Development and Use of Novel Multispecific Tumor Inhibitors

Abstract

The present disclosure relates to antibodies containing structural domains that bind to GPRC5D and their use. The GPRC5D×CD3 bispecific antibody has a strong killing effect on low-expression cells and is at the same time milder to T cell activation, and the GPRC5D×BCMA×CD3 trispecific antibody + can kill both BCMA + cells and GPRC5D cells, helps to overcome the resistance that occurs with a single target, and brings about a more profound and long-lasting tumor inhibitory effect in clinical applications.

Description

Technical Field

[0001] The present disclosure relates to antibodies comprising a structural domain that binds to GPRC5D and their use, and in particular to multispecific antibodies comprising a structural domain that binds to GPRC5D and their use.

Background Art

[0002] New therapies that have revolutionized the treatment options for multiple myeloma, such as Blenrep (belantamab mafodotin), an ADC therapy targeting B cell maturation antigen (BCMA), approved for sale by the FDA in 2020, Abecma (idecabtagene vicleucel), a CAR-T therapy targeting BCMA, approved for sale in 2021, and Carvykti (ciltacabtagene autoleucel), a recently approved CAR-T therapy targeting BCMA, have been successively approved for sale. However, multiple myeloma is still considered an incurable disease, and ultimately most patients relapse. The disappearance or low expression of the BCMA antigen during treatment may lead to tumor recurrence and drug resistance (van de Donk et al., CAR T-cell therapy for multiple myeloma: state of the art and prospects, Lancet Haematol. 2021 Jun;8(6):e446-e461; Gazeau N et al., Effective anti-BCMA retreatment in multiple myeloma, Blood Adv (2021) 5 (15); Da Via MC et al., Homozygous BCMA gene deletion in response to anti-BCMA CAR T cells in a patient with multiple myeloma, Nat Med. 2021 Apr;27(4)). Therefore, developing other novel targeted therapies that do not rely on BCMA, or combination therapies based on different targets, is a reasonable strategy to reduce the recurrence and drug resistance of multiple myeloma due to antigen loss.

[0003] GPRC5D (G-protein-coupled receptor family C group 5 member D) is an orphan G protein-coupled receptor with a seven-transmembrane structure (Brauner-Osborne H et al., Cloning and characterization of a human orphan family C G-protein coupled receptor GPRC5D, Biochim Biophys Acta. 2001 Apr 16;1518(3)), and this gene is expressed only in cells that produce hard keratin in the hair follicle part of the skin (Inoue S et al., The RAIG family member, GPRC5D, is associated with hard-keratinized structures, J Invest Dermatol.2004 Mar;122(3)). In addition, analysis of gene expression in myeloma patients found that GPRC5D was specifically highly expressed in multiple myeloma cells and was associated with poor prognosis (Atamaniuk J et al., Overexpression of G protein-coupled receptor 5D in the bone marrow is associated with poor prognosis in patients with multiple myeloma, Eur J Clin Invest.2012 Sep;42(9); Cohen Y et al., GPRC5D is a promising marker for monitoring the tumor load and to target multiple myeloma cells, Hematology.2013 Nov;18(6)).Smith EL et al. further demonstrated that the GPRC5D receptor protein is selectively overexpressed in malignant myeloid plasmacytoma, independent of BCMA expression, and is a novel immunotherapy target (Smith EL et al., PRC5D is a target for the immunotherapy of multiple myeloma with rationally designed CAR T cells, Sci Transl Med. 2019 Mar 27;11(485)).

[0004] Currently, bispecific antibodies and CAR-T therapy targeting GPRC5D have shown remarkable therapeutic effects on multiple myeloma in preclinical trials.

[0005] Talquetamab (JNJ-64407564) is a GPRC5D × CD3 bispecific antibody developed by Janssen. + It has been shown that it can mediate effective killing of myeloma cells by T cells, and a significant positive correlation was observed between the killing effect and the cellular GPRC5D expression level (Pillarisetti K et al., A T-cell-redirecting bispecific G-protein-coupled receptor class 5 member D×CD3 antibody to treat multiple myeloma, Blood.2020 Apr 9;135(15);Verkleij CPM et al., Preclinical activity and determinants of response of the GPRC5D×CD3 bispecific antibody Talquetamab in multiple myeloma, Blood Adv.2021 Apr 27;5(8)).

[0006] Kodama T et al. also reported a new GPRC5D×CD3 bispecific antibody, but since it does not recognize the cynomolgus monkey GPRC5D receptor, another method is needed to evaluate preclinical safety (Kodama T et al., Anti-GPRC5D / CD3 Bispecific T-Cell-Redirecting Antibody for the Treatment of Multiple Myeloma, Mol Cancer Ther. 2019 Sep;18(9)).

[0007] In US20210054094A1, Fertig et al. described two anti-GPRC5D antibodies, 5E11 and 5F11, obtained from rat immunization. However, this anti-GPRC5D antibody binds to stably transfected cells with high affinity but does not bind to cynomolgus monkey GPRC5D + stably transfected cells. The other two antibodies, 10B10 and B72, bind to cynomolgus monkey GPRC5D, but the binding level is low and the mediated TDCC activity is weak. + Fernandez de Larrea C et al. reported that CAR-T therapy targeting both BCMA and GPRC5D was shown to prevent relapse due to BCMA escape in a mouse xenograft tumor model (Fernandez de Larrea C et al., Defining an Optimal Dual-Targeted CAR T-cell Therapy Approach Simultaneously Targeting BCMA and GPRC5D to Prevent BCMA Escape-Driven Relapse in Multiple Myeloma, Blood Cancer Discov. 2020 Sep;1(2))

[0008]

[0009] ​Despite the preclinical efficacy shown by the above-mentioned drug developed based on an antibody targeting GPRC5D, there remains a need to develop more excellent treatment means clinically to reduce recurrence due to antigen escape and drug resistance, and to reduce side effects such as cytokine release syndrome and neurotoxicity during the treatment process.

Summary of the Invention

[0010] In this study, the inventors newly discovered a plurality of anti-GPRC5D monoclonal antibodies that can bind to human and cynomolgus monkey GPRC5D receptors on the cell membrane with high affinity using different mouse immunization strategies. Additionally, a novel humanized GPRC5D×CD3 bispecific antibody constructed based on the anti-CD3e antibody SP34 (Pessano et al., The T3 / T cell receptor complex: antigenic distinction between the two 20-kd T3 (T3-delta and T3-epsilon) subunits, EMBO J. 1985 Feb;4(2)) has strong killing power against low-expression cells and is gentler on T cell activation, which unexpectedly discovered that it may lead to clinically excellent efficacy and a better safety profile. The inventors further constructed a GPRC5D×BCMA×CD3 trispecific antibody, which showed better efficacy than the BCMA×CD3 bispecific antibody and the potential to bring about a more sustained response in a preclinical mouse model.

[0011] In one aspect, the present disclosure provides an antibody that binds to GPRC5D or an antigen-binding portion thereof.

[0012] In another aspect, the present disclosure provides a nucleic acid encoding the aforementioned antibody or an antigen-binding portion thereof.

[0013] In another aspect, the present disclosure provides a multispecific antibody or an antigen-binding portion thereof that includes at least (a) the aforementioned antibody or an antigen-binding fragment thereof that binds to the GPRC5D antigen, and (b) an antibody or an antigen-binding fragment thereof that binds to the CD3 antigen, and includes the following. (a) The aforementioned antibody or an antigen-binding fragment thereof that binds to the GPRC5D antigen is a first antigen-binding portion, the GPRC5D antigen is a first antigen, the first antigen-binding portion includes a first heavy-chain variable region and a first light-chain variable region, and the first antigen-binding portion includes a first binding structural domain that binds to the first antigen. (b) The antibody or an antigen-binding fragment thereof that binds to the CD3 antigen is a second antigen-binding portion, the CD3 antigen is a second antigen, the second antigen-binding portion includes a second heavy-chain variable region and a second light-chain variable region, and the second antigen-binding portion includes a second binding structural domain that binds to the second antigen. More preferably, the multispecific antibody or an antigen-binding portion thereof further includes (c) an antibody or an antigen-binding fragment thereof that binds to the BCMA antigen. (c) The antibody or an antigen-binding fragment thereof that binds to the BCMA antigen is a third antigen-binding portion, the BCMA antigen is a third antigen, the third antigen-binding portion includes a third heavy-chain variable region and a third light-chain variable region, and the third antigen-binding portion includes a third binding structural domain that binds to the third antigen.

[0014] In another aspect, the present disclosure provides a nucleic acid encoding the aforementioned multispecific antibody or an antigen-binding portion thereof.

[0015] In another aspect, the present disclosure provides a vector encoding the aforementioned nucleic acid.

[0016] In another aspect, the present disclosure provides a cell containing the aforementioned vector.

[0017] In another aspect, the present disclosure provides a composition comprising the aforementioned antibody or antigen-binding portion thereof that binds to GPRC5D, the aforementioned multispecific antibody or antigen-binding portion thereof, the aforementioned nucleic acid, and / or the aforementioned cell.

[0018] In another aspect, the present disclosure provides a kit comprising the aforementioned antibody or antigen-binding portion thereof that binds to GPRC5D, the aforementioned multispecific antibody or antigen-binding portion thereof, the aforementioned nucleic acid, the aforementioned cell, and / or the aforementioned composition.

[0019] In another aspect, the present disclosure provides the use of the aforementioned antibody or antigen-binding portion thereof that binds to GPRC5D, the aforementioned multispecific antibody or antigen-binding portion thereof, the aforementioned nucleic acid, the aforementioned cell, and / or the aforementioned composition in the preparation of a medicament or kit for use in the diagnosis, treatment, or prevention of cancer or autoimmune diseases.

Brief Description of the Drawings

[0020]

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Modes for Carrying Out the Invention

[0021] I. Definitions In the present disclosure, scientific and technical terms used herein have the meanings generally understood by those skilled in the art, unless otherwise specified. Further, terms and experimental procedures related to protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, and immunology used herein are terms and ordinary procedures widely used in their respective fields. Also, to better understand the present disclosure, definitions and explanations of related terms are provided below.

[0022] Provided herein are antibodies (e.g., monoclonal antibodies) that specifically bind to GPRC5D and antigen-binding fragments thereof. In a specific embodiment, provided herein is a monoclonal anti-GPRC5D antibody that specifically binds to GPRC5D, and the anti-GPRC5D antibody includes variants of the parental antibody. In a specific embodiment, provided herein is an antibody that specifically binds to GPRC5D (e.g., human GPRC5D). In certain embodiments, provided herein is an anti-GPRC5D antibody that includes modifications at one or more amino acid residues (e.g., 1 to 3 amino acid substitutions in the framework region of the heavy chain variable region), and this antibody retains the affinity for the antigen as compared to the parental antibody that does not include such modifications.

[0023] As used herein, unless otherwise specified, the term "about" or "approximately" means within ±10% of a given value or range. When an integer is required, this term means within ±10% of the given value or range and is rounded up or down to the nearest integer.

[0024] In the case of an antibody chain polypeptide sequence, the phrase "substantially identical" is understood to mean an antibody chain that exhibits at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity to the reference polypeptide sequence. In the case of a nucleic acid sequence, this term is understood to mean a nucleotide sequence that exhibits at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity to the reference nucleic acid sequence.

[0025] "Identity" or "sameness" of sequences has the meaning recognized in the art, and the percentage of sequence identity between two nucleic acid or polypeptide molecules or regions can be calculated using the disclosed techniques. Sequence identity can be measured along the entire length of a polynucleotide or polypeptide or along a region of the molecule. There are numerous methods for measuring identity between two polynucleotides or polypeptides, but the term "identity" is well known to those skilled in the art (Carrillo, H. & Lipman, D., SIAM J Applied Math 48:1073 (1988)).

[0026] A "substitution-type" variant is a variant in which at least one amino acid residue in the native sequence has been removed and a different amino acid has been inserted at the same position. The substitution can be a single substitution in which only one amino acid is substituted within the molecule, or a multiple substitution in which two or more amino acids are substituted within the same molecule. Multiple substitutions can be located at contiguous sites. Similarly, one amino acid can be substituted with two or more residues, and such variants include both substitutions and insertions. An "insertion-type" variant is a variant in which one or more amino acids have been inserted adjacent to a particular position in the native sequence. Adjacent amino acids mean being linked to the α-carboxy or α-amino group of the amino acid. A "deletion-type" variant is a variant in which one or more amino acids have been removed from the native amino acid sequence. Usually, deletion-type variants lack one or two amino acids in a particular region of the molecule.

[0027] With respect to the variable structural domain of an antibody, the term "variable" refers to the portion of the target molecule that is used for the specific recognition and binding of a particular antibody to a specific target and that has extensive sequence differences among antibodies. However, the variability is not evenly distributed throughout the entire variable domain of the antibody. The variability is concentrated in three segments known as complementarity-determining regions (CDRs; CDR1, CDR2, CDR3) or hypervariable regions, which are located in both the variable structural domains of the light and heavy chains. The more highly conserved portions of the variable structural domain are called framework (FR) regions or framework sequences. Each variable structural domain of the native heavy and light chains consists of four FR regions, which mainly adopt a β-sheet structure, and they are linked by three CDRs, forming loops that link the β-sheet structures and in some cases form part of the β-sheet structures. The CDRs of each chain are usually closely associated by the FR regions and contribute to the formation of the antibody target-binding site (epitope or determinant) with the help of the CDRs of the other chain. As used herein, the numbering of immunoglobulin amino acid residues is based on the Kabat et al. immunoglobulin amino acid residue numbering system, unless otherwise specified. One CDR may have the ability to specifically bind to the associated epitope.

[0028] As used herein, an "antibody fragment" or "antigen-binding fragment" of an antibody refers to any portion of a full-length antibody that is less than full-length but contains at least a portion of the variable region of the antibody that binds to an antigen (e.g., one or more CDRs and / or one or more antibody-binding sites), and thus retains at least a portion of the binding specificity and the specific binding ability of the full-length antibody. Thus, an antigen-binding fragment refers to an antibody fragment that contains an antigen-binding portion that binds to the same antigen as the antibody from which the antibody fragment is derived. Antibody fragments include antibody derivatives produced by enzymatic treatment of a full-length antibody, derivatives produced by synthesis such as recombinant derivatives, etc. Antibodies include antibody fragments. Examples of antibody fragments include, but are not limited to, Fab, Fab’, F(ab’)2, single-chain Fv (scFv), Fv, dsFv, diabody, Fd and Fd’ fragments, and other fragments including modified fragments (see, for example, Methods in Molecular Biology, Vol 207: Recombinant Antibodies for Cancer Therapy Methods and Protocols (2003); Chapter 1; p3-25, Kipriyanov). The fragments can include, for example, multiple chains linked by disulfide bonds and / or peptide linkers. Antibody fragments generally contain at least about 50 amino acids and typically contain at least about 200 amino acids. An antigen-binding fragment, when inserted into an antibody framework (e.g., by replacing the corresponding region), binds immunospecifically to an antigen (i.e., at least, at least about 10 7 ~10 8 M -1Any antibody fragment that obtains an antibody showing Ka of ) is included. A "functional fragment" or "anti-GPRC5D antibody analog" is a fragment or analog that blocks or substantially reduces the ability of the receptor to bind to a ligand or initiate signal transduction. As used herein, a functional fragment generally has the same meaning as an "antibody fragment" and, in the case of an antibody, can refer to a fragment such as Fv, Fab, F(ab’)2, etc. that blocks or substantially reduces the ability of the receptor to bind to a ligand or initiate signal transduction. An "Fv" fragment consists of a dimer (VH-VL dimer) formed by non-covalent binding of one heavy chain variable structural domain and one light chain variable structural domain. In this configuration, the three CDRs of each variable structural domain interact with each other to determine the target binding site on the surface of the VH-VL dimer, similar to the case of an intact antibody. The six CDRs together confer the target binding specificity of the intact antibody. However, even a single variable structural domain (or half of the Fv containing only three target-specific CDRs) can have the ability to recognize and bind a target.

[0029] As used herein, "monoclonal antibody" refers to a population of identical antibodies, indicating that individual antibody molecules in the monoclonal antibody population are identical to other antibody molecules. This feature is in contrast to that of a polyclonal antibody population containing antibodies with multiple different sequences. Monoclonal antibodies can be prepared by many 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, for example, by fusing them with myeloma cells to produce a hybridoma cell line or by infecting B cells with a virus such as EBV. Recombinant techniques can also be used to prepare antibodies from a clonal population of host cells by transforming the host cells in vitro with a plasmid having an artificial sequence of nucleotides encoding the antibody.

[0030] As used herein, a full-length antibody is an antibody that has two full-length heavy chains (e.g., VH-CH1-CH2-CH3 or VH-CH1-CH2-CH3-CH4) and two full-length light chains (VL-CL) as well as a hinge region, and includes, for example, antibodies naturally produced by antibody-secreting B cells and synthetically produced antibodies having the same structural domains.

[0031] The term "chimeric antibody" refers to an antibody in which the variable region sequence is derived from one species and the constant region sequence is derived from another species, for example, an antibody in which the variable region sequence is derived from a mouse antibody and the constant region sequence is derived from a human antibody.

[0032] A "humanized" antibody is a chimeric immunoglobulin, immunoglobulin chain or fragment thereof (e.g., Fv, Fab, Fab’, F(ab’)2 or other antigen-binding subsequence of an antibody) that means a non-human (e.g., mouse) antibody form that includes a minimal sequence derived from a non-human immunoglobulin. Preferably, a humanized antibody is a human immunoglobulin (recipient antibody), and the residues of the complementarity-determining regions (CDRs) of the recipient antibody are replaced with the residues of the CDRs of a non-human species (donor antibody) such as a mouse, rat, rabbit, etc. having the desired specificity, affinity, and capacity.

[0033] Furthermore, in humanization, it is also possible to mutate amino acid residues within the CDR1, CDR2, and / or CDR3 regions of VH and / or VL, thereby improving one or more binding properties (e.g., affinity) of the antibody. For example, mutations can be introduced by mutagenesis via PCR, and their effects on antibody binding or other functional properties can be evaluated using the in vitro or in vivo assays described herein. Usually, conservative mutations are introduced. Such mutations can be amino acid substitutions, additions, or deletions. Furthermore, the mutations within the CDRs are usually one or two or less. Thus, the humanized antibodies described in the present disclosure also include antibodies that contain one or two two-amino acid mutations within the CDRs.

[0034] "Conservative amino acid substitution" can include substituting a native amino acid residue with a non-native residue such that the polarity or charge of the amino acid residue at that position is minimally or not at all affected. Examples of conservative substitutions include substituting non-polar (hydrophobic) amino acid residues such as isoleucine, valine, leucine, n-leucine, alanine, methionine with another, for example, substituting polar (hydrophilic) amino acid residues such as between arginine and lysine, between glutamine and asparagine, between glycine and serine with another, substituting basic amino acid residues such as lysine, arginine or histidine with another, or substituting acidic residues such as aspartic acid or glutamic acid with another. The phrase "conservative amino acid substitution" also includes substituting a non-derivatized residue with a chemically derivatized residue, provided that the polypeptide exhibits the desired biological activity. Other exemplary amino acid substitutions that can be used in accordance with the present invention are shown in Table 1 below.

