of O:170 or any variant thereof, and the amino acid sequence SEQ. D NO:168 or any variant thereof.

[0095] According to the antibody or antigen-binding portion thereof of any of the above embodiments, the second binding domain comprises , amino acid sequences SEQ ID NO: 114, 115, 116, 119, 122, 125 , 128, 131, 134, 135, or any variant thereof; and / or the amino acid sequences SEQ ID NOs: 95, 96, 97, 102, 103, 108 , 111, or any variant thereof.

[0096] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N O: A second heavy chain CDR1 selected from 114, 119, 134 or any variant thereof; a second amino acid sequence selected from SEQ ID NO: 115, 135 or any variant thereof; Heavy chain CDR2, amino acid sequences SEQ ID NOs: 116, 122, 125, 128, 131 or any variant thereof, and / or a second heavy chain CDR3 and / or an amino acid sequence thereof. A second light chain CD44 selected from SEQ ID NO: 95, 102 or any variant thereof. R1, amino acid sequence SEQ ID NO: 96, 103, 111 or any variant thereof a second light chain CDR2 selected from the amino acid sequence SEQ ID NO: 91, 108 or and a second light chain CDR3 selected from any variant of:

[0097] In some embodiments, the heavy chain CDR1, CDR2 and CDR3 of the second binding domain The R3 sequence is the second heavy chain C of the amino acid sequence SEQ ID NO: 114, 115, 116. DR1, CDR2 and CDR3 sequences, amino acid sequences SEQ ID NO: 119, 115 , the second heavy chain CDR1, CDR2 and CDR3 sequences of 116, the amino acid sequence SEQ ID The second heavy chain CDR1, CDR2 and CDR3 sequences of NOs: 119, 115 and 122, The second heavy chain CDR1, CDR2 of amino acid sequence SEQ ID NO: 119, 115, 125 2 and CDR3 sequences, the second of amino acid sequences SEQ ID NO: 119, 115, 128 Heavy chain CDR1, CDR2 and CDR3 sequences of amino acid sequence SEQ ID NO:119 , 115, 131, the second heavy chain CDR1, CDR2 and CDR3 sequences, amino acid sequence SE The second heavy chain CDR1, CDR2 and CDR3 of Q ID NOs: 134, 135 and 116 and the light chain CDR1, CDR2 and CDR3 sequences of the second binding domain The second light chain CDR1, CDR2, CDR3, CDR4, CDR5, CDR6, CDR7, CDR8, CDR9, CDR10, CDR11, CDR12, CDR13, CDR14, CDR15, CDR16, CDR17, CDR18, CDR19, CDR110, CDR111, CDR12, CDR13, CDR14, CDR15, CDR16, 2 and CDR3 sequences, the second of the amino acid sequences SEQ ID NO: 102, 103, 97 Light chain CDR1, CDR2 and CDR3 sequences, amino acid sequence SEQ ID NO:95, 9 6, 108 second light chain CDR1, CDR2 and CDR3 sequences, amino acid sequence SEQ I D From the second light chain CDR1, CDR2 and CDR3 sequences of NOs: 95, 111, 108 Be chosen.

[0098] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N O: the second heavy chain CDR1, CDR2 and CDR3 sequences of 114, 115, 116, and The second light chain CDR1, CDR2 and CDR3 of the amino acid sequences SEQ ID NO: 95, 96, 97 and CDR3 sequences.

[0099] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N O: the second heavy chain CDR1, CDR2 and CDR3 sequences of 114, 115, 116, and The second light chain CDR1, CDR2 of amino acid sequence SEQ ID NO: 102, 103, 97 2 and CDR3 sequences.

[0100] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N O: the second heavy chain CDR1, CDR2 and CDR3 sequences of 114, 115, 116, and Second light chain CDR1, CDR2 of amino acid sequence SEQ ID NO: 95, 96, 108 and CDR3 sequences.

[0101] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N O: the second heavy chain CDR1, CDR2 and CDR3 sequences of 114, 115, 116, and The second light chain CDR1, CDR2 of amino acid sequence SEQ ID NO: 95, 111, 108 2 and CDR3 sequences.

[0102] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N O:112, 117, 120, 123, 126, 129, 132, 136, 138, 14 a second heavy chain variable region selected from the group consisting of the amino acid sequence S EQ ID NO: 93, 98, 100, 104, 106, 109 or any variant thereof and a second light chain variable region selected from:

[0103] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N The second heavy chain variable region of O:136 or any variant thereof, and the amino acid sequence SEQ. D NO:93 or any variant thereof.

[0104] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N The second heavy chain variable region of O:138 or any variant thereof, and the amino acid sequence SEQ. D NO:93 or any variant thereof.

[0105] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N The second heavy chain variable region of O:138 or any variant thereof, and the amino acid sequence SEQ. D NO:98 or any variant thereof.

[0106] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N The second heavy chain variable region of O:138 or any variant thereof, and the amino acid sequence SEQ. The second light chain variable region of D NO:100 or any variant thereof.

[0107] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N The second heavy chain variable region of O:112 or any variant thereof, and the amino acid sequence SEQ. D NO:106 or any variant thereof.

[0108] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N The second heavy chain variable region of O:136 or any variant thereof, and the amino acid sequence SEQ. D NO:106 or any variant thereof.

[0109] In some embodiments, the second binding domain has the amino acid sequence SEQ ID N The second heavy chain variable region of O:138 or any variant thereof, and the amino acid sequence SEQ. D NO:106 or any variant thereof.

[0110] According to any of the above-described embodiments of the antibody or antigen-binding portion thereof, the first antigen-binding portion The first light chain is a kappa-type light chain and the second light chain of the second antigen-binding moiety is a lambda-type light chain.

[0111] In some embodiments, the second light chain variable region of the second antigen-binding portion comprises Gln4 0Glu mutation (VλCD3: Gln40Glu), and the second antigen-binding portion The heavy chain variable region of this antibody has a Gln39Lys mutation (VHCD3:Gln39Lys). do.

[0112] In some embodiments, the first light chain variable region of the first antigen-binding portion comprises Gln4 In some embodiments, the VκBCMA polypeptide has a Gln42Lys mutation (VκBCMA: Gln42Lys). wherein the first heavy chain variable region of the first antigen-binding portion has a Gln39Glu mutation (VH BCMA:Gln39Glu).

[0113] In some embodiments, the first antigen-binding portion and the second antigen-binding portion of the bispecific antibody The Fc portion of the binding moiety adopts a knob-into-hole structure. In a preferred embodiment, the Fc portion comprises a human IgG4 knob-into-hole structure. To hire.

[0114] In some embodiments, the first heavy chain is selected from the group consisting of SEQ ID NOs: 174, 178, or wherein the first light chain comprises a heavy chain selected from any variant thereof, and the second light chain comprises a heavy chain selected from SEQ ID NO: 17 2, 176 or any variant thereof.

[0115] In some preferred embodiments, the first heavy chain is SEQ ID NO: 174 or wherein the first light chain comprises a heavy chain selected from any variant thereof, and the second light chain comprises a heavy chain selected from SEQ ID NO: 17 2 or any variant thereof.

[0116] In some preferred embodiments, the first heavy chain is SEQ ID NO: 174 or wherein the first light chain comprises a heavy chain selected from any variant thereof, and the second light chain comprises a heavy chain selected from SEQ ID NO: 17 6 or any variant thereof.

[0117] In some preferred embodiments, the first heavy chain is SEQ ID NO: 178 or wherein the first light chain comprises a heavy chain selected from any variant thereof, and the second light chain comprises a heavy chain selected from SEQ ID NO: 17 2 or any variant thereof.

[0118] In some preferred embodiments, the first heavy chain is SEQ ID NO: 178 or wherein the first light chain comprises a heavy chain selected from any variant thereof, and the second light chain comprises a heavy chain selected from SEQ ID NO: 17 6 or any variant thereof.

[0119] In some embodiments, the second heavy chain has the sequence of SEQ ID NO: 182 or any of its derivatives. The first heavy chain comprises a heavy chain of any variant thereof, and the second light chain comprises a heavy chain of SEQ ID NO: 180 or any variant thereof. Contains variant light chains. In some preferred embodiments, the first binding domain of the bispecific antibody is an amino acid sequence. a first heavy chain variable region of SEQ ID NO: 187 or any variant thereof; and , the first light chain variable region of amino acid sequence SEQ ID NO: 188 or any variant thereof and the second binding domain comprises the amino acid sequence SEQ ID NO: 138, or any of its and a variant second heavy chain variable region of the amino acid sequence SEQ ID NO: 106 or and a second light chain variable region of any variant of

[0120] In some preferred embodiments, the first binding domain of the bispecific antibody is an amino acid sequence. a first heavy chain variable region of SEQ ID NO: 145 or any variant thereof; and , the first light chain variable region of amino acid sequence SEQ ID NO: 142 or any variant thereof and the second binding domain comprises the amino acid sequence SEQ ID NO: 138, or any of its and a variant second heavy chain variable region of the amino acid sequence SEQ ID NO: 106 or and a second light chain variable region of any variant of

[0121] In some preferred embodiments, the first binding domain of the bispecific antibody is an amino acid sequence. a first heavy chain variable region of SEQ ID NO: 150 or any variant thereof; and , the first light chain variable region of amino acid sequence SEQ ID NO: 148 or any variant thereof and the second binding domain comprises the amino acid sequence SEQ ID NO: 138, or any of its and a variant second heavy chain variable region of the amino acid sequence SEQ ID NO: 106 or and a second light chain variable region of any variant of

[0122] In some preferred embodiments, the first binding domain of the bispecific antibody is an amino acid sequence. a first heavy chain variable region of SEQ ID NO: 154 or any variant thereof; and , the first light chain variable region of amino acid sequence SEQ ID NO: 152 or any variant thereof and the second binding domain comprises the amino acid sequence SEQ ID NO: 138, or any of its and a variant second heavy chain variable region of the amino acid sequence SEQ ID NO: 106 or and a second light chain variable region of any variant of

[0123] In some preferred embodiments, the first binding domain of the bispecific antibody is an amino acid sequence. a first heavy chain variable region of SEQ ID NO: 158 or any variant thereof; and , the first light chain variable region of amino acid sequence SEQ ID NO: 156 or any variant thereof and the second binding domain comprises the amino acid sequence SEQ ID NO: 138, or any of its and a variant second heavy chain variable region of the amino acid sequence SEQ ID NO: 106 or The second light chain variable region of any variant of

[0124] In some preferred embodiments, the first binding domain of the bispecific antibody comprises an amino acid sequence. a first heavy chain variable region of SEQ ID NO: 162 or any variant thereof; and , the first light chain variable region of amino acid sequence SEQ ID NO: 160 or any variant thereof and the second binding domain comprises the amino acid sequence SEQ ID NO: 138, or any of its and a variant second heavy chain variable region of the amino acid sequence SEQ ID NO: 106 or The second light chain variable region of any variant of

[0125] In some preferred embodiments, the first binding domain of the bispecific antibody comprises an amino acid sequence. a first heavy chain variable region of SEQ ID NO: 166 or any variant thereof; and , the first light chain variable region of amino acid sequence SEQ ID NO: 164 or any variant thereof and the second binding domain comprises the amino acid sequence SEQ ID NO: 138, or any of its and a variant second heavy chain variable region of the amino acid sequence SEQ ID NO: 106 or The second light chain variable region of any variant of

[0126] In some preferred embodiments, the first binding domain of the bispecific antibody comprises an amino acid sequence. a first heavy chain variable region of SEQ ID NO: 170 or any variant thereof; and , the first light chain variable region of amino acid sequence SEQ ID NO: 168 or any variant thereof and the second binding domain comprises the amino acid sequence SEQ ID NO: 138, or any of its and a variant second heavy chain variable region of the amino acid sequence SEQ ID NO: 106 or and a second light chain variable region of any variant of

[0127] In one specific embodiment, the first heavy chain comprises SEQ ID NO: 174. wherein the first light chain of the bispecific antibody comprises SEQ ID NO: 172 and the second heavy chain comprises The second light chain comprises SEQ ID NO: 180. .

[0128] In one specific embodiment, the first heavy chain comprises SEQ ID NO: 174. wherein the first light chain of the bispecific antibody comprises SEQ ID NO: 176 and the second heavy chain comprises The second light chain comprises SEQ ID NO: 180. .

[0129] In one specific embodiment, the first heavy chain comprises SEQ ID NO: 178. wherein the first light chain of the bispecific antibody comprises SEQ ID NO: 172 and the second heavy chain comprises The second light chain comprises SEQ ID NO: 180. .

[0130] In one specific embodiment, the first heavy chain comprises SEQ ID NO: 178. wherein the first light chain of the bispecific antibody comprises SEQ ID NO: 176 and the second heavy chain comprises The second light chain comprises SEQ ID NO: 180. .

[0131] In one embodiment, the present disclosure provides an antibody that binds to BCMA, comprising the first antigen-binding portion. The antibody or an antigen-binding portion thereof is provided.

[0132] In some embodiments, the antibody or antigen-binding portion thereof that binds to BCMA is at least 60% or more, 65% or more, with the antibody or antigen-binding portion thereof of any embodiment; Over 70%, Over 75%, Over 80%, Over 85%, Over 90%, Over 95%, Over 96% , greater than 97%, greater than 98%, greater than 99% or higher sequence identity.

[0133] In one aspect, the present disclosure provides an antibody or antigen-binding portion thereof according to any of the preceding aspects. a nucleic acid encoding, or at least 60%, 65%, 70%, 75% thereof Over 80%, Over 85%, Over 90%, Over 95%, Over 96%, Over 97%, 9 Nucleic acid molecules having greater than 8%, greater than 99% or higher sequence identity are provided.

[0134] In some embodiments, the encoding nucleic acid of the first heavy chain variable region of the first antigen-binding portion are the nucleotide sequences SEQ ID NOs: 11, 25, 35, 42, 49, 54, 64 , 70, 80, 146, 151, 155, 159, 163, 167, 171, 189 or and / or a variant thereof of the first light chain variable region of the first antigen-binding portion. The encoding nucleic acid is selected from the nucleotide sequences SEQ ID NO: 16, 21, 30, 39, 46, 52, 59, 68, 75, 82, 86, 91, 143, 149, 153, 157, 161 , 165, 169, 190 or any variant thereof. In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 189, and the first light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:190.

[0135] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 11, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:16.

[0136] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 11, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:21.

[0137] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO:25, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:30.

[0138] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 35, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:39.

[0139] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 42, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:46.

[0140] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 49, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:52.

[0141] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 54, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:59.

[0142] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 11, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:39.

[0143] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO:64, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:68.

[0144] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 70, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:75.

[0145] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 80, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:82.

[0146] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 11, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:86.

[0147] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 11, and the first light chain variable The coding nucleic acid for the region is the nucleotide sequence SEQ ID NO:91.

[0148] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 146, and the first light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:143.

[0149] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 151, and the first light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:149.

[0150] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 155, and the first light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:153.

[0151] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 159, and the first light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:157.

[0152] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 163, and the first light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:161.

[0153] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 167, and the first light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:165.

[0154] In some preferred embodiments, the first heavy chain variable region of the first antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 171, and the first light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:169.