Table 1

[0035] As used herein, the term "CDR" refers to complementarity-determining region, and it is known that there are three in each heavy and light chain of an antibody molecule. CDRs are also called hypervariable regions and are present in the variable regions of each heavy and light chain of an antibody, having sites with very high variability in the primary structure of the CDRs. Herein, the CDRs of the heavy chain are represented as CDR1, CDR2, CDR3 from the amino terminus of the amino-terminal sequence of the heavy chain, and the CDRs of the light chain are represented as CDR1, CDR2, CDR3 from the amino terminus of the amino-terminal sequence of the light chain. These sites are close to each other in the tertiary structure and determine the specificity of the antigen to which the antibody binds.

[0036] As used herein, the term "epitope" refers to any antigenic determinant on an antigen to which the complementary site of an antibody binds. Epitope determinants usually include chemically active surface portions of molecules such as amino acids or sugar side chains, and usually have specific three-dimensional structural features and specific charge features.

[0037] As used herein, with respect to an antibody or an antigen-binding fragment thereof, "specifically binds" or "immunologically specifically binds" are used interchangeably herein, and refers to the ability of an antibody or antigen-binding fragment to form one or more non-covalent bonds with a homologous antigen through non-covalent interactions between the antibody and the antibody-binding site of the antigen. The antigen may be an isolated antigen or may be present in tumor cells. Usually, an antibody that immunologically specifically binds (or specifically binds) to an antigen binds to the antigen with an affinity constant Ka of about 1×10 7 M -1 or 1×10 8 M -1 or higher (or a dissociation constant (Kd) of 1×10 -7 M or 1×10 -8 M or lower). The affinity constant can be determined by standard kinetic methods for antibody reactions, such as immunoassays, surface plasmon resonance (SPR) (Rich and Myszka (2000) Curr.Opin.Biotechnol 11:54; Englebienne (1998) Analyst.123:1599), isothermal titration calorimetry (ITC), or other kinetic interaction assays known in the art (e.g., Paul, ed., Fundamental Immunology, 2nd ed., Raven Press, New York, pages 332-336 (1989), and see also U.S. Patent No. 7,229,619, which describes exemplary SPR and ITC methods for calculating the binding affinity of an antibody). Apparatus and methods for detecting and monitoring binding rates in real time are known and commercially available (see Malmqvist (2000) Biochem.Soc.Trans.27:335).

[0038] As used herein, the terms "polynucleotide" and "nucleic acid molecule" generally refer to an oligomer or polymer consisting of at least two linked nucleotides or nucleotide derivatives, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) that are linked together by phosphodiester bonds. As used herein, the term "nucleic acid molecule" is intended to include DNA molecules and RNA molecules. The nucleic acid molecule may be single-stranded or double-stranded and may be cDNA.

[0039] Alternatively, in another embodiment, for example, random mutations can be introduced by saturation mutagenesis along all or part of the anti-GPRC5D antibody coding sequence, and the resulting modified anti-GPRC5D antibody can be screened for improved binding activity.

[0040] As used herein, "expression" refers to the process of producing a polypeptide by transcription and translation of a polynucleotide. The expression level of a polypeptide can be evaluated using any method known in the art, such as a method for determining the amount of polypeptide produced from a host cell. Such methods include, but are not limited to, quantification of polypeptide in cell lysates by ELISA, Coomassie blue staining after gel electrophoresis, Lowry protein assay, Bradford protein assay, etc.

[0041] As used herein, a "host cell" is a cell that is used to receive, retain, replicate, and amplify a vector. A host cell may also be used to express a polypeptide encoded by a vector. When the host cell divides, the nucleic acid contained in the vector is replicated, thereby amplifying the nucleic acid. The host cell may be a eukaryotic cell or a prokaryotic cell. Suitable host cells include, but are not limited to, HEK cells such as CHO cells, various COS cells, HeLa cells, and HEK 293 cells.

[0042] As used herein, "vector" is a replicable nucleic acid, and when the vector is transfected into a suitable host cell, one or more heterologous proteins can be expressed therefrom. With respect to vectors, those vectors into which nucleic acids encoding polypeptides or fragments thereof can usually be introduced by restriction digestion and ligation are included. Vectors also include those vectors containing nucleic acids encoding polypeptides. Vectors are used to introduce nucleic acids encoding polypeptides into host cells, to amplify nucleic acids, or to express / display polypeptides encoded by nucleic acids. Vectors are usually free, but can be designed to integrate a gene or a part thereof into a genomic chromosome. Vectors of artificial chromosomes, such as yeast artificial vectors and mammalian artificial chromosomes, are also conceivable. The selection and use of such mediators are known to those skilled in the art.

[0043] As used herein, vectors also include "viral vectors" or "viral vectors". A viral vector is an engineered virus that is operably linked to an exogenous gene for introducing the exogenous gene into a cell (as a mediator or shuttle).

[0044] As used herein, an "expression vector" includes a vector capable of expressing DNA operably linked to regulatory sequences that can affect the expression of that DNA fragment, such as a promoter region. Such additional fragments may include promoter and terminator sequences and optionally may include one or more origins of replication, one or more selectable markers, enhancers, polyadenylation signals, etc. Expression vectors generally are derived from plasmid or viral DNA or may contain elements of both. Thus, an expression vector refers to a recombinant DNA or RNA construct such as a plasmid, phage, recombinant virus, or other vector that, when introduced into a suitable host cell, results in the expression of cloned DNA. Suitable expression vectors are known to those of skill in the art and include expression vectors that are replicable in eukaryotic and / or prokaryotic cells and expression vectors that are free or integrated into the host cell genome.

[0045] As used herein, "treating" an individual having a disease or disease state means that the symptoms of the individual are partially or completely alleviated or do not change after treatment. Thus, treatment includes prevention, therapy, and / or cure. Prevention refers to the prevention of a potential disease and / or the prevention of symptom exacerbation or disease progression. Treatment also includes any pharmaceutical use of any antibody or antigen-binding fragment thereof provided and any composition provided herein.

[0046] As used herein, a "therapeutic effect" refers to an effect resulting from the treatment of an individual that modifies, usually improves or ameliorates, or cures the symptoms of a disease or disease state.

[0047] As used herein, a "therapeutically effective amount" or "therapeutically effective dosage" refers to an amount of a substance, compound, material, or composition containing a compound that is at least sufficient to produce a therapeutic effect upon administration to a subject. Thus, it is the amount necessary to prevent, cure, ameliorate, arrest, or partially arrest the symptoms of a disease or disorder.

[0048] As used herein, "prophylactically effective amount" or "prophylactically effective dosage" refers to the amount of a substance, compound, material, or composition containing a compound that, when administered to a subject, prevents or delays the onset or recurrence of a desired prophylactic effect, e.g., a disease or condition, and reduces the likelihood of the onset or recurrence of the disease or condition. A complete prophylactically effective amount does not necessarily occur upon a single administration and may occur only after a series of administrations. Thus, a prophylactically effective amount can be administered in one or multiple doses.

[0049] As used herein, the terms "patient" or "subject" refer to mammals such as humans.

[0050] As used herein, unless otherwise specified, the terms "comprising," "including," "having," and "containing" are generally understood to be open-ended and non-limiting, including their grammatical equivalents, and do not exclude other unlisted elements or steps, for example.

[0051] II. MODE FOR CARRYING OUT THE INVENTION In one aspect, the present disclosure is (1) comprising a heavy chain variable region and a light chain variable region, wherein (i) the heavy chain variable region comprises the following (a) to (c): (a) an HCDR1 sequence having the following formula: TSYNMH (SEQ ID NO: 5), and (b) an HCDR2 sequence having the following formula: YIYPGNGGTNYNQX1FX2X3 (SEQ ID NO: 242), where X1 is K or N, X2 is K or Q, X3 is A or G, and (c) an HCDR3 sequence having the following formula: GYGYRYWYFDV (SEQ ID NO: 7), and (ii) the light chain variable region comprises the following (a) to (c): (a) an LCDR1 sequence having the following formula: X4ASSSVX5X6MH (SEQ ID NO: 243), where X4 is S or R, X5 is S or R, X6 is Y or F, and (b) An LCDR2 array having the following formula: DTSKX7AS (SEQ ID NO: 244), where X7 is V or L, and (c) An LCDR3 array having the following formula: QQWX8X9X 10 PX 11 T (SEQ ID NO: 245), where X8 is S or N, X9 is S or N, X 10 is N, Y or K, X 11 is L or P, or (2) Comprising a heavy chain variable region and a light chain variable region, where (i) The heavy chain variable region comprises the following (a) to (c), (a) An HCDR1 array having the following formula: TGYTMN (SEQ ID NO: 25), and (b) An HCDR2 array having the following formula: LINPYNGGX 12 X 13 YNX 14 KFX 15 G (SEQ ID NO: 246), where X 12 is S or T, X 13 is R, S or N, X 14 is Q or L, X 15 is K or Q, and (c) An HCDR3 array having the following formula: X 16 X 17 X 18 RX 19 X 20 MDY (SEQ ID NO: 247), where X 16 is W or L, X 17 is A, P or G, X 18 is I or L, X 19 is Y or N, X20 is A or G, and (ii) the light chain variable region contains the following (a) to (c): (a) an LCDR1 array having the following formula: X 21 ASQX 22 X 23 X 24 X 25 NX 26 X 27 (SEQ ID NO: 248), where X 21 is K, R or Q, X 22 is N or S, X 23 is V or I, X 24 is G or S, X 25 is T or N, X 26 is V or L, X 27 is A or H, and (b) an LCDR2 array having the following formula: X 28 ASX 29 X 30 X 31 S (SEQ ID NO: 249), where X 28 is F, Y or S, X 29 is Y, L or Q, X 30 is R, L or S, X 31 is N or I, and (c) an LCDR3 array having the following formula: QQX 32 NSX 33 X 34 X 35 T (SEQ ID NO: 250), where X 32 is Y or S, X 33 is Y or W, X 34 is L or P, X 35 is L or Q, or (3) contains a heavy chain variable region and a light chain variable region, where (i) the heavy chain variable region contains the following (a) to (c): (a) HCDR1 sequence having the following formula: TNFPIE (SEQ ID NO: 42), and (b) HCDR2 sequence having the following formula: NFHPYNDVTKYNEKFX 36 G (SEQ ID NO: 251), where X 36 is K or Q, and (c) HCDR3 sequence having the following formula: ITRGY (SEQ ID NO: 44), and (ii) The light chain variable region includes the following (a) - (c): (a) LCDR1 sequence having the following formula: KSSQSLLYSRNQKNSLA (SEQ ID NO: 47), and (b) LCDR2 sequence having the following formula: WASTRES (SEQ ID NO: 48), and (c) LCDR3 sequence having the following formula: QQYYSYPLT (SEQ ID NO: 49), or (4) Comprising a heavy chain variable region and a light chain variable region, where (i) The heavy chain variable region includes the following (a) - (c): (a) HCDR1 sequence having the following formula: TSYX 37 MY (SEQ ID NO: 252), where X 37 is Y or T, and (b) HCDR2 sequence having the following formula: X 38 INPSX 39 GX 40 TX 41 YNX 42 KFX 43 X 44 (SEQ ID NO: 253), where X 38 is G or Y, X 39 is N or S, X 40 is G or Y, X 41 is K or N, X 42 is Q or E, X 43is K or Q, and X 44 is S, D or G, and (c) an HCDR3 sequence having the following formula: X 45 X 46 X 47 X 48 X 49 X 50 X 51 YX 52 X 53 X 54 X 55 DY (SEQ ID NO: 254), where X 45 is G or W, and X 46 is G or S, and X 47 is G or L, and X 48 is R or N, and X 49 is E or S, and X 50 is Y or R, and X 51 is Y or L, and X 52 is A or Y, and X 53 is Y or M, and X 54 is A or absent, and X 55 is M or absent, and (ii) the light chain variable region includes the following (a) to (c): (a) an LCDR1 sequence having the following formula: X 56 ASX 57 X 58 X 59 X 60 X 61 YX 62 X 63 (SEQ ID NO: 255), where X 56 is R, Q or S, and X 57 is Q or S, and X 58 is D or S, and X 59 is I or V, and X 60 is S or absent, and X 61 is N or S, and X 62 is L or M, and X 63 is N or Y, and (b) The LCDR2 array having the following formula: X 64 TSX 65 LX 66 S (SEQ ID NO: 256), where X 64 is Y or G, and X 65 is R or N, and X 66 is H or A, and (c) The LCDR3 array having the following formula: QQX 67 X 68 SX 69 PX 70 T (SEQ ID NO: 257), where X 67 is R or G, and X 68 is Y or S, and X 69 is L or Y, and X 70 is Y or L, or (5) Comprising a heavy chain variable region and a light chain variable region, where (i) The heavy chain variable region comprises the following (a) to (c): (a) The HCDR1 array having the following formula: TX 71 YVMX 72 (SEQ ID NO: 258), where X 71 is S or N, and X 72 is Y or H, and (b) The HCDR2 array having the following formula: YINPYNDGTKYNEX 73 FX 74 G (SEQ ID NO: 259), where X 73 is K or R, and X 74 is K, T or Q, and (c) The HCDR3 array having the following formula: GGX 75 X 76 X 77 X 78 X 79 X 80 X 81 X 82 X 83Y (SEQ ID NO: 260), where X 75 is M or V, and X 76 is L or R, and X 77 is T or R, and X 78 is T or Y, and X 79 is R or F, and X 80 is S or absent, and X 81 is L or absent, and X 82 is F or absent, and X 83 is A, T, V, L or D, and (ii) The light chain variable region includes the following (a) to (c): (a) An LCDR1 sequence having the following formula: RASQX 84 IX 85 X 86 X 87 X 88 X 89 (SEQ ID NO: 261), where X 84 is R, E or D, and X 85 is G or S, and X 86 is G or S, and X 87 is Y or N, and X 88 is L or F, and X 89 is N or S, and (b) An LCDR2 sequence having the following formula: AX 90 SX 91 LDS (SEQ ID NO: 262), where X 90 is A or T, and X 91 is T or S, and (c) An LCDR3 sequence having the following formula: LQYAX 92 X 93 PX 94 T (SEQ ID NO: 263), where X 92 is S or N, and X 93 is Y or F, and X 94 is P or Y, or (6) Comprising a heavy chain variable region and a light chain variable region, wherein (i) The heavy chain variable region comprises the following (a) to (c): (a) An HCDR1 sequence having the following formula: SSYGMS (SEQ ID NO: 58), and (b) An HCDR2 sequence having the following formula: TISSGGSYTYYPDSVKG (SEQ ID NO: 59), and (c) An HCDR3 sequence having the following formula: HX 95 X 96 X 97 X 98 X 99 X 100 X 101 X 102 DY (SEQ ID NO: 264), where X 95 is G or E, X 96 is G or A, X 97 is S or R, X 98 is S or A, X 99 is S or T, X 100 is Y or F, X 101 is V or absent, X 102 is M or absent, and (ii) The light chain variable region comprises the following (a) to (c): (a) An LCDR1 sequence having the following formula: RSSKSLLHSNGNTYLY (SEQ ID NO: 63), and (b) An LCDR2 sequence having the following formula: RMSNLAS (SEQ ID NO: 64), and (c) An LCDR3 sequence having the following formula: MQHLEYPFT (SEQ ID NO: 65), or (7) Comprising a heavy chain variable region and a light chain variable region, wherein (i) The heavy chain variable region comprises the following (a) to (c): (a) An HCDR1 sequence having the following formula: TRYWMH (SEQ ID NO: 15), and (b) An HCDR2 sequence having the following formula: EFNPSNGRINYNEKFX 103 X 104 (SEQ ID NO: 265), where X 103 is K or Q, X 104 is N or G, and (c) An HCDR3 sequence having the following formula: GFAY (SEQ ID NO: 17), and (ii) The light chain variable region includes the following (a) to (c), (a) An LCDR1 sequence having the following formula: X 105 ASSTVSYIH (SEQ ID NO: 266), where X 105 is S or R, and (b) An LCDR2 sequence having the following formula: DTSKLAS (SEQ ID NO: 21), and (c) An LCDR3 sequence having the following formula: QQWTTNPWT (SEQ ID NO: 22), Disclosed is an antibody or an antigen-binding fragment thereof that binds to the GPRC5D antigen.

[0052] In another aspect, the present disclosure provides an antibody or an antigen-binding fragment thereof that binds to GPRC5D, comprising a heavy chain CDR1 selected from the amino acid sequences shown in SEQ ID NO: 5, 15, 25, 42, 58, 68, 98, 108, 119, 241 or any variants thereof, a heavy chain CDR2 selected from the amino acid sequences shown in SEQ ID NO: 6, 16, 26, 35, 43, 52, 59, 69, 78, 89, 99, 115, 133, 147 or variants thereof, a heavy chain CDR3 selected from the amino acid sequences shown in SEQ ID NO: 7, 17, 27, 44, 60, 70, 79, 90, 100, 109, 116, 120, 134, 139, 148 or any variants thereof, and a light chain CDR1 selected from the amino acid sequences shown in SEQ ID NO: 10, 20, 30, 38, 47, 63, 73, 82, 93, 103, 112, 123, 130, 167, 173, 179, 190, 208 or any variants thereof, a light chain CDR2 selected from the amino acid sequences shown in SEQ ID NO: 11, 21, 31, 48, 64, 74, 83, 94, 104, 124, 151, 209 or variants thereof, and a light chain CDR3 selected from the amino acid sequences shown in SEQ ID NO: 12, 22, 32, 39, 49, 55, 65, 75, 84, 95, 105, 125, 144 or any variants thereof.

[0053] In another aspect, the present disclosure provides an antibody or an antigen-binding fragment thereof that binds to GPRC5D, comprising a combination of heavy chain CDRs and light chain CDRs selected from the following: (1) A heavy chain CDR1, CDR2, and CDR3 sequence comprising the amino acid sequences shown in SEQ ID NO: 5, 6, 7, respectively, and a light chain CDR1, CDR2, and CDR3 sequence comprising the amino acid sequences shown in SEQ ID NO: 10, 11, 12, respectively. (2) A heavy chain CDR1, CDR2, and CDR3 sequence comprising the amino acid sequences shown in SEQ ID NO: 15, 16, 17, respectively, and a light chain CDR1, CDR2, and CDR3 sequence comprising the amino acid sequences shown in SEQ ID NO: 20, 21, 22, respectively. (3) The heavy-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 26, and 27, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 30, 31, and 32, respectively, (4) The heavy-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 5, 35, and 7, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 38, 21, and 39, respectively, (5) The heavy-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 42, 43, and 44, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 47, 48, and 49, respectively, (6) The heavy-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 5, 52, and 7, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 38, 21, and 55, respectively, (7) The heavy-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 58, 59, and 60, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 63, 64, and 65, respectively, (8) The heavy-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 68, 69, and 70, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 73, 74, and 75, respectively, (9) The heavy-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 241, 78, and 79, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 82, 83, and 84, respectively, (10) The heavy-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 42, 43, and 44, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 47, 48, and 49, respectively, (11) Heavy chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 25, 89, and 90 respectively, and light chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 93, 94, and 95 respectively, (12) Heavy chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 98, 99, and 100 respectively, and light chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 103, 104, and 105 respectively, (13) Heavy chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 108, 99, and 109 respectively, and light chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 112, 104, and 105 respectively, (14) Heavy chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 98, 115, and 116 respectively, and light chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 112, 104, and 105 respectively, (15) Heavy chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 119, 99, and 120 respectively, and light chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 123, 124, and 125 respectively, (16) Heavy chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 108, 99, and 116 respectively, and light chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 130, 104, and 105 respectively, (17) Heavy chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 108, 133, and 134 respectively, and light chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 112, 104, and 105 respectively, (18) A heavy-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 58, 59, and 139, respectively, and a light-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 63, 64, and 65, respectively. (19) A heavy-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 98, 99, and 109, respectively, and a light-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 103, 104, and 105, respectively. (20) A heavy-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 98, 99, and 109, respectively, and a light-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 103, 104, and 144, respectively. (21) A heavy-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 25, 147, and 148, respectively, and a light-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 93, 151, and 95, respectively. (22) A heavy-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 15, 164, and 17, respectively, and a light-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 167, 21, and 22, respectively. (23) A heavy-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 25, 170, and 27, respectively, and a light-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 173, 31, and 32, respectively. (24) A heavy-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 5, 176, and 7, respectively, and a light-chain CDR1, CDR2, and CDR3 sequence containing the amino acid sequences shown in SEQ ID NOs: 179, 21, and 39, respectively. (25) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 42, 182, and 44, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 47, 48, and 49, respectively, (26) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 68, 187, and 70, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 190, 74, and 75, respectively, (27) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 119, 193, and 120, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 123, 124, and 125, respectively, (28) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 170, and 27, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 208, 209, and 32, respectively, (29) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 170, and 27, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 173, 209, and 32, respectively, (30) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 170, and 27, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 208, 31, and 32, respectively.