[0155] In some embodiments, the encoding nucleic acid for the first heavy chain of the first antigen-binding moiety is The nucleic acid sequence is selected from SEQ ID NO: 175, 179 or any variant thereof, and and / or the encoding nucleic acid of the first light chain of the first antigen-binding portion has the nucleotide sequence S Selected from EQ ID NO: 173, 177 or any variant thereof.

[0156] In some preferred embodiments, the encoding nucleic acid of the first heavy chain of the first antigen-binding portion is the nucleotide sequence SEQ ID NO:175, and the first sequence of the first antigen-binding portion The coding nucleic acid for the light chain is the nucleotide sequence SEQ ID NO:173.

[0157] In some preferred embodiments, the encoding nucleic acid of the first heavy chain of the first antigen-binding portion is the nucleotide sequence SEQ ID NO:175, and the first sequence of the first antigen-binding portion The coding nucleic acid for the light chain is the nucleotide sequence SEQ ID NO:177.

[0158] In some preferred embodiments, the encoding nucleic acid of the first heavy chain of the first antigen-binding portion is the nucleotide sequence SEQ ID NO:179, and the first The coding nucleic acid for the light chain is the nucleotide sequence SEQ ID NO:173.

[0159] In some preferred embodiments, the encoding nucleic acid of the first heavy chain of the first antigen-binding portion is the nucleotide sequence SEQ ID NO:179, and the first The coding nucleic acid for the light chain is the nucleotide sequence SEQ ID NO:177.

[0160] In some embodiments, the encoding nucleic acid of the second heavy chain variable region of the second antigen-binding portion are the nucleotide sequences SEQ ID NOs: 113, 118, 121, 124, 127, 130, 133, 137, 139, 141 or any variant thereof; and / or , the encoding nucleic acid of the second light chain variable region of the second antigen-binding portion is represented by the nucleotide sequence SEQ Selected from ID NOs: 94, 99, 101, 105, 107, 110 or any variant thereof Be found out.

[0161] In some preferred embodiments, the second heavy chain variable region of the second antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 137, and the second light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:94.

[0162] In some preferred embodiments, the second heavy chain variable region of the second antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 139, and the second light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:94.

[0163] In some preferred embodiments, the second heavy chain variable region of the second antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 139, and the second light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:99.

[0164] In some preferred embodiments, the second heavy chain variable region of the second antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 139, and the second light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:101.

[0165] In some preferred embodiments, the second heavy chain variable region of the second antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 113, and the second light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:107.

[0166] In some preferred embodiments, the second heavy chain variable region of the second antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 137, and the second light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:107.

[0167] In some preferred embodiments, the second heavy chain variable region of the second antigen-binding portion The code nucleic acid has the nucleotide sequence SEQ ID NO: 139, and the second light chain The coding nucleic acid for the variable region is the nucleotide sequence SEQ ID NO:107.

[0168] In some embodiments, the encoding nucleic acid for the second heavy chain of the second antigen-binding moiety is The nucleic acid sequence is SEQ ID NO: 183 or any variant thereof, and / or The coding nucleic acid for the second light chain of the antigen-binding portion of SEQ ID NO: 181 or any variant thereof.

[0169] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The encoding nucleic acid for the heavy chain variable region is the nucleotide sequence SEQ ID NO: 189, and and the encoding nucleic acid of the first light chain variable region is the nucleotide sequence SEQ ID NO:190 and the encoding nucleic acid of the second heavy chain variable region of the second antigen-binding portion is the nucleotide sequence S EQ ID NO: 139, and the encoding nucleic acid of the second light chain variable region is The sequence is SEQ ID NO:107. In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The encoding nucleic acid for the heavy chain variable region is the nucleotide sequence SEQ ID NO: 146, and and the encoding nucleic acid of the first light chain variable region is the nucleotide sequence SEQ ID NO:143 and the encoding nucleic acid of the second heavy chain variable region of the second antigen-binding portion is the nucleotide sequence S EQ ID NO: 139, and the encoding nucleic acid of the second light chain variable region is The sequence is SEQ ID NO:107.

[0170] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The encoding nucleic acid for the heavy chain variable region is the nucleotide sequence SEQ ID NO: 151, and and the encoding nucleic acid of the first light chain variable region is the nucleotide sequence SEQ ID NO:149 and the encoding nucleic acid of the second heavy chain variable region of the second antigen-binding portion is the nucleotide sequence S EQ ID NO: 139, and the encoding nucleic acid of the second light chain variable region is The sequence is SEQ ID NO:107.

[0171] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The encoding nucleic acid for the heavy chain variable region is the nucleotide sequence SEQ ID NO: 155, and and the encoding nucleic acid of the first light chain variable region is the nucleotide sequence SEQ ID NO:153 and the encoding nucleic acid of the second heavy chain variable region of the second antigen-binding portion is the nucleotide sequence S EQ ID NO: 139, and the encoding nucleic acid of the second light chain variable region is The sequence is SEQ ID NO:107.

[0172] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The encoding nucleic acid for the heavy chain variable region is the nucleotide sequence SEQ ID NO: 159, and and the encoding nucleic acid of the first light chain variable region is the nucleotide sequence SEQ ID NO:157 and the encoding nucleic acid of the second heavy chain variable region of the second antigen-binding portion is the nucleotide sequence S EQ ID NO: 139, and the encoding nucleic acid of the second light chain variable region is The sequence is SEQ ID NO:107.

[0173] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The encoding nucleic acid for the heavy chain variable region is the nucleotide sequence SEQ ID NO: 163, and and the encoding nucleic acid of the first light chain variable region is the nucleotide sequence SEQ ID NO:161 and the encoding nucleic acid of the second heavy chain variable region of the second antigen-binding portion is the nucleotide sequence S EQ ID NO: 139, and the encoding nucleic acid of the second light chain variable region is The sequence is SEQ ID NO:107.

[0174] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The encoding nucleic acid for the heavy chain variable region is the nucleotide sequence SEQ ID NO: 167, and and the encoding nucleic acid of the first light chain variable region is represented by the nucleotide sequence SEQ ID NO: 165. and the encoding nucleic acid of the second heavy chain variable region of the second antigen-binding portion is Q ID NO: 139, and the encoding nucleic acid of the second light chain variable region is The sequence is SEQ ID NO:107.

[0175] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The encoding nucleic acid for the heavy chain variable region is the nucleotide sequence SEQ ID NO: 171, and and the encoding nucleic acid of the first light chain variable region is the nucleotide sequence SEQ ID NO:169 and the encoding nucleic acid of the second heavy chain variable region of the second antigen-binding portion is the nucleotide sequence S EQ ID NO: 139, and the encoding nucleic acid of the second light chain variable region is The sequence is SEQ ID NO:107.

[0176] In some preferred embodiments, the encoding nucleic acid for the second heavy chain of the second antigen-binding portion is the nucleotide sequence SEQ ID NO: 183, and the second antigen-binding portion The coding nucleic acid for the second light chain is the nucleotide sequence SEQ ID NO:181.

[0177] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The coding nucleic acid for the heavy chain is the nucleotide sequence SEQ ID NO: 175, and the first The coding nucleic acid for the first light chain of the antigen-binding portion of 73, and the encoding nucleic acid of the second heavy chain of the second antigen-binding portion is the nucleotide sequence SEQ ID NO: 183, and the encoding nucleic acid of the second light chain of the second antigen-binding portion is The nucleotide sequence is SEQ ID NO:181.

[0178] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The coding nucleic acid for the heavy chain is the nucleotide sequence SEQ ID NO: 175, and the first The coding nucleic acid for the first light chain of the antigen-binding portion of 77, and the encoding nucleic acid of the second heavy chain of the second antigen-binding portion is the nucleotide sequence SEQ ID NO: 183, and the encoding nucleic acid of the second light chain of the second antigen-binding portion is The nucleotide sequence is SEQ ID NO:181.

[0179] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The coding nucleic acid for the heavy chain is the nucleotide sequence SEQ ID NO: 179, and the first The coding nucleic acid for the first light chain of the antigen-binding portion of 73, and the encoding nucleic acid of the second heavy chain of the second antigen-binding portion is the nucleotide sequence SEQ ID NO: 183, and the encoding nucleic acid of the second light chain of the second antigen-binding portion is The nucleotide sequence is SEQ ID NO:181.

[0180] In some preferred embodiments, the first antigen-binding portion of the bispecific antibody The coding nucleic acid for the heavy chain is the nucleotide sequence SEQ ID NO: 179, and the first The coding nucleic acid for the first light chain of the antigen-binding portion of 77, and the encoding nucleic acid of the second heavy chain of the second antigen-binding portion is the nucleotide sequence SEQ ID NO: 183, and the encoding nucleic acid of the second light chain of the second antigen-binding portion is The nucleotide sequence is SEQ ID NO:181.

[0181] In one embodiment, the present disclosure provides an antibody that binds to BCMA, comprising the first antigen-binding portion. The antibody or an antigen-binding portion thereof is provided.

[0182] In one aspect, the present disclosure provides a vector comprising the nucleic acid.

[0183] In one aspect, the present disclosure provides a cell comprising the nucleic acid or vector.

[0184] In one aspect, the present disclosure provides a method for producing a bispecific antibody or an antigen-binding portion thereof, a nucleic acid, a vector, or a combination thereof. The present invention provides compositions comprising the target and / or cells.

[0185] In one aspect, the present disclosure provides a method for the preparation of a bispecific antibody or an antibody thereof covalently linked to a therapeutic moiety. Antibody-drug conjugates are provided that include an original binding moiety.

[0186] Preferably, the therapeutic moiety is a cytotoxic moiety, a chemotherapeutic agent, a cellular factor, an immunosuppressant, an immunostimulatory agent, or the like. The compound is selected from a stimulant, a degradative peptide, or a radioisotope.

[0187] The antibodies of the present disclosure may be used as therapeutic or diagnostic tools for various diseases in which BCMA is adversely expressed or manifested. It is useful as a tool.

[0188] In one embodiment of a BCMA-associated disease, in cells of the diseased tissue or organ The expression of BCMA in the tissue or organ is increased compared to that in a healthy tissue or organ. 10% or more, especially 20% or more, 50% or more, 100% or more, 200% or more, 500% or more It means an increase of 1000% or more, 10000% or more, or more. In embodiments, expression is found only in diseased tissue and not in corresponding healthy tissue. According to the present disclosure, BCMA-associated diseases include, for example, B cell disorders, This includes B cell disorders such as plasma cell disorders and / or autoimmune diseases.

[0189] In some embodiments, the disease is cancer. Cancer is a BCMA-associated cancer, including multiple myeloma, malignant plasmacytoma, Hodgkin lymphoma, and ovarian cancer. Nodal lymphocyte-predominant Hodgkin's lymphoma, Kahler's disease and myeloid leukemia, plasma cell leukemia, Plasmacytoma, B-cell prolymphocytic leukemia, hairy cell leukemia, B-cell non-Hodgkin's lymphoma (N HL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), acute lymphocytic Allergic leukemia (ALL), chronic myeloid leukemia (CML), follicular lymphoma, Burkitt lymphoma lymphoma, marginal zone lymphoma, mantle cell lymphoma, large cell lymphoma, precursor B lymphocytic lymphoma Myeloid leukemia, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma Follicular lymphoma, marginal zone lymphoma, mucosa-associated lymphoid tissue lymphoma, small cell lymphoma Lymphoid lymphoma, mantle cell lymphoma, Burkitt lymphoma, primary mediastinal (thymic) lymphoma B-cell lymphoma, lymphoplasmacytic lymphoma, Waldenstrom's macroglobulin Lymphocyte leukemia, nodal marginal zone B-cell lymphoma, splenic marginal zone lymphoma, intravascular large B-cell lymphoma lymphoma, primary effusion lymphoma, lymphomatoid granulomatosis, T cell / histiocytocyte-rich large B cell Lymphoma, primary central nervous system lymphoma, primary cutaneous diffuse large B-cell lymphoma (leg type), elderly EBV-positive diffuse large B-cell lymphoma, inflammation-associated diffuse large B-cell lymphoma Cell lymphoma, intravascular large B-cell lymphoma, ALK-positive large B-cell lymphoma, Large B-cell lymphoma arising in interstitial lymphoma, HHV8-associated multicentric Castleman disease It has characteristics intermediate between diffuse large B-cell lymphoma, diffuse large B-cell lymphoma, and Burkitt lymphoma. Unclassified B-cell lymphoma, diffuse large B-cell lymphoma, and classical Hodgkin lymphoma Unclassified B-cell lymphoma with features intermediate between those of other B-cell-related lymphomas It is the B-cell-related cancer of choice.

[0190] In plasma cell disorders, a single plasma cell clone replicates uncontrollably, resulting in: The clone produces large amounts of a single (monoclonal) antibody called M-protein. In some cases, for example, when people have monoclonal gammopathy, The antibodies produced are incomplete and consist of only light or heavy chains. The antibodies produced by these antibodies are usually limited to one type. , multiple myeloma, plasmacytoma, plasma cell leukemia, macroglobulinemia, amyloidosis Waldenstrom's macroglobulinemia, solitary plasmacytoma of bone, extramedullary plasmacytoma , osteosclerotic myeloma, heavy chain disease, monoclonal gammopathy of undetermined significance, and schizophrenia. The present invention is selected from the group consisting of bronchial and rectal multiple myeloma.

[0191] In some embodiments, the disease is, for example, systemic lupus erythematosus or rheumatoid arthritis. autoimmune diseases such as

[0192] In some embodiments, the therapeutic agent is an antibody that specifically binds to an activating T cell antigen. Including the body.

[0193] In one embodiment, the therapeutic agent is an antibody that specifically binds to CD3, particularly CD3ε. Includes:

[0194] The methods of treating diseases and conditions using bispecific antibodies according to the present disclosure include administering to a mammal the therapeutic an effective amount of the antibody or antigen-binding fragment thereof, or nucleic acid molecule, or vector according to any of the preceding embodiments; The method includes administering a vector, or cell, or pharmaceutical composition to the subject.

[0195] In some embodiments, the present disclosure provides a method for treating a patient with a serum level of 40 μg / ml or greater. and administering to the patient an antibody capable of binding to BCMA to treat or prevent a cancer disease. In another embodiment, the method for preventing the inflammatory bowel disease is provided. or more, 300 μg / ml or more, 400 μg / ml or more, or 500 μg / ml or more of serum levels In another embodiment, the antibody is administered at a concentration of 800 μg / ml or less, 00μg / ml or less, 600μg / ml or less, 550μg / ml or less, or 500μg In one embodiment, the antibody is administered to achieve a serum level of 0.1 mg / ml or less. The serum level to be achieved is 40 μg / ml to 700 μg / ml, preferably 40 μg / ml ~600μg / ml, preferably 50μg / ml to 500μg / ml, for example For example, 150μg / ml to 500μg / ml, or 300μg / ml to 500μg / ml As used herein, the term "serum level" refers to the level in serum of the substance under consideration. In one embodiment, the concentration refers to a concentration that provides serum levels for 7 days or more or 14 days or more. In one embodiment, the method provides a dose of 300 mg / m2 or more, for example, 600 mg / m2 or more. m2 or more, and preferably 1500 mg / m2 or less, 1200 mg / m2 or less, or This includes administering an antibody dose of 1000 mg / m2 or less.

[0196] In some embodiments, the present disclosure provides a method for administering a dose of 300 mg / m2 or more to a patient, e.g., 60 mg / m2 or more. 0 mg / m² or more, and preferably 1500 mg / m² or less, 1200 mg / m² or less or administering an antibody capable of binding to BCMA at a dose of 1000 mg / m2 or less. The present invention provides a method for treating or preventing cancer diseases.

[0197] In some embodiments, the present disclosure provides a method for administering to a patient an antibody capable of binding to BCMA. A method for treating or preventing a cancer disease, comprising: 60% or more, 70% or more, 80% or more, or 90% or more of the cancer cells are BCMA positive and / or 40% or more, preferably 50% or more or 60% or more of the cancer cells in the patient The present invention provides a method for treating or preventing cancer diseases in which BCMA surface expression is positive. The present disclosure provides: a. 50% or more, preferably 60% or more, 70% or more, 80% or more, or 9 0% or more BCMA-positive cancer cells, and / or 40% or more, preferably 50% To identify patients with cancer cells that are positive for BCMA surface expression in ≥ 60% or ≥ 60% of patients. and b. a method for treating or treating a cancer disease, comprising administering to a patient an antibody capable of binding to BCMA. Further provided is a method for preventing the disease. In one embodiment, the disease is treated in more than 95% or 98% of patients. In one embodiment, 70% or more of the patients have BCMA-positive cancer cells. More than 0% or more than 90% of cancer cells are positive for BCMA surface expression.

[0198] In one embodiment of the method according to any aspect of the present invention, the outcome of treating the cancer disease is: In one embodiment, stabilization of the disease state is achieved for at least two months. or more than 3 months or more than 6 months.

[0199] In some embodiments, the present disclosure provides a method for administering to a patient an antibody capable of binding to BCMA. In one embodiment, the present invention provides a method for stabilizing the condition of a cancer patient, the method comprising: Stabilization of the condition can last for 2 months or more, 3 months or more, or 6 months or more.

[0200] In one embodiment of the method according to any aspect described herein, a single dose or administering the antibody in multiple doses

[0201] In some embodiments, the present disclosure provides a method for administering to a patient a plurality of antibodies capable of binding to BCMA. The present invention provides a method for treating or preventing cancer disease, comprising administering to a subject a therapeutically effective amount of a compound comprising the steps of:

[0202] According to the present disclosure, when the antibody is administered in multiple doses, preferably three or more doses, four or more doses are administered. ≥ 1 dose, ≥ 5 doses, ≥ 6 doses, ≥ 7 doses, ≥ 8 doses, ≥ 9 doses More than one dose, or more than 10 doses, and preferably not more than 30 doses, not more than 25 doses the antibody is administered in one dose, 20 or fewer doses, 15 or fewer doses, or 10 or fewer doses. Preferably, the time interval is 7 days or more, 10 days or more, 14 days or more, or 20 days or more. The antibody dose is administered over a period of preferably 7 to 30 days, 10 to 20 days, and preferably about 1 Doses of antibody are administered at 4 day time intervals.

[0203] In one embodiment, the antibody is administered to achieve a serum level of 40 μg / ml or greater. In different embodiments, the concentration is 50 μg / ml or more, 150 μg / ml or more, 300 μg / ml or more to achieve serum levels of ≥ 400 μg / ml, ≥ 400 μg / ml, or ≥ 500 μg / ml. In different embodiments, the antibody is administered at a concentration of 800 μg / ml or less, 700 μg / ml or less. Blood levels of 600 μg / ml or less, 550 μg / ml or less, or 500 μg / ml or less In one embodiment, the antibody is administered to provide serum levels of , 40 μg / ml to 700 μg / ml, preferably 40 μg / ml to 600 μg / ml, Preferably, 50 μg / ml to 500 μg / ml, for example, 150 μg / ml to 500 μg In one embodiment, the concentration is 7. In one embodiment, the method provides serum levels for 3 days or more, or 14 days or more. 00 mg / m² or more, for example, 600 mg / m² or more, and preferably 150 Use of antibodies at 0 mg / m² or less, 1200 mg / m² or less, or 1000 mg / m² or less This includes administering in an amount.

[0204] any of the preceding in the preparation of a medicament for treating a BCMA-associated disease in a mammal. Antibody or antigen-binding fragment thereof, nucleic acid molecule, vector, cell, or drug according to an embodiment Use of the product composition is also provided.

[0205] According to any of the above aspects, optionally the antibody is conjugated to another drug, such as , labeled or cytotoxic conjugates.

[0206] In one aspect, the present disclosure provides, for example, an antibody according to the present disclosure, a fragment thereof, a homolog thereof, Derivatives, e.g., labeled or cytotoxic conjugates, and antibody instructions The present invention further includes kits containing the above-mentioned instructions, conjugates for killing specific types of cells, etc. The document provides guidance for the in vitro, in vivo or ex vivo use of antibodies, conjugates, etc. The antibody may be in liquid or solid form, and is usually lyophilized. The kit may further include other suitable reagents such as buffers, reconstitution solutions, and other reagents depending on the intended use. The reagents are packaged in predetermined amounts and may contain other ingredients necessary for their therapeutic use. Combination with instructions for use, such as for use in a diagnostic assay, is also contemplated. If the substrate is labeled, for example, with an enzyme, the kit may include a substrate and A substrate precursor that provides a cofactor required by the enzyme (e.g., a detectable chromophore or fluorophore). In addition, stabilizers, buffers (e.g., blocking buffers or lysis buffers) may be included. Other additives such as buffer solutions may also be included. The relative amounts of the various reagents may be varied to increase the concentration of the reagent solutions. This allows for flexibility for the user, space savings, and reagent savings. These reagents can also be provided in the form of dry powder, usually lyophilized. It is supplied as a premixed solution and contains excipients that, when dissolved, provide a solution of the appropriate concentration of the reagent. It can be provided.

[0207] The antibody or functional fragment thereof, or the nucleic acid molecule, or the vector according to any of the above embodiments, or The present invention relates to the preparation of a reagent for inhibiting BCMA binding in a cell, a drug composition, or a kit. The present invention provides for use in

[0208] The antibodies of the present disclosure are also useful in immunoassays, purification methods, and immunoglobulin or fragments thereof. Such uses are well known to those skilled in the art.

[0209] Correspondingly, the present disclosure further provides a composition comprising an anti-BCMA antibody or fragment thereof according to the present disclosure. The antibody is conveniently combined with a pharmaceutically acceptable carrier, diluent or excipient. This can be done by means known in the art.

[0210] The term "drug composition" as used in this disclosure refers to a formulation of various preparations. The formulation containing a therapeutically effective amount of the multivalent antibody may be a sterile liquid solution, liquid suspension, or lyophilized form. and may optionally contain stabilizers or excipients.

[0211] The antibodies of the present disclosure may be used as a composition to be administered alone or in combination with other active agents. It may also be used.

[0212] In some embodiments, the humanized antibodies of the present disclosure are The therapeutic moiety may be, for example, a cytotoxin, a chemotherapeutic agent, a cytokine, It may be an immunosuppressant, an immunostimulant, a degraded peptide or a radioisotope. The conjugates are also referred to herein as "antibody-drug conjugates" or "ADCs." .

[0213] In some embodiments, the antibody is conjugated to a cytotoxic moiety. The toxic moieties are taxol, cytochalasin B, gramicidin D, ethidium bromide, emethidium bromide, Cephaeline, mitomycin, etoposide, teniposide, vincristine vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxybenzoates Tubulin inhibitors such as transenedione, maytansine or analogues or derivatives thereof , monomethyl auristatin E or F or analogs or derivatives thereof inhibitors, dolastatin 10 or 15 or analogs thereof, irinotecan or analogs thereof, Mitoxantrone, mithramycin, actinomycin D ), 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, Docaine, propranolol, puromycin, calicheamicin or its analogues or derivatives, methotrexate, 6-mercaptopurine, 6-thio Guanine, cytarabine, fludarabine, 5-fluorouracil, decarbazine, hydroxybenzoates antimetabolites such as urea, asparaginase, gemcitabine or cladribine, e.g. Mechlorethamine, thiopurine, chlorambucil, melphalan , carmustine (BSNU), lomustine (CCNU), cyclophosphamide, Busulf Antihistamine, dibromomannitol, streptozotocin, dacarbazine (DTIC), procaine alkylating agents such as rubazine and mitomycin C, cisplatin or carboplatin Platinum derivatives, duocarmycin A, duocarmycin SA, and rachelmycin (CC -1065) or its analogs or derivatives, actinomycin, bleomycin, Dow Norubicin, doxorubicin, idarubicin, mithramycin, mitomycin, mitoxin Antibiotics such as santron, primycin, and anthramycin (AMC) Biosubstances, pyrrolo[2,1-c][1,4]-benzodiazepine (PDB), diphtheria toxin Ricin and related molecules such as the diphtheria A chain and its active fragments and hybrid molecules, or deglycosylated ricin A chain toxins, ricin such as cholera toxin, SLT I, SLT I Shiga-like toxins such as SLT I, SLT IIV, LT toxins, C3 toxins, Shiga toxins, pertussis toxins, Tangius toxin, Bowman-Birk soybean protease inhibitor, Pseudomonas exotoxin, Allorin , Saporin, modeccin, geranin, abrin A chain, modeccin A chain, α-sarcin, Aleurites fordii ) proteins, dianthin proteins, PAPI, PAPII and PAP-S Phytolacca americana protein, bitter melon (momordica charantia) inhibitors, curcin, crotin (croti n), saponaria con (sapaonaria officinalis) inhibitor, gero nin, mitogellin, restrictocin, phenomycin and Enomycin toxin, ribonuclease (RNase), DNase I, Staphylococcus aureus Dotoxin A, pokeweed antiviral protein, diphtheria toxin, pseudomonas It may be selected from eggplant endotoxins.

[0214] In some embodiments, the antibody is an auristatin or a peptide analog thereof, derivative Auristatins are conjugated to a steroid or prodrug. Affects GTP hydrolysis and nuclear and cell division, and has anticancer and antifungal activity For example, auristatin E is a compound derived from p-acetylbenzoic acid or benzoyl benzoate. It can react with gallic acid to produce AEB and AEVB, respectively. Statin derivatives include AFP, MMAF (monomethyl auristatin F), and MMAE (monomethyl auristatin E). Suitable auristatins and auristatin analogs Suitable for auristatin-Ab conjugates, derivatives and prodrugs Linkers may be used in conjunction with any of the methods described, for example, in U.S. Pat. Nos. 5,635,483, 5,780,588, and 6 ,214,345 and International Patent Application Publications WO02088172 and WO20040109 57, WO2005081711, WO2005084390, WO200613267 0, WO03026577, WO200700860, WO207011968 and WO It is described in 205082023.

[0215] In some embodiments, the antibody is a pyrrolo[2,1-c][1,4]-benzodiazepine. Conjugated to zepine (PDB), its peptide analogs, derivatives or prodrugs Suitable PDBs, PDB derivatives and related techniques are described, for example, in Hartley JAet al., Cancer Res 2010, 70(17):6849-6858, An Tonow D. et al., Cancer J 2008, 14(3):154-1 69, Howard PWet al.,Bioorg Med Chem Let t 2009;19:6463-6466, and Sagnou et al., Bioo rg Med Chem Lett 2000;10(18):2083-2086 It is listed.

[0216] In some embodiments, the antibody is an anthracycline antibiotic, mertansine, Calicheamicin, duocarmycin, rachelmycin (CC-1065), dolastatin 10, dolastatin 15, irinotecan, monomethyl auristatin E, monomethyl auristatin Uristatin F, PDB, or any analog, derivative, or prodrug thereof Conjugated to a cytotoxic moiety of choice.

[0217] In some embodiments, the antibody is an anthracycline antibiotic or analog thereof, In some embodiments, the antibody is conjugated to a derivative or prodrug. Conjugated to mertansine or its analogues, derivatives or prodrugs. In one embodiment, the antibody is a calicheamicin or an analog, derivative, or prodrug thereof. In some embodiments, the antibody is conjugated to a duocarmycin or Conjugated to an analog, derivative, or prodrug thereof. The antibody may be rachelmycin (CC-1065) or its analogs, derivatives, or prodrugs. In some embodiments, the antibody is conjugated to dolastatin 10 or Conjugated to an analog, derivative, or prodrug thereof. The antibody is conjugated to dolastatin 15 or an analog, derivative, or prodrug thereof. In some embodiments, the antibody is monomethyl auristatin E or a derivative thereof. In some embodiments, the antibody is conjugated to an analog, derivative, or prodrug. The compound is conjugated to monomethyl auristatin F or its analogs, derivatives, or prodrugs. In some embodiments, the antibody is pyrrolo[2,1-c][1,4]- Conjugated to a benzodiazepine or its analogue, derivative or prodrug. In some embodiments, the antibody is irinotecan or an analog, derivative, or prodrug thereof. conjugate to

[0218] In some embodiments, the antibody inhibits cytokines (e.g., IL-2, IL-4, IL-6, IL-7, IL-10, IL-12, IL-13, IL-15, IL-18, IL-23, IL-24, IL-27, IL-28a, IL-28b, IL-29, KG F, IFNα, IFNβ, IFNγ, GM-CSF, CD40L, Flt3 ligand, stem Conjugated to cellular factors, ancestim and TNFα.

[0219] In some embodiments, the antibody is coupled to a radioisotope or a chelate containing a radioisotope. For example, the antibody is conjugated to a chelating linker (e.g., DOTA, DTPA). or tiuxetan). The antibody may further be conjugated to Alternatively, it may contain one or more radiolabeled amino acids or other radiolabeled molecules. Non-limiting examples of radioisotopes include 3H, 14C, 15N, 35S, 90Y, 99Tc, 125I, 131I, 186Re, 213Bi, 2 For therapeutic purposes, radioactive materials that emit beta or alpha particle radiation include 25Ac and 227Th. Radioactive isotopes, such as 131I, 90Y, 211At, 212Bi, 67Cu, 186R e, 188Re and 212Pb may also be used.

[0220] Techniques for conjugating molecules to antibodies are well known in the art. , nucleic acid molecules containing N-hydroxysuccinimide ester or maleimide functional groups, respectively The engineered cysteine ​​is covalently linked to a lysine or cysteine ​​in the antibody via a Conjugation methods using or incorporating unnatural amino acids are conjugation methods. It has been reported that the identity of the acyl donor glutamate can be improved by the use of acyl donors such as glutamate. Gin-containing tags (e.g., Gin-containing peptide tags or Q tags) or polypeptide engineering (e.g., by amino acid deletion, insertion, substitution, or mutation of the polypeptide) One could envision an Fc-containing polypeptide engineered with endogenous glutamines that are made reactive. , wherein the transglutaminase contains or is coupled to an amine donating agent (e.g., a reactive amine). The amine donor is covalently linked to the acyl donor glutamine-containing tag. or Fc-containing polypeptides via accessible / exposed / reactive endogenous glutamines Stable and homogeneous engineered Fc-containing polypeptide conjugates site-specifically conjugated with It is possible to form a uniform population (WO2012059882).