[0054] In another aspect, the present disclosure provides an antibody or an antigen-binding fragment thereof that binds to GPRC5D, comprising a heavy chain variable region selected from the amino acid sequences shown in SEQ ID NOs: 3, 13, 23, 33, 40, 50, 56, 66, 76, 87, 96, 106, 113, 117, 126, 131, 140, 145, 162, 168, 174, 180, 185, 191, 202, 232 or any variants thereof, and a light chain variable region selected from the amino acid sequences shown in SEQ ID NOs: 8, 18, 28, 36, 45, 53, 61, 71, 80, 85, 91, 101, 110, 121, 128, 135, 142, 149, 165, 171, 177, 183, 188, 194, 206, 212, 216, 220, 230 or any variants thereof.

[0055] In a preferred embodiment, the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 3 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 8 or any variant thereof.

[0056] In a preferred embodiment, the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 13 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 18 or any variant thereof.

[0057] In a preferred embodiment, the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 23 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 28 or any variant thereof.

[0058] In a preferred embodiment, the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 33 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 36 or any variant thereof.

[0059] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 40 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 45 or any variant thereof.

[0060] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 50 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 53 or any variant thereof.

[0061] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 56 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 61 or any variant thereof.

[0062] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 66 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 71 or any variant thereof.

[0063] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 76 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 80 or any variant thereof.

[0064] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 40 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 85 or any variant thereof.

[0065] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 87 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 91 or any variant thereof.

[0066] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy-chain variable region of the amino acid sequence shown in SEQ ID NO: 96 or any variant thereof, and a light-chain variable region of the amino acid sequence shown in SEQ ID NO: 101 or any variant thereof.

[0067] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy-chain variable region of the amino acid sequence shown in SEQ ID NO: 106 or any variant thereof, and a light-chain variable region of the amino acid sequence shown in SEQ ID NO: 110 or any variant thereof.

[0068] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy-chain variable region of the amino acid sequence shown in SEQ ID NO: 113 or any variant thereof, and a light-chain variable region of the amino acid sequence shown in SEQ ID NO: 110 or any variant thereof.

[0069] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy-chain variable region of the amino acid sequence shown in SEQ ID NO: 117 or any variant thereof, and a light-chain variable region of the amino acid sequence shown in SEQ ID NO: 121 or any variant thereof.

[0070] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy-chain variable region of the amino acid sequence shown in SEQ ID NO: 126 or any variant thereof, and a light-chain variable region of the amino acid sequence shown in SEQ ID NO: 128 or any variant thereof.

[0071] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy-chain variable region of the amino acid sequence shown in SEQ ID NO: 131 or any variant thereof, and a light-chain variable region of the amino acid sequence shown in SEQ ID NO: 135 or any variant thereof.

[0072] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy-chain variable region of the amino acid sequence shown in SEQ ID NO: 137 or any variant thereof, and a light-chain variable region of the amino acid sequence shown in SEQ ID NO: 61 or any variant thereof.

[0073] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 140 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 101 or any variant thereof.

[0074] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 140 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 142 or any variant thereof.

[0075] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 145 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 149 or any variant thereof.

[0076] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 162 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 165 or any variant thereof.

[0077] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 168 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 171 or any variant thereof.

[0078] In a preferred embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 174 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 177 or any variant thereof.

[0079] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 180 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 183 or any variant thereof.

[0080] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 185 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 188 or any variant thereof.

[0081] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 191 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 194 or any variant thereof.

[0082] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 206 or any variant thereof.

[0083] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 212 or any variant thereof.

[0084] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 216 or any variant thereof.

[0085] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 220 or any variant thereof.

[0086] In a preferred embodiment, the antibody or its antigen-binding fragment comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 232 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 230 or any variant thereof.

[0087] In a preferred embodiment, the antibody or its antigen-binding fragment is humanized.

[0088] In another aspect, the present disclosure provides a nucleic acid encoding the aforementioned antibody or its antigen-binding fragment.

[0089] In a preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleic acid sequence selected from the nucleotide sequences shown in SEQ ID NOs: 4, 14, 24, 34, 41, 51, 57, 67, 77, 88, 97, 107, 114, 118, 127, 132, 138, 141, 146, 163, 169, 175, 181, 186, 192, 203 or any variants thereof, and a light chain variable region nucleic acid sequence selected from the nucleotide sequences shown in SEQ ID NOs: 9, 19, 29, 37, 46, 54, 62, 72, 81, 86, 92, 102, 111, 122, 129, 136, 143, 150, 166, 172, 178, 184, 189, 195, 207, 213, 217, 221 or any variants thereof.

[0090] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 4 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 9 or any variant thereof.

[0091] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 14 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 19 or any variant thereof.

[0092] In a more preferred embodiment, the nucleic acid comprises a nucleotide sequence of a heavy chain variable region of the nucleotide sequence shown in SEQ ID NO: 24 or any variant thereof, and a nucleotide sequence of a light chain variable region of the nucleotide sequence shown in SEQ ID NO: 29 or any variant thereof.

[0093] In a more preferred embodiment, the nucleic acid comprises a nucleotide sequence of a heavy chain variable region of the nucleotide sequence shown in SEQ ID NO: 34 or any variant thereof, and a nucleotide sequence of a light chain variable region of the nucleotide sequence shown in SEQ ID NO: 37 or any variant thereof.

[0094] In a more preferred embodiment, the nucleic acid comprises a nucleotide sequence of a heavy chain variable region of the nucleotide sequence shown in SEQ ID NO: 41 or any variant thereof, and a nucleotide sequence of a light chain variable region of the nucleotide sequence shown in SEQ ID NO: 46 or any variant thereof.

[0095] In a more preferred embodiment, the nucleic acid comprises a nucleotide sequence of a heavy chain variable region of the nucleotide sequence shown in SEQ ID NO: 51 or any variant thereof, and a nucleotide sequence of a light chain variable region of the nucleotide sequence shown in SEQ ID NO: 54 or any variant thereof.

[0096] In a more preferred embodiment, the nucleic acid comprises a nucleotide sequence of a heavy chain variable region of the nucleotide sequence shown in SEQ ID NO: 57 or any variant thereof, and a nucleotide sequence of a light chain variable region of the nucleotide sequence shown in SEQ ID NO: 62 or any variant thereof.

[0097] In a more preferred embodiment, the nucleic acid comprises a nucleotide sequence of a heavy chain variable region of the nucleotide sequence shown in SEQ ID NO: 67 or any variant thereof, and a nucleotide sequence of a light chain variable region of the nucleotide sequence shown in SEQ ID NO: 72 or any variant thereof.

[0098] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 77 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 81 or any variant thereof.

[0099] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 41 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 86 or any variant thereof.

[0100] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 88 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 92 or any variant thereof.

[0101] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 97 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 102 or any variant thereof.

[0102] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 107 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 111 or any variant thereof.

[0103] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 114 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 111 or any variant thereof.

[0104] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 118 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 122 or any variant thereof.

[0105] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 127 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 129 or any variant thereof.

[0106] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 132 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 136 or any variant thereof.

[0107] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 138 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 62 or any variant thereof.

[0108] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 141 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 102 or any variant thereof.

[0109] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 141 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 143 or any variant thereof.

[0110] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 146 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 150 or any variant thereof.

[0111] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 163 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 166 or any variant thereof.

[0112] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 169 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 172 or any variant thereof.

[0113] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 175 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 178 or any variant thereof.

[0114] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 181 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 184 or any variant thereof.

[0115] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 186 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 189 or any variant thereof.

[0116] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 192 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 195 or any variant thereof.

[0117] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 203 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 207 or any variant thereof.

[0118] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 203 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 213 or any variant thereof.

[0119] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 203 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 217 or any variant thereof.

[0120] In a more preferred embodiment, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 203 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 221 or any variant thereof.

[0121] In another aspect, the present disclosure provides a multispecific antibody or an antigen-binding fragment thereof comprising at least (a) an antibody or an antigen-binding fragment thereof that binds to the GPRC5D antigen according to any one of claims 1 to 4, and (b) an antibody or an antigen-binding fragment thereof that binds to the CD3 antigen. (a) The antibody or antigen-binding fragment thereof that binds to the GPRC5D antigen according to any one of claims 1 to 4 is the first antigen-binding portion, the GPRC5D antigen is the first antigen, the first antigen-binding portion includes a first heavy-chain variable region and a first light-chain variable region, and the first antigen-binding portion includes a first binding structural domain that binds to the first antigen.

[0122] In a preferred embodiment, the first binding structural domain includes a first heavy-chain CDR selected from the amino acid sequences shown in SEQ ID NO: 25, 170, 27 or any variant thereof, and a first light-chain CDR selected from the amino acid sequences shown in SEQ ID NO: 31, 32, 173, 208, 209 or any variant thereof, and (b) The antibody or antigen-binding fragment thereof that binds to the CD3 antigen is the second antigen-binding portion, the CD3 antigen is the second antigen, the second antigen-binding portion includes a second heavy-chain variable region and a second light-chain variable region, and the second antigen-binding portion includes a second binding structural domain that binds to the second antigen, wherein the second binding structural domain includes a second heavy-chain CDR1 selected from the amino acid sequence shown in SEQ ID NO: 159 or a variant thereof, a second heavy-chain CDR2 selected from the amino acid sequence shown in SEQ ID NO: 160 or a variant thereof, a second heavy-chain CDR3 selected from the amino acid sequence shown in SEQ ID NO: 161 or a variant thereof, and a second light-chain CDR1 selected from the amino acid sequence shown in SEQ ID NO: 154 or any variant thereof, a second light-chain CDR2 selected from the amino acid sequence shown in SEQ ID NO: 155 or any variant thereof, and a second light-chain CDR3 selected from the amino acid sequence shown in SEQ ID NO: 156 or any variant thereof.

[0123] In a preferred embodiment, the second binding structural domain includes a second heavy-chain variable region selected from the amino acid sequence shown in SEQ ID NO: 157 or any variant thereof, and a second light-chain variable region selected from the amino acid sequence shown in SEQ ID NO: 152 or any variant thereof.

[0124] In a preferred embodiment, the second heavy chain of the second antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 222, and the second light chain of the second antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 224.

[0125] In a preferred embodiment, the first binding structure domain comprises a first heavy chain CDR1 selected from the amino acid sequence shown in SEQ ID NO: 25 or any variant thereof, a first heavy chain CDR2 selected from the amino acid sequence shown in SEQ ID NO: 170 or any variant thereof, a first heavy chain CDR3 selected from the amino acid sequence shown in SEQ ID NO: 27 or any variant thereof, and a first light chain CDR1 selected from the amino acid sequence shown in SEQ ID NOs: 173, 208 or any variant thereof, a first light chain CDR2 selected from the amino acid sequence shown in SEQ ID NOs: 31, 209 or any variant thereof, and a first light chain CDR3 selected from the amino acid sequence shown in SEQ ID NO: 32 or any variant thereof.

[0126] In a preferred embodiment, the first light chain CDRs of the first binding structure domain are selected from the first light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 173, 31, and 32, respectively, the first light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 208, 209, and 32, respectively, the first light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 173, 209, and 32, respectively, and the first light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 208, 31, and 32, respectively, and the first heavy chain CDRs of the first binding structure domain are selected from the first heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 170, and 27, respectively.

[0127] In a preferred embodiment, the first binding structure domain comprises a first heavy chain variable region selected from the amino acid sequences shown in SEQ ID NOs: 168, 202, 232 or any variants thereof, and a first light chain variable region selected from the amino acid sequences shown in SEQ ID NOs: 171, 206, 212, 216, 220, 230 or any variants thereof.

[0128] In a preferred embodiment, the first binding structure domain comprises a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 168 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 171 or any variant thereof.

[0129] In a preferred embodiment, the first binding structure domain comprises a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 206 or any variant thereof.

[0130] In a preferred embodiment, the first binding structure domain comprises a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 212 or any variant thereof.

[0131] In a preferred embodiment, the first binding structure domain comprises a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 216 or any variant thereof.

[0132] In a preferred embodiment, the first binding structure domain comprises a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 220 or any variant thereof.

[0133] In a preferred embodiment, the first binding structure domain comprises a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 232 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 230 or any variant thereof.

[0134] In a preferred embodiment, the first heavy chain of the first antigen-binding portion is selected from the amino acid sequences shown in SEQ ID NOs: 196 and 200, and the first light chain of the first antigen-binding portion is selected from the amino acid sequences shown in SEQ ID NOs: 198, 204, 210, 214 and 218. More preferably, the first heavy chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 196, and the first light chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 198. More preferably, the first heavy chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 200, and the first light chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 204. More preferably, the first heavy chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 200, and the first light chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 210. More preferably, the first heavy chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 200, and the first light chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 214. More preferably, the first heavy chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 200, and the first light chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 218.

[0135] In a more preferred embodiment, the multispecific antibody or antigen-binding fragment thereof further comprises (c) an antibody or antigen-binding fragment thereof that binds to the BCMA antigen: (c) An antibody or antigen-binding fragment thereof that binds to the BCMA antigen is the third antigen-binding portion, the BCMA antigen is the third antigen, the third antigen-binding portion includes a third heavy-chain variable region and a third light-chain variable region, and the third antigen-binding portion includes a third binding structural domain that binds to the third antigen.

[0136] In a preferred embodiment, the third binding structural domain includes a third heavy-chain variable region selected from the amino acid sequence shown in SEQ ID NO: 268 or any variant thereof, and a third light-chain variable region selected from the amino acid sequence shown in SEQ ID NO: 267 or any variant thereof.

[0137] In a preferred embodiment, the third antigen-binding portion includes a third heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 233 or any variant thereof, and a third light chain consisting of the amino acid sequence shown in SEQ ID NO: 226 or any variant thereof.

[0138] In a preferred embodiment, the multispecific antibody is a trispecific antibody, wherein any two of the first heavy-chain variable region, the second heavy-chain variable region, and the third heavy-chain variable region are linked to a constant region, and the remaining heavy-chain variable region is not linked to a constant region, and the heavy-chain variable region not linked to a constant region is linked to any one of the two heavy-chain variable regions linked to the constant region, preferably linked by a linker, and preferably, the heavy-chain variable region not linked to the constant region is linked by a linker to its corresponding light-chain variable region.

[0139] In a preferred embodiment, the first heavy chain variable region or the first light chain variable region of the first antigen-binding portion is linked by a linker to the second heavy chain variable region or the second light chain variable region of the second antigen-binding portion. Preferably, the first heavy chain variable region and the first light chain variable region of the first antigen-binding portion are linked by a linker. More preferably, the first light chain variable region of the first antigen-binding portion is linked to the second heavy chain variable region of the second antigen-binding portion.

[0140] In a preferred embodiment, the first heavy chain variable region or the first light chain variable region of the first antigen-binding portion is linked by a linker to the third heavy chain variable region or the third light chain variable region of the third antigen-binding portion. Preferably, the first heavy chain variable region and the first light chain variable region of the first antigen-binding portion are linked to each other by a linker.

[0141] In a preferred embodiment, the linker is a flexible linker.

[0142] In a preferred embodiment, the linker is (GGGGS) n , where n = 2 - 5. For example, GGGGSGGGGSGGGGS (SEQ ID NO: 231), GSTSGSGKSSEGKG (SEQ ID NO: 273), GSGGGSG (SEQ ID NO: 274), GGGSGGSG (SEQ ID NO: 275).

[0143] In a more preferred embodiment, the trispecific antibody comprises light chain 1, heavy chain 1, light chain 2, and heavy chain 2.

[0144] In a more preferred embodiment, the light chain 1 includes the third light chain variable region of the amino acid sequence shown in SEQ ID NO: 267, the heavy chain 1 includes the first light chain variable region of the amino acid sequence shown in SEQ ID NO: 230, the first heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 232, and the third heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 268, the light chain 2 includes the second light chain variable region of the amino acid sequence shown in SEQ ID NO: 152, the heavy chain 2 includes the second heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 157. More preferably, the light chain 1 includes the amino acid sequence shown in SEQ ID NO: 226, the heavy chain 1 includes the amino acid sequence shown in SEQ ID NO: 228, the light chain 2 includes the amino acid sequence shown in SEQ ID NO: 224, and the heavy chain 2 includes the amino acid sequence shown in SEQ ID NO: 222.

[0145] In a more preferred embodiment, the light chain 1 includes the third light chain variable region of the amino acid sequence shown in SEQ ID NO: 267, the heavy chain 1 includes the first heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 232, the first light chain variable region of the amino acid sequence shown in SEQ ID NO: 230, and the third heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 268, the light chain 2 includes the second light chain variable region of the amino acid sequence shown in SEQ ID NO: 152, the heavy chain 2 includes the second heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 157. More preferably, the light chain 1 includes the amino acid sequence shown in SEQ ID NO: 226, the heavy chain 1 includes the amino acid sequence shown in SEQ ID NO: 234, the light chain 2 includes the amino acid sequence shown in SEQ ID NO: 224, and the heavy chain 2 includes the amino acid sequence shown in SEQ ID NO: 222.

[0146] In a more preferred embodiment, the light chain 1 comprises the second light chain variable region of the amino acid sequence shown in SEQ ID NO: 152, the heavy chain 1 comprises the first light chain variable region of the amino acid sequence shown in SEQ ID NO: 230, the first heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 232, and the second heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 157, the light chain 2 comprises the third light chain variable region of the amino acid sequence shown in SEQ ID NO: 267, the heavy chain 2 comprises the third heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 268, and more preferably, the light chain 1 comprises the amino acid sequence shown in SEQ ID NO: 224, the heavy chain 1 comprises the amino acid sequence shown in SEQ ID NO: 236, the light chain 2 comprises the amino acid sequence shown in SEQ ID NO: 226, and the heavy chain 2 comprises the amino acid sequence shown in SEQ ID NO: 233.

[0147] In a more preferred embodiment, the light chain 1 comprises the second light chain variable region of the amino acid sequence shown in SEQ ID NO: 152, the heavy chain 1 comprises the first heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 232, the first light chain variable region of the amino acid sequence shown in SEQ ID NO: 230, and the second heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 157, the light chain 2 comprises the third light chain variable region of the amino acid sequence shown in SEQ ID NO: 267, the heavy chain 2 comprises the third heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 268, and more preferably, the light chain 1 comprises the amino acid sequence shown in SEQ ID NO: 224, the heavy chain 1 comprises the amino acid sequence shown in SEQ ID NO: 239, the light chain 2 comprises the amino acid sequence shown in SEQ ID NO: 226, and the heavy chain 2 comprises the amino acid sequence shown in SEQ ID NO: 233.