[0221] The therapeutic agents of the embodiments may be formulated to provide improved translocation, delivery, tolerance, etc. and administered with appropriate pharmaceutically acceptable carriers, excipients and other agents incorporated therein. It is understood that many suitable formulations are described in pharmacopoeias known to medicinal chemists. For example, Remington's Pharmaceutical Sciences ences (15th ed., Mack Publishing Company, East n, Pa. (1975), especially Chapter 87 by Blaug and Seymour. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, Lipids, lipid (cationic or anionic)-containing carriers (e.g., Lipofectin™), D NA conjugates, anhydrous slurries, oil-in-water and water-in-oil emulsions, emulsions Polyethylene glycol (polyethylene glycol of various molecular weights), semi-solid gel, and polyethylene glycol. The active ingredient in the formulation is not inactivated by the formulation, is physiologically compatible, and tolerates the route of administration. This applies to the treatment or method of treatment according to the present invention, provided that:

[0222] In one embodiment, the antibodies can be used as therapeutic agents. is typically a disease associated with aberrant BCMA expression, activity, and / or signaling in a subject. Therapeutic regimens are used to treat, ameliorate, and / or prevent a disease or condition. In a method, a subject, e.g., a subject with aberrant BCMA expression, activity, and / or signaling, a disease or disorder associated with (or at risk of or developing) a BCMA-associated disorder, e.g., This can be achieved by identifying a human patient suffering from a disease (such as a rheumatoid arthritis). administering to a subject an antibody preparation having high specificity and high affinity for a target antigen; and Generally, antibodies exert their effect by binding to a target. The administered antibody binds to the target (e.g., BC Eliminate, inhibit, or interfere with the expression, activity, and / or signaling function of MA The administered antibody may bind to the target (e.g., BCMA) naturally. It may eliminate, inhibit, or interfere with the binding of endogenous ligands to the target protein. For example, the antibody binds to a target and modulates BCMA expression, activity, and / or signaling; Inhibit, suppress, reduce, antagonize, neutralize, and / or otherwise interfere. In certain embodiments, the heavy and light chains may be combined to treat a disease or disorder associated with aberrant BCMA expression. An antibody having the chain CDRs can be administered to a subject.

[0223] In another embodiment, the antibody to BCMA is a BCMA locus known in the art. used in methods related to localization and / or quantification (e.g., in suitable biological samples) Determination of BCMA and / or levels of BCMA in specimens, diagnostic methods, protein imaging - Patent Application 20070122997 In certain embodiments, BCMA or a derivative, fragment, or analog thereof is Antibodies containing an antigen-binding domain derived from an antibody specific for a mimetic or homologue are pharmaceutically active. These compounds are used as therapeutic agents (hereinafter referred to as "therapeutic agents").

[0224] In another embodiment, immunoaffinity, chromatography, or immunoprecipitation, etc. BCMA polypeptides are isolated using antibodies specific for BCMA by standard techniques. Antibodies (or fragments thereof) against the BCMA protein can be used to detect the protein in biological samples. In some embodiments, for example, a particular therapeutic regimen may be used to detect the quality of the BCMA in biological samples as part of clinical trial procedures to determine efficacy of Detecting. Conjugating (i.e., physically linking) the antibody to a detectable substance Detectable substances include various enzymes, prosthetic groups, and fluorescent materials. Examples of suitable enzymes include horseradish peroxidase, luminescent materials, bioluminescent materials, and radioactive materials. peroxidase, alkaline phosphatase, β-galactosidase or acetylcholine Examples of suitable prosthetic group complexes include streptavidin / biotin. Examples of suitable fluorescent substances include umbelliferone, fluoroquinone, and avidin / biotin. Fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinyl a luminescent material comprising aminofluorescein, dansylamide chloride, or phycoerythrin; Examples of bioluminescent materials include luciferase, fluorescein, and the like. Examples of suitable radioactive materials include 125I, 131I, 35S, and aequorin. , or contains 3H.

[0225] In another embodiment, the antibodies of the present disclosure detect BCMA or its proteins in a sample. In some embodiments, the protein fragments can be used as reagents to detect the presence of the protein fragments. In the method, the antibody comprises a detectable label. The antibody is a polyclonal antibody, or more preferably a or monoclonal antibodies. Intact antibodies or fragments thereof (e.g., Fab, s The term "label" in reference to an antibody is used to refer to a detectable F(ab')2. Conjugating (i.e., physically binding) a suitable substance to an antibody directly binds the antibody. and indirectly labeling the antibody by reaction with another directly labeled reagent. Examples of indirect labeling include detection of a primary antibody with a fluorescently labeled secondary antibody; and biotin to allow detection with fluorescently labeled streptavidin. The term "biological sample" refers to a tissue sample isolated from a subject. , cells, and biological fluids, as well as tissues, cells, and fluids present within a subject's body. Thus, the term "biological sample" includes serum, plasma, or lymph. , blood and fractions or components in blood. In other words, the detection method according to the embodiment , in vitro and in vivo, in biological samples, analytes mRNA, protein, or is used to detect genomic DNA. For example, in vitro detection techniques for mRNA as an analyte are also used. Techniques include Northern hybridization and in situ hybridization. As an in vitro detection technique for analyte proteins, enzyme-linked immunosorbent assay (ELI) SA), Western blotting, immunoprecipitation, and immunofluorescence. In vitro techniques for detection of genomic DNA include Southern hybridizations. The implementation procedure for ELISA is, for example, "ELISA: Theory and Practice: Methods in Molecular Biology”, Volume 42, JR Crowther (ed.) Human Press, Totowa, NJ, 19 95, "Immunoassay", E. Diamandis & T. Christop Oulus, Academic Press, Inc., San Diego, C alif., 1996, and "Practice and Theory of E zyme immunoassays”, P. Tijssen, Elsevier S. Science Publishers, Amsterdam, 1985. In addition, in vivo detection techniques for analyte proteins include the use of labeled anti-analyte proteins. For example, the antibody may be labeled with a radiolabel and then administered to the subject. The presence and location of the radiolabel in vivo is detected by standard imaging techniques.

[0226] The antibodies and derivatives, fragments, analogs and homologs thereof described herein may be administered in any suitable pharmaceutical composition. The principles, considerations, and methods involved in the preparation of such compositions are described. Guidelines for selecting compositional ingredients are well known in the art and are described, for example, in Remi et al. ngton's Pharmaceutical Sciences: The Sci ence And Practice Of Pharmacy 19th Editi on (edited by Alfonso R. Gennaro et al.) Mack Pub. Co., Eas ton, Pa.: 1995;Drug Absorption Enhanceme nt: Concepts, Possibilities, Limitations , And Trends, Harwood Academic Publisher s, Langhorne, Pa., 1994; and Peptide And Pr otein Drug Delivery(Advances In Parenter (Al Sciences, Vol. 4), 1991, M. Dekker, New York Refer to

[0227] Such compositions typically comprise an antibody and a pharmaceutically acceptable carrier. If present, preferably with minimal inhibition of specific binding to the binding domain of the target protein. For example, a fragment of an antibody that has the ability to bind to a target protein sequence based on the variable region sequence of the antibody. It is possible to design peptide molecules that retain their strength. Such peptides can be synthesized chemically and and / or can be produced by recombinant DNA techniques (see, e.g., Marasco et al. al.,Proc.Natl.Acad.Sci.USA,90:7889-7893 (1993)).

[0228] As used herein, the term "pharmaceutically acceptable carrier" refers to a substance that is acceptable for pharmaceutical administration. Any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption agents that may be used Suitable pharmaceutically acceptable carriers are intended to include agents such as acetaminophen, ... It is described in Mington's Pharmaceutical Sciences This is a standard reference work in the field and is incorporated herein by reference. Preferred examples of such carriers or diluents include water, saline, Ringer's solution, dextrose, liposomes and Non-aqueous vehicles, such as fixed oils, can also be used. The use of antibodies for therapeutically active substances is well known in the art. Its use in compositions other than conventional media or reagents is contemplated.

[0229] The pharmaceutical compositions of the above embodiments are formulated to be compatible with their intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., , inhalation), transdermal (i.e., topical), transmucosal, and rectal administration. As a solution or suspension for administration, sterile injectable diluents such as water, saline, fixed oils, polyethylene glycol, propylene glycol, glycerol, propylene glycol or other synthetic solvents, antibacterial agents, e.g. benzyl alcohol or methyl p-hydroxybenzoate, antioxidants such as aspartame, corbic acid or sodium bisulfite, chelating agents such as ethylenediaminetetraacetic acid ( EDTA), buffers, e.g., acetate, citrate or phosphate, and osmolality adjusters, e.g. For example, sodium chloride or dextrose. pH can be determined by the presence of an acid or base, e.g., hydrochloric acid or It can be prepared with sodium hydroxide. Parenteral preparations are packaged in ampoules, disposable syringes, glass or It can be packaged in a plastic multi-dose vial.

[0230] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions. Suitable for intravenous administration. Pharmaceutically acceptable carriers for this purpose include saline, sterile water, and Cremophor ELT. M (BASF, Parsippany, NJ) or phosphate buffered saline (PBS In all cases, the composition must be sterile and fluid for easy syringability. It must also be stable under the conditions of preparation and storage and must be bacterially The carrier must be able to prevent the contaminating action of microorganisms such as bacteria and fungi. polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol) The solvent or dispersion medium may be a solvent or dispersion medium containing ethanol, etc., and suitable mixtures thereof. A coating, such as lecithin, may be used to maintain a desired particle size in dispersions; In addition, surfactants can be used to maintain proper fluidity. , parahydroxybenzoic acid ester, chlorobutanol, phenol, ascorbic acid, thime This can be achieved with various antibacterial and antifungal agents such as rosal. , polyols (e.g., mannitol, sorbitol), sodium chloride, and other isotonic agents. It is preferable to include in the composition. Prolonged absorption of injectable compositions can be achieved by, for example, adding monostearate. This can be achieved by including in the composition an agent that delays absorption, for example, aluminum or gelatin.

[0231] Optionally, the antibody may be combined with one or a combination of the above-listed ingredients (if desired) in the required amounts. Prepare a sterile injectable solution by incorporating the compound in the appropriate solvent (if necessary) and filtering to sterilize. Typically, the antibody is dispersed in a basic dispersion medium containing other necessary ingredients from those listed above. Dispersions are prepared by incorporating the compound into a sterile vehicle, such as a sterile powder for preparing a sterile injectable solution. The preparation method involves the addition of the active ingredient and any additional ingredients from a sterile filtered solution of the ingredients. A vacuum dried and freeze dried powder containing the desired components is obtained.

[0232] For inhalation administration, a pressurized container or syringe containing a suitable supply of propellant, e.g., a gas such as carbon dioxide, is used. delivers the compound in the form of an aerosol spray from a dispenser or nebulizer.

[0233] Systemic administration may also be by transmucosal or transdermal means. For transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Penetrants are well known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acids. Transmucosal administration can be accomplished using nasal sprays or suppositories. For this purpose, one or more antibodies may be administered in the form of a paste, ointment, gel, or other formulation known in the art. can be formulated into a cream.

[0234] The compounds may further be used in conventional suppository bases (e.g., cocoa butter or other glycerides). The active ingredient can be prepared and delivered rectally in the form of a suppository (with a suction cup) or retention enema.

[0235] In one embodiment, the antibody is administered via an implant or microencapsulated delivery system. Use a carrier that prevents rapid excretion from the body, such as a sustained / controlled release formulation containing As a biodegradable, biocompatible polymer, for example, , ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoester The method for preparing such a formulation may include: This is obvious to those skilled in the art.

[0236] It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. As used herein, a unit dosage form refers to a dosage form suitable for a single dose for the subject to be treated. refers to physiologically discrete units, each unit being a desired therapeutic agent in combination with the required pharmaceutical carrier. The antibody or antibodies contain a specific amount calculated to produce a therapeutic effect. The specifications for each unit dosage form depend on the inherent properties of the antibody and the specific therapeutic effect to be achieved, as well as the The limitations inherent in the techniques for preparing such antibodies for treating individuals are dictated by the Directly depended on.

[0237] The pharmaceutical compositions may be contained in a container, pack, or dispenser together with instructions for administration. That's fine.

[0238] The formulations herein may contain more than one of the above antibodies depending on the particular condition to be treated. The antibodies may further comprise antibodies that preferably have complementary activities but do not adversely affect each other. Additionally, or alternatively, the composition may contain an agent that enhances the function of the composition, such as a cytotoxin, a scavenger, or the like. Such molecules may include cytokines, chemotherapeutic agents, or growth inhibitors. For example, they may be combined in a kit. They may be present in combination when used.

[0239] In one embodiment, one or more of the antibodies are used in combination therapy, i.e., with other agents, For example, therapeutic agents (for treating pathological conditions or disorders, e.g., various forms of cancer, autoimmune diseases, Autoimmune and inflammatory disorders). As used herein, the term "in combination" refers to It refers to administering the reagents substantially simultaneously, simultaneously, or sequentially. Sequential administration is preferred. Or, when the administration of the second compound begins, the first compound in the two compounds is still curative. In some cases, "in combination" refers to a combination of the compounds of the present disclosure in a kit. The antibody and other therapeutic agent may be included simultaneously.

[0240] For example, a combination therapy may include one or more antibodies according to the present disclosure and one or more additional therapeutic agents. therapeutic agents (e.g., one or more cytokine and growth factor inhibitors, immunosuppressants, anti-inflammatory agents, metabolic inhibitors, enzyme inhibitors and / or cytotoxins or cell proliferation inhibitors, as described in more detail below Such combination therapy includes the co-preparation and / or co-administration of the administered The availability of low doses of selected therapeutic agents reduces the potential for complications associated with various monotherapies. Toxicity or complications can be avoided.

[0241] In one embodiment, the administration of the anti-BCMA antibody and the administration of the therapeutic agent is The number of cells can be reduced.

[0242] In one embodiment, the treatment regimen does not administer a corresponding anti-BCMA antibody. The subject's cytokine profile regarding administration of the T cell activating therapeutic agent compared to the treatment regimen This can effectively reduce the carbon dioxide emissions.

[0243] For the purposes of clarity and conciseness, technical features may be presented in the present invention as part of the same or separate embodiments. Although described in the specification, the scope of the invention does not include all or any combination of the features described. It is understood that combinations of embodiments may be included.

[0244] Example

[0245] Example 1 Antigen Expression and Construction of Stably Transfected Cells Human BCMA (Sino biological, HG10620-M), Rhesus monkey BCMA (Sino biological, CG90103-G, Cynomolgus monkey) The extracellular domain sequence of human BCM was used as a template. Amplified rhesus BCMA (Met1-Asn53) or rhesus BCMA (Met1-Asn52). , C-terminally fused to mouse IgG2a Fc, and transfected into HEK293E cells. and expressing it to produce human or cynomolgus monkey BCMA-mFc bivalent recombinant protein (A). The amino acid sequences are shown in SEQ ID NOs. 1-2.) or human IgG Fc (knob) and human IgG Fc (hole) into HEK293E cells and co-transfected with human or cynomolgus BCMA-hFc monovalent recombinant protein (amino acid sequence shown in SEQ ID NO: Figure 1 shows the results of human and cynomolgus monkey BCMA-mFc bivalent recombinant antibodies. SDS-PAGE results of proteins and BCMA-hFc kih monovalent recombinant protein showed.