[0148] In another aspect, the present disclosure provides a nucleic acid encoding the aforementioned multispecific antibody or an antigen-binding fragment thereof.

[0149] In a preferred embodiment, the multispecific antibody is a bispecific antibody. The nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody is selected from the nucleotide sequences shown in SEQ ID NO: 169 and SEQ ID NO: 203, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion is selected from the nucleotide sequences shown in SEQ ID NO: 172, SEQ ID NO: 207, SEQ ID NO: 213, SEQ ID NO: 217, and SEQ ID NO: 221. The nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO: 153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody is selected from the nucleotide sequences shown in SEQ ID NO: 197 and SEQ ID NO: 201, and the nucleic acid encoding the first light chain of the first antigen-binding portion is selected from the nucleotide sequences shown in SEQ ID NO: 199, SEQ ID NO: 205, SEQ ID NO: 211, SEQ ID NO: 215, and SEQ ID NO: 219. The nucleic acid encoding the second heavy chain of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO: 225.

[0150] In a more preferred embodiment, the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 169, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 172. The nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 197, and the nucleic acid encoding the first light chain of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 199. The nucleic acid encoding the second heavy chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 225.

[0151] In a more preferred embodiment, the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 203, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 207. The nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 201, and the nucleic acid encoding the first light chain of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 205. The nucleic acid encoding the second heavy chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 225.

[0152] In a more preferred embodiment, the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 203, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 213. The nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 201, the nucleic acid encoding the first light chain of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 211, the nucleic acid encoding the second heavy chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 225.

[0153] In a more preferred embodiment, the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 203, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 217, the nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 201, the nucleic acid encoding the first light chain of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 215, the nucleic acid encoding the second heavy chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 225.

[0154] In a more preferred embodiment, the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 203, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 221, the nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 201, the nucleic acid encoding the first light chain of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 219, the nucleic acid encoding the second heavy chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 225.

[0155] In a preferred embodiment, the multispecific antibody is a trispecific antibody, and the trispecific antibody comprises nucleic acids encoding a light chain 1, a heavy chain 1, a light chain 2, and a heavy chain 2.

[0156] In a more preferred embodiment, the nucleic acid encoding the light chain 1 comprises the nucleotide sequence shown in SEQ ID NO: 269, the nucleic acid encoding the heavy chain 1 comprises the nucleotide sequences shown in SEQ ID NO: 271, SEQ ID NO: 272, and SEQ ID NO: 270, the nucleic acid encoding the light chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 153, the nucleic acid encoding the heavy chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 158, and more preferably, the nucleic acid encoding the light chain 1 of the trispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 227, the nucleic acid encoding the heavy chain 1 comprises the nucleotide sequence shown in SEQ ID NO: 229, the nucleic acid encoding the light chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 225, and the nucleic acid encoding the heavy chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 223.

[0157] In a more preferred embodiment, the nucleic acid encoding the light chain 1 comprises the nucleotide sequence shown in SEQ ID NO: 269, the nucleic acid encoding the heavy chain 1 comprises the nucleotide sequences shown in SEQ ID NO: 272, SEQ ID NO: 271, and SEQ ID NO: 270, the nucleic acid encoding the light chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 153, the nucleic acid encoding the heavy chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 158, and more preferably, the nucleic acid encoding the light chain 1 of the multispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 227, the nucleic acid encoding the heavy chain 1 comprises the nucleotide sequence shown in SEQ ID NO: 235, the nucleic acid encoding the light chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 225, and the nucleic acid encoding the heavy chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 223.

[0158] In a more preferred embodiment, the nucleic acid encoding the light chain 1 comprises the nucleotide sequence shown in SEQ ID NO: 153, the nucleic acid encoding the heavy chain 1 comprises the nucleotide sequences shown in SEQ ID NO: 271, SEQ ID NO: 272, and SEQ ID NO: 158, the nucleic acid encoding the light chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 269, the nucleic acid encoding the heavy chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 270, and more preferably, the nucleic acid encoding the light chain 1 of the multispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 225, the nucleic acid encoding the heavy chain 1 comprises the nucleotide sequence shown in SEQ ID NO: 237, the nucleic acid encoding the light chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 227, and the nucleic acid encoding the heavy chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 238.

[0159] In a more preferred embodiment, the nucleic acid encoding the light chain 1 comprises the nucleotide sequence shown in SEQ ID NO: 153, the nucleic acid encoding the heavy chain 1 comprises the nucleotide sequences shown in SEQ ID NO: 272, SEQ ID NO: 271, and SEQ ID NO: 158, the nucleic acid encoding the light chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 269, the nucleic acid encoding the heavy chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 270, and more preferably, the nucleic acid encoding the light chain 1 of the multispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 225, the nucleic acid encoding the heavy chain 1 comprises the nucleotide sequence shown in SEQ ID NO: 240, the nucleic acid encoding the light chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 227, and the nucleic acid encoding the heavy chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 238.

[0160] In another aspect, the present disclosure provides a vector comprising the aforementioned nucleic acid.

[0161] In another aspect, the present disclosure provides a cell comprising the nucleic acid or the vector.

[0162] In another aspect, the present disclosure provides a composition comprising the GPRC5D antibody or antigen-binding fragment thereof described in the foregoing claims, the aforementioned multispecific antibody or antigen-binding fragment thereof, the aforementioned nucleic acid, the aforementioned vector, and / or the aforementioned cell.

[0163] In another aspect, the present disclosure provides a kit comprising the GPRC5D antibody or antigen-binding portion thereof described above, the aforementioned multispecific antibody or antigen-binding portion thereof, the aforementioned nucleic acid, the aforementioned vector, the aforementioned cell, and / or the aforementioned composition.

[0164] In another aspect, the present disclosure provides the use of the GPRC5D antibody or antigen-binding portion thereof described above, the aforementioned multispecific antibody or antigen-binding portion thereof, the aforementioned nucleic acid, the aforementioned vector, the aforementioned cell, and / or the aforementioned composition in the preparation of a medicament or kit for use in the diagnosis, treatment, or prevention of cancer or autoimmune diseases.

[0165] In a preferred embodiment, the cancer is selected from multiple myeloma, malignant plasma cell tumor, Hodgkin lymphoma, nodular lymphocyte-predominant Hodgkin lymphoma, Kahlers disease, and myeloid leukemia, plasma cell leukemia, plasmacytoma, B-cell prolymphocytic leukemia, hairy cell leukemia, B-cell non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL), chronic myeloid leukemia (CML), follicular lymphoma, Burkitt lymphoma, marginal zone lymphoma, mantle cell lymphoma, large cell lymphoma, precursor B-lymphoblastic lymphoma, myeloid leukemia, Waldenström macroglobulinemia, diffuse large B-cell lymphoma, follicular lymphoma, marginal zone lymphoma, mucosa-associated lymphoid tissue lymphoma, small cell lymphocytic lymphoma, mantle cell lymphoma, Burkitt lymphoma, primary mediastinal (thymic) large B-cell lymphoma, lymphoplasmacytic lymphoma, Waldenström macroglobulinemia, nodal marginal zone B-cell lymphoma, splenic marginal zone lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, lymphomatoid granulomatosis, T-cell / histiocyte-rich large B-cell lymphoma, primary central nervous system lymphoma, primary cutaneous diffuse large B-cell lymphoma (lower extremity type), EBV-positive diffuse large B-cell lymphoma in the elderly, inflammation-associated diffuse large B-cell lymphoma, intravascular large B-cell lymphoma, ALK-positive large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma occurring in HHV8-associated multicentric Castleman disease, unclassified B-cell lymphoma having intermediate characteristics between diffuse large B-cell lymphoma and Burkitt lymphoma, unclassified B-cell lymphoma having intermediate characteristics between diffuse large B-cell lymphoma and classical Hodgkin lymphoma, and other B-cell-related lymphomas, which is a B-cell-related cancer.

[0166] In a preferred embodiment, the autoimmune disease is selected from immune kidney disease, systemic lupus erythematosus or rheumatoid arthritis.

[0167] In one embodiment of the present disclosure, the antibody is administered in a single dose or multiple doses.

[0168] When the antibody is administered in multiple doses according to the present disclosure, preferably, the antibody is administered in at least 3 doses, at least 4 doses, at least 5 doses, at least 6 doses, at least 7 doses, at least 8 doses, at least 9 doses, or at least 10 doses, preferably at most 30 doses, 25 doses, 20 doses, 15 doses, or 10 doses. Preferably, the doses of the antibody are administered at intervals of at least 7 days, at least 10 days, at least 14 days or at least 20 days. Preferably, the doses of the antibody are administered at intervals of 7 to 30 days, 10 to 20 days, preferably about 14 days.

[0169] In one aspect, the present disclosure provides the use of a therapeutic agent in the preparation of a medicament for the treatment of a cancer-related disease in a subject, wherein the therapeutic agent is selected from a single therapeutic agent or a combination therapeutic agent.

[0170] In some embodiments, the single-agent therapeutic agent is selected from the aforementioned antibody or antigen-binding portion thereof, nucleic acid, vector, cell, and / or composition.

[0171] In one aspect, the present disclosure provides a kit comprising the aforementioned antibody or antigen-binding portion thereof, nucleic acid molecule, vector, cell, and / or composition.

[0172] The kits described herein include the antibodies of the present disclosure, fragments thereof, homologs, derivatives thereof, etc., such as conjugates having labels or cytotoxicity, instruction manuals for the antibodies, conjugates that kill specific types of cells, etc. The instruction manuals include instructions for using the antibodies, conjugates, etc. in vitro, in vivo, or ex vivo. The antibodies may be in liquid or solid form, and are usually lyophilized. The kits can include other suitable reagents, such as buffers, reconstitution solutions, and other components necessary for the intended use. Combinations of reagents packaged in predetermined amounts and their instruction manuals are contemplated, and the use is, for example, for performing therapeutic uses or diagnostic assays. When the antibody is labeled, for example, labeled with an enzyme, the kit includes substrates and cofactors necessary for the enzyme (e.g., to provide a substrate precursor for detecting a chromophore or fluorophore). Additionally, other additives such as stabilizers, buffers (e.g., blocking buffers, or lysis buffers) may be included. The relative amounts of the various reagents can be varied to provide concentrates of the reagent solutions, which provides flexibility to the user, space savings, reagent savings, etc. These reagents can also be supplied in the form of dry powders, usually lyophilized, containing excipients, and upon dissolution, provide reagent solutions at appropriate concentrations.

[0173] In one aspect, the present disclosure provides the use of the antibody or its antigen-binding portion, nucleic acid molecule, vector, cell, and / or resistant composition in the preparation of a medicament or kit for use in the diagnosis, treatment, or prevention of cancer-related diseases.

[0174] Furthermore, the antibodies of the present disclosure can be used in immunoassays, purification methods, and other methods using immunoglobulins or fragments thereof. Such uses are well known in the art.

[0175] Accordingly, the present disclosure also provides a composition comprising the anti-GPRC5D antibody of the present disclosure or a fragment thereof, which is conveniently combined with a pharmaceutically acceptable carrier, diluent, or excipient, which is a conventional means in the art.

[0176] As used herein, the term "pharmaceutical composition" refers to formulations of a variety of preparations. It is in the form of a sterile liquid solution, a liquid suspension or a lyophilized form, optionally containing stabilizers or excipients.

[0177] It should be understood that the therapeutic agents according to the described embodiments are administered in a pharmaceutically acceptable suitable carrier, excipient, and formulation together with other reagents that improve properties such as mobility, delivery, and tolerance. Many suitable formulations are described in the pharmacopoeia known to all pharmaceutical chemists: Remington’s Pharmaceutical Sciences (15th Edition, Mack Publishing Company, Easton, Pa. (1975)), particularly in Chapter 87 by Blaug, Seymour. These formulations include, for example, pastes, plasters, gels, waxes, oils, lipids, lipid-containing (cationic or anionic) carriers (e.g., Lipofectin TM ), DNA conjugates, dry powders for inhalation, water-in-oil and oil-in-water emulsions, emulsions of polyglycols (polyglycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing polyglycols. Provided that the active ingredient in the formulation is not inactivated by the formulation, the formulation is physiologically compatible, and the formulation is resistant to the route of administration, any of the aforementioned mixtures is suitable for use in the treatment or therapy according to the present disclosure.

[0178] For inhaled administration, the compound is delivered as an aerosol spray from a pressurized container, dispenser, or nebulizer containing a gas such as carbon dioxide, which is a suitable propellant.

[0179] Alternatively, it can also be administered systemically by transmucosal or transdermal means. In the case of transmucosal or transdermal administration, a penetration enhancer suitable for penetrating the barrier is used in the formulation. Such penetration enhancers are generally known in the art and include detergents, bile salts, and fusidic acid derivatives used for transmucosal administration. Transmucosal administration can be achieved by the use of nasal sprays or suppositories. In the case of transdermal administration, one or more of the antibodies can be formulated as an ointment, a cream, a gel, or a lotion, as is generally known in the art.

[0180] The compound can also be formulated for rectal delivery in the form of a suppository (having a conventional suppository base such as cocoa butter or other glycerides) or a retention enema.

[0181] In one embodiment, the antibody can be formulated with a carrier that prevents rapid elimination from the body, such as a sustained-release / controlled-release formulation including, for example, implants and microencapsulation delivery systems. For example, biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyglycollate, and polylactic acid can be used. Methods for preparing such formulations will be apparent to those skilled in the art.

[0182] In particular, formulating the parenteral composition in dosage units is advantageous for ease of administration and consistency of dosage. As used herein, the term dosage unit form refers to a physically discrete unit suitable as a unit dose to be administered to a subject to be treated. Each unit contains a predetermined quantity of one or more of the antibodies calculated to be combined with the desired pharmaceutical carrier to produce the desired therapeutic effect. The specifications for the dosage unit forms of the described embodiments are dictated by and directly dependent on the unique characteristics of the antibody and the specific therapeutic effect to be achieved, as well as the limitations inherent in the art of formulating such antibodies for use in treating individuals.

[0183] The pharmaceutical composition can be placed in a container, a package, or a dispenser together with instructions for administration.

[0184] The formulations described herein may also include one or more of the foregoing antibodies, preferably antibodies having complementary activities that do not adversely affect each other, depending on the specific condition being treated. Alternatively or in addition to these, the composition may include agents that enhance their functions, such as, for example, cytotoxic agents, cytokines, chemotherapeutic agents, or growth inhibitors. Such molecules are present in an effective amount for the intended purpose in a suitable combination. For example, they can be presented in combination in a kit or presented in combination at the time of use.

[0185] In one embodiment, one or more of the antibodies can be administered in combination therapy, i.e., in combination with other reagents such as therapeutic agents (which can be used to treat pathological conditions or disorders such as various forms of cancer and inflammatory diseases). As used herein, the term "in combination" means substantially simultaneous, parallel, or sequential administration of the reagents. When administered sequentially, at the time of initiation of administration of the second compound, the first of the two compounds is preferably detected at an effective concentration at the site of treatment. In one embodiment, "in combination" can also mean including both the antibodies of the present disclosure and other therapeutic agents in a kit.

[0186] For example, combination therapy can include co-formulation and / or co-administration of one or more of the antibodies described herein with one or more additional therapeutic agents (e.g., one or more cytokines and growth factor inhibitors, immunosuppressive agents, anti-inflammatory agents, metabolic inhibitors, enzyme inhibitors, and / or cytotoxins or cell growth inhibitors, described in more detail below). Such combination therapy can advantageously reduce the dosage of the therapeutic agents administered, thereby avoiding toxicities or complications that may be associated with various monotherapy regimens.

[0187] The anti-GPRC5D antibodies provided by the present disclosure bind to the human GPRC5D receptor with high affinity, while experimentally proven to bind to cynomolgus GPRC5D with an affinity comparable to that of the human GPRC5D receptor. The novel humanized GPRC5D×CD3 bispecific antibody constructed using the anti-GPRC5D antibody provided by the present disclosure has a strong killing effect on low-expressing cells, is milder to T cell activation, and in a mouse transplanted tumor model, the GPRC5D×BCMA×CD3 trispecific antibody is BCMA + cells and GPRC5D + cells, and is significantly superior to the BCMA×CD3 or GPRC5D×CD3 bispecific antibody in killing myeloma cells, helps to overcome the resistance caused by a single target, and brings about a more profound and long-lasting tumor inhibitory effect in clinical applications.

[0188] For clarity and brevity, some features are described herein as part of the same or separate embodiments, but it will be understood that the scope of the present disclosure may include numerous embodiments having all or a combination of some of the described features.

[0189] Examples Example 1 Production of Anti-GPRC5D Antibody (1) Construction of GPRC5D Stable Transfection Cell Line Using human GPRC5D cDNA (manufactured by Sino Biological, product number: HG24447-UT, the amino acid sequence of which is shown in SEQ ID NO: 1) and cynomolgus monkey GPRC5D cDNA (synthesized by Suzhou Genechem Co., Ltd., the amino acid sequence of which is shown in SEQ ID NO: 2) as templates respectively, lentiviral vector plasmids containing the full-length sequences of human and monkey GPRC5D were constructed. The constructed lentiviral plasmids and packaging plasmids were co-transfected into HEK293T cells (ATCC CRL-11268) according to the instructions of the lentiviral packaging kit (Lenti-Pac HIV Expression Packaging Kit, Gene Copoeia, product number: HPK-LvTR-20) to perform lentiviral packaging. 48 hours after transfection, the medium was collected, centrifuged at 500*g for 10 minutes to remove cell debris, and the culture supernatant containing lentiviral particles was obtained. After filtering this with a 0.45 μm PES membrane, it was dispensed into 1.5 mL EP tubes, 10 μL was taken from each, and used for infection of 1×10 6 HEK293T cells or CHO cells (ATCC CCL-61). 48 hours later, 8 μg / ml puromycin was added for pressure selection to screen for stably transfected cells, and monoclonal cell lines HEK293-hGPRC5D (clone 2H3), HEK293-cyGPRC5D (clone 1F11), CHO-hGPRC5D (clone 2D10), and CHO-cyGPRC5D (clone 3E7) that highly express human or cynomolgus monkey GPRC5D were obtained. Hereinafter, they are abbreviated as HEK293-hGPRC5D, HEK293-cyGPRC5D, CHO-hGPRC5D, and CHO-cyGPRC5D respectively, and the results are shown in FIGS. 1 and 2.

[0190] The amino acid sequence of human GPRC5D is shown in SEQ ID NO: 1, [Chemical formula] The amino acid sequence of cynomolgus monkey GPRC5D is shown in SEQ ID NO: 2. [Chemical formula]

[0191] (2) Immunization of Animals and Preparation of Hybridomas Female Balb / C mice at 4 - 6 weeks of age were selected. A eukaryotic expression plasmid containing the CMV promoter was constructed using the full - length sequences of human or cynomolgus monkey GPRC5D, and the GPRC5D DNA of human and cynomolgus monkey was alternately used to immunize mice (20 μg of plasmid per mouse) using a gene gun. After 4 - 5 immunizations, mice showing a serum titer meeting the requirements were shock - immunized with CHO - hGPRC5D stable transfection cells (5×10 6 cells / mouse). Three days later, the mice were sacrificed by cervical dislocation, and the spleens and some peripheral lymph nodes of the mice were collected, ground in DMEM medium to obtain a suspension rich in B cells. An appropriate amount of the suspension of lymph node and spleen cells was taken, mixed with SP2 / 0, and cell fusion was carried out using an electrofusion apparatus (BTX). After electrofusion, the cells were plated using ClonaCell (registered trademark) - HY Medium D (manufactured by STEMCELL Technologies), and IgG + positive monoclonal clones were selected and transplanted into 96 - well plates. After 2 - 3 days of culture, the supernatant was taken, and hybridoma clones that bind to HEK293 - hGPRC5D and HEK293 - cyGPRC5D cells were screened.