[0246] Lenti vectors and packaging plates containing the full-length human and cynomolgus monkey BCMA sequences The plasmid (Genecoepia) was co-transfected into HEK293T cells. 48 hours after transfection, the pseudovirus-containing culture supernatant was collected and 10 μL was The resulting vector was then used to infect 1×10 CHO cells with different concentrations of puromycin. cin) was added and subjected to an anti-inflammatory screening, and finally human or cynomolgus BCMA was detected. Highly expressing stably transfected CHO-hBCMA cells (clone 2C2) and CHO Figure 2 shows the results of flow cytometry analysis of BCMA-positive cells. The results showed the detection of human and monkey BCMA stably transfected cells, and the human BCMA stably transfected Transfected cells (CHO-hBCMA-2C2) and cynomolgus monkey BCMA stable transfectants Both transfected cells (CHO-cynoBCMA-1A2) expressed high levels of BCM. A was expressed.

[0247] Construction of human or cynomolgus monkey CD3εγ heterodimers: Human CD3γ (UniProt P09693, Gln23-Asn116) and CD3ε (UniProt P077 The extracellular domain nucleotide sequence (Gln23-Asp126) was synthesized and the C-terminus was Each was fused to a human IgG Fc hole or Fc knob and expressed to form Human CD3εγ-Fc heterodimer (human CD3γ IgG Fc(hole) amino acid sequence) The amino acid sequence is shown in SEQ ID NO. 6 and corresponds to the amino acid sequence of human CD3ε IgG Fc (knob). The amino acid sequence is shown in SEQ ID NO. 7. ot Q95LI7,Gln23-Asn110), and CD3ε (UniProt Q95LI5, Gln22-Asp117) was synthesized and the C-terminus was modified to cynomolgus monkey IgG F Cynomolgus monkey CD3 expressed and fused to either the Fc hole or the Fc knob Amino acids of εγ-Fc heterodimer (cynomolgus monkey CD3γ IgG Fc(hole) The sequence is shown in SEQ ID NO. 8 and corresponds to cynomolgus monkey CD3γ IgG Fc (kno The amino acid sequence of b) is shown in SEQ ID NO. 9. CD3γ-Fc and CD3ε The recombinant plasmid expressing -Fc was transfected with PEI (Polysciences, #24765 -2) 3 mg / mL, and mixed with HEK293E cells (medium OPM-293 CD03 D PM) and cultured at 37°C, 120 rpm, 5% CO2 for 7 days. The culture supernatant was collected and purified by Protein A affinity chromatography. The human or cynomolgus monkey CD3εγ-Fc recombinant protein was obtained and analyzed by SDS-PAGE. The purity was greater than 95% (Figure 3).

[0248] Example 2. Animal immunization and preparation of BCMA antibodies BCMA-mFc recombinant protein or CHO-BCMA stably transfected cells The immunogen was an immunoadjuvant (Titermax, Sigma-Aldric). h) in equal volume, and 6-week-old Balb / c female mice (Beijing Vital After the first immunization, the mice were given the same dose of Booster immunizations were given every two weeks, with human and monkey antigen immunizations alternately. After vaccination, BCMA antibody titers were detected in mouse serum at levels exceeding 1:10,000. After three days of booster immunization with the antigen via the tail vein, the spleen and peripheral lymph nodes of the mice were A portion of the segment was collected, total RNA was extracted, and reverse transcription was performed using IgG-specific primers. The cDNA of the target gene was obtained, and the variable region fragments of the light and heavy chains were amplified, respectively, and transformed into phage Fa b) The library was constructed, and the number of transformed colonies (library volume) was approximately 1. It was 2×1010.

[0249] 1. Phage antibody library screening Using the classical panning method, 1 mL of BCMA immune library phage was diluted in 2% BSA After mixing with 3.5 mL of PBS solution, human or cynomolgus monkey BCMA recombinant protein Maxisorp Immunotube (Nunc) was added to coat the cells, and the cells were allowed to stand for 1 hour. After combining, the mixture was washed multiple times with PBS-Tween (0.5% v / v) and PBS, and then nonspecific Phages bound with high or low affinity were removed and then purified with 100 mM triethylamine. Affinity-bound phages were eluted, neutralized, and then infected into TG1 cells. After screening, a single clone was selected and inoculated into a U-shaped 96-well plate. IPTG was added to induce expression, and the induced supernatant containing Fab was collected and used for ELISA detection. Screening was performed using the method described above, and 265 positive clones were obtained in the primary screening. .

[0250] After re-induction of expression, 10 μg / ml of biotinylated human BCMA recombinant protein was added. was immobilized on the SA probe, and the binding time was 200 s and the dissociation time was 300 s. We then detected 59 high affinity markers (Octet, Fortebio). Clones were selected and transfected with human and cynomolgus BCMA-CHO stably transfected cells. Binding to cell membrane surface receptors was detected, and 52 of the clones were found to be human and It was able to cross-recognize cynomolgus monkey BCMA-CHO stably transfected cells.

[0251] 2. Antibody affinity measurement using Biacore A portion of the high-affinity antibody clones was selected, the plasmid was extracted, and the clone was transformed into TG1 cells. Induce expression overnight at 30 °C with 0.1 mM IPTG and extract the recombinant vector from the periplasmic space of the bacteria. The expressed Fab was extracted and purified by Ni-NTA affinity chromatography. The Fab antibody was obtained. Biacore affinity measurement was performed using Protein A. The captured human BCMA-hFc recombinant tag was obtained using a GE Healthcare The protein (0.5 μg / mL) was set at 200 RU, and after the baseline was stabilized, Gradient diluted antibody Fab (2-fold dilutions from 10 μg / mL, 7 gradient dilutions) was added to 30 The chip was run at a flow rate of 1 μL / min, the binding time was 350 s, and the dissociation time was 60 The time was 0 seconds. The kinetic constants were calculated by fitting using the ngmuir 1:1 kinetics model. The results of the affinity measurements are shown in Table 1. [Table 1]

[0252] The BCMA antibody variable region sequences are shown below. [Table 2]

[0253] 3G11 The amino acid sequence of the 3G11 heavy chain VH is shown in SEQ ID NO. 10, and its encoding nucleic acid is shown in SEQ ID NO. 11, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 12, 13 and 14, respectively. [ka] Nucleic acid sequence [ka]

[0254] The amino acid sequence of the 3G11 light chain VK is shown in SEQ ID NO. 15, and its encoding nucleic acid is is shown in SEQ ID NO. 16, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 17, 18 and 19, respectively. [ka] Nucleic acid sequence [ka]

[0255] 6G5 The amino acid sequence of the 6G5 heavy chain VH is shown in SEQ ID NO. 10, and its encoding nucleic acid is The HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO. 11, respectively. These are shown in SEQ ID NOs. 12, 13 and 14. [ka] Nucleic acid sequence [ka]

[0256] The amino acid sequence of the 6G5 light chain VK is shown in SEQ ID NO. 20, and its encoding nucleic acid is The HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO. 21, respectively. These are shown in SEQ ID NOs. 22, 23 and 19. [ka] Nucleic acid sequence [ka]

[0257] 15G9 The amino acid sequence of the 15G9 heavy chain VH is shown in SEQ ID NO. 24, and its encoding nucleic acid is shown in SEQ ID NO. 25, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 26, 27 and 28, respectively. [ka] Nucleic acid sequence [ka]

[0258] The amino acid sequence of the 15G9 light chain VK is shown in SEQ ID NO. 29, and its encoding nucleic acid is is shown in SEQ ID NO. 30, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 31, 32 and 33, respectively. [ka] Nucleic acid sequence [ka]

[0259] 15H10 The amino acid sequence of the 15H10 heavy chain VH is shown in SEQ ID NO. 34, and its coding sequence is The acid is shown in SEQ ID NO. 35, and its HCDR1, HCDR2 and HCDR3 are shown in They are shown in SEQ ID NOs. 12, 36, and 37, respectively. [ka] Nucleic acid sequence [ka]

[0260] The amino acid sequence of the 15H10 light chain VK is shown in SEQ ID NO. 38, and its coding sequence is The acid is shown in SEQ ID NO. 39, and its HCDR1, HCDR2 and HCDR3 are shown in They are shown in SEQ ID NOs. 17, 40, and 19, respectively. [ka] Nucleic acid sequence [ka]

[0261] 16B9 The amino acid sequence of the 16B9 heavy chain VH is shown in SEQ ID NO. 41, and its encoding nucleic acid is shown in SEQ ID NO. 42, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 12, 43 and 44, respectively. [ka] Nucleic acid sequence [ka]

[0262] The amino acid sequence of the 16B9 light chain VK is shown in SEQ ID NO. 45, and its encoding nucleic acid is is shown in SEQ ID NO. 46, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 47, 18 and 19, respectively. [ka] Nucleic acid sequence [ka]

[0263] 23F8 The amino acid sequence of 23F8 heavy chain VH is shown in SEQ ID NO. 48, and its encoding nucleic acid is shown in SEQ ID NO. 49, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 12, 50 and 14, respectively. [ka] Nucleic acid sequence [ka]

[0264] The amino acid sequence of the 23F8 light chain VK is shown in SEQ ID NO. 51, and its encoding nucleic acid is is shown in SEQ ID NO. 52, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 17, 40 and 19, respectively. [ka] Nucleic acid sequence [ka]

[0265] 23G9 The amino acid sequence of the 23G9 heavy chain VH is shown in SEQ ID NO. 53, and its encoding nucleic acid is shown in SEQ ID NO. 54, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 55, 56 and 57, respectively. [ka] Nucleic acid sequence [ka]

[0266] The amino acid sequence of the 23G9 light chain VK is shown in SEQ ID NO. 58, and its encoding nucleic acid is is shown in SEQ ID NO. 59, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 60, 61 and 62, respectively. [ka] Nucleic acid sequence [ka]

[0267] 29A11 The amino acid sequence of the 29A11 heavy chain VH is shown in SEQ ID NO. 10, and its coding sequence is The acid is shown in SEQ ID NO. 11, and its HCDR1, HCDR2 and HCDR3 are shown in They are shown in SEQ ID NOs. 12, 13, and 14, respectively. [ka] Nucleic acid sequence [ka]

[0268] The amino acid sequence of the 29A11 light chain VK is shown in SEQ ID NO. 38, and its coding sequence is The acid is shown in SEQ ID NO. 39, and its HCDR1, HCDR2 and HCDR3 are shown in They are shown in SEQ ID NOs. 17, 40, and 19, respectively. [ka] Nucleic acid sequence [ka]

[0269] 34G2 The amino acid sequence of the 34G2 heavy chain VH is shown in SEQ ID NO. 63, and its encoding nucleic acid is shown in SEQ ID NO. 64, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 65, 66 and 14, respectively. [ka] Nucleic acid sequence [ka]

[0270] The amino acid sequence of the 34G2 light chain VK is shown in SEQ ID NO. 67, and its encoding nucleic acid is shown in SEQ ID NO. 68, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 17, 40 and 19, respectively. [ka] Nucleic acid sequence [ka]

[0271] 38E2 The amino acid sequence of the 38E2 heavy chain VH is shown in SEQ ID NO. 69, and its encoding nucleic acid is shown in SEQ ID NO. 70, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 71, 72 and 73, respectively. [ka] Nucleic acid sequence [ka]

[0272] The amino acid sequence of the 38E2 light chain VK is shown in SEQ ID NO. 74, and its encoding nucleic acid is shown in SEQ ID NO. 75, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 76, 77 and 78, respectively. [ka] Nucleic acid sequence [ka]

[0273] 38D9 The amino acid sequence of the 38D9 heavy chain VH is shown in SEQ ID NO. 79, and its encoding nucleic acid is shown in SEQ ID NO. 80, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 12, 13 and 14, respectively. [ka] Nucleic acid sequence [ka]

[0274] The amino acid sequence of the 38D9 light chain VK is shown in SEQ ID NO. 81, and its encoding nucleic acid is shown in SEQ ID NO. 82, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 83, 84 and 19, respectively. [ka] Nucleic acid sequence [ka]

[0275] 38F9 The amino acid sequence of 38F9 heavy chain VH is shown in SEQ ID NO. 10, and its encoding nucleic acid is shown in SEQ ID NO. 11, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 12, 13 and 14, respectively. [ka] Nucleic acid sequence [ka]

[0276] The amino acid sequence of the 38F9 light chain VK is shown in SEQ ID NO. 85, and its encoding nucleic acid is is shown in SEQ ID NO. 86, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 87, 88 and 89, respectively. [ka] Nucleic acid sequence [ka]

[0277] 40C6 The amino acid sequence of the 40C6 heavy chain VH is shown in SEQ ID NO. 10, and its encoding nucleic acid is shown in SEQ ID NO. 11, and its HCDR1, HCDR2 and HCDR3 are They are shown in SEQ ID NOs. 12, 13 and 14, respectively. [ka] Nucleic acid sequence [ka]

[0278] The amino acid sequence of the 40C6 light chain VK is shown in SEQ ID NO. 90, and its encoding nucleic acid is shown in SEQ ID NO. 91, and its LCDR1, LCDR2 and LCDR3 are They are shown in SEQ ID NOs. 92, 23 and 19, respectively. [ka] Nucleic acid sequence [ka]

[0279] Example 3. Construction of BCMAxCD3 bispecific antibodies 1. CD3 humanized antibody Mouse-derived hybridoma CD3 antibody (EMBO J. 1985.4(2): 337 -344; J.Immunol.1986,137(4): 1097-100; J .Exp.Med.1991,174: 319-326; J.Immunol.19 91,147(9): 3047-52) to identify CD3 receptors in humans and cynomolgus monkeys. The sequence of the recognized nucleotide sequence was as follows:

[0280] The light chain amino acid sequence of the anti-CD3 mouse monoclonal antibody is set forth in SEQ ID NO. 184. showed. [ka]

[0281] The heavy chain amino acid sequence of the anti-CD3 mouse monoclonal antibody is set forth in SEQ ID NO. 185. showed. [ka]

[0282] Anti-CD3 mouse monoclonal antibody was humanized to identify the most identical human germline gene I. MGT_hVL7-43 was selected, light chain CDR grafted, and human IGLJ3 was used as FM4. *02, human IMGT_hVH3-73 was selected, heavy chain CDR grafting was performed, and FM4 Human IGHJ4*01 was used as the target gene. Different heavy and light chain variants were designed and obtained. It was.

[0283] The sequence of the light chain variable region of the CD3 humanized antibody was as follows: [Table 3]

[0284] The amino acid sequence of hVL1 is shown in SEQ ID NO. 93, and its encoding nucleic acid is shown in SEQ The LCDR1, LCDR2 and LCDR3 are shown in ID NO. 94, and the LCDR1, LCDR2 and LCDR3 are shown in SE Shown in Q ID Nos. 95, 96 and 97. [ka]

[0285] The amino acid sequence of hVL2 is shown in SEQ ID NO. 98, and its encoding nucleic acid is shown in SEQ The LCDR1, LCDR2 and LCDR3 are shown in ID NO. 99, and the LCDR1, LCDR2 and LCDR3 are shown in SE Shown in Q ID Nos. 95, 96 and 97. [ka]

[0286] The amino acid sequence of hVL3 is shown in SEQ ID NO. 100, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO. 101, and its LCDR1, LCDR2 and LCDR3 are respectively These are shown in SEQ ID NOs. 102, 103 and 97. [ka]

[0287] The amino acid sequence of hVL4 is shown in SEQ ID NO. 104, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO.105, and its LCDR1, LCDR2 and LCDR3 are respectively These are shown in SEQ ID NOs. 102, 103 and 97. [ka]

[0288] The amino acid sequence of hVL5 is shown in SEQ ID NO. 106, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO.107, and its LCDR1, LCDR2 and LCDR3 are respectively These are shown in SEQ ID NOs. 95, 96 and 108. [ka]

[0289] The amino acid sequence of hVL6 is shown in SEQ ID NO. 109, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO.110, and its LCDR1, LCDR2 and LCDR3 are respectively These are shown in SEQ ID NOs. 95, 111 and 108. [ka]

[0290] The sequence of the heavy chain variable region of the CD3 humanized antibody was as follows: [Table 4]

[0291] The amino acid sequence of hVH1 is shown in SEQ ID NO. 112, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO. 113, and its HCDR1, HCDR2 and HCDR3 are respectively These are shown in SEQ ID NOs. 114, 115 and 116. [ka]

[0292] The amino acid sequence of hVH2 is shown in SEQ ID NO. 117, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO. 118, and its HCDR1, HCDR2 and HCDR3 are shown in These are shown in SEQ ID NOs. 119, 115 and 116. [ka]

[0293] The amino acid sequence of hVH3 is shown in SEQ ID NO. 120, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO. 121, and its HCDR1, HCDR2 and HCDR3 are shown in These are shown in SEQ ID NOs. 119, 115 and 122. [ka]

[0294] The amino acid sequence of hVH4 is shown in SEQ ID NO. 123, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO. 124, and its HCDR1, HCDR2 and HCDR3 are shown in These are shown in SEQ ID NOs. 119, 115 and 125. [ka]

[0295] The amino acid sequence of hVH5 is shown in SEQ ID NO. 126, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO. 127, and its HCDR1, HCDR2 and HCDR3 are shown in These are shown in SEQ ID NOs. 119, 115 and 128. [ka]

[0296] The amino acid sequence of hVH6 is shown in SEQ ID NO. 129, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO. 130, and its HCDR1, HCDR2 and HCDR3 are shown in These are shown in SEQ ID NOs. 119, 115 and 131. [ka]

[0297] The amino acid sequence of hVH7 is shown in SEQ ID NO. 132, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO. 133, and its HCDR1, HCDR2 and HCDR3 are shown in These are shown in SEQ ID NOs. 134, 135 and 116. [ka]

[0298] The amino acid sequence of hVH8 is shown in SEQ ID NO. 136, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO. 137, and its HCDR1, HCDR2 and HCDR3 are respectively These are shown in SEQ ID NOs. 114, 115 and 116. [ka]

[0299] The amino acid sequence of hVH9 is shown in SEQ ID NO. 138, and its encoding nucleic acid is shown in SEQ ID NO. Q ID NO. 139, and its HCDR1, HCDR2 and HCDR3 are shown in These are shown in SEQ ID NOs. 114, 115 and 116. [ka]

[0300] The amino acid sequence of hVH10 is shown in SEQ ID NO. 140, and its encoding nucleic acid is SEQ ID NO. EQ ID NO. 141, and its HCDR1, HCDR2 and HCDR3 are respectively These are shown in SEQ ID NOs. 114, 115 and 116. [ka]

[0301] The entire sequences of the light and heavy chain humanized mutants were synthesized, and then the antibody λ light chain constant region or human Ig G4 heavy chain constant regions CH1-CH3 were cloned into a eukaryotic expression vector containing HEK Co-transfected 293E cells and cultured at 37°C, 120 rpm, and 5% CO for 5–6 days. After culturing, the culture supernatant was collected and purified using a Protein A chromatography column. Made.

[0302] 2. Measurement of affinity by ELISA Human CD3εγ protein was coated overnight at 4°C. After that, CD3 antibody was added to each well at different dilutions and incubated for 1 hour. HPR-labeled goat anti-human IgG Fc was added as a secondary antibody, and the color was developed with TMB solution. The reaction was stopped with concentrated sulfuric acid, and the absorbance was read at 450 nm (the results are shown in Figure 4).

[0303] Figure 4 shows a combination of anti-CD3 humanized antibodies (aC D3-hVH8 / VL1, aCD3-hVH9 / VL1, aCD3-hVH9 / VL2, aCD3-hVH9 / VL3, aCD3-hVH1 / VL5, aCD3-hVH8 / VL 5, including aCD3-hVH9 / VL5), CD3 humanized antibodies have high affinity and inhibit CD3εγ bound to the recombinant protein.

[0304] 3. Measurement of affinity for Jurkat T cells Jurkat cells in the logarithmic growth phase were blocked with 3% BSA for 30 minutes and then incubated at 5 × 1 0 4Cells / well were added to a U-shaped 96-well plate, centrifuged, and the supernatant was discarded. Gradient diluted antibody (antibody concentration: 30 μg / mL, diluted 3-fold, 5 gradient dilutions) After washing with the primary antibody, 50 μL of the diluted solution was added and incubated at 4°C for 1 hour. Alexa Fluro647-conjugated goat anti-human I diluted at 1:300 as the secondary antibody gG Fc(Jackson ImmunoResearch,109-606-170 ) was added to each well, and the wells were incubated at 4°C for 45 minutes. After washing, 5 mL of PBS was added to each well. The cells were resuspended in 0 μL and subjected to FACS (iQue, Intellicyt) detection (Results (shown in Figure 5).

[0305] Figure 5 shows a combination of anti-CD3 humanized antibodies that bind to Jurkat cells. The CD3 humanized antibody hVH9 / VL5 (aCD3-hVH9 / VL5) has intermediate affinity. Binding to Jurkat cells.

[0306] 4. Screening of BCMA antibody and CD3 antibody combinations Select a subset of high-affinity BCMA clones and compare them with the most homologous human germline gene. These antibodies were then CDR-grafted to produce the BCMA humanized antibodies h38E2, h23F8, and h4 0C6, h38D9, h29A11, h15H10 and h34G2 (Table 5) were obtained. See Schaefer W et al. (PNAS, 2011) for a humanized BCMA antibody and CD3 Antibody aCD3-hVH9 / VL5 (light chain variable region amino acid sequence is SEQ ID NO. 1) The heavy chain variable region amino acid sequence is shown in SEQ ID NO. 138) The humanized BCMA antibody light chain variable region was screened to contain the kappa light chain constant region. The heavy chain variable region was cloned into a eukaryotic expression vector containing the human IgG4 constant domain containing the mutation. The light chain variable region of the humanized CD3 antibody was cloned into a eukaryotic expression vector, and the λ light chain constant region was expressed as The eukaryotic expression vector containing the cloning region, the heavy chain hinge region, and the constant regions CH2 and CH3 was The heavy chain variable region of the CD3 antibody was cloned into a eukaryotic expression vector containing the human CH1 constant region. The four plasmids encoding the light and heavy chain genes were mixed in a 1:1:1:1 ratio. The resulting mixture was transfected into HEK293E cells, and the cells were expressed for 5-6 days. The culture supernatant was then collected. After one-step purification with Protein A, SEC-HPLC revealed that the monomer content The activity was >55% and was used for the relative activity evaluation. CD3 humanized antibody and BCMA50-C were synthesized according to S201213678247. A BCMA50×CD3 (B) bispecific antibody was constructed and used as a reference or as described in the literature. iTE) was expressed. [Table 5]

[0307] h38E2 The amino acid sequence of h38E2 light chain VK is shown in SEQ ID NO. 142. The nucleic acid is shown in SEQ ID NO. 143, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs. 144, 77 and 78, respectively. [ka] Nucleic acid sequence [ka]

[0308] The amino acid sequence of h38E2 heavy chain VH is shown in SEQ ID NO. 145, and its code The nucleic acid is shown in SEQ ID NO. 146, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs. 71, 147, and 73, respectively. [ka] Nucleic acid sequence JPEG2026027375000080.jpg15125

[0309] h23F8 The amino acid sequence of h23F8 light chain VK is shown in SEQ ID NO. 148. The nucleic acid is shown in SEQ ID NO. 149, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs. 17, 40, and 19, respectively. [ka] Nucleic acid sequence JPEG2026027375000082.jpg15125

[0310] The amino acid sequence of h23F8 heavy chain VH is shown in SEQ ID NO. 150, and its code The nucleic acid is shown in SEQ ID NO. 151, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs. 12, 50, and 14, respectively. [ka] Nucleic acid sequence [ka]

[0311] h40C6 The amino acid sequence of the h40C6 light chain VK is shown in SEQ ID NO. 152, and its code The nucleic acid is shown in SEQ ID NO. 153, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs. 92, 23, and 19, respectively. [ka] Nucleic acid sequence [ka]

[0312] The amino acid sequence of h40C6 heavy chain VH is shown in SEQ ID NO. 154, and its code The nucleic acid is shown in SEQ ID NO. 155, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs. 12, 13, and 14, respectively. [ka] Nucleic acid sequence [ka]

[0313] h38D9 The amino acid sequence of the h38D9 light chain VK is shown in SEQ ID NO. 156, and its code The nucleic acid is shown in SEQ ID NO. 157, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs. 83, 84 and 19, respectively. [ka] Nucleic acid sequence [ka]

[0314] The amino acid sequence of the h38D9 heavy chain VH is shown in SEQ ID NO. 158. The nucleic acid is shown in SEQ ID NO. 159, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs. 12, 13, and 14, respectively. [ka] Nucleic acid sequence [ka]

[0315] h29A11 The amino acid sequence of the h29A11 light chain VK is shown in SEQ ID NO. 160. The LCDR1, LCDR2 and LCDR3 sequences are shown in SEQ ID NO. 161. 3 are shown in SEQ ID NOs. 17, 40, and 19, respectively. [ka] Nucleic acid sequence JPEG2026027375000094.jpg14125

[0316] The amino acid sequence of the h29A11 heavy chain VH is shown in SEQ ID NO. 162. The nucleic acid is shown in SEQ ID NO. 163, and its HCDR1, HCDR2 and HCDR 3 are shown in SEQ ID NOs. 12, 13 and 14, respectively. [ka] Nucleic acid sequence [ka]

[0317] h15H10 The amino acid sequence of the h15H10 light chain VK is shown in SEQ ID NO. 164. The LCDR1, LCDR2 and LCDR3 sequences are shown in SEQ ID NO. 165. 3 are shown in SEQ ID NOs. 17, 40, and 19, respectively. [ka] Nucleic acid sequence [ka]

[0318] The amino acid sequence of the h15H10 heavy chain VH is shown in SEQ ID NO. 166. The nucleic acid is shown in SEQ ID NO. 167, and its HCDR1, HCDR2 and HCDR 3 are shown in SEQ ID NOs. 12, 36 and 37, respectively. [ka] Nucleic acid sequence [ka]

[0319] h34G2 The amino acid sequence of the h34G2 light chain VK is shown in SEQ ID NO. 168. The nucleic acid is shown in SEQ ID NO. 169, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs. 17, 40, and 19, respectively. [ka] Nucleic acid sequence JPEG2026027375000102.jpg14125

[0320] The amino acid sequence of h34G2 heavy chain VH is shown in SEQ ID NO. 170. The nucleic acid is shown in SEQ ID NO. 171, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs. 65, 66, and 14, respectively. [ka] Nucleic acid sequence [ka]

[0321] 5. T cell dependent cytotoxicity test (TDCC) Freshly isolated PBMCs and logarithmically growing RPMI-8226 cells (effector The antibody was mixed with the antibody (initial concentration 40 μg / Add 50 μL of (mL, 10-fold dilution, 7 gradients) and incubate at 37°C in 5% CO for 24 hours. After the incubation, the cells were centrifuged at 1000 rpm for 3 minutes, and the supernatant was collected to detect LDH release. Then, 50 μL of the supernatant was transferred to a black immunoplate and 50 μL of LDH detection substrate was added per well. The reaction was stopped after 10 minutes and detected (Biotek Synergy HT). The killing rate was calculated using the mathematical formula: After discarding the supernatant, the remaining cells in the wells were washed twice with 4% bovine serum. After washing, 100 μg / mL of human IgG was added to block the T cell activity for 10 minutes. The early-labeled CD69 and late-labeled CD25 were added to the cells, and antibodies (CD25-PE, CD4- APC, CD69-FITC, and CD8-APC) and incubated on ice for 20 minutes. After the incubation, 200 μL of 4% bovine serum was added to each well, and the plate was then resuspended in 10 ml of PBS. Wash, discard the supernatant, and add 60 μL / well of propidium iodide (PI) solution (1:200 dilution). Add the cells, incubate on ice for 5 minutes, and analyze by flow cytometry (BD FACS The mathematical formula for the kill rate was as follows:

number

[0322] The killing activity of the bispecific antibody combinations is shown in Table 6. [Table 6]

[0323] 6. Construction of BCMA×CD3κλ humanized bispecific antibody To optimize the humanized sequence, clone 29A11, which has a high killing efficiency, and its sequence The clone 6G5, which has homology with mouse IGHV9-3, was selected. The light chain variable region is derived from mouse IGKV3-12, and the light chain and heavy chain CDRs were transplanted into the human germline genes IGHV7-4-1*02 and IGVK_7_3, respectively. The designed humanized BCMA antibody sequence was obtained. A light chain-containing humanized CD3 antibody arm was engineered to contain a charge mutant (VκBCMA: Gln42Lys; VH BCMA:Gln39Glu;VλCD3:Gln40Glu;VHCD3:Gln39 A novel BCMA-CD3κλ humanized bispecific antibody with native IgG configuration was developed by introducing Lys To achieve heterodimeric pairing, the Fc of the bispecific antibody was The portion adopts the human IgG4 knob-into-hole structure (Atwell et al., 1997), mutations Ser228Pro, Leu235Glu and Pro329Ala This maintained the stability of the hinge region and reduced interactions with Fcγ receptors and C1q.