[0192] Example 2 Screening of Hybridoma Clones That Bind to Human and Cynomolgus Monkey GPRC5D 5×10 in each well of a 96 - well U - shaped plate 4Add individual HEK293-hGPRC5D cells or HEK293-cyGPRC5D cells, centrifuge to discard the supernatant, add 100 μL / well of hybridoma culture supernatant, and incubate on ice for 60 minutes. After washing away the unbound primary antibody, add 50 μL of secondary antibody solution (Anti-mouse IgG, Fc specific-AF647, Jackson Immuno Research) to each well and incubate on ice for 30 minutes. Wash away the excess secondary antibody, add PBS solution to resuspend the cells, and detect the binding between the antibody and the cells using a flow cytometer (Intellicyte iQue). In the first screening, a total of 290 hybridoma clones specifically binding to hGPRC5D and cyGPRC5D stable transfection cells were obtained. After concentrating and purifying the supernatant of the above positive clones, GPRC5D + Clones that bind to tumor cells were further screened, and among them, 77 clones showed high binding signals to myeloma cells MM.1S and MM.1R (both purchased from ATCC, catalog numbers CRL-2974 and CRL-2975, respectively).

[0193] Extract RNA from hybridoma candidate monoclonal cells by the TRNzol lysis method, reverse transcribe to synthesize single-stranded cDNA, use this as a template for amplification, and obtain the nucleotide sequence of the antibody variable region in the hybridoma monoclonal cells. Clone the light chain variable region of the above antibody into a vector containing the human light chain constant region, clone the heavy chain variable region into a vector containing the human heavy chain constant region, co-transfect the plasmid into CHO-S cells, secrete intact IgG antibody into the medium by expression, express at 37 °C, 5% CO2, 125 rpm for 7 days, then collect the supernatant, purify by Protein A chromatography to obtain chimeric IgG, and measure the IgG concentration with a spectrophotometer. The control antibody GC5B596 was synthesized according to Patent Document US10562968B2, expressed in CHO cells, and then purified with Protein A.

[0194] The binding of the above antibodies to human and cynomolgus monkey GPRC5D stable transfection cells was detected by flow cytometry. The CHO-hGPRC5D and CHO-cyGPRC5D stable transfection cells prepared in Example 1 were taken, and the concentration of the cell suspension was adjusted to 5×10 5 cells / ml. 100 μl / well of the cell suspension was added to a 96-well U-bottom plate, centrifuged at 300 g for 5 minutes, the supernatant was discarded, 100 μL of gradient-diluted antibody (starting concentration 200 nM, 3-fold dilution, 12 gradients) was added to each well, and incubated at 4°C for 45 minutes. The secondary antibody, Alexa Fluro647-labeled goat anti-human IgG Fc (1:300 dilution), was added at 50 μL / well, incubated on ice for 45 minutes, and detected by flow cytometry.

[0195] Figure 3 is a diagram showing the binding of the anti-GPRC5D chimeric antibody to the GPRC5D stable transfection cell line. Among them, Figure 3A shows the detection results by flow cytometry of the chimeric antibody and human GPRC5D stable transfection cells. The binding activity of all chimeric antibodies was significantly superior to that of GC5B596. Figure 3B shows the detection results by flow cytometry of the chimeric antibody and cynomolgus monkey GPRC5D stable transfection cells. Among them, clones 2E1, 2F1, 4B1, 4D3, 8D1, 10B2, 10E8, 15D1, and 19E2, etc., had stronger binding ability. GC5B596 weakly bound to cynomolgus monkey GPRC5D stable transfection cells only at the highest concentration of 200 nM, and the results are shown in Table 1.

Table 2

[0196] The variable region sequences of the above anti-GPRC5D monoclonal antibodies are as follows.

Table 3

[0197] 1C1 The amino acid sequence of VH is shown in SEQ ID NO: 3, its coding nucleic acid is shown in SEQ ID NO: 4, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 5, 6, 7 respectively.

Chemical Structure

[0198] Nucleic acid sequence

Chemical Structure

[0199] The amino acid sequence of VL is shown in SEQ ID NO: 8, its coding nucleic acid is shown in SEQ ID NO: 9, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 10, 11, 12 respectively.

Chemical Structure

[0200] Nucleic acid sequence

Chemical Structure

[0201] 2E1 The amino acid sequence of VH is shown in SEQ ID NO: 13, its coding nucleic acid is shown in SEQ ID NO: 14, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 15, 16, 17 respectively.

Chemical Structure

[0202] Nucleic acid sequence

Chemical Structure

[0203] The amino acid sequence of VL is shown in SEQ ID NO: 18, its coding nucleic acid is shown in SEQ ID NO: 19, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 20, 21, 22 respectively.

Chem.

[0204] Nucleic acid sequence

Chem.

[0205] 2F1 The amino acid sequence of VH is shown in SEQ ID NO: 23, its coding nucleic acid is shown in SEQ ID NO: 24, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 25, 26, 27 respectively.

Chem.

[0206] Nucleic acid sequence

Chem.

[0207] The amino acid sequence of VL is shown in SEQ ID NO: 28, its coding nucleic acid is shown in SEQ ID NO: 29, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 30, 31, 32 respectively.

Chem.

[0208] Nucleic acid sequence

Chem.

[0209] 4B1 The amino acid sequence of VH is shown in SEQ ID NO: 33, its encoding nucleic acid is shown in SEQ ID NO: 34, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 5, 35, 7 respectively.

Chem.

[0210] Nucleic acid sequence

Chem.

[0211] The amino acid sequence of VL is shown in SEQ ID NO: 36, its encoding nucleic acid is shown in SEQ ID NO: 37, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 38, 21, 39 respectively.

Chem.

[0212] Nucleic acid sequence

Chem.

[0213] 4D3 The amino acid sequence of VH is shown in SEQ ID NO: 40, its encoding nucleic acid is shown in SEQ ID NO: 41, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 42, 43, 44 respectively.

Chem.

[0214] Nucleic acid sequence

Chem.

[0215] The amino acid sequence of VL is shown in SEQ ID NO: 45, its coding nucleic acid is shown in SEQ ID NO: 46, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 47, 48, 49 respectively.

Chemical formula

[0216] Nucleic acid sequence

Chemical formula

[0217] 6H1 The amino acid sequence of VH is shown in SEQ ID NO: 50, its coding nucleic acid is shown in SEQ ID NO: 51, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 5, 52, 7 respectively.

Chemical formula

[0218] Nucleic acid sequence

Chemical formula

[0219] The amino acid sequence of VL is shown in SEQ ID NO: 53, its coding nucleic acid is shown in SEQ ID NO: 54, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 38, 21, 55 respectively.

Chemical formula

[0220] Nucleic acid sequence

Chemical formula

[0221] 8C2 The amino acid sequence of VH is shown in SEQ ID NO: 56, its coding nucleic acid is shown in SEQ ID NO: 57, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NOs: 58, 59, and 60, respectively.

Chemical formula

[0222] Nucleic acid sequence

Chemical formula

[0223] The amino acid sequence of VL is shown in SEQ ID NO: 61, its coding nucleic acid is shown in SEQ ID NO: 62, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NOs: 63, 64, and 65, respectively.

Chemical formula

[0224] Nucleic acid sequence

Chemical formula

[0225] 8D1 The amino acid sequence of VH is shown in SEQ ID NO: 66, its coding nucleic acid is shown in SEQ ID NO: 67, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NOs: 68, 69, and 70, respectively.

Chemical formula

[0226] Nucleic acid sequence

Chemical formula

[0227] The amino acid sequence of VL is shown in SEQ ID NO: 71, its coding nucleic acid is shown in SEQ ID NO: 72, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NOs: 73, 74, and 75, respectively.

Chemical Structure

[0228] Nucleic acid sequence

Chemical Structure

[0229] 10B1 The amino acid sequence of VH is shown in SEQ ID NO: 76, its coding nucleic acid is shown in SEQ ID NO: 77, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NOs: 241, 78, and 79, respectively.

Chemical Structure

[0230] Nucleic acid sequence

Chemical Structure

[0231] The amino acid sequence of VL is shown in SEQ ID NO: 80, its coding nucleic acid is shown in SEQ ID NO: 81, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NOs: 82, 83, and 84, respectively.

Chemical Structure

[0232] Nucleic acid sequence

Chemical Structure

[0233] 10B2 The amino acid sequence of VH is shown in SEQ ID NO: 40, its encoding nucleic acid is shown in SEQ ID NO: 41, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs: 42, 43, 44 respectively.

Chemical formula

[0234] Nucleic acid sequence

Chemical formula

[0235] The amino acid sequence of VL is shown in SEQ ID NO: 85, its encoding nucleic acid is shown in SEQ ID NO: 86, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs: 47, 48, 49 respectively.

Chemical formula

[0236] Nucleic acid sequence

Chemical formula

[0237] 10E8 The amino acid sequence of VH is shown in SEQ ID NO: 87, its encoding nucleic acid is shown in SEQ ID NO: 88, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs: 25, 89, 90 respectively.

Chemical formula

[0238] Nucleic acid sequence

Chemical formula

[0239] The amino acid sequence of VL is shown in SEQ ID NO: 91, its coding nucleic acid is shown in SEQ ID NO: 92, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 93, 94, 95 respectively.

Chemical Structure

[0240] Nucleic acid sequence

Chemical Structure

[0241] 12A4 The amino acid sequence of VH is shown in SEQ ID NO: 96, its coding nucleic acid is shown in SEQ ID NO: 97, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 98, 99, 100 respectively.

Chemical Structure

[0242] Nucleic acid sequence

Chemical Structure

[0243] The amino acid sequence of VL is shown in SEQ ID NO: 101, its coding nucleic acid is shown in SEQ ID NO: 102, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 103, 104, 105 respectively.

Chemical Structure

[0244] Nucleic acid sequence

Chemical Structure

[0245] 14D1 The amino acid sequence of VH is shown in SEQ ID NO: 106, its encoding nucleic acid is shown in SEQ ID NO: 107, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 108, 99, 109 respectively.

Chemical Structure

[0246] Nucleic acid sequence

Chemical Structure

[0247] The amino acid sequence of VL is shown in SEQ ID NO: 110, its encoding nucleic acid is shown in SEQ ID NO: 111, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 112, 104, 105 respectively.

Chemical Structure

[0248] Nucleic acid sequence

Chemical Structure

[0249] 15A6 The amino acid sequence of VH is shown in SEQ ID NO: 113, its encoding nucleic acid is shown in SEQ ID NO: 114, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 98, 115, 116 respectively.

Chemical Structure

[0250] Nucleic acid sequence

Chemical Structure

[0251] The amino acid sequence of VL is shown in SEQ ID NO: 110, its coding nucleic acid is shown in SEQ ID NO: 111, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NOs: 112, 104, and 105, respectively.

Chemical formula

[0252] Nucleic acid sequence

Chemical formula

[0253] 15D1 The amino acid sequence of VH is shown in SEQ ID NO: 117, its coding nucleic acid is shown in SEQ ID NO: 118, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NOs: 119, 99, and 120, respectively.

Chemical formula

[0254] Nucleic acid sequence

Chemical formula

[0255] The amino acid sequence of VL is shown in SEQ ID NO: 121, its coding nucleic acid is shown in SEQ ID NO: 122, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NOs: 123, 124, and 125, respectively.

Chemical formula

[0256] Nucleic acid sequence

Chemical formula

[0257] 15H5 The amino acid sequence of VH is shown in SEQ ID NO: 126, its encoding nucleic acid is shown in SEQ ID NO: 127, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO: 108, 99, and 116, respectively.

Chemical Structure

[0258] Nucleic acid sequence

Chemical Structure

[0259] The amino acid sequence of VL is shown in SEQ ID NO: 128, its encoding nucleic acid is shown in SEQ ID NO: 129, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO: 130, 104, and 105, respectively.

Chemical Structure

[0260] Nucleic acid sequence

Chemical Structure

[0261] 17D12 The amino acid sequence of VH is shown in SEQ ID NO: 131, its encoding nucleic acid is shown in SEQ ID NO: 132, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO: 108, 133, and 134, respectively.

Chemical Structure

[0262] Nucleic acid sequence

Chemical Structure

[0263] The amino acid sequence of VL is shown in SEQ ID NO: 135, its coding nucleic acid is shown in SEQ ID NO: 136, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NOs: 112, 104, and 105, respectively.

Chemical Structure

[0264] Nucleic acid sequence

Chemical Structure

[0265] 18C3 The amino acid sequence of VH is shown in SEQ ID NO: 137, its coding nucleic acid is shown in SEQ ID NO: 138, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NOs: 58, 59, and 139, respectively.

Chemical Structure

[0266] Nucleic acid sequence

Chemical Structure

[0267] The amino acid sequence of VL is shown in SEQ ID NO: 61, its coding nucleic acid is shown in SEQ ID NO: 62, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NOs: 63, 64, and 65, respectively.

Chemical Structure

[0268] Nucleic acid sequence

Chemical Structure

[0269] 18E7 The amino acid sequence of VH is shown in SEQ ID NO: 140, its encoding nucleic acid is shown in SEQ ID NO: 141, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NOs: 98, 99, and 109, respectively.

Chemical formula

[0270] Nucleic acid sequence

Chemical formula

[0271] The amino acid sequence of VL is shown in SEQ ID NO: 101, its encoding nucleic acid is shown in SEQ ID NO: 102, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NOs: 103, 104, and 105, respectively.

Chemical formula

[0272] Nucleic acid sequence

Chemical formula

[0273] 19E2 The amino acid sequence of VH is shown in SEQ ID NO: 140, its encoding nucleic acid is shown in SEQ ID NO: 141, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NOs: 98, 99, and 109, respectively.

Chemical formula

[0274] Nucleic acid sequence

Chemical formula

[0275] The amino acid sequence of VL is shown in SEQ ID NO: 142, its coding nucleic acid is shown in SEQ ID NO: 143, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NOs: 103, 104, and 144, respectively.

Chemical formula

[0276] Nucleic acid sequence

Chemical formula

[0277] 22C1 The amino acid sequence of VH is shown in SEQ ID NO: 145, its coding nucleic acid is shown in SEQ ID NO: 146, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NOs: 25, 147, and 148, respectively.

Chemical formula

[0278] Nucleic acid sequence

Chemical formula

[0279] The amino acid sequence of VL is shown in SEQ ID NO: 149, its coding nucleic acid is shown in SEQ ID NO: 150, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NOs: 93, 151, and 95, respectively.

Chemical formula

[0280] Nucleic acid sequence

Chemical formula

[0281] Example 3 Construction of GPRC5D×CD3κλ Bispecific Antibody Clones 2E1, 2F1, 4B1, 4D3, 8D1, and 15D1 with high binding affinity to GPRC5D were selected, and their CDRs were transplanted into highly homologous human germline genes to construct bispecific antibodies with humanized anti-CD3e antibody VH9VL5. The Fc portion of the bispecific antibody adopted the knob-into-hole structure of human IgG4 to achieve heterodimer pairing (Atwell et al., Stable Heterodimers from Remodeling the Domain Interface of a Homodimer using a Phage Display Library, J. Mol. Biol. (1997) 270). The hinge region maintained stability by mutating Ser228Pro, Leu235Glu, and Pro329Ala, completely eliminating the interaction with Fcγ receptor and C1q.

[0282] The sequence information of the humanized anti-CD3e antibody VH9VL5 is as follows.

[0283] The amino acid sequence of VL is shown in SEQ ID NO: 152, its coding nucleic acid is shown in SEQ ID NO: 153, and its LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO: 154, 155, and 156 respectively.

Chemical formula

[0284] Nucleic acid sequence

Chemical formula

[0285] The amino acid sequence of VH is shown in SEQ ID NO: 157, its coding nucleic acid is shown in SEQ ID NO: 158, and its HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO: 159, 160, and 161 respectively.

Chemical formula

[0286] Nucleic acid sequence

Chem.

Table 4

[0287] Humanized antibody 2E1 The amino acid sequence of VH is shown in SEQ ID NO: 162, its coding nucleic acid is shown in SEQ ID NO: 163, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 15, 164, 17 respectively.

Chem.

[0288] Nucleic acid sequence

Chem.

[0289] The amino acid sequence of VL is shown in SEQ ID NO: 165, its coding nucleic acid is shown in SEQ ID NO: 166, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 167, 21, 22 respectively.

Chem.

[0290] Nucleic acid sequence

Chem.

[0291] Humanized antibody 2F1 The amino acid sequence of VH is shown in SEQ ID NO: 168, its coding nucleic acid is shown in SEQ ID NO: 169, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 25, 170, 27 respectively.

Chem.

[0292] Nucleic acid sequence

Chem.

[0293] The amino acid sequence of VL is shown in SEQ ID NO: 171, its coding nucleic acid is shown in SEQ ID NO: 172, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs: 173, 31, 32 respectively.

Chem.

[0294] Nucleic acid sequence

Chem.

[0295] Humanized antibody 4B1 The amino acid sequence of VH is shown in SEQ ID NO: 174, its coding nucleic acid is shown in SEQ ID NO: 175, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs: 5, 176, 7 respectively.

Chem.

[0296] Nucleic acid sequence

Chem.

[0297] The amino acid sequence of VL is shown in SEQ ID NO: 177, its coding nucleic acid is shown in SEQ ID NO: 178, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs: 179, 21, 39 respectively.

Chem.

[0298] Nucleic acid sequence [Chemical]

[0299] Humanized antibody 4D3 The amino acid sequence of VH is shown in SEQ ID NO: 180, its coding nucleic acid is shown in SEQ ID NO: 181, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 42, 182, 44 respectively. [Chemical]

[0300] Nucleic acid sequence [Chemical]

[0301] The amino acid sequence of VL is shown in SEQ ID NO: 183, its coding nucleic acid is shown in SEQ ID NO: 184, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 47, 48 and 49 respectively. [Chemical]

[0302] Nucleic acid sequence [Chemical]

[0303] Humanized antibody 8D1 The amino acid sequence of VH is shown in SEQ ID NO: 185, its coding nucleic acid is shown in SEQ ID NO: 186, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 68, 187, 70 respectively. [Chemical]

[0304] Nucleic acid sequence [Chemical]

[0305] The amino acid sequence of VL is shown in SEQ ID NO: 188, its coding nucleic acid is shown in SEQ ID NO: 189, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs: 190, 74, 75 respectively.

Chemical Structure

[0306] Nucleic acid sequence

Chemical Structure

[0307] Humanized antibody 15D1 The amino acid sequence of VH is shown in SEQ ID NO: 191, its coding nucleic acid is shown in SEQ ID NO: 192, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs: 119, 193, 120 respectively.

Chemical Structure

[0308] Nucleic acid sequence

Chemical Structure

[0309] The amino acid sequence of VL is shown in SEQ ID NO: 194, its coding nucleic acid is shown in SEQ ID NO: 195, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs: 123, 124, 125 respectively.

Chemical Structure

[0310] Nucleic acid sequence

Chemical Structure

[0311] Plasmids encoding the corresponding antibody fragments were mixed at a ratio of κ light chain: λ light chain: heavy chain 1: heavy chain 1 = 2:2:1:1, mixed with 3 mg / mL of PEI, and co-transfected into CHO-S cells. The cells were cultured in 500 mL of CD CHO AGT medium (Gibco #12490-001) at 37 °C, 5% CO2, and 150 rpm. 4% CHO Feed C+ supplement (Gibco #A25031-05) was added on the 2nd, 4th, and 6th days of transient transfection respectively. When the cell viability decreased to about 85%, the fermentation broth was harvested and purified by Protein A affinity chromatography. The monomer content showed more than 90% by SEC-HPLC. The control GPRC5D×CD3 bispecific antibody GCDB72 was synthesized and assembled according to Patent Document US10562968B2. The isotype control anti-KLH×CD3, anti-KLH antibody, and anti-CD3e antibody VH9VL5 were constructed according to the method of WO2022042661A1.

[0312] Example 4 Binding Activity and TDCC Killing Activity of GPRC5D×CD3κλ Bispecific Antibody (1) Binding of GPRC5D×CD3κλ Bispecific Antibody to GPRC5D + Binding to Tumor Cells Logarithmically growing MM.1R tumor cells were collected, 200 μg / mL of mouse IgG (Jackson ImmunoResearch, 115-005-03) was added, and after blocking in an ice bath for 30 minutes, the concentration of the cell suspension was adjusted to 5×10 5Adjusted to cells / mL, 100 μL per well was added to a 96-well U-bottom plate, centrifuged at 300 g for 5 minutes, and the supernatant was discarded. 100 μL of gradient-diluted antibody (starting concentration 200 nM, 3-fold dilution, 10 gradients) was added per well and incubated at 4 °C for 60 minutes. The primary antibody was washed away, Alexa Fluro647-labeled goat anti-human IgG Fc (1:300 dilution) was added at 50 μL / well, incubated for 20 minutes, washed once, then PI was added at 50 μL / well, incubated for 5 minutes, and detected by flow cytometry. As a result, as shown in Figure 4, the GPRC5D×CD3κλ bispecific antibody recognized all MM.1R tumor cells. Among them, the binding activity of GPRC5D×CD3κλ bispecific antibodies such as 2E1×CD3, 2F1×CD3, 4B1×CD3, 4D3×CD3, and 8D1×CD3 to tumor cells was significantly stronger than that of GCDB72. The mean fluorescence intensity of MM.1R binding at the maximum concentration of 200 nM was about 2.60 - 4.4 times that of GCDB72 (Table 4).

[0313] (2) TDCC effect mediated by GPRC5D×CD3κλ bispecific antibody Freshly isolated PBMC were collected, mixed with MM.1R tumor cells in the logarithmic growth phase, with effector cell:target cell = 10:1, and 50 μL of gradient-diluted antibody was added to each well (antibody concentration was 10-fold dilution from 66.7 nM, 7 gradients), and incubated at 37 °C, 5% CO2 for 24 hours. After incubation, 50 μL of the supernatant was transferred to a new black enzyme plate, 50 μL / well of LDH detection substrate was added, the reaction was terminated after 10 minutes, and LDH release was detected. As a result, as shown in Figure 5, the 2F1 GPRC5D×CD3κλ bispecific antibody had strong killing activity against tumor cells (EC50 = 0.65 pM) and was significantly superior to GCDB72 (Table 4).

Table 5

[0314] Example 5 Optimization of 2F1 GPRC5D×CD3κλ humanized bispecific antibody (1) Construction of 2F1 GPRC5D×CD3κλ bispecific antibody variants The 2F1 GPRC5D×CD3κλ bispecific antibody has strong TDCC activity. To optimize the affinity, surface charge distribution, and isoelectric point of the humanized 2F1 antibody, different VK1 and VH1 human germline family genes (such as VK1-33, VK1-39, VH1-46, VH1-69, etc.) were selected to obtain different 2F1×CD3κλ bispecific antibody variants (see Tables 5 and 6). The light chain plasmid and heavy chain plasmid encoding the GPRC5D antibody and CD3 antibody were mixed at a ratio of κ light chain:λ light chain:heavy chain 1:heavy chain 2 = 2:2:1:1, mixed with 3 mg / mL of PEI, and co-transfected into CHO-S cells. Cultured in CD CHO AGT medium at 37 °C, 5% CO2, and 150 rpm, and 4% CHO Feed C+ supplement was added on the 2nd, 4th, and 6th days after transient transfection. When the cell viability decreased to about 85%, the fermentation broth was harvested, filtered, and initially purified by Protein A affinity chromatography, and it was shown by SEC-HPLC that the monomer content was over 90%. Further purification by Capto S ImpAct ion exchange chromatography increased the monomer content to over 98 - 99%.

Table 6

Table 7-1

Table 7-2

Table 7-3

[0315] (2) Binding of 2F1 GPRC5D×CD3κλ bispecific antibody and its variants to GPRC5D + Binding to tumor cell lines Take MM.1R cells in the logarithmic growth phase, adjust the concentration of the cell suspension to 5×10 5 cells / mL, add 100 μL per well to a 96-well U-bottom plate, centrifuge at 300 g for 5 minutes, discard the supernatant, add 100 μL of serially diluted antibody (starting concentration 1800 nM, 3-fold dilution, 12 gradients) to each well, and incubate at 4°C for 60 minutes. Wash away the primary antibody, add Alexa Fluro647-labeled goat anti-human IgG Fc (1:300 dilution) at 50 μL / well, incubate on ice for 20 minutes, wash once, then add PI at 50 μL / well, incubate for 5 minutes, and detect with a flow cytometer. As a result, as shown in Figure 6, both the 2F1 GPRC5D×CD3κλ bispecific antibody and its mutants recognized MM.1R cells, and their affinity, although showing some differences, was slightly weaker compared to the parental 2F1×CD3 bispecific antibody, but both showed higher binding ability than GCDB72.

[0316] (3) Binding of 2F1 GPRC5D×CD3κλ bispecific antibody mutants to human and cynomolgus monkey GPRC5D stable transfection cell lines Take HEK293-hGPRC5D and HEK293-cyGPRC5D stable transfection cells in the logarithmic growth phase, and adjust the cell concentration to 5×10 5Adjusted to cells / mL, added 100 μL / well of cell suspension to a 96-well U-bottom plate, centrifuged at 300 g for 5 minutes, discarded the supernatant, added 100 μL of serially diluted 2F1 GPRC5D×CD3κλ bispecific antibody variants H4K5 and 4K7 and GCDB72 (starting concentration 1800 nM, 3-fold dilution, 12 gradients) to each well, and incubated at 4°C for 60 minutes. Added Alexa Fluro647-labeled goat anti-human IgG Fc (1:300 dilution) at 50 μL / well as the secondary antibody, incubated on ice for 20 minutes, washed once, then added PI solution (1:300) at 50 μL / well, incubated for 5 minutes, and detected with a flow cytometer. As a result, as shown in Figure 7, the 2F1 GPRC5D×CD3κλ bispecific antibody variants H4K5 and H4K7 bound to cells with similar affinity (<5-fold) to human GPRC5D (Figure 7A) and cynomolgus monkey GPRC5D (Figure 7B), so cynomolgus monkeys could be used as relevant species for the study. However, the binding of GCDB72 to cynomolgus monkey GPRC5D stable transfection cells was weak (Table 6).

[0317] (4) Binding of 2F1 GPRC5D×CD3κλ bispecific antibody variants to Jurkat cells Collected human leukemia T cell line Jurkat cells in the logarithmic growth phase, added 200 μg / mL of mouse IgG (Jackson ImmunoResearch, 115-005-03) to the cells, and applied an ice bath for 30 minutes. Adjusted the cell concentration to 5×10 with 4% fetal bovine serum 5Adjust to cells / mL, add 100 μL per well to a 96-well U-bottom plate, centrifuge at 300 g, discard the supernatant, add 100 μL each of gradient-diluted 2F1 GPRC5D×CD3κλ bispecific antibody variants H4K5 and H4K7 and GCDB72 to each well (starting concentration 1,800 nM, 3-fold dilution, 10 gradients), and incubate at 4°C for 60 minutes. Add Alexa Fluro647-labeled goat anti-human IgG Fc (1:300 dilution) as the secondary antibody at 50 μL / well, incubate on ice for 20 minutes, wash once, then add PI at 50 μL / well, incubate for 5 minutes, and detect with a flow cytometer. As shown in Fig. 8, the results of the detection indicated that the 2F1 GPRC5D×CD3κλ bispecific antibody variant had equivalent binding ability to T cells, but was significantly weaker than GCDB72, reduced the retention of the GPRC5D×CD3κλ bispecific antibody in peripheral lymphoid organs, increased the enrichment at the tumor site, gently activated T cells, and served to reduce cytokine release.

[0318] (5) TDCC mediated by 2F1 GPRC5D×CD3κλ bispecific antibody variant Freshly isolated PBMC were collected and mixed with MM.1R tumor cells in logarithmic growth phase at an effector cell:target cell ratio of 10:1. 50 μL of serially diluted antibody was added to each well (antibody concentration was 10-fold dilution from 66.7 nM, 7 gradients), and incubated at 37 °C, 5% CO2 for 24 hours. After incubation, 50 μL of the supernatant was transferred to a new black enzyme plate, 50 μL / well of LDH detection substrate was added, the reaction was terminated after 10 minutes, and LDH release was detected. The cells remaining in the well were washed twice with 4% calf blood, then 100 μg / mL human IgG was added and incubated for 10 minutes, T cell activation detection antibodies (CD25-PE, CD4-APC, CD69-FITC, CD8-APC) were added and incubated on ice for 20 minutes. After washing, the supernatant was discarded, 60 μL / well of PI was added and left standing on ice for 5 minutes, and detected by flow cytometer. As a result of the detection, as shown in Figure 9, the 2F1 GPRC5D×CD3κλ bispecific antibody variant can effectively kill tumor cells by mediating TDCC (Figure 9A). During the killing process, the expression of T cell activation markers CD69 and CD25 increased (Figure 9B), and it was shown that both CD8 + T cells and CD4 + T cells were effectively activated.

[0319] Example 6 Activation of the T cell activation pathway by the GPRC5D×CD3κλ bispecific antibody variant The luciferase reporter plasmid pGL4.30[luc2P / NFAT-RE / Hygro] (Promega, E8481) was transfected into Jurkat cells, and screening was performed by adding 200 μg / ml hygromycin B to obtain Jurkat-NFAT luc stable transfection reporter cells. GPRC5D +The target cells, GPRC5D×CD3 antibody, and Jurkat-NFAT-luc reporter cells were co-incubated. After the GPRC5D×CD3 antibody bound to the target cells, the other end bound to the CD3 receptor on the surface of Jurkat cells, causing aggregation of the CD3 intracellular signal motif, transmitting the activation signal to the nucleus via the NFAT pathway, and ultimately increasing the expression of the reporter gene luciferase.

[0320] Jurkat-NFAT-luc reporter cells and MM.1R cells in the logarithmic growth phase were collected, centrifuged, and the supernatant was discarded, and then resuspended at 2×10 6 cells / ml. 50 μl / well of target cells were inoculated into a 96-well plate, centrifuged at 300 g for 5 minutes, and the supernatant was discarded. Jurkat-NFAT-luc reporter cells were inoculated into a 96-well plate at 50 μl / well, and gradient-diluted GPRC5D×CD3κλ bispecific antibody variant or control antibody was added to each well at 50 μL each (starting concentration 20 μg / ml, 10-fold dilution, 10 gradients), and incubated at 5% CO2, 37 °C for 6 hours. After the incubation, according to the instructions of the ONE-Glo Luciferase Assay System, 100 μL of the detection reagent was added to each well, and left at room temperature for 3 minutes to detect the fluorescence signal (Biotek Synergy HT). As shown in Figure 10, the activation of Jurkat cells by the GPRC5D×CD3κλ bispecific antibody variant was dependent on the presence of target cells, and the EC50 was about 0.66 - 0.77 nM, both of which were weaker than the activation of Jurkat T cells by GCDB72 (EC50 = 0.066 nM). The in vitro activity results of the GPRC5D×CD3κλ bispecific antibody variant are summarized in Table 7.

Table 8

[0321] Example 7 GPRC5D×CD3κλ bispecific antibody variant against subcutaneous MM.1S transplanted tumor model in immunodeficient mice Six- to eight-week-old female B-NDG mice (manufactured by Biocytogen) were selected, and 6×106 Individual MM.1S cells were subcutaneously inoculated, and when the tumors grew to 130 - 140 mm 3 they were randomly grouped at the time of growth. They were divided into a drug administration group at 1.0 mg / kg, 0.1 mg / kg, 0.01 mg / kg, and a negative control group of KLH×CD3 at 1 mg / kg, respectively. 1×10 7 PBMC cells were injected into the tail vein of each mouse. Six days later, the first administration to the mice was started, and the administration interval was once every 5 days for a total of 2 administrations. The tumor volume and body weight of the mice were monitored. At the end of the experiment, the mice were killed by cervical dislocation, and the tumors were collected and weighed and recorded.

[0322] As a result, as shown in Figure 11, the in vivo efficacy of the GPRC5D×CD3κλ bispecific antibody variant showed a dose - correlation. The tumors were completely inhibited or regressed in the high - dose group and the medium - dose group. h2F1 H4K5 and h2F1 H4K7 showed a stronger tumor - inhibiting effect than GCDB72 at high, medium, and low concentrations. Figure 12 shows that the tumor - bearing mice tolerated the above doses well without adverse reactions such as weight loss.

[0323] Example 8 Construction of GPRC5D×BCMA×CD3 triTCE trispecific antibody (1) Construction of GPRC5D×BCMA×CD3 triTCE antibody The variable regions of the light and heavy chains of the humanized GPRC5D antibody 2F1 H4K5 were linked via a GGGGSGGGGSGGGGS flexible polypeptide chain to the heavy - chain N - terminus of the BCMA arm or the CD3 arm of the BCMA×CD3κλ005 bispecific antibody (see Chinese Patent Application CN114573703A) to construct a novel GPRC5D×BCMA×CD3 triTCE trispecific antibody that can simultaneously recognize two tumor antigens and recruit T cells. The structure of GPRC5D×BCMA×CD3 triTCE is shown in Figure 13.

[0324] In Tables 8 and 9, the amino acid sequence of the light chain of BCMA is shown in SEQ ID NO: 226, its coding nucleic acid is shown in SEQ ID NO: 227, its light chain variable region is shown in SEQ ID NO: 267, its light chain variable region coding nucleic acid is shown in SEQ ID NO: 269, the amino acid sequence of its heavy chain is shown in SEQ ID NO: 233, its coding nucleic acid is shown in SEQ ID NO: 238, its heavy chain variable region is shown in SEQ ID NO: 268, and its heavy chain variable region coding nucleic acid is shown in SEQ ID NO: 270.

[0325] In Tables 8 and 9, the amino acid sequence of the variable light chain region (GPRC5D-VL) of GPRC5D is shown in SEQ ID NO: 230, its coding nucleic acid is shown in SEQ ID NO: 271, the amino acid sequence of the variable heavy chain region (GPRC5D-VH) of GPRC5D is shown in SEQ ID NO: 232, and its coding nucleic acid is shown in SEQ ID NO: 272. [Table 9] [Table 10-1] [Table 10-2] [Table 10-3] [Table 10-4]

[0326] (2) Expression and purification of GPRC5D×BCMA×CD3 triTCE antibody Plasmids encoding the CD3-GPRC5D scFv arm and the BCMA arm (or plasmids encoding the BCMA-GPRC5D scFv arm and the CD3 arm) were mixed at a ratio of 1:2:1:2 (heavy chain: light chain = 1:2), transfected into CHO-S cells, and allowed to express at 37°C for 5 - 6 days. After that, the culture supernatant was collected, subjected to Protein A and Capto S ImpAct ion exchange chromatography, eluted with a gradient of phosphate-citrate / 0.5 M NaCl pH 7.5, and the elution peaks with monomer purity >90% were combined, concentrated, and exchanged into 10 mM acetic acid / 150 mM arginine pH 4.8 buffer. It was confirmed by mass spectrometry LC-MS (Agilent 1290 Infinity II; Q-TOF 6545XT) to be a GPRC5D×BCMA×CD3 triTCE trispecific antibody.

[0327] (3) Binding activity of the GPRC5D×BCMA×CD3 triTCE antibody (i) BLI affinity measurement The affinity of the GPRC5D×BCMA×CD3 triTCE antibody for the BCMA and CD3 recombinant antigens was detected using the biofilm interference method (BLI / Gator prime). The biotinylated BCMA antigen or CD3e antigen was captured with an SA probe, and the binding and dissociation signals of the probe were detected with the triTCE antibody as the analyte. As a result, even when the GPRC5D scFv was fused and expressed at the N-terminus of the BCMA arm or the CD3 arm, there was little effect on the binding of BCMA or CD3 (Table 10).

[0328] (ii) Binding to cell lines stably transfected with GPRC5D and BCMA Using the full-length human BCMA sequence (Sino biological, HG10620-M) as a template, a lentiviral vector plasmid containing the full-length human BCMA sequence was constructed. The constructed lentiviral plasmid and packaging plasmid were co-transfected into HEK293T cells (ATCC CRL-11268) according to the instructions of the lentiviral packaging kit (Lenti-Pac HIV Expression Packaging Kit, Gene Copoeia, product number: HPK-LvTR-20) to perform lentiviral packaging. 10 μL of the supernatant containing lentiviral particles was used to infect 1×10 6 HEK293T cells. After 48 hours, 8 μg / ml of puromycin was added for pressure screening, and finally, stable transfection cells HEK-hBCMA with high expression of human BCMA were isolated. HEK293-hGPRC5D and HEK293-hBCMA in the logarithmic growth phase, which were stably transfected cells, were taken, and the concentration of the cell suspension was adjusted to 5×10 5 cells / ml. The cell suspension was added to a 96-well U-bottom plate at 100 μL / well, centrifuged at 300 g for 5 minutes, the supernatant was discarded, 100 μL of gradient-diluted antibody was added to each well, and incubated at 4°C for 60 minutes. As the secondary antibody, Alexa Fluro647-labeled goat anti-human IgG Fc (1:300 dilution) was added at 50 μL / well, incubated on ice for 20 minutes, washed once, then PI solution was added at 50 μL / well, incubated for 5 minutes, and detected by a flow cytometer. As a result, the GPRC5D×BCMA×CD3 triTCE trispecific antibody bound to both the GPRC5D receptor and the BCMA receptor on the cell membrane (Figures 14 and 15), and its affinity (EC50) was approximately 2.7 - 14.1 nM (Table 10).

[0329] (iii) Binding to Jurkat cells Jurkat cells in the logarithmic growth phase were collected, 200 μg / mL of mouse IgG was added to the cells, and an ice bath was applied for 30 minutes. The cells were washed with 4% fetal bovine serum and adjusted to 5×10 5Adjusted to cells / mL, added 100 μL per well to a 96-well U-bottom plate, centrifuged at 300 g, discarded the supernatant, added 100 μL of serially diluted antibody to each well, and incubated at 4 °C for 60 minutes. Alexa Fluro647-labeled goat anti-human IgG Fc (1:300 dilution) was added at 50 μL / well as the secondary antibody, incubated on ice for 20 minutes, washed once, then PI was added at 50 μL / well, incubated for 5 minutes, and detected with a flow cytometer. As a result, the GPRC5D×BCMA×CD3 triTCE trispecific antibody bound to Jurkat cells with low affinity, and the binding did not reach a plateau even at the highest concentration of 200 nM (Figure 16).