[0324] Plasmids encoding the corresponding antibody fragments were mixed in a 2:2 ratio of κ light chain:λ light chain:heavy chain 1:heavy chain 2. The mixture was mixed at a ratio of 1:1 with 3 mg / mL PEI and then cotransfected with CHO-S cells. The cells were transfected with CD CHO AGT medium (Gibco #12490-001) for 500 ml. 1 mL, and incubated at 37°C, 5% CO2, and 150 rpm. Then, on days 2, 4, and 6, 4% CHO Feed C+ feed (Gibco When the cell viability decreased to about 85%, the fermentation liquid was After harvesting and filtering, the cells were purified by Protein A affinity chromatography. Purification was performed using SEC-HPLC, which showed that the monomer content was over 92%. After pAct ion exchange chromatography, 50-300 mM NaCl / phosphate The elution peaks were combined and the monomer content was further reduced to 98-99%. This improvement was greater than that (Table 8). [Table 7] JPEG2026027375000108.jpg174125JPEG2026027375000109.jpg22125 [Table 8]

[0325] Example 4. Binding activity of BCMAxCD3κλ bispecific antibodies BCMA recombinant protein, overexpressing stably transfected cells, or BC MA + By detecting binding to tumor cells, the parental antibody BCMA antigen arm of the bispecific antibody was Determine the total titer and CD3 recombinant antigen, Jurkat cells, or freshly isolated peripheral blood T cells The affinity of the bispecific antibody CD3 arms was determined by detecting binding to

[0326] 1. Measurement of antigen affinity of BCMA×CD3κλ bispecific antibodies 10 μg of human or cynomolgus monkey BCMA or CD3εγ recombinant antigen by amino group / mL was coupled to a CM5 chip (GE healthcare), and the amount of antigen binding was measured. The concentration was controlled at approximately 200 RU. After the baseline was stabilized, gradient diluted antibody (10 μg / mL The solution (diluted 2-fold from 0.01 to 0.01, diluted 2-fold and diluted in 7 gradients) was run through the chip at a flow rate of 30 μL / min. The binding time was 350 seconds and the dissociation time was 600 seconds. The parent was fitted with a 1:1 binding model using 200 evaluation software. The affinity constants were obtained. The results of the affinity measurements are shown in Table 9. [Table 9]

[0327] 2. Binding of BCMA × CD3κλ bispecific antibodies to BCMA stably transfected cells If CHO-human BCMA stably transfected cells prepared in Example 1 were in the logarithmic growth phase. Cells and CHO-cynomolgus monkey BCMA stably transfected cells were cultured in 4% bovine serum ( Hyclone, SH30626.06) to 5 × 10 cells 5 Adjust to cells / ml 100 μl / well of the cell suspension was added to a U-shaped 96-well plate and centrifuged at 300 g for 5 minutes. Centrifuge, discard the supernatant, and add gradient diluted antibodies (initial concentration 1800 nM, 3-fold dilution, 1 100 μL of the secondary antibody (0 gradient) was added and incubated at 4°C for 60 minutes. lexa Fluro647-labeled goat anti-human IgG Fc (1:300 dilution) 50 μL / The wells were then incubated on ice for 20 min, washed once, and then resuspended in PI solution (1:30 0) Add 50 μL / well, incubate for 5 minutes, and analyze by flow cytometry. As shown in Figure 6 and Table 10, the BCMAxCD3κλ bispecific antibody It bound with high affinity to human and cynomolgus monkey BCMA stably transfected cells. [Table 10]

[0328] 3. BCMA × CD3κλ bispecific antibody + Binding to tumor cells NCI-H929 and RPMI-8226 cells in the logarithmic growth phase were cultured using mouse IgG. (Jackson Immuno Research, 115-005-03)200μ Add 5 × 10 cells to 5 × 10 cells / mL and block in an ice bath for 30 min with 4% bovine serum. 5 individual cells Adjust the volume to 1 / mL, add 100 μL per well to a U-shaped 96-well plate, and heat at 300 g for 5 min. Centrifuge for 1 minute, discard the supernatant, and add gradient diluted antibody (initial concentration 1800 nM, 3-fold diluted) to each well. 100 μL of the solution (10 gradients) was added and incubated at 4°C for 60 minutes. Remove the primary antibody and inoculate with Alexa Fluro647-conjugated goat anti-human IgG Fc (1:3 50 μL / well of 10000 dilutions of 10 ... Afterwards, 50 μL / well of PI was added and incubated for 5 minutes. Flow cytometry was then performed. The detection results are shown in Figure 7 and Table 11. The body has high affinity for BCMA +Binds to tumor cells NCI-H929 and RPMI-8226 Ta. [Table 11]

[0329] 4. Binding of BCMA×CD3κλ bispecific antibodies to Jurkat cells Jurkat cells in the logarithmic growth phase were isolated and incubated with mouse IgG (Jackson Immunoglobulin G). Add 200 μg / mL of chlorine-containing acetone (No. Research, 115-005-03) and in an ice bath for 30 minutes. The cells were blocked with 4% bovine serum for 5 min. 5 Adjust to 96 cells / mL. Add 100 μL / well to a well plate, centrifuge at 300 g, discard the supernatant, and Add 100 μL of diluted antibody (initial concentration 1800 nM, 3-fold dilution, 10 gradient) to each well. The secondary antibody, Alexa Fluro6, was added and incubated at 4°C for 60 minutes. Add 50 μL / well of 47-labeled goat anti-human IgG Fc (1:300 dilution) and incubate on ice. After incubating for 20 minutes and washing once, 50 μL / well of PI was added and incubated for 5 minutes. The results were then detected by flow cytometry (BD C6). As shown in Tables 8 and 12, the BCMA×CD3κλ bispecific antibody inhibits human leukemia T with intermediate affinity. It bound to the cell line Jurkat cells. [Table 12]

[0330] 5. Binding of BCMA×CD3κλ bispecific antibodies to peripheral blood T cells Fresh human or cynomolgus monkey peripheral blood was collected and immobilized on Ficoll-Paque Plus (G PBMCs were isolated in 4% bovine serum ( Hyclone, SH30626.06) to 5 × 10 cells 5 Adjust to cells / mL Add 100 μL / well to a U-shaped 96-well plate, centrifuge, discard the supernatant, and Add 100 μL / well of diluted antibody (initial concentration 1800 nM, 3-fold dilutions, 10 gradients) The secondary antibody was Alexa Fluro647. Add 50 μL / well of labeled goat anti-human IgG Fc (1:300 dilution) and incubate in an ice bath for 20 min. After blocking for 1 minute and washing once, 50 μL / well of PI was added and incubated for 5 minutes. The results were analyzed by flow cytometry (BD C6). 6, the control antibody REGN5458 was synthesized and prepared.

[0331] The detection results are shown in Figure 9 and Table 13. The BCMA x CD3κλ bispecific antibody inhibited the proliferation of human peripheral blood. It recognizes blood CD4+ T and CD8+ T cells, and its affinity for human T cells is approximately 60-97nM. Both of these have weaker binding affinity to the BCMA receptor than the BCMA antigen arm, and are bispecific. This favored preferential recruitment of the antibody to tumor cells. [Table 13]

[0332] Example 5. BCMA×CD3κλ bispecific antibody-mediated TDCC Freshly isolated PBMCs were taken and the target cells NCI-H92, which were in the logarithmic growth phase, were cultured. 9 and RPMI-8226 cells were mixed and diluted gradiently at an effector cell / target cell ratio of 8:1. Diluted antibody (antibody concentration: 66.7 nM diluted 10-fold, 7 gradient dilutions) 50 μL 1 / well and cultured at 5% CO2 and 37°C for 24 hours. After the culture was completed, 50 μL of supernatant was Transfer the plate to a new black immunoplate, add 50 μL / well of LDH detection substrate, and incubate for 10 minutes. After 2 hours, the reaction was terminated and LDH release was detected. The remaining cells in the wells were incubated with 4% bovine serum for 2 hours. After washing twice, 100 μg / mL of human IgG was added and incubated for 10 minutes. Antibodies for detecting cell activation (CD25-PE, CD4-APC, CD69-FITC, and CD8 -APC) was added and incubated on ice for 20 minutes. Add 60 μL / well of PI, place on ice and incubate for 5 minutes. BCMA×CD3κλ bispecific antibodies were detected by immunohistochemistry. The results show that the specific antibodies kill NCI-H929 cells and activate T cells. 1A and 11B are the BCMA×CD3 κλ bispecific antibody RPMI-8226, respectively. BCMA expression in tumors with different levels of BCMA was shown to be a potent inhibitor of T cell killing and T cell activation. BCMA×CD3κλ bispecific antibody against cells NCI-H929 and RPMI-8226 Both of these antibodies can mediate effective killing by T cells and have potent killing activity. It corresponded to the control antibody.

[0333] Example 6: Activation of T cell activation pathways by BCMA x CD3κλ bispecific antibodies Luciferase reporter plasmid pGL4.30 [luc2P / NFAT-RE / Hygro] (Promega, E8481) was transfected into Jurkat cells. 200 μg / ml of hygromycin B was added, and the cells were screened using Jurkat- NFAT luc stably transfected reporter cells were obtained. Jurs in logarithmic growth phase were used. The kat-NFAT-luc reporter cells and target cells (RPMI-8226) were taken and centrifuged. Separate and discard the supernatant. 6 The cells were resuspended at 50 cells / ml. μl / well was inoculated into a 96-well plate, centrifuged at 300 g for 5 minutes, and the supernatant was discarded. Plate 50 μl / well of Jurkat-NFAT-luc reporter cells in 96-well plates. Each well was inoculated with gradient-diluted BCMA × CD3κλ bispecific antibody or control antibody. Add 50 μL of the antibody KLH×CD3 (initial concentration 20 μg / ml, 10-fold dilution, 10 gradients). After incubation, the cells were incubated at 37°C in 5% CO2 for 6 hours. Add detection reagent to each well according to the instructions in the Ciferase Assay System manual. Add 100 μL of the solution, let it stand at room temperature for 3 minutes, and then read it on a microplate reader (Biotek The detection results are shown in Figure 12. The polyspecific antibody inhibited the NFAT It can activate the signaling pathway and, in the absence of target cells, inhibit the NFAT signaling pathway. Did not activate.

[0334] Example 7: Nonspecific activation of PBMCs by BCMA×CD3κλ bispecific antibodies Freshly isolated PBMCs were taken and gradient diluted antibodies (antibody concentration 66.7 nM) were added to each well. Add 50 μL of the diluted solution (10-fold dilution from 1 to 7 gradient dilutions) and incubate at 37°C in 5% CO2. After incubation for 24 hours, 50 μL of the supernatant was transferred to a new black immunoplate. Add 50 μL / well of LDH detection substrate and wait 10 minutes to terminate the reaction and release of LDH. After detecting the IgG, the remaining cells in the wells were washed twice with 4% bovine serum, and then 100 μL of human IgG was added. Add 100 μg / mL of the antibody for detecting T cell activation (CD25-P) and incubate for 10 minutes. E, CD4-APC, CD69-FITC, and CD8-APC) were added and incubated on ice. After incubation for 20 minutes, the plate was washed, the supernatant was discarded, and 60 μL / well of PI was added. The mixture was incubated on ice for 5 minutes and then detected by flow cytometry. The results are shown in Figure 13, and in the absence of target cells, the BCMA × CD3κλ bispecific antibody , had no activating effect on peripheral blood T cells and was equivalent to the negative control KLH×CD3.

[0335] Example 8. Binding of BCMAxCD3κλ bispecific antibodies to Fc receptors His-Tag antibody (50 μg / ml) was coupled to the CM5 chip via the amino group. , His-tagged FcγRI and FcγRIIA, respectively H131 and FcγRIIIA V 158 The recombinant protein was captured at a flow rate of 10 μL / min for a capture time of 40 seconds. After the line stabilized, gradient dilution of the antibody (starting at 37.5 μg / mL, diluted 2-fold) was performed. The mixture was passed through the chip at a flow rate of 30 μL / min, the binding time was 120 seconds, and the dissociation time was 100 μL. The time interval was 200 seconds, and the fitting was performed using the Biacore evaluation software. The results are shown in Figure 14. The specific antibodies bind to activating receptors FcγRI and FcγRIIA. H131 and FcγRIIIA V 158 The wild-type IgG4 and control antibodies did not bind to FcγRI and FcγRII, respectively. A H131 or FcγRIIIA V158 was bonded to.

[0336] Example 9. BCMA x CD3κλ bispecific antibody binds to BCMA receptor ligands Binding disruption Prepare human BCMA recombinant antigen at 1 μg / mL in carbonate buffer at pH 9.6. The plate was coated with 5% skim milk / P After blocking with PBS for 1 hour at room temperature, each well was filled with antibody (diluted 4-fold from 30 μg / mL). 50 μL of the diluted solution (a total of 10 gradients) was added and incubated at room temperature for 30 minutes. Biotin-labeled human APRIL recombinant protein (Novopr) at a concentration of 240 ng / mL 50 μL of cellulose acetate (CU89) was added, followed by incubation for 30 minutes. 0.05% PBST (PBS + 0.05% Tween-20) and PBS, respectively. After washing three times, 100 μL of SA-HRP (1:8000 dilution) was added to each well. The mixture was incubated at room temperature for 45 minutes. After washing, 100 μL of TMB coloring solution was added to each well. After 10 minutes, Add 50 μL of sulfuric acid to terminate the reaction, and measure the OD450nm using a microplate reader. A three-parameter fitting curve was generated using Graphpad software. The detection results are shown in Figure 15. The BCMA × CD3κλ bispecific antibody binds to the BCMA receptor. It effectively blocked binding to APRIL.

[0337] Example 10: Immune reconstitution mouse subcutaneous NCI-H929 tumor transplant model Six to eight-week-old female B-NGD mice (Biocytogen Pharmaceutic als Co., Ltd.) was selected and subcutaneously injected with 2 × 10 6 NCI-H929 cells (M Atrigel (mixed 1:1) was inoculated into the mice, and the tumors were 60 mm 3 When it reaches The subjects were divided into three groups: 3.0 mg / kg, 0.6 mg / kg, and 0.12 mg / kg. The negative control group was KLH×CD3 at 3 mg / kg. MC cells 1 x 10 7 The mice were injected into the tail vein, and the first administration was started three days later. The tumor volume and body weight of the mice were measured every two days. After the experiment was completed, the mice were sacrificed by cervical dislocation and the tumors were weighed and recorded. The results are shown in Figure 16 and demonstrate that the in vivo efficacy of the BCMAxCD3κλ bispecific antibody varies with dose. The tumor inhibition rates at low, medium and high doses were 55%, 95% and 1%, respectively. 08% (BCMA×CD3κλ005), and 46%, 94%, and 108% (BCMA×C D3κλ006), and its therapeutic effect was far superior to that of the control antibody. The dose was well tolerated by the mice, and no adverse effects such as weight loss were observed. Ta.

Claims

1. (a) a first antigen-binding portion or antigen-binding fragment thereof that binds to a BCMA antigen on the surface of a target cell, the first antigen-binding portion comprising a first heavy chain and a first light chain, the first antigen-binding portion comprising a first binding domain that binds to the first antigen, the first binding domain being a combination of heavy and light chain complementarity-determining regions (CDRs); the first heavy chain CDR1, CDR2, and CDR3 comprise amino acid sequences having 100% sequence identity to SEQ ID NOs: 12, 186, and 14, respectively, and the first light chain CDR1, CDR2, and CDR3 comprise amino acid sequences having 100% sequence identity to SEQ ID NOs: 17, 40, and 19, respectively; or a first antigen-binding portion or antigen-binding fragment thereof, wherein the first heavy chain CDR1, CDR2, and CDR3 comprise amino acid sequences having 100% sequence identity to SEQ ID NOs: 12 and 1314, respectively, and the first light chain CDR1, CDR2, and CDR3 comprise amino acid sequences having 100% sequence identity to SEQ ID NOs: 17, 40, and 19, respectively; and (b) a second antigen-binding portion or antigen-binding fragment thereof that binds to a CD3 antigen on the surface of a T cell, wherein the second antigen-binding portion comprises a second heavy chain and a second light chain, and the second antigen-binding portion comprises a second binding domain that binds to a second antigen; Including, Bispecific antibodies or antigen-binding portions thereof.