[0330] From the binding data of BLI and FACS, the binding affinities of the GPRC5D×BCMA×CD3 triTCE trispecific antibody to BCMA, GPRC5D, and CD3 were basically the same as those of the GPRC5D×CD3κλ bispecific antibody and the BCMA×CD3κλ bispecific antibody (Table 10).

Table 11

[0331] Example 9 Killing of GPRC5D + and BCMA + against cell lines Freshly isolated PBMCs were collected and mixed with target cells in logarithmic growth phase, NCI-H929 (purchased from ATCC; catalog number CRL-9068), MM.1S, HEK293-hGPRC5D, or HEK293-hBCMA that had been stably transfected, respectively, at an effector cell:target cell ratio of 10:1. 50 μL of serially diluted antibody was added to each well (antibody concentrations were diluted 10-fold from 66.7 nM, 8 gradients), and the cells were incubated at 37 °C in 5% CO2 for 24 hours. After incubation, 50 μL of the supernatant was transferred to a new black enzyme plate, 50 μL / well of LDH detection substrate was added, the reaction was terminated after 10 minutes, and LDH release was detected. As a result, as shown in Figure 17, the GPRC5D×BCMA×CD3 triTCE trispecific antibody was specific for BCMA + single positive target cells (Figure 17A), GPRC5D +It can effectively kill both single positive target cells (Figure 17B) and myeloma cell lines MM.1S (Figure 17C) and NCI-H929 (Figure 17D) that express both GPRC5D and BCMA receptors simultaneously (Verkleij et al., Preclinical activity and determinants of response of the GPRC5DxCD3 bispecific antibody talquetamab in multiple myeloma, Blood Adv. 2021 Apr 27;5(8); Pillarisetti et al., A T-cell-redirecting bispecific G-protein-coupled receptor class 5 member D x CD3 antibody to treat multiple myeloma, Blood.2020;135(15); Pillarisetti et al., Teclistamab is an active T cell-redirecting bispecific antibody against B-cell maturation antigen for multiple myeloma, Blood Adv.2020 Sep 22;4(18)). Among them, the killing activity of GPRC5D×BCMA×CD3 triTCE-v1 and v2 against tumor cells was significantly superior to that of the parental GPRC5D×CD3 bispecific antibody and BCMA×CD3 bispecific antibody (Table 11).

Table 12

Claims

1. An antibody or antigen-binding fragment thereof that binds to the GPRC5D antigen, wherein (1) it comprises a heavy chain variable region and a light chain variable region, where (i) the heavy chain variable region comprises the following (a) to (c): (a) an HCDR1 sequence having the following formula: TSYNM H (SEQ ID NO: 5), and (b) an HCDR2 sequence having the following formula: YIYPGNGGTNYNQX １ FX ２ X ３ (SEQ ID NO: 242), where X １ is K or N, and X ２ is K or Q, and X ３ is A or G, and (c) an HCDR3 sequence having the following formula: GYGYRYWYFDV (SEQ ID NO: 7), and (ii) the light chain variable region comprises the following (a) to (c): (a) an LCDR1 sequence having the following formula: X ４ ASSSVX ５ X ６ MH (SEQ ID NO: 243), where X ４ is S or R, and X ５ is S or R, and X ６ is Y or F, and (b) an LCDR2 sequence having the following formula: DTSKX ７ AS (SEQ ID NO: 244), wherein X ７ is V or L, and (c) an LCDR3 sequence having the following formula: QQWX ８ X ９ X １０ PX １１ T (array number 245), where X ８ is S or N, and X ９ is S or N, and X １０ is N, Y or K, and X １１ is L or P, or (2) it comprises a heavy chain variable region and a light chain variable region, where (i) the heavy chain variable region comprises the following (a) to (c): (a) an HCDR1 sequence having the following formula: TG YTM N (SEQ ID NO: 25), and (b) an HCDR2 sequence having the following formula: LINPYNGGX １２ X １３ YNX １４ KFX １５ G (SEQ ID NO: 246), where X １２ is S or T, and X １３ is R, S or N, and X １４ is Q or L, and X １５ is K or Q, and (c) an HCDR3 sequence having the following formula: X １６ X １７ X １８ RX １９ X ２０ MDY (SEQ ID NO: 247), where X １６ is either W or L, and X １７ is either A, P or G, and X １８ is either I or L, and X １９ is either Y or N, and X ２０ is either A or G, and (ii) the light chain variable region comprises the following (a) to (c): (a) an LCDR1 sequence having the following formula: X ２１ ASQX ２２ X ２３ X ２４ X ２５ NX ２６ X ２７ (Sequence number 248), where X ２１ is K, R or Q, and X ２２ is N or S, and X ２３ is V or I, and X ２４ is G or S, and X ２５ is T or N, and X ２６ is V or L, and X ２７ is A or H, and (b) an LCDR2 sequence having the following formula: X ２８ ASX ２９ X ３０ X ３１ S (SEQ ID NO: 249), where X ２８ is F, Y or S, and X ２９ is Y, L or Q, and X ３０ is R, L or S, and X ３１ is N or I, and (c) an LCDR3 sequence having the following formula: QQX ３２ NSX ３３ X ３４ X ３５ T (sequence number 250), where X ３２ is Y or S, and X ３３ is Y or W, and X ３４ is L or P, and X ３５ is L or Q, or (3) it comprises a heavy chain variable region and a light chain variable region, where (i) the heavy chain variable region comprises the following (a) to (c): (a) an HCDR1 sequence having the following formula: TNFPI E (SEQ ID NO: 42), and (b) an HCDR2 sequence having the following formula: NFHPYNDTKYNEKFXX ３６ G (SEQ ID NO: 251), where X ３６ is K or Q, and (c) an HCDR3 sequence having the following formula: ITRGY (SEQ ID NO: 44), and (ii) the light chain variable region comprises the following (a) to (c): (a) an LCDR1 sequence having the following formula: KSSQSL LYS RNQKNSLA (SEQ ID NO: 47), and (b) an LCDR2 sequence having the following formula: WASTRES (SEQ ID NO: 48), and (c) an LCDR3 sequence having the following formula: QQYYSYPLT (SEQ ID NO: 49), or (4) it comprises a heavy chain variable region and a light chain variable region, where (i) the heavy chain variable region comprises the following (a) to (c): (a) an HCDR1 sequence having the following formula: TSYX ３７ MY (SEQ ID NO: 252), wherein X ３７ is Y or T, and (b) an HCDR2 sequence having the following formula: X ３８ INPSX ３９ GX ４０ TX ４１ YNX ４２ KFX ４３ X ４４ (Array No. 253), where X ３８ is G or Y, and X ３９ is N or S, and X ４０ is G or Y, and X ４１ is K or N, and X ４２ is Q or E, and X ４３ is K or Q, and X ４４ is S, D or G, and (c) an HCDR3 sequence having the following formula: X ４５ X ４６ X ４７ X ４８ X ４９ X ５０ X ５１ YX ５２ X ５３ X ５４ X ５５ DY (array number 254), where X ４５ is G or W, and X ４６ is G or S, and X ４７ is G or L, and X ４８ is R or N, and X ４９ is E or S, and X ５０ is Y or R, and X ５１ is Y or L, and X ５２ is A or Y, and X ５３ is Y or M, and X ５４ is A or does not exist, and X ５５ is M or does not exist, and (ii) the light chain variable region comprises the following (a) to (c): (a) an LCDR1 sequence having the following formula: X ５６ ASX ５７ X ５８ X ５９ X ６０ X ６１ YX ６２ X ６３ (Array No. 255), where X ５６ is R, Q, or S, and X ５７ is Q or S, and X ５８ is D or S, and X ５９ is I or V, and X ６０ is S or does not exist, and X ６１ is N or S, and X ６２ is L or M, and X ６３ is N or Y, and (b) an LCDR2 sequence having the following formula: X ６４ TSX ６５ LX ６６ S (SEQ ID NO: 256), where X ６４ is Y or G, and X ６５ is R or N, and X ６６ is H or A, and (c) an LCDR3 sequence having the following formula: QQX ６７ X ６８ SX ６９ PX ７０ T (array number 257), where X ６７ is R or G, and X ６８ is Y or S, and X ６９ is L or Y, and X ７０ is Y or L, or (5) it comprises a heavy chain variable region and a light chain variable region, where (i) The heavy chain variable region includes the following (a) to (c): (a) An HCDR1 sequence having the following formula: TX ７１ YVMX ７２ (SEQ ID NO: 258), where X ７１ is S or N, and X ７２ is Y or H, and (b) An HCDR2 sequence having the following formula: YINPYNDGTKYNE X ７３ FX ７４ G (SEQ ID NO: 259), where X ７３ is K or R, and X ７４ is K, T or Q, and (c) An HCDR3 sequence having the following formula: GGX ７５ X ７６ X ７７ X ７８ X ７９ X ８０ X ８１ X ８２ X ８３ Y (array number 260), where X ７５ is M or V, and X ７６ is L or R, and X ７７ is T or R, and X ７８ is T or Y, and X ７９ is R or F, and X ８０ is S or does not exist, and X ８１ is L or does not exist, and X ８２ is F or does not exist, and X ８３ is A, T, V, L or D, and (ii) The light chain variable region includes the following (a) to (c): (a) An LCDR1 sequence having the following formula: RASQX ８４ IX ８５ X ８６ X ８７ X ８８ X ８９ (SEQ ID NO: 261), where X ８４ is R, E or D, and X ８５ is G or S, and X ８６ is G or S, and X ８７ is Y or N, and X ８８ is L or F, and X ８９ is N or S, and (b) An LCDR2 sequence having the following formula: AX ９０ SX ９１ LDS (SEQ ID NO: 262), where X ９０ is A or T, and X ９１ is T or S, and (c) An LCDR3 sequence having the following formula: LQYAX ９２ X ９３ PX ９４ T (SEQ ID NO: 263), where X ９２ is S or N, and X ９３ is Y or F, and X ９４ is P or Y, or (6) Comprising a heavy chain variable region and a light chain variable region, wherein (i) The heavy chain variable region includes the following (a) to (c): (a) An HCDR1 sequence having the following formula: SSYGMS (SEQ ID NO: 58), and (b) An HCDR2 sequence having the following formula: TISSGGSYYPDSSVK (SEQ ID NO: 59), and (c) An HCDR3 sequence having the following formula: HX ９５ X ９６ X ９７ X ９８ X ９９ X １００ X １０１ X １０２ DY (array number 264), where X ９５ is G or E, X ９６ is G or A, X ９７ is S or R, X ９８ is S or A, X ９９ is S or T, X １００ is Y or F, X １０１ is V or does not exist, X １０２ is M or does not exist, and (ii) The light chain variable region includes the following (a) to (c): (a) An LCDR1 sequence having the following formula: RSSKSL LH SN GNTYL Y (SEQ ID NO: 63), and (b) An LCDR2 sequence having the following formula: RMSNLA S (SEQ ID NO: 64), and (c) An LCDR3 sequence having the following formula: MQHLEY PFT (SEQ ID NO: 65), or (7) Comprising a heavy chain variable region and a light chain variable region, wherein (i) The heavy chain variable region includes the following (a) to (c): (a) An HCDR1 sequence having the following formula: TRYWMH (SEQ ID NO: 15), and (b) An HCDR2 sequence having the following formula: EFNPSNGRINYNEKF X １０３ X １０４ (SEQ ID NO: 265), where X １０３ is K or Q, and X １０４ is N or G, and (c) An HCDR3 sequence having the following formula: GFAY (SEQ ID NO: 17), and (ii) The light chain variable region includes the following (a) to (c): (a) An LCDR1 sequence having the following formula: X １０５ ASSTVSYIH (SEQ ID NO: 266), where X １０５ is S or R, and (b) An LCDR2 sequence having the following formula: DTSKLA S (SEQ ID NO: 21), and (c) An LCDR3 sequence having the following formula: QQWTTN PWT (SEQ ID NO: 22), An antibody or an antigen-binding fragment thereof.

2. The heavy chain CDR1 selected from the amino acid sequences shown in SEQ ID NOs: 5, 15, 25, 42, 58, 68, 98, 108, 119, 241 or any variants thereof, the heavy chain CDR2 selected from the amino acid sequences shown in SEQ ID NOs: 6, 16, 26, 35, 43, 52, 59, 69, 78, 89, 99, 115, 133, 147 or variants thereof, the heavy chain CDR3 selected from the amino acid sequences shown in SEQ ID NOs: 7, 17, 27, 44, 60, 70, 79, 90, 100, 109, 116, 120, 134, 139, 148 or any variants thereof, and the light chain CDR1 selected from the amino acid sequences shown in SEQ ID NOs: 10, 20, 30, 38, 47, 63, 73, 82, 93, 103, 112, 123, 130, 167, 173, 179, 190, 208 or any variants thereof, the light chain CDR2 selected from the amino acid sequences shown in SEQ ID NOs: 11, 21, 31, 48, 64, 74, 83, 94, 104, 124, 151, 209 or any variants thereof, the light chain CDR3 selected from the amino acid sequences shown in SEQ ID NOs: 12, 22, 32, 39, 49, 55, 65, 75, 84, 95, 105, 125, 144 or any variants thereof, the antibody according to claim 1 or an antigen-binding fragment thereof.

3. (1) The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 5, 6, and 7, respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 10, 11, and 12, respectively. (2) The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 15, 16, and 17, respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 20, 21, and 22, respectively. (3) The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 26, and 27, respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 30, 31, and 32, respectively. (4) The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 5, 35, and 7, respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 38, 21, and 39, respectively. (5) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 42, 43, and 44, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 47, 48, and 49, respectively, (6) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 5, 52, and 7, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 38, 21, and 55, respectively, (7) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 58, 59, and 60, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 63, 64, and 65, respectively, (8) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 68, 69, and 70, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 73, 74, and 75, respectively, (9) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 241, 78, and 79, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 82, 83, and 84, respectively, (10) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 42, 43, and 44, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 47, 48, and 49, respectively, (11) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 89, and 90, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 93, 94, and 95, respectively, (12) Heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 98, 99, and 100, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 103, 104, and 105, respectively, The heavy-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 108, 99, and 109, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 112, 104, and 105, respectively, The heavy-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 98, 115, and 116, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 112, 104, and 105, respectively, The heavy-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 119, 99, and 120, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 123, 124, and 125, respectively, The heavy-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 108, 99, and 116, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 130, 104, and 105, respectively, The heavy-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 108, 133, and 134, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 112, 104, and 105, respectively, The heavy-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 58, 59, and 139, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 63, 64, and 65, respectively, The heavy-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 98, 99, and 109, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 103, 104, and 105, respectively, The heavy-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 98, 99, and 109, respectively, and the light-chain CDR1, CDR2, and CDR3 sequences containing the amino acid sequences shown in SEQ ID NOs: 103, 104, and 144, respectively, The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 147, and 148 respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 93, 151, and 95 respectively, The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 15, 164, and 17 respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 167, 21, and 22 respectively, The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 170, and 27 respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 173, 31, and 32 respectively, The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 5, 176, and 7 respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 179, 21, and 39 respectively, The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 42, 182, and 44 respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 47, 48, and 49 respectively, The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 68, 187, and 70 respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 190, 74, and 75 respectively, The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 119, 193, and 120 respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 123, 124, and 125 respectively, The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 170, and 27 respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 208, 209, and 32 respectively, The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 170, and 27, respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 173, 209, and 32, respectively, The heavy chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 25, 170, and 27, respectively, and the light chain CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences shown in SEQ ID NOs: 208, 31, and 32, respectively, The antibody or antigen-binding fragment thereof according to claim 1 or 2, comprising a combination of a heavy chain CDR and a light chain CDR selected from the above.

4. Comprising a heavy chain variable region selected from the amino acid sequences shown in SEQ ID NOs: 3, 13, 23, 33, 40, 50, 56, 66, 76, 87, 96, 106, 113, 117, 126, 131, 140, 145, 162, 168, 174, 180, 185, 191, 202, 232 or any variant thereof, and a light chain variable region selected from the amino acid sequences shown in SEQ ID NOs: 8, 18, 28, 36, 45, 53, 61, 71, 80, 85, 91, 101, 110, 121, 128, 135, 142, 149, 165, 171, 177, 183, 188, 194, 206, 212, 216, 220, 230 or any variant thereof, Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 3 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 8 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 13 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 18 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 23 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 28 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 33 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 36 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 40 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 45 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 50 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 53 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 56 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 61 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 66 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 71 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 76 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 80 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 40 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 85 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 87 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 91 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 96 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 101 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 106 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 110 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 113 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 110 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 117 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 121 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 126 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 128 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 131 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 135 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 137 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 61 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 140 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 101 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 140 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 142 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 145 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 149 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 162 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 165 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 168 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 171 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 174 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 177 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 180 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 183 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 185 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 188 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 191 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 194 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 206 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 212 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 216 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 220 or any variant thereof. Preferably, it comprises a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 232 or any variant thereof, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 230 or any variant thereof. Preferably, the antibody or its antigen-binding fragment is humanized. The antibody or its antigen-binding fragment according to any one of claims 1 to 3.

5. A nucleic acid encoding the antibody or its antigen-binding fragment according to any one of claims 1 to 4, Preferably, the nucleic acid comprises a heavy chain variable region nucleic acid sequence selected from the nucleotide sequences shown in SEQ ID NO: 4, 14, 24, 34, 41, 51, 57, 67, 77, 88, 97, 107, 114, 118, 127, 132, 138, 141, 146, 163, 169, 175, 181, 186, 192, 203 or any variants thereof, and a light chain variable region nucleic acid sequence selected from the nucleotide sequences shown in SEQ ID NO: 9, 19, 29, 37, 46, 54, 62, 72, 81, 86, 92, 102, 111, 122, 129, 136, 143, 150, 166, 172, 178, 184, 189, 195, 207, 213, 217, 221 or any variants thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 4 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 9 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 14 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 19 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 24 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 29 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 34 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 37 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 41 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 46 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 51 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 54 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 57 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 62 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 67 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 72 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 77 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 81 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 41 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 86 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 88 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 92 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 97 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 102 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 107 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 111 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 114 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 111 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 118 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 122 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 127 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 129 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 132 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 136 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 138 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 62 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 141 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 102 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 141 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 143 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 146 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 150 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 163 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 166 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 169 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 172 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 175 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 178 or any variant thereof. More preferably, the nucleic acid comprises a heavy chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 181 or any variant thereof, and a light chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 184 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 186 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 189 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 192 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 195 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 203 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 207 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 203 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 213 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 203 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 217 or any variant thereof. More preferably, the nucleic acid comprises a heavy-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 203 or any variant thereof, and a light-chain variable region nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 221 or any variant thereof.