2. the first binding domain comprises a first heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 187 and a first light chain variable region comprising the amino acid sequence of SEQ ID NO: 188; or the first binding domain comprises a first heavy chain variable region comprising an amino acid sequence that exhibits at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 162, and a first light chain variable region comprising an amino acid sequence that exhibits at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 160; 2. The bispecific antibody or antigen-binding portion thereof of claim 1, comprising:

3. 3. The bispecific antibody or antigen-binding portion thereof of claim 1, wherein the second binding domain comprises, as second heavy chain CDR1, CDR2, and CDR3, amino acid sequences that exhibit 100% sequence identity with the amino acid sequences SEQ ID NOs: 114, 115, and 116, and as second light chain CDR1, CDR2, and CDR3, amino acid sequences that exhibit 100% sequence identity with the amino acid sequences SEQ ID NOs: 95, 96, and 108.

4. the second binding domain comprises a second heavy chain variable region comprising an amino acid sequence that exhibits at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence SEQ ID NO: 138; 4. The bispecific antibody or antigen-binding portion thereof of claim 1 , comprising a second light chain variable region comprising an amino acid sequence that exhibits at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to ID NO:

106.

5. 5. The bispecific antibody or antigen-binding portion thereof of claim 1, wherein the first light chain of the first antigen-binding portion is a -type light chain and the second light chain of the second antigen-binding portion is a -type light chain.

6. the second light chain variable region of the second antigen-binding portion has a Gln40Glu mutation (VbD3: Gln40Glu); and / or the second heavy chain variable region of the second antigen-binding portion is Gln ３９ Lys mutation (VHCD3: Gln ３９ Lys), and / or the first light chain variable region of the first antigen-binding portion is Gln ４２ Lys mutation (VαCMA: Gln ４２ Lys), and / or the first heavy chain variable region of the first antigen-binding portion is Gln ３９ Glu mutation (VHBCMA: Gln ３９ Glu), and / or the Fc regions of the first antigen-binding portion and the second antigen-binding portion of the bispecific antibody adopt a knob-into-hole architecture; and / or 6. The bispecific antibody or antigen-binding portion thereof according to claim 1 , wherein the Fc regions of the first antigen-binding portion and the second antigen-binding portion of the bispecific antibody adopt a human IgG4 knob-into-hole conformation.

7. the first binding domain of the bispecific antibody comprises a first heavy chain variable region of amino acid sequence SEQ ID NO: 187 and a first light chain variable region of amino acid sequence SEQ ID NO: 188, and the second binding domain comprises a second heavy chain variable region of amino acid sequence SEQ ID NO: 138 and a second light chain variable region of amino acid sequence SEQ ID NO: 106; or 7. The bispecific antibody or antigen-binding portion thereof of claim 1 , wherein the first binding domain of the bispecific antibody comprises a first heavy chain variable region exhibiting at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 162 and a first light chain variable region exhibiting at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 160, and the second binding domain comprises a second heavy chain variable region of amino acid sequence SEQ ID NO: 138 and a second light chain variable region of amino acid sequence SEQ ID NO:

106.

8. 8. The bispecific antibody or antigen-binding portion thereof of claim 1 , wherein the first heavy chain comprises a heavy chain selected from SEQ ID NOs: 174, 178 and the first light chain comprises a light chain selected from SEQ ID NOs:

172.

9. the first heavy chain comprises a heavy chain selected from SEQ ID NO: 174 and the first light chain comprises a light chain selected from SEQ ID NO: 172; or 9. The bispecific antibody or antigen-binding portion thereof of claim 1 , wherein the first heavy chain comprises a heavy chain selected from SEQ ID NO: 178 and the first light chain comprises a light chain selected from SEQ ID NO:

172.

10. 10. The bispecific antibody or antigen-binding portion thereof of claim 1, wherein the second heavy chain comprises the heavy chain of SEQ ID NO: 182 and the second light chain comprises the light chain of SEQ ID NO:

180.

11. the first heavy chain of the bispecific antibody comprises the heavy chain of SEQ ID NO: 174, the first light chain comprises the light chain of SEQ ID NO: 172, the second heavy chain comprises the heavy chain of SEQ ID NO: 182, and the second light chain comprises the light chain of SEQ ID NO: 180; or 11. The bispecific antibody or antigen-binding portion thereof of claim 1, wherein the first heavy chain of the bispecific antibody comprises the heavy chain of SEQ ID NO: 178, the first light chain comprises the light chain of SEQ ID NO: 172, the second heavy chain comprises the heavy chain of SEQ ID NO: 182, and the second light chain comprises the light chain of SEQ ID NO:

180.

12. An antibody or antigen-binding portion thereof that binds to BCMA, comprising the first antigen-binding portion of any one of claims 1 to 11.

13. A nucleic acid encoding the bispecific antibody or antigen-binding portion thereof of any one of claims 1 to 11, or the antibody or antigen-binding portion thereof that binds to BCMA of claim 12.

14. The nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion has the nucleotide sequence of SEQ ID NO: 189, and the nucleic acid encoding the first light chain variable region has the nucleotide sequence of SEQ ID NO: the nucleotide sequence of ID NO: 190, and / or The nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion has the nucleotide sequence of SEQ ID NO: 163, and the nucleic acid encoding the first light chain variable region has the nucleotide sequence of SEQ ID NO:

164. the nucleotide sequence of ID NO: 161, and / or the nucleic acid encoding the first heavy chain of the first antigen-binding moiety is selected from the nucleotide sequence of SEQ ID NO: 179, 175, and / or the nucleic acid encoding the first light chain of the first antigen-binding moiety is selected from the nucleotide sequence of SEQ ID NO: 173, and / or the nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion has the nucleotide sequence of SEQ ID NO: 139 and the nucleic acid encoding the second light chain variable region has the nucleotide sequence of SEQ ID NO: 107; and / or 14. The nucleic acid of claim 13, wherein the nucleic acid encoding the second heavy chain of the second antigen-binding portion is the nucleotide sequence of SEQ ID NO: 183, and / or the nucleic acid encoding the second light chain of the second antigen-binding portion is the nucleotide sequence of SEQ ID NO:

181.

15. the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody has the nucleotide sequence of SEQ ID NO: 163, the nucleic acid encoding the first light chain variable region has the nucleotide sequence of SEQ ID NO: 161, the nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion has the nucleotide sequence of SEQ ID NO: 139, and the nucleic acid encoding the second light chain variable region has the nucleotide sequence of SEQ ID NO: 107; and / or the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody has the nucleotide sequence of SEQ ID NO: 179, the nucleic acid encoding the first light chain of the first antigen-binding portion has the nucleotide sequence of SEQ ID NO: 173, the nucleic acid encoding the second heavy chain of the second antigen-binding portion has the nucleotide sequence of SEQ ID NO: 183, and the nucleic acid encoding the second light chain of the second antigen-binding portion has the nucleotide sequence of SEQ ID NO: 181; and / or 15. The nucleic acid of claim 14, wherein the nucleic acid encoding the first heavy chain of the second antigen-binding portion of the bispecific antibody is the nucleotide sequence of SEQ ID NO: 175, the nucleic acid encoding the first light chain of the first antigen-binding portion is the nucleotide sequence of SEQ ID NO: 173, the nucleic acid encoding the second heavy chain of the second antigen-binding portion is the nucleotide sequence of SEQ ID NO: 183, and the nucleic acid encoding the second light chain of the second antigen-binding portion is the nucleotide sequence of SEQ ID NO:

181.

16. A vector comprising the nucleic acid according to any one of claims 13 to 15.

17. A cell comprising the nucleic acid according to any one of claims 13 to 15 or the vector according to claim 16.

18. 18. A composition comprising a bispecific antibody or antigen-binding portion thereof according to any one of claims 1 to 11, or an antibody or antigen-binding portion thereof that binds to BCMA according to claim 12, or a nucleic acid according to any one of claims 13 to 15, or a vector according to claim 16, or a cell according to claim 17.

19. 13. An antibody-drug conjugate comprising the bispecific antibody, or antigen-binding portion thereof, of any one of claims 1 to 11, covalently linked to a therapeutic moiety, or the antibody, or antigen-binding portion thereof, that binds to BCMA, of claim 12.

20. 20. The antibody-drug conjugate of claim 19, wherein the therapeutic moiety is selected from a cytotoxic moiety, a chemotherapeutic agent, a cytokine, an immunosuppressant, an immunostimulatory agent, a degradative peptide, and a radioisotope.

21. The cytotoxic moiety may be selected from the group consisting of taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine (cephaline), mitomycin, etoposide, teniposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracenedione, tubulin inhibitors, mitotic inhibitors, dolastatin 10 or 15, irinotecan, mitoxantrone, mithramycin, actinomycin D D), 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, calicheamicin, antimetabolites, alkylating agents, cisplatin or carboplatin, duocarmycin A, duocarmycin SA, rachelmycin (CC-1065), antibiotics, pyrrolo[2,1-c][1,4]-benzodiazepine (PDB), diphtheria toxin and related molecules, ricin toxin, cholera toxin, shiga-like toxin, pertussis toxin, tetanus toxin, Bowman-Birk soybean protease inhibitor, Pseudomonas exotoxin, allorin, saporin, modeccin, geranin, abrin A chain, modeccin A chain, sarcin, Aleurites fordii protein, dianthin protein, Phytolacca americana protein, momordica charantia inhibitor, curcin, crotin, saponaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin and enomycin toxins, ribonuclease (RNase), DNase I, Staphylococcus aureus endotoxin A, pokeweed antiviral protein, diphtheria toxin, Pseudomonas endotoxin, The cytokine is selected from IL-2, IL-4, IL-6, IL-7, IL-10, IL-12, IL-13, IL-15, IL-18, IL-23, IL-24, IL-27, IL-28a, IL-28b, IL-29, KGF, IFNγ, GM-CSF, CD40L, Flt3 ligand, stem cell factor, ancestim, and TNF; The radioisotope is ３ H. １４ C. １５ N. ３５ S. ６７ Cu, ９０ Y. ９９ Tc, １２５ I, １３１ I, １８６ Re, １８８ Re, ２１１ At, ２１２ Bi, ２１２ Pb, ２１３ Bi, ２２５ Ac and ２２７ The antibody-drug conjugate of claim 20, wherein the antibody-drug conjugate is selected from Th.

22. the tubulin inhibitor comprises maytansine; the mitotic inhibitor is selected from monomethyl auristatin E and F; the antimetabolite is selected from methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, fludarabine, 5-fluorouracil, decarbazine, hydroxyurea, asparaginase, gemcitabine, and cladribine; The alkylating agent may be mechlorethamine, thiopurine, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, or dacarbazine. (DTIC), procarbazine, mitomycin C; the antibiotic is selected from actinomycin, bleomycin, daunorubicin, doxorubicin, idarubicin, mithramycin, mitomycin, primycin, and anthramycin (AMC); the diphtheria toxin and related molecules are selected from the diphtheria A chain and active fragments thereof, and hybrid molecules; the ricin is selected from ricin A and deglycosylated ricin A chain toxins; the Shiga-like toxin is selected from SLT I, SLT II, ​​SLT IIV, LT toxin, C3 toxin, and Shiga toxin; 22. The antibody-drug conjugate of claim 21, wherein the Phytolacca americana protein is selected from PAPI, PAPII, and PAP-S.

23. A kit comprising a specific antibody or antigen-binding portion thereof according to any one of claims 1 to 12, or an antibody or antigen-binding portion thereof that binds to BCMA according to claim 12, or a nucleic acid molecule according to any one of claims 13 to 15, or a vector according to claim 16, or a cell according to claim 17, or a composition according to claim 18, and / or an antibody-drug conjugate according to any one of claims 19 to 22.

24. Use of the specific antibody or antigen-binding portion thereof according to any one of claims 1 to 11, the antibody or antigen-binding portion thereof that binds to BCMA according to claim 12, the nucleic acid molecule according to any one of claims 13 to 15, the vector according to claim 16, the cell according to claim 17, the composition according to claim 18, and / or the antibody-drug conjugate according to any one of claims 19 to 22 for the diagnosis, treatment or prevention of a BCMA-associated disease, including a B-cell disease, with the proviso that humans are excluded as subjects for said diagnosis, treatment or prevention.

25. 25. The use of the specific antibody or antigen-binding portion thereof, antibody or antigen-binding portion thereof that binds to BCMA, nucleic acid molecule, vector, cell, composition, and / or antibody-drug conjugate of claim 24, The diseases include multiple myeloma, malignant plasmacytoma, Hodgkin's lymphoma, nodular lymphocyte-predominant Hodgkin's lymphoma, Kahler's disease and myeloid leukemia, plasma cell leukemia, plasmacytoma, B-cell prolymphocytic leukemia, hairy cell leukemia, B-cell non-Hodgkin's lymphoma (NHL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL), chronic myeloid leukemia (CML), follicular lymphoma, and Burkitt's lymphoma. , marginal zone lymphoma, mantle cell lymphoma, large cell lymphoma, precursor B-lymphocytic lymphoma, myeloid leukemia, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma, follicular lymphoma, marginal zone lymphoma, mucosa-associated lymphoid tissue lymphoma, small cell lymphocytic lymphoma, mantle cell lymphoma, Burkitt's lymphoma, primary mediastinal (thymic) large B-cell lymphoma, lymphoplasmacytic lymphoma, Waldenstrom's Macroglobulinemia, nodal marginal zone B-cell lymphoma, splenic marginal zone lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, lymphomatoid granulomatosis, T-cell / histiocytocyte-rich large B-cell lymphoma, primary central nervous system lymphoma, primary cutaneous diffuse large B-cell lymphoma (leg type), elderly EBV-positive diffuse large B-cell lymphoma, inflammation-associated diffuse large B-cell lymphoma, intravascular large B-cell lymphoma, ALK-positive B-cell-related cancers selected from large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma arising in HHV8-associated multicentric Castleman disease, unclassified B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma, unclassified B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma, and other B-cell-related lymphomas; and / or the B-cell disorder is selected from multiple myeloma, plasmacytoma, plasma cell leukemia, macroglobulinemia, amyloidosis, Waldenström's macroglobulinemia, solitary plasmacytoma of bone, extramedullary plasmacytoma, osteosclerosing myeloma, heavy chain disease, monoclonal gammopathy of undetermined significance, and smoldering multiple myeloma; and / or The disease comprises an autoimmune disease such as systemic lupus erythematosus or rheumatoid arthritis.

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