6. A multispecific antibody or an antigen-binding fragment thereof, comprising at least (a) an antibody or an antigen-binding fragment thereof that binds to the GPRC5D antigen according to any one of claims 1 to 4, and (b) an antibody or an antigen-binding fragment thereof that binds to the CD3 antigen. The antibody or an antigen-binding fragment thereof that binds to the GPRC5D antigen according to any one of claims 1 to 4 is a first antigen-binding portion, the GPRC5D antigen is a first antigen, the first antigen-binding portion comprises a first heavy-chain variable region and a first light-chain variable region, and the first antigen-binding portion comprises a first binding structural domain that binds to the first antigen. Preferably, the first binding structure domain includes a first heavy chain CDR selected from the amino acid sequences shown in SEQ ID NO: 25, 170, 27 or any variants thereof, and a first light chain CDR selected from the amino acid sequences shown in SEQ ID NO: 31, 32, 173, 208, 209 or any variants thereof, and (b) The antibody or antigen-binding fragment thereof that binds to the CD3 antigen is a second antigen-binding portion, the CD3 antigen is a second antigen, the second antigen-binding portion includes a second heavy chain variable region and a second light chain variable region, the second antigen-binding portion includes a second binding structure domain that binds to the second antigen, wherein the second binding structure domain includes a second heavy chain CDR1 selected from the amino acid sequence shown in SEQ ID NO: 159 or a variant thereof, a second heavy chain CDR2 selected from the amino acid sequence shown in SEQ ID NO: 160 or a variant thereof, a second heavy chain CDR3 selected from the amino acid sequence shown in SEQ ID NO: 161 or a variant thereof, and a second light chain CDR1 selected from the amino acid sequence shown in SEQ ID NO: 154 or any variant thereof, a second light chain CDR2 selected from the amino acid sequence shown in SEQ ID NO: 155 or any variant thereof, a second light chain CDR3 selected from the amino acid sequence shown in SEQ ID NO: 156 or any variant thereof, Preferably, the second binding structure domain includes a second heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 157 or any variant thereof, and a second light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 152 or any variant thereof, Preferably, the second heavy chain of the second antigen-binding portion includes the amino acid sequence shown in SEQ ID NO: 222, and the second light chain of the second antigen-binding portion includes the amino acid sequence shown in SEQ ID NO: 224, Preferably, the first binding structure domain includes a first heavy chain CDR1 selected from the amino acid sequence shown in SEQ ID NO: 25 or any variant thereof, a first heavy chain CDR2 selected from the amino acid sequence shown in SEQ ID NO: 170 or any variant thereof, a first heavy chain CDR3 selected from the amino acid sequence shown in SEQ ID NO: 27 or any variant thereof, and a first light chain CDR1 selected from the amino acid sequences shown in SEQ ID NOs: 173 and 208 or any variant thereof, a first light chain CDR2 selected from the amino acid sequences shown in SEQ ID NOs: 31 and 209 or any variant thereof, and a first light chain CDR3 selected from the amino acid sequence shown in SEQ ID NO: 32 or any variant thereof, Preferably, the first light chain CDRs of the first binding structure domain are respectively a first light chain CDR1, CDR2, and CDR3 sequence including the amino acid sequences shown in SEQ ID NOs: 173, 31, and 32, a first light chain CDR1, CDR2, and CDR3 sequence including the amino acid sequences shown in SEQ ID NOs: 208, 209, and 32, a first light chain CDR1, CDR2, and CDR3 sequence including the amino acid sequences shown in SEQ ID NOs: 173, 209, and 32, a first light chain CDR1, CDR2, and CDR3 sequence including the amino acid sequences shown in SEQ ID NOs: 208, 31, and 32, and are selected therefrom, and the first heavy chain CDRs of the first binding structure domain are selected from the first heavy chain CDR1, CDR2, and CDR3 sequences respectively including the amino acid sequences shown in SEQ ID NOs: 25, 170, and 27, Preferably, the first binding structure domain includes a first heavy chain variable region selected from the amino acid sequences shown in SEQ ID NOs: 168, 202, and 232 or any variants thereof, and a first light chain variable region selected from the amino acid sequences shown in SEQ ID NOs: 171, 206, 212, 216, 220, and 230 or any variants thereof, Preferably, the first binding structure domain includes a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 168 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 171 or any variant thereof, Preferably, the first binding structure domain includes a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 206 or any variant thereof. Preferably, the first binding structure domain includes a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 212 or any variant thereof. Preferably, the first binding structure domain includes a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 216 or any variant thereof. Preferably, the first binding structure domain includes a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 202 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 220 or any variant thereof. Preferably, the first binding structure domain includes a first heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 232 or any variant thereof, and a first light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 230 or any variant thereof. Preferably, the first heavy chain of the first antigen-binding portion is selected from the amino acid sequences shown in SEQ ID NOs: 196 and 200, and the first light chain of the first antigen-binding portion is selected from the amino acid sequences shown in SEQ ID NOs: 198, 204, 210, 214, and 218. More preferably, the first heavy chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 196, and the first light chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO:

198. More preferably, the first heavy chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 200, and the first light chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO:

204. More preferably, the first heavy chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 200, and the first light chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO:

210. More preferably, the first heavy chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 200, and the first light chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO:

214. More preferably, the first heavy chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO: 200, and the first light chain of the first antigen-binding portion comprises the amino acid sequence shown in SEQ ID NO:

218. More preferably, the multispecific antibody or its antigen-binding fragment further comprises (c) an antibody or its antigen-binding fragment that binds to the BCMA antigen. (c) The antibody or its antigen-binding fragment that binds to the BCMA antigen is a third antigen-binding portion, the BCMA antigen is a third antigen, the third antigen-binding portion comprises a third heavy chain variable region and a third light chain variable region, and the third antigen-binding portion comprises a third binding structure domain that binds to the third antigen. Preferably, the third binding structure domain comprises a third heavy chain variable region selected from the amino acid sequence shown in SEQ ID NO: 268 or any variant thereof, and a third light chain variable region selected from the amino acid sequence shown in SEQ ID NO: 267 or any variant thereof. Preferably, the third antigen-binding portion comprises a third heavy chain comprising the amino acid sequence shown in SEQ ID NO: 233 or any variant thereof, and a third light chain comprising the amino acid sequence shown in SEQ ID NO: 226 or any variant thereof. Preferably, the multispecific antibody is a trispecific antibody, wherein any two of the first heavy chain variable region, the second heavy chain variable region, and the third heavy chain variable region are linked to a constant region, and the remaining heavy chain variable region is not linked to a constant region, and the heavy chain variable region not linked to a constant region is linked to any one of the two heavy chain variable regions linked to the constant region, preferably by a linker, and preferably, the heavy chain variable region not linked to a constant region is linked to its corresponding light chain variable region by a linker. Preferably, the first heavy chain variable region or the first light chain variable region of the first antigen-binding portion is linked to the second heavy chain variable region or the second light chain variable region of the second antigen-binding portion by a linker, preferably, the first heavy chain variable region and the first light chain variable region of the first antigen-binding portion are linked by a linker, and more preferably, the first light chain variable region of the first antigen-binding portion is linked to the second heavy chain variable region of the second antigen-binding portion. Preferably, the first heavy chain variable region or the first light chain variable region of the first antigen-binding portion is linked to the third heavy chain variable region or the third light chain variable region of the third antigen-binding portion by a linker, preferably, the first heavy chain variable region and the first light chain variable region of the first antigen-binding portion are linked to each other by a linker. Preferably, the linker is a flexible linker, and more preferably, the linker is (GGGGGS) ｎ , n = 2-5, specifically, GGGGGGSGGGGGSGGGGGS (SEQ ID NO: 231), GSTSGSGKSSEGKGG (SEQ ID NO: 273), GSGGGGSG (SEQ ID NO: 274), GGGSGGSG (SEQ ID NO: 275), More preferably, the trispecific antibody comprises light chain 1, heavy chain 1, light chain 2, and heavy chain 2. More preferably, the light chain 1 includes the third light chain variable region of the amino acid sequence shown in SEQ ID NO: 267, the heavy chain 1 includes the first light chain variable region of the amino acid sequence shown in SEQ ID NO: 230, the first heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 232, and the third heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 268, the light chain 2 includes the second light chain variable region of the amino acid sequence shown in SEQ ID NO: 152, the heavy chain 2 includes the second heavy chain variable region of the amino acid sequence shown in SEQ ID NO:

157. More preferably, the light chain 1 includes the amino acid sequence shown in SEQ ID NO: 226, the heavy chain 1 includes the amino acid sequence shown in SEQ ID NO: 228, the light chain 2 includes the amino acid sequence shown in SEQ ID NO: 224, and the heavy chain 2 includes the amino acid sequence shown in SEQ ID NO:

222. More preferably, the light chain 1 includes the third light chain variable region of the amino acid sequence shown in SEQ ID NO: 267, the heavy chain 1 includes the first heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 232, the first light chain variable region of the amino acid sequence shown in SEQ ID NO: 230, and the third heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 268, the light chain 2 includes the second light chain variable region of the amino acid sequence shown in SEQ ID NO: 152, the heavy chain 2 includes the second heavy chain variable region of the amino acid sequence shown in SEQ ID NO:

157. More preferably, the light chain 1 includes the amino acid sequence shown in SEQ ID NO: 226, the heavy chain 1 includes the amino acid sequence shown in SEQ ID NO: 234, the light chain 2 includes the amino acid sequence shown in SEQ ID NO: 224, and the heavy chain 2 includes the amino acid sequence shown in SEQ ID NO:

222. More preferably, the light chain 1 includes the second light chain variable region of the amino acid sequence shown in SEQ ID NO: 152, the heavy chain 1 includes the first light chain variable region of the amino acid sequence shown in SEQ ID NO: 230, the first heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 232, and the second heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 157, the light chain 2 includes the third light chain variable region of the amino acid sequence shown in SEQ ID NO: 267, the heavy chain 2 includes the third heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 268, more preferably, the light chain 1 includes the amino acid sequence shown in SEQ ID NO: 224, the heavy chain 1 includes the amino acid sequence shown in SEQ ID NO: 236, the light chain 2 includes the amino acid sequence shown in SEQ ID NO: 226, and the heavy chain 2 includes the amino acid sequence shown in SEQ ID NO:

233. More preferably, the light chain 1 includes the second light chain variable region of the amino acid sequence shown in SEQ ID NO: 152, the heavy chain 1 includes the first heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 232, the first light chain variable region of the amino acid sequence shown in SEQ ID NO: 230, and the second heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 157, the light chain 2 includes the third light chain variable region of the amino acid sequence shown in SEQ ID NO: 267, the heavy chain 2 includes the third heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 268, more preferably, the light chain 1 includes the amino acid sequence shown in SEQ ID NO: 224, the heavy chain 1 includes the amino acid sequence shown in SEQ ID NO: 239, the light chain 2 includes the amino acid sequence shown in SEQ ID NO: 226, and the heavy chain 2 includes the amino acid sequence shown in SEQ ID NO: 233, a multispecific antibody or an antigen-binding fragment thereof.

7. A nucleic acid encoding the multispecific antibody or an antigen-binding fragment thereof according to claim 6, Preferably, the multispecific antibody is a bispecific antibody, and the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody is selected from the nucleotide sequences shown in SEQ ID NO: 169 and SEQ ID NO: 203, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion is selected from the nucleotide sequences shown in SEQ ID NO: 172, SEQ ID NO: 207, SEQ ID NO: 213, SEQ ID NO: 217 and SEQ ID NO:

221. The nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO:

153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody is selected from the nucleotide sequences shown in SEQ ID NO: 197 and SEQ ID NO: 201, and the nucleic acid encoding the first light chain of the first antigen-binding portion is selected from the nucleotide sequences shown in SEQ ID NO: 199, SEQ ID NO: 205, SEQ ID NO: 211, SEQ ID NO: 215 and SEQ ID NO:

219. The nucleic acid encoding the second heavy chain of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO:

225. More preferably, the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 169, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

172. The nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 197, and the nucleic acid encoding the first light chain of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

199. The nucleic acid encoding the second heavy chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

225. More preferably, the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 203, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

207. The nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 201, and the nucleic acid encoding the first light chain of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

205. The nucleic acid encoding the second heavy chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

225. More preferably, the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 203, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

213. The nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 201, the nucleic acid encoding the first light chain of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 211, the nucleic acid encoding the second heavy chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

225. More preferably, the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 203, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

217. The nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 201, the nucleic acid encoding the first light chain of the first antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 215, the nucleic acid encoding the second heavy chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion comprises the nucleotide sequence shown in SEQ ID NO:

225. More preferably, the nucleic acid encoding the first heavy chain variable region of the first antigen-binding portion of the bispecific antibody includes the nucleotide sequence shown in SEQ ID NO: 203, and the nucleic acid encoding the first light chain variable region of the first antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO:

221. The nucleic acid encoding the second heavy chain variable region of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO: 158, and the nucleic acid encoding the second light chain variable region of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO:

153. More preferably, the nucleic acid encoding the first heavy chain of the first antigen-binding portion of the bispecific antibody includes the nucleotide sequence shown in SEQ ID NO: 201, the nucleic acid encoding the first light chain of the first antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO: 219, the nucleic acid encoding the second heavy chain of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO: 223, and the nucleic acid encoding the second light chain of the second antigen-binding portion includes the nucleotide sequence shown in SEQ ID NO:

225. Preferably, the multispecific antibody is a trispecific antibody, and the trispecific antibody includes a light chain 1, a heavy chain 1, a light chain 2, and a heavy chain 2. More preferably, the nucleic acid encoding the light chain 1 includes the nucleotide sequence shown in SEQ ID NO: 269, the nucleic acid encoding the heavy chain 1 includes the nucleotide sequences shown in SEQ ID NO: 271, SEQ ID NO: 272, and SEQ ID NO: 270, the nucleic acid encoding the light chain 2 includes the nucleotide sequence shown in SEQ ID NO: 153, and the nucleic acid encoding the heavy chain 2 includes the nucleotide sequence shown in SEQ ID NO:

158. More preferably, the nucleic acid encoding the light chain 1 of the trispecific antibody includes the nucleotide sequence shown in SEQ ID NO: 227, the nucleic acid encoding the heavy chain 1 includes the nucleotide sequence shown in SEQ ID NO: 229, the nucleic acid encoding the light chain 2 includes the nucleotide sequence shown in SEQ ID NO: 225, and the nucleic acid encoding the heavy chain 2 includes the nucleotide sequence shown in SEQ ID NO:

223. More preferably, the nucleic acid encoding the light chain 1 includes the nucleotide sequence shown in SEQ ID NO: 269, the nucleic acid encoding the heavy chain 1 includes the nucleotide sequences shown in SEQ ID NO: 272, SEQ ID NO: 271, and SEQ ID NO: 270, the nucleic acid encoding the light chain 2 includes the nucleotide sequence shown in SEQ ID NO: 153, the nucleic acid encoding the heavy chain 2 includes the nucleotide sequence shown in SEQ ID NO: 158, and more preferably, the nucleic acid encoding the light chain 1 of the multispecific antibody includes the nucleotide sequence shown in SEQ ID NO: 227, the nucleic acid encoding the heavy chain 1 includes the nucleotide sequence shown in SEQ ID NO: 235, the nucleic acid encoding the light chain 2 includes the nucleotide sequence shown in SEQ ID NO: 225, and the nucleic acid encoding the heavy chain 2 includes the nucleotide sequence shown in SEQ ID NO:

223. More preferably, the nucleic acid encoding the light chain 1 includes the nucleotide sequence shown in SEQ ID NO: 153, the nucleic acid encoding the heavy chain 1 includes the nucleotide sequences shown in SEQ ID NO: 271, SEQ ID NO: 272, and SEQ ID NO: 158, the nucleic acid encoding the light chain 2 includes the nucleotide sequence shown in SEQ ID NO: 269, the nucleic acid encoding the heavy chain 2 includes the nucleotide sequence shown in SEQ ID NO: 270, and more preferably, the nucleic acid encoding the light chain 1 of the multispecific antibody includes the nucleotide sequence shown in SEQ ID NO: 225, the nucleic acid encoding the heavy chain 1 includes the nucleotide sequence shown in SEQ ID NO: 237, the nucleic acid encoding the light chain 2 includes the nucleotide sequence shown in SEQ ID NO: 227, and the nucleic acid encoding the heavy chain 2 includes the nucleotide sequence shown in SEQ ID NO:

238. More preferably, the nucleic acid encoding the light chain 1 comprises the nucleotide sequence shown in SEQ ID NO: 153, the nucleic acid encoding the heavy chain 1 comprises the nucleotide sequences shown in SEQ ID NO: 272, SEQ ID NO: 271, and SEQ ID NO: 158, the nucleic acid encoding the light chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 269, the nucleic acid encoding the heavy chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 270, and more preferably, the nucleic acid encoding the light chain 1 of the multispecific antibody comprises the nucleotide sequence shown in SEQ ID NO: 225, the nucleic acid encoding the heavy chain 1 comprises the nucleotide sequence shown in SEQ ID NO: 240, the nucleic acid encoding the light chain 2 comprises the nucleotide sequence shown in SEQ ID NO: 227, and the nucleic acid encoding the heavy chain 2 comprises the nucleotide sequence shown in SEQ ID NO:

238.

8. A vector comprising the nucleic acid according to claim 5 or 7.

9. A cell comprising the nucleic acid according to claim 5 or 7 or the vector according to claim 8.

10. A composition comprising the GPRC5D antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, the multispecific antibody or antigen-binding fragment thereof according to claim 6, the nucleic acid according to claim 5 or 7, the vector according to claim 8, and / or the cell according to claim 9.

11. A kit comprising the GPRC5D antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, the multispecific antibody or antigen-binding fragment thereof according to claim 6, the nucleic acid according to claim 5 or 7, the vector according to claim 8, the cell according to claim 9, and / or the composition according to claim 10.

12. Use of the GPRC5D antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, the multispecific antibody or antigen-binding fragment thereof according to claim 6, the nucleic acid according to claim 5 or 7, the vector according to claim 8, the cell according to claim 9, and / or the composition according to claim 10 in the preparation of a medicament or kit for use in the diagnosis, treatment, or prevention of cancer or autoimmune diseases, Preferably, the cancer is a B-cell related cancer selected from multiple myeloma, malignant plasmacytoma, Hodgkin lymphoma, nodular lymphocyte-predominant Hodgkin lymphoma, Kahlers disease and myeloid leukemia, plasmacytic leukemia, plasmacytoma, B-cell prolymphocytic leukemia, hairy cell leukemia, B-cell non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL), chronic myeloid leukemia (CML), follicular lymphoma, Burkitt lymphoma, marginal zone lymphoma, mantle cell lymphoma, large cell lymphoma, precursor B-lymphoblastic lymphoma, myeloid leukemia, Waldenström macroglobulinemia, diffuse large B-cell lymphoma, follicular lymphoma, marginal zone lymphoma, mucosa-associated lymphoid tissue lymphoma, small cell lymphocytic lymphoma, mantle cell lymphoma, Burkitt lymphoma, primary mediastinal (thymic) large B-cell lymphoma, lymphoplasmacytic lymphoma, Waldenström macroglobulinemia, nodal marginal zone B-cell lymphoma, splenic marginal zone lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, lymphomatoid granulomatosis, T-cell / histiocyte-rich large B-cell lymphoma, primary central nervous system lymphoma, primary cutaneous diffuse large B-cell lymphoma (lower extremity type), EBV-positive diffuse large B-cell lymphoma in the elderly, inflammation-related diffuse large B-cell lymphoma, intravascular large B-cell lymphoma, ALK-positive large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma arising from HHV8-related multicentric Castleman disease, unclassified B-cell lymphoma with intermediate features between diffuse large B-cell lymphoma and Burkitt lymphoma, unclassified B-cell lymphoma with intermediate features between diffuse large B-cell lymphoma and classical Hodgkin lymphoma, and other B-cell related lymphomas, Preferably, the autoimmune disease is selected from immune renal disease, systemic lupus erythematosus or rheumatoid arthritis for use.

